益肾活血胶囊对动脉粥样硬化的免疫调节作用及斑块稳定性的影响
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摘要
第一部分益肾活血胶囊对载脂蛋白E基因敲除小鼠主动脉粥样硬化的免疫调节作用及斑块稳定性的影响
研究背景
动脉粥样硬化(Atherosclerosis,AS)是严重危害人类健康的常见病,其发病率有逐年增加的趋势。众多研究表明,体液免疫和细胞免疫参与了AS形成的全过程。而且,免疫介导和炎症反应在AS的发生机制和冠心病心血管突发事件的发生中起重要作用。因此免疫调节和抗炎治疗将有望成为防治AS的重要手段之一。白细胞分化抗原40(Cell differentiation antigen 40,CD40)与其配体(CD40 ligand,CD40L,又称CD154)是一对互补跨膜糖蛋白,分别属于肿瘤坏死因子(Tumornecrosis factor,TNF)受体和TNF超家族成员,是免疫系统细胞间信息传递的主要介质,其相互作用显著影响AS相关细胞的功能,并且与斑块的发生发展及斑块的稳定性密切相关。研究发现CD40通路很可能是动脉粥样硬化发生发展过程中免疫炎症调节的上层机制。致AS抗原如氧化修饰的低密度脂蛋白(Oxidized lowdensity lipoprotein,oxLDL),热休克蛋白60(Heat shock protein 60,HSP60)等可促进CD40/CD40L的激活与表达,CD40L与CD40结合,可诱导内皮细胞(Endothelialcells,ECs)、血管平滑肌细胞(Vascular smooth muscle cells,VSMCs),巨噬细胞(Macrophage,MФ)等表达产生血管细胞粘附分子-1(Vascular adhesion molecule-1,VCAM-1)、细胞间黏附分子-1(Intercellular adhesion molecule-1,ICAM-1)和E-选择素(E selectin)及化学趋化因子白介素-8(Interleukin-8,IL-8)、单核细胞炎性蛋白-1和单核细胞趋化蛋白-1(Monocyte chemoattractant protein-1,MCP-1)等,促进白细胞进入血管壁及平滑肌细胞向内膜的迁移与增殖。而且,CD40/CD40L相互作用还可诱导ECs、VSMCs、MΦ表达和释放基质金属蛋白酶和组织因子,促进斑块不稳定、破裂及局部血栓形成。
益肾活血胶囊(Yishenhuoxue Capsule,YC)是根据导师的经验方研制而成,是临床治疗AS的有效的中药复方制剂。我们以前的研究表明其能够降低动脉粥样硬化指数,降低高同型半胱氨酸血症引起的炎症因子的高表达。但是其对CD40信号系统及斑块稳定性的影响尚不明了,本研究从体内试验探讨了益肾活血胶囊对动脉粥样硬化过程中免疫炎症因子oxLDL、CD40、基质金属蛋白酶-9(Matrixmetalloproteinase-9,MMP-9)、VCAM-1的调节作用及对动脉粥样硬化斑块稳定性的影响,以期为该药及益肾活血法治疗AS提供科学依据。
目的
观察益肾活血胶囊对载脂蛋白E基因敲除(Apolipoprotein E-knockout,apoE~(-/-))小鼠血脂,血清oxLDL,主动脉壁的组织学及超微结构、血管壁免疫炎症因子CD40、VCAM-1和MMP-9水平以及斑块内部构成(如脂质,胶原纤维,MΦ,SMCs的含量等)的变化,探讨益肾活血胶囊对动脉粥样硬化的免疫调节作用及对动脉粥样硬化斑块稳定性的影响。
方法
8周龄雄性apoE~(-/-)小鼠45只,同种属C57BL/6J小鼠9只。apoE~(-/-)小鼠给予高脂饮食喂养(15%脂肪,0.25%胆固醇,84.75%基础饲料),C57BL/6J小鼠给予普通饲料喂养,于高脂喂养12周末,将apoE~(-/-)小鼠随机分为模型组、益肾活血胶囊小剂量组(小剂量组)、益肾活血胶囊大剂量组(大剂量组)、辛伐他汀对照组(辛伐他汀组)和三七总皂苷对照组(三七总皂苷组),每组9只。C57BL/6J小鼠为正常组,共6组。小剂量组和大剂量组分别给予YC 500 mg/kg,2500 mg/kg灌胃,1次/d;辛伐他汀组给予辛伐他汀3.3 mg/kg灌胃,1次/d;三七总皂苷组给予三七总皂苷60 mg/kg灌胃,1次/d;灌胃时间为12周,模型组和正常组给予生理盐水0.2 ml/只,1次/d,时间12周。于试验24周末,处死各组试验鼠,摘眼球取血1 ml,氧化酶法测定各实验组动物血清总胆固醇(Total cholesterol,TC)、低密度脂蛋白胆固醇(Low density lipoprotein cholesterol,LDL-C)、甘油三脂(Triglyceride,TG)浓度,酶联免疫吸附试验(Enzyme linked immunosorbent assay,ELISA)测定动物血清中oxLDL含量。用磷酸盐缓冲液(Phosphate buffer saline,PBS)从左心室逆行灌注主动脉,自主动脉根部起至胸主动脉离断主动脉,主动脉弓部约1 mm迅速投入3%戊二醛中制备透射电镜标本,余主动脉根部及主动脉弓投入4%多聚甲醛中固定,制备冰冻切片,分别行HE染色,天狼星红染色,油红O染色及SMCs、MΦ和免疫炎症因子CD40、VCAM-1、MMP-9免疫组化染色。胸主动脉迅速投入液氮中。采用实时定量RT-PCR技术检测胸主动脉CD40、VCAM-1和MMP-9的mRNA表达。采用SPSS 11.5统计软件进行统计分析。
结果
1.小鼠血脂测定
与正常对照组相比,模型组小鼠血清TC、TG、LDL-C浓度明显升高(P均＜0.001);大剂量组、辛伐他汀组及三七总皂苷组小鼠血脂浓度与模型组比较均有明显程度下降,其中三七总皂苷组与大剂量组下降最为显著(P均＜0.001),其次为辛伐他汀组;小剂量组与模型组相比,除TG外,TC、LDL-C浓度与模型组相比差异无显著性(P均＞0.05);小剂量组和大剂量组相比具有非常显著性差异(P均＜0.001)。
2.小鼠血清oxLDL含量比较
与正常对照组相比,模型组小鼠血清oxLDL含量明显升高(P＜0.001);各个药物处理组小鼠oxLDL含量与模型组比较均有明显程度下降,其中大剂量组oxLDL含量下降最为显著(P＜0.001),其次为辛伐他汀组,三七总皂苷组和小剂量组(P＜0.001,或P＜0.01)。
3.apoE~(-/-)小鼠AS病变面积检测
正常组主动脉壁光滑,完整,无AS斑块形成。apoE~(-/-)小鼠均见AS斑块形成,管腔呈不同程度的狭窄。斑块面积/管腔面积分别是32.54±5.13%(模型组),26.22±4.74%(小剂量组),14.16±5.18%(大剂量组),14.43±4.29%(辛伐他汀组),12.04±5.02%(三七总皂苷组)。与模型组相比,各用药组斑块面积/管腔面积比值有不同程度降低,其中大剂量组,辛伐他汀组,三七总皂苷组与小剂量组相比差异有统计学意义(P＜0.05或P＜0.001)。
4.apoE~(-/-)小鼠AS斑块内部构成的变化大剂量组,辛伐他汀组和三七总皂苷组斑块内MΦ浸润及脂质含量低于模型组(P均＜0.001),同时斑块内VSMCs含量高于模型组(P均＜0.001)。小剂量组与模型组相比,斑块内脂质含量降低(P＜0.05),MΦ浸润水平及VSMCs含量经统计无显著差异(P＞0.05)。大剂量组与小剂量组相比,差异有显著性(P＜0.001,或P＜0.01)。大剂量组与辛伐他汀组,三七总皂苷组之间差异无显著性(P＞0.05)。
5.主动脉内皮细胞超微结构改变
正常组血管EC为单层扁平细胞,连续性好,胞膜平整,胞浆均匀,紧贴内弹性膜,线粒体形态结构正常。内弹性膜连续,厚度均匀。模型组EC明显肿胀,连续性中断,有明显的内皮细胞脱落现象,胞核内异染色质明显增多,边集于核膜下,线粒体明显肿胀,脊肿胀、溶解。内弹性膜有中断,厚度不均匀,可见VSMC自缺口伸入内膜。各药物处理组与模型组相比,EC形态有不同程度的改善,连续性较好,胞核形态结构基本正常,线粒体数目略增多,形态结构基本正常。内弹性膜连续,厚度均匀。
6.主动脉CD40、VCAM-1、MMP-9蛋白表达水平比较
(1)CD40蛋白表达比较:正常主动脉CD40表达很少,模型组小鼠血管壁CD40蛋白表达显著升高(P＜0.001);各药物处理组CD40蛋白表达与模型组相比,有非常显著性差异(P＜0.001);辛伐他汀组和大剂量组CD40蛋白表达明显低于小剂量组(P＜0.05)。
(2)VCAM-1蛋白表达比较:与正常对照组相比,模型组小鼠血管壁VCAM-1蛋白表达显著升高(P＜0.001);与模型组相比,各药物处理组VCAM-1蛋白表达均明显下降(P＜0.01或P＜0.05);大剂量组与小剂量组比较,差异有统计学意义(P＜0.05)。
(3)MMP-9蛋白表达比较:与正常对照组相比,模型组小鼠血管壁MMP-9蛋白表达显著升高(P＜0.001);与模型组相比,各药物处理组MMP-9蛋白表达均明显下降(P＜0.001或P＜0.01或P＜0.05);大剂量组、辛伐他汀组、三七总皂苷组MMP-9蛋白表达较小剂量组明显下降(P＜0.01或P＜0.05)。
7.主动脉CD40、VCAM-1、MMP-9 mRNA表达水平比较
(1)CD40 mRNA表达比较:与正常对照组相比,模型组小鼠血管壁CD40mRNA表达显著升高(P＜0.001);各药物处理组CD40 mRNA表达较模型组均有不同程度下降,其中辛伐他汀组下降最为显著(P＜0.001),其次为三七总皂苷组和大剂量组(P＜0.001);小剂量组与大剂量组、辛伐他汀组、三七总皂苷组比较有显著差异(P＜0.001)。
(2)VCAM-1 mRNA表达比较:与正常对照组相比,模型组小鼠血管壁VCAM-1mRNA表达显著升高(P＜0.001);与模型组相比,各药物处理组VCAM-1mRNA表达均显著下降(P＜0.05或P＜0.001);大剂量组、辛伐他汀组VCAM-1表达低于三七总皂苷组和小剂量组(P＜0.001)。
(3)MMP-9 mRNA表达比较:与正常对照组相比,模型组小鼠血管壁MMP-9mRNA表达明显升高(P＜0.001);与模型组相比,各药物处理组MMP-9 mRNA表达均显著下降(P＜0.001);大剂量组、辛伐他汀组、三七总皂苷组与小剂量组比较,差异具有统计学意义(P＜0.001)。
结论
1.益肾活血胶囊能够降低apoE~(-/-)小鼠血脂水平。
2.益肾活血胶囊能够降低apoE~(-/-)小鼠血清oxLDL含量。
3.益肾活血胶囊抑制了免疫炎症因子CD40、MMP-9、VCAM-1的mRNA和蛋白表达。
4.益肾活血胶囊对AS具有免疫调节作用,能够抑制AS的进展,抑制AS斑块由稳定向不稳定发展。其机制与降低血脂,下调oxLDL及CD40、MMP-9、VCAM-1的表达有关。
第二部分益肾活血提取液对oxLDL诱导的人脐静脉内皮细胞CD40、MMP-9及VCAM-1表达的影响
目的
观察益肾活血提取液对oxLDL诱导的人脐静脉内皮细胞(Human umbilicalvein endothelial cells,HUVECs)CD40、MMP-9及VCAM-1表达的影响,从体外试验探讨益肾活血胶囊抗AS的机制。
方法
采用水煎醇沉法制备益肾活血提取液。采用胰蛋白酶液灌注消化法分离获取HUVECs并进行体外培养,取3-5代细胞进行试验。试验共分7组,空白组不予oxLDL刺激及药物处理;oxLDL刺激组(刺激组)细胞培养液中加入oxLDL(终浓度为50μg/ml)培养24 h;oxLDL+益肾活血提取液小剂量组(剂量组)、oxLDL+益肾活血提取液中剂量组(中剂量组)、oxLDL+益肾活血提取液大剂量组(大剂量组)细胞培养液中加入益肾活血提取液(终浓度分别为100μg/ml,500μg/ml,1mg/ml)培养24 h,oxLDL+辛伐他汀组(辛伐他汀组)细胞培养液中加入辛伐他汀(终浓度为10μmol/1)培养24 h,oxLDL+三七总皂苷组(三七总皂苷组)细胞培养液中加入三七总皂苷(终浓度为100μg/ml)培养24 h,后5组于药物培养24 h后加入oxLDL(终浓度为50μg/ml)继续培养24 h进行检测。采用实时定量RT-PCR检测VCAM-1的mRNA表达,免疫印迹法(Western blot)检测CD40、MMP-9的蛋白表达。试验重复3次,实验数据采用统计学方法进行分析。
结果
1.VCAM-1 mRNA表达水平的比较。与空白组相比,刺激组VCAM-1 mRNA表达明显增加(P＜0.001)。与刺激组相比,各药物处理组VCAM-1 mRNA表达均显著下降(P＜0.05或P＜0.001)。其中辛伐他汀组下降最为显著,其次是大剂量组、三七总皂苷组。辛伐他汀组、大剂量组与小剂量组相比,差异有统计学意义(P＜0.05)。
2.CD40蛋白表达水平的比较。与空白组相比,刺激组CD40蛋白表达明显增加(P＜0.001)。与刺激组相比,除小剂量组外余各药物处理组CD40蛋白表达均显著下降(P＜0.05或P＜0.01)。其中辛伐他汀组和大剂量组下降最为显著,其次是中剂量组和三七总皂苷组。
3.MMP-9蛋白表达水平的比较。与空白组相比,刺激组MMP-9的蛋白表达显著增高(P＜0.001)。与刺激组相比,各药物处理组MMP-9的蛋白表达均显著下降(P＜0.01或P＜0.001)。大剂量组、辛伐他汀组与中剂量组、三七总皂苷组和小剂量组比较,差异有统计学意义(P＜0.05,P＜0.01或P＜0.001)。
结论
1.oxLDL可诱导HUVECs高表达VCAM-1、CD40和MMP-9。
2.给予益肾活血提取液、辛伐他汀及三七总皂苷预处理后,再给予oxLDL刺激,VCAM-1的mRNA表达降低,CD40、MMP-9的蛋白表达下降。
3.随着益肾活血提取液浓度的增加,VCAM-1、CD40和MMP-9的表达呈下降趋势。提示益肾活血提取液对oxLDL诱导的免疫炎症因子的调节作用在一定范围内具有剂量依赖性。
Part 1 Effect of Yishenhuoxue capsule on regulating immune response and plaque stability in apolipoprotein E-knockout mice
Background
Atherosclerosis(AS) is a common disease which done great harm to human health.Multitude studies had indicated that both humoral and cellular immunity participate the processes of AS.So regulating the immune response will become an important method in treatment of AS.
Cell differentiation antigen 40(CD40) and CD40 ligand(CD40L) is a pair of transmembrane glycoprotein which belongs to tumor necrosis factor receptor(TNFR) and tumor necrosis factor(TNF) superfamilies.It has been shown that the costimulatory signal delivered by CD40-CD40L interactions participate the immune inflammatory response caused by oxLDL.CD40-CD40L interactions can induce endothelial cells(ECs),vascular smooth muscle cells(VSMCs) and macrophage(MΦ) expressing adhesion molecules such as vascular adhesion molecule-1(VCAM-1), intercellular adhesion molecule-1(ICAM-1),E selectin and chemotatic factors such as interleukin-8(IL-8),monocyte inflammatory protein-1(MIP-1) and monecyte chemoattractant protein-1(MCP-1),et al.These adhesion molecules and chemotatic factors can promote the leukocyte dipping into the vascular wall and the migration and proliferation of VSMCs in intima.In addition,CD40-CD40L interactions can promote ECs,VSMCs and MΦexpressing matrix metalloproteinase and tissue factor, thus can cause plaque unstability,rupture and thrombopoiesis.
Yishenhuoxue Capsule(YC),an extract of traditional Chinese medicine,has been used as an antiatherosclerotic agent for years in clinic.Previous studies have shown that YC can lower atherosclerotic index,down-regulate the expression of inflammatory factors caused by high hyperhomocysteinemia,but the effect of YC on modulating the immune response and plaque stability has not been clarified.In this study,we observed the effects of YC on serum levels of lipid and oxLDL and the expression of CD40,matrix metalloproteinase-9(MMP-9) and VCAM-1 in atherosclerotic plaques and on the plaque stability in vivo.The study coule provide scientific evidence for the YC on treatment of AS in clinic.
Aim
To investigate the possible mechanisms of YC on modulating immune response in atherosclerosis and on plaque stability through observing the plasma oxLDL level, changes of the morphology and the ultrastructure and detecting the variation of CD40, VCAM-1 and MMP-9 mRNA and protein expression and the composition of plaques.
Methods
Mail 45 apolipoprotein E-knockout(apoE~(-/-)) mice at 8 weeks of age were fed a high fat high cholesterol diet including 15%fat and 0.25%cholesterol.Mail 9 C57BL/6J mice at 8 weeks of age were fed a normal chow diet.At 20 weeks of age, apoE~(-/-) mice were randomly divided into five groups,the ApoE-KO group,the apoE~(-/-) +YSHXH group(YSHXH group),the ApoE~(-/-)+ YSHXL group(YSHXL group),the apoE~(-/-) + total panax notoginsenoside group(TPNS group) and the apoE~(-/-)+ simvastain group(Simvastain group)(n=9 for each group).C57BL/6J mice were the normal control group(Control group).Mice in apoE~(-/-)+YSHXH and apoE~(-/-)+YSHXL group were given YC at 2500 mg/kg,500 mg/kg,respectively.Mice in TPNS group were given TPNS at 60 mg/kg.And mice in Simvastain group were given Simvastain at 3.3 mg/kg.All drugs were given orally,once a day for 12 weeks.Mice in ApoE-KO and control groups were given 0.2 ml distilled water,once a day for 12 weeks.After administered for 12 weeks,all mice were sacrificed.1 ml blood was obtained from each mouse by removing eyeball.Serum levels of total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) and trigliyceride(TG) was detected by oxidation enzyme method.Serum oxLDL level was mas determined by enzyme linked immunosorbent assay(ELISA).The aorta was perfused with sterile phosphatebuffered saline(PBS) by cardiac puncture.The aortic arch about 1mm was fixed in 3% glutaraldehyde to make transmission electron microscope sample,and the surplus aortic arch and the root of aorta was fixed in 4%paraformaldehyde overnight for histology and immunohistochemistry.The thoracic aorta was rapidly frozen in liquid nitrogen for later detection of CD40,VCAM-1,MMP-9 mRNA expression.
Results
1.Comparison of serum lipid concentration
The serum TC,TG and LDL-C concentration was significantly higher in apoE~(-/-) group than that in Comal group(P<0.001).Lipid levels in YSHXH,Simvastain and TPNS group were significantly lower compared with that in apoE~(-/-) group(P<0.001). The TC,LDL-C levels were no difference between YSHXL and apoE~(-/-) group mice(P >0.05),while the TG concentration in YSHXL group was lower than that in apoE~(-/-) group(P<0.05).High dose YC was superior to low dose YC in lowering serum lipid level(P<0.001).
2.Comparison of serum oxLDL concentration
The serum oxLDL concentration in apoE~(-/-) group was significantly higher than that in Control group(P<0.001).OxLDL levels in drugs treated groups were lower than that in apoE~(-/-) group(P<0.001 or P<0.01).There were significant difference between YSHXH and YSHXL group(P<0.001).
3.Effect of YC on areas of atherosclerotic lesion in apoE~(-/-) mice
A normal aorta wall was observed in Control group mice.Atherosclerotic lesions were observed in all apoE~(-/-) mice and the lumens were narrowed in a different degree. The ratio of lesion area to vessel area in apoE~(-/-) group was higher than that in YSHXH, TPNS and Simvastain group(32.54±5.13%vs 14.16±5.18%,12.04±5.02%,14.43±4.29%,P<0.05 or P<0.01).No difference between YSHXL and apoE~(-/-) group (32.54±5.13%vs 26.22±4.74%,P>0.05).
4.Effect of YC on plaque composition in apoE~(-/-) mice
Compared with the apoE~(-/-) group,MΦand lipid infiltration in plaques were lower and VSMCs in plaques were higher in the YSHXH,Simvastain and TPNS group(P<0.001).Lipid infiltration in the YSHXL group was also lower than the apoE~(-/-) group(P<0.05),but the level of MΦand VSMCs in plaques of the two groups was no significant difference(P>0.05).These indicated that YC could promote plaque stability,and high dose YC was superior to low dose YC.
5.Ultrastructural observation of the ECs in aorta In the Control group,the ECs exhibited characteristics of monolayer pavement cells with a good succession,the cell membrane was intacted and kytoplasm was uniformity.Morphous of mitochondria is normal.The membrana elastica interna was succession and uniformity.In the apoE~(-/-) group,the ECs had obviously engorgement and the succession between ECs was interruption.ECs amotic phenomenon was observed and nuclei heterochromatin obviously increased.The mitochondria obviously swelled with the ridge swelling and dissolved.The membrana elastica interna was uneven and interrupted and VSMCs migrated into intima from the gap. However,these changes were ameliorated when treated with YC,Simvastain and TPNS.The nuclei had slight heterochromatin.The numbers of mitochondria with slight increasing,a consecutive membrana elastica intema was observed.
6.Comparison of expression of CD40,VCAM-1 and MMP-9 protein in aorta wall
(1) Comparison of expression of CD40 protein:The expression of CD40 protein in the apoE~(-/-) group was higher than that in the Control group(P<0.001).The expression of CD40 protein was significantly decreased after treatment with YC, simvastain and TPNS(P<0.001).The expression of CD40 protein was decreased significantly in the Simvastain group,the high dose YC group compared with that in the low dose group(P<0.05).
(2) Comparison of expression of VCAM-1 protein:The expression of VCAM-1 protein in the apoE~(-/-) group was higher than that in the Control group(P<0.001). After treatment with YC,simvastain and TPNS,the expression of VCAM-1 protein in each group was decreased in a different degree compared with the apoE~(-/-) group(P< 0.01 or P<0.05).The expression of VCAM-1 protein was decreased significantly in the YSHXH group compared with that in the YSHXL group(P<0.05).
(3) Comparison of expression of MMP-9 protein:The expression of MMP-9 protein in the apoE~(-/-) group was higher than that in the Control group(P<0.001).The expression of VCAM-1 protein in drugs treated groups was significantly decreased compared with the apoE~(-/-) group(P<0.001 or P<0.01 or P<0.05).The expression of VCAM-1 protein was decreased significantly in the YSHXH group,the Simvastain group and the TPNS group compared with that in the YSHXL group(P<0.01 or P< 0.05).
7.Comparison of expression of CD40,VCAM-1 and MMP-9 mRNA in aorta wall
(1) Comparison of expression of CD40 mRNA:The expression of CD40 mRNA in the apoE~(-/-) group was higher than that in the normal group(P<0.001). After treatment with YC,simvastain and TPNS,CD40 mRNA was decreased in a different degree compared with the apoE~(-/-) group,and among those treatment groups, the Simvastain group had the lowest CD40 mRNA expression(P<0.001),next was the TPNS group and the YSHXH group.The expression of CD40 mRNA in the YSHXH group,the Simvastain group and the TPNS group were lower than that in the YSHXL group(all P<0.001).
(2) Comparison expression of VCAM-1 mRNA:The expression of VCAM-1 mRNA in the apoE~(-/-) group was higher than that in the normal group(P<0.001). VCAM-1 mRNA was decreased in a different degree in drugs treatment groups compared with the apoE~(-/-) group.The expression of VCAM-1 mRNA in the YSHXH group and the Simvastain group were lower than that in the TPNS group and YSHXL group(P<0.001).
(3) Comparison expression of MMP-9 mRNA:The expression of MMP-9 mRNA in the apoE~(-/-) group was higher than that in the normal group(P<0.001).VCAM-1 mRNA was decreased significantly in drugs treatment groups compared with the apoE~(-/-) group(P<0.001).The expression of VCAM-1 mRNA in the YSHXH group, the TPNS group and the Simvastain group were lower than that in the YSHXL group (P<0.001).
Conclusion
1.YC could decrease blood lipid level in apoE~(-/-) mice.
2.YC could decrease serum oxLDL concentration in apoE~(-/-) mice.
3.YC could down-regulated the mRNA and protein expression of CD40,MMP-9 and VCAM-1.
4.YC could regulate immune response in AS,inhibit the development of AS and prevent transformation from stable plaque to unstable plaque.The mechanisms were possibly associated with the suppressive effect of YC on blood levels of lipid and oxLDL and CD40、MMP-9 and VCAM-1 expression in aortas of apoE~(-/-) mice.
Part 2 Effect of Yishenhuoxue extraction on CD40,MMP-9 and VCAM-1 expression in human umbilical vein endothelial cells induced by oxLDL
Aim
To observe the Yishenhuoxue extraction on the CD40、MMP-9 and VCAM-1 expression in human umbilical vein endothelial cells(HUVECs) induced by oxLDL. Investigate the effect of Yishenhuoxue extraction on atherosclerosis in vitro.
Methods
The Yishenhuoxue extraction was prepared by decocting with distilled water and precipitating with ethanol.The HUVECs were obtained by perfusing trypsinase liquid in umbilical cord and cultured in vitro.The 3-5 generation cells were used in the study. Cells were divided into 7 groups,the normal group,the oxLDL stimulated group (stimulate group),the oxLDL+low dose Yishenhuoxue extraction group(low dose group),the oxLDL+middle dose Yishenhuoxue extraction group(middle dose group), the oxLDL+high dose Yishenhuoxue extraction group(high dose group),the oxLDL +simvastain group(simvastain group) and the oxLDL+total panax notoginsenosidum group(TPNS group).Cells in stimulate group were admoved oxLDL at 50μg/ml into cell culture fluid and cultured for 24 h.Cells in low dose group、middle dose group and high dose group were pretreated with Yishenhuoxue extraction at 100μg/ml,500μg/ml and 1 mg/ml,respectively for 24 h,after 24 h, cells were given oxLDL at 50μg/ml and cultured for another 24 h.Cells in simvastain group and TPNS group were pretreated with simvastain at 10μmol/l and TPNS at 100μg/ml for 24 h,after 24 h,cells were given oxLDL at 50μg/ml and cultured for another 24 h.The expression of VCAM-1 mRNA was detected by real time RT-PCR, the expression of CD40,MMP-9 protein was detected by western blot.All experimental data analyzed by statistics.
Results
1.Comparison of VCAM-1 mRNA:The expression of VCAM-1 mRNA in stimulate group was significantly higher than that in the normal group(P<0.001). The VCAM-1 mRNA expression was significantly lower in lose dose group,middle dose group,high dose group,simvastain group and TPNS group compared with that in stimulate group(P<0.05 or P<0.001).Among those five groups,the simvastain group had the lowest expression of VCAM-1 mRNA,next was the high dose group and TPNS group.The VCAM-1 mRNA was significantly lower in simvastain group and high dose group compared with low dose group(P<0.05).
2.Comparison of CD40 protein:The expression of CD40 protein in stimulate group was significantly higher than that in the normal group(P<0.001).HUVECs treated with Yishenhuoxue extraction,simvastain and TPNS for 24 hours indicated a decrease of CD40 protein expression.CD40 protein was the lowest in simvastain and high dose group,high dose Yishenhuoxue extraction and middle dose Yishenhuoxue extraction was superior to low dose Yishenhuoxue extraction(P<0.05 or P<0.01).
3.Comparison of MMP-9 protein:The expression of MMP-9 protein in stimulate group was significantly higher than that in the normal group(P<0.001).The MMP-9 protein expression was significantly lower in lose dose group,middle dose group, high dose group,simvastain group and TPNS group compared with that in stimulate group(P<0.01 or P<0.001).The MMP-9 protein was significantly lower in high dose group and simvastain group compared with that in middle dose group,TPNS group and low dose group(P<0.05,P<0.01 or P<0.001).
Conclusion
1.oxLDL could induce the expression of CD40、VCAM-1 and MMP-9 in HUVECs at a high level.
2.Pretreatment with Yishenhuoxue extraction,simvastain and TPNS could down-regnlate the expression of CD40、VCAM-1 and MMP-9 in HUVECs induced by oxLDL.
3.The expression of CD40、VCAM-1 and MMP-9 in HUVECs had a descending tendency along with the increasing concentration of Yishenhuoxue extraction.It indicated that the effect of Yishenhuoxue extraction on modulating immune inflammatory factors in HUVECs induced by oxLDL was in a dose dependent manner.
研究背景
动脉粥样硬化(Atherosclerosis,AS)是严重危害人类健康的常见病,其发病率有逐年增加的趋势。众多研究表明,体液免疫和细胞免疫参与了AS形成的全过程。而且,免疫介导和炎症反应在AS的发生机制和冠心病心血管突发事件的发生中起重要作用。因此免疫调节和抗炎治疗将有望成为防治AS的重要手段之一。白细胞分化抗原40(Cell differentiation antigen 40,CD40)与其配体(CD40 ligand,CD40L,又称CD154)是一对互补跨膜糖蛋白,分别属于肿瘤坏死因子(Tumornecrosis factor,TNF)受体和TNF超家族成员,是免疫系统细胞间信息传递的主要介质,其相互作用显著影响AS相关细胞的功能,并且与斑块的发生发展及斑块的稳定性密切相关。研究发现CD40通路很可能是动脉粥样硬化发生发展过程中免疫炎症调节的上层机制。致AS抗原如氧化修饰的低密度脂蛋白(Oxidized lowdensity lipoprotein,oxLDL),热休克蛋白60(Heat shock protein 60,HSP60)等可促进CD40/CD40L的激活与表达,CD40L与CD40结合,可诱导内皮细胞(Endothelialcells,ECs)、血管平滑肌细胞(Vascular smooth muscle cells,VSMCs),巨噬细胞(Macrophage,MФ)等表达产生血管细胞粘附分子-1(Vascular adhesion molecule-1,VCAM-1)、细胞间黏附分子-1(Intercellular adhesion molecule-1,ICAM-1)和E-选择素(E selectin)及化学趋化因子白介素-8(Interleukin-8,IL-8)、单核细胞炎性蛋白-1和单核细胞趋化蛋白-1(Monocyte chemoattractant protein-1,MCP-1)等,促进白细胞进入血管壁及平滑肌细胞向内膜的迁移与增殖。而且,CD40/CD40L相互作用还可诱导ECs、VSMCs、MΦ表达和释放基质金属蛋白酶和组织因子,促进斑块不稳定、破裂及局部血栓形成。
益肾活血胶囊(Yishenhuoxue Capsule,YC)是根据导师的经验方研制而成,是临床治疗AS的有效的中药复方制剂。我们以前的研究表明其能够降低动脉粥样硬化指数,降低高同型半胱氨酸血症引起的炎症因子的高表达。但是其对CD40信号系统及斑块稳定性的影响尚不明了,本研究从体内试验探讨了益肾活血胶囊对动脉粥样硬化过程中免疫炎症因子oxLDL、CD40、基质金属蛋白酶-9(Matrixmetalloproteinase-9,MMP-9)、VCAM-1的调节作用及对动脉粥样硬化斑块稳定性的影响,以期为该药及益肾活血法治疗AS提供科学依据。
目的
观察益肾活血胶囊对载脂蛋白E基因敲除(Apolipoprotein E-knockout,apoE~(-/-))小鼠血脂,血清oxLDL,主动脉壁的组织学及超微结构、血管壁免疫炎症因子CD40、VCAM-1和MMP-9水平以及斑块内部构成(如脂质,胶原纤维,MΦ,SMCs的含量等)的变化,探讨益肾活血胶囊对动脉粥样硬化的免疫调节作用及对动脉粥样硬化斑块稳定性的影响。
方法
8周龄雄性apoE~(-/-)小鼠45只,同种属C57BL/6J小鼠9只。apoE~(-/-)小鼠给予高脂饮食喂养(15%脂肪,0.25%胆固醇,84.75%基础饲料),C57BL/6J小鼠给予普通饲料喂养,于高脂喂养12周末,将apoE~(-/-)小鼠随机分为模型组、益肾活血胶囊小剂量组(小剂量组)、益肾活血胶囊大剂量组(大剂量组)、辛伐他汀对照组(辛伐他汀组)和三七总皂苷对照组(三七总皂苷组),每组9只。C57BL/6J小鼠为正常组,共6组。小剂量组和大剂量组分别给予YC 500 mg/kg,2500 mg/kg灌胃,1次/d;辛伐他汀组给予辛伐他汀3.3 mg/kg灌胃,1次/d;三七总皂苷组给予三七总皂苷60 mg/kg灌胃,1次/d;灌胃时间为12周,模型组和正常组给予生理盐水0.2 ml/只,1次/d,时间12周。于试验24周末,处死各组试验鼠,摘眼球取血1 ml,氧化酶法测定各实验组动物血清总胆固醇(Total cholesterol,TC)、低密度脂蛋白胆固醇(Low density lipoprotein cholesterol,LDL-C)、甘油三脂(Triglyceride,TG)浓度,酶联免疫吸附试验(Enzyme linked immunosorbent assay,ELISA)测定动物血清中oxLDL含量。用磷酸盐缓冲液(Phosphate buffer saline,PBS)从左心室逆行灌注主动脉,自主动脉根部起至胸主动脉离断主动脉,主动脉弓部约1 mm迅速投入3%戊二醛中制备透射电镜标本,余主动脉根部及主动脉弓投入4%多聚甲醛中固定,制备冰冻切片,分别行HE染色,天狼星红染色,油红O染色及SMCs、MΦ和免疫炎症因子CD40、VCAM-1、MMP-9免疫组化染色。胸主动脉迅速投入液氮中。采用实时定量RT-PCR技术检测胸主动脉CD40、VCAM-1和MMP-9的mRNA表达。采用SPSS 11.5统计软件进行统计分析。
结果
1.小鼠血脂测定
与正常对照组相比,模型组小鼠血清TC、TG、LDL-C浓度明显升高(P均＜0.001);大剂量组、辛伐他汀组及三七总皂苷组小鼠血脂浓度与模型组比较均有明显程度下降,其中三七总皂苷组与大剂量组下降最为显著(P均＜0.001),其次为辛伐他汀组;小剂量组与模型组相比,除TG外,TC、LDL-C浓度与模型组相比差异无显著性(P均＞0.05);小剂量组和大剂量组相比具有非常显著性差异(P均＜0.001)。
2.小鼠血清oxLDL含量比较
与正常对照组相比,模型组小鼠血清oxLDL含量明显升高(P＜0.001);各个药物处理组小鼠oxLDL含量与模型组比较均有明显程度下降,其中大剂量组oxLDL含量下降最为显著(P＜0.001),其次为辛伐他汀组,三七总皂苷组和小剂量组(P＜0.001,或P＜0.01)。
3.apoE~(-/-)小鼠AS病变面积检测
正常组主动脉壁光滑,完整,无AS斑块形成。apoE~(-/-)小鼠均见AS斑块形成,管腔呈不同程度的狭窄。斑块面积/管腔面积分别是32.54±5.13%(模型组),26.22±4.74%(小剂量组),14.16±5.18%(大剂量组),14.43±4.29%(辛伐他汀组),12.04±5.02%(三七总皂苷组)。与模型组相比,各用药组斑块面积/管腔面积比值有不同程度降低,其中大剂量组,辛伐他汀组,三七总皂苷组与小剂量组相比差异有统计学意义(P＜0.05或P＜0.001)。
4.apoE~(-/-)小鼠AS斑块内部构成的变化大剂量组,辛伐他汀组和三七总皂苷组斑块内MΦ浸润及脂质含量低于模型组(P均＜0.001),同时斑块内VSMCs含量高于模型组(P均＜0.001)。小剂量组与模型组相比,斑块内脂质含量降低(P＜0.05),MΦ浸润水平及VSMCs含量经统计无显著差异(P＞0.05)。大剂量组与小剂量组相比,差异有显著性(P＜0.001,或P＜0.01)。大剂量组与辛伐他汀组,三七总皂苷组之间差异无显著性(P＞0.05)。
5.主动脉内皮细胞超微结构改变
正常组血管EC为单层扁平细胞,连续性好,胞膜平整,胞浆均匀,紧贴内弹性膜,线粒体形态结构正常。内弹性膜连续,厚度均匀。模型组EC明显肿胀,连续性中断,有明显的内皮细胞脱落现象,胞核内异染色质明显增多,边集于核膜下,线粒体明显肿胀,脊肿胀、溶解。内弹性膜有中断,厚度不均匀,可见VSMC自缺口伸入内膜。各药物处理组与模型组相比,EC形态有不同程度的改善,连续性较好,胞核形态结构基本正常,线粒体数目略增多,形态结构基本正常。内弹性膜连续,厚度均匀。
6.主动脉CD40、VCAM-1、MMP-9蛋白表达水平比较
(1)CD40蛋白表达比较:正常主动脉CD40表达很少,模型组小鼠血管壁CD40蛋白表达显著升高(P＜0.001);各药物处理组CD40蛋白表达与模型组相比,有非常显著性差异(P＜0.001);辛伐他汀组和大剂量组CD40蛋白表达明显低于小剂量组(P＜0.05)。
(2)VCAM-1蛋白表达比较:与正常对照组相比,模型组小鼠血管壁VCAM-1蛋白表达显著升高(P＜0.001);与模型组相比,各药物处理组VCAM-1蛋白表达均明显下降(P＜0.01或P＜0.05);大剂量组与小剂量组比较,差异有统计学意义(P＜0.05)。
(3)MMP-9蛋白表达比较:与正常对照组相比,模型组小鼠血管壁MMP-9蛋白表达显著升高(P＜0.001);与模型组相比,各药物处理组MMP-9蛋白表达均明显下降(P＜0.001或P＜0.01或P＜0.05);大剂量组、辛伐他汀组、三七总皂苷组MMP-9蛋白表达较小剂量组明显下降(P＜0.01或P＜0.05)。
7.主动脉CD40、VCAM-1、MMP-9 mRNA表达水平比较
(1)CD40 mRNA表达比较:与正常对照组相比,模型组小鼠血管壁CD40mRNA表达显著升高(P＜0.001);各药物处理组CD40 mRNA表达较模型组均有不同程度下降,其中辛伐他汀组下降最为显著(P＜0.001),其次为三七总皂苷组和大剂量组(P＜0.001);小剂量组与大剂量组、辛伐他汀组、三七总皂苷组比较有显著差异(P＜0.001)。
(2)VCAM-1 mRNA表达比较:与正常对照组相比,模型组小鼠血管壁VCAM-1mRNA表达显著升高(P＜0.001);与模型组相比,各药物处理组VCAM-1mRNA表达均显著下降(P＜0.05或P＜0.001);大剂量组、辛伐他汀组VCAM-1表达低于三七总皂苷组和小剂量组(P＜0.001)。
(3)MMP-9 mRNA表达比较:与正常对照组相比,模型组小鼠血管壁MMP-9mRNA表达明显升高(P＜0.001);与模型组相比,各药物处理组MMP-9 mRNA表达均显著下降(P＜0.001);大剂量组、辛伐他汀组、三七总皂苷组与小剂量组比较,差异具有统计学意义(P＜0.001)。
结论
1.益肾活血胶囊能够降低apoE~(-/-)小鼠血脂水平。
2.益肾活血胶囊能够降低apoE~(-/-)小鼠血清oxLDL含量。
3.益肾活血胶囊抑制了免疫炎症因子CD40、MMP-9、VCAM-1的mRNA和蛋白表达。
4.益肾活血胶囊对AS具有免疫调节作用,能够抑制AS的进展,抑制AS斑块由稳定向不稳定发展。其机制与降低血脂,下调oxLDL及CD40、MMP-9、VCAM-1的表达有关。
第二部分益肾活血提取液对oxLDL诱导的人脐静脉内皮细胞CD40、MMP-9及VCAM-1表达的影响
目的
观察益肾活血提取液对oxLDL诱导的人脐静脉内皮细胞(Human umbilicalvein endothelial cells,HUVECs)CD40、MMP-9及VCAM-1表达的影响,从体外试验探讨益肾活血胶囊抗AS的机制。
方法
采用水煎醇沉法制备益肾活血提取液。采用胰蛋白酶液灌注消化法分离获取HUVECs并进行体外培养,取3-5代细胞进行试验。试验共分7组,空白组不予oxLDL刺激及药物处理;oxLDL刺激组(刺激组)细胞培养液中加入oxLDL(终浓度为50μg/ml)培养24 h;oxLDL+益肾活血提取液小剂量组(剂量组)、oxLDL+益肾活血提取液中剂量组(中剂量组)、oxLDL+益肾活血提取液大剂量组(大剂量组)细胞培养液中加入益肾活血提取液(终浓度分别为100μg/ml,500μg/ml,1mg/ml)培养24 h,oxLDL+辛伐他汀组(辛伐他汀组)细胞培养液中加入辛伐他汀(终浓度为10μmol/1)培养24 h,oxLDL+三七总皂苷组(三七总皂苷组)细胞培养液中加入三七总皂苷(终浓度为100μg/ml)培养24 h,后5组于药物培养24 h后加入oxLDL(终浓度为50μg/ml)继续培养24 h进行检测。采用实时定量RT-PCR检测VCAM-1的mRNA表达,免疫印迹法(Western blot)检测CD40、MMP-9的蛋白表达。试验重复3次,实验数据采用统计学方法进行分析。
结果
1.VCAM-1 mRNA表达水平的比较。与空白组相比,刺激组VCAM-1 mRNA表达明显增加(P＜0.001)。与刺激组相比,各药物处理组VCAM-1 mRNA表达均显著下降(P＜0.05或P＜0.001)。其中辛伐他汀组下降最为显著,其次是大剂量组、三七总皂苷组。辛伐他汀组、大剂量组与小剂量组相比,差异有统计学意义(P＜0.05)。
2.CD40蛋白表达水平的比较。与空白组相比,刺激组CD40蛋白表达明显增加(P＜0.001)。与刺激组相比,除小剂量组外余各药物处理组CD40蛋白表达均显著下降(P＜0.05或P＜0.01)。其中辛伐他汀组和大剂量组下降最为显著,其次是中剂量组和三七总皂苷组。
3.MMP-9蛋白表达水平的比较。与空白组相比,刺激组MMP-9的蛋白表达显著增高(P＜0.001)。与刺激组相比,各药物处理组MMP-9的蛋白表达均显著下降(P＜0.01或P＜0.001)。大剂量组、辛伐他汀组与中剂量组、三七总皂苷组和小剂量组比较,差异有统计学意义(P＜0.05,P＜0.01或P＜0.001)。
结论
1.oxLDL可诱导HUVECs高表达VCAM-1、CD40和MMP-9。
2.给予益肾活血提取液、辛伐他汀及三七总皂苷预处理后,再给予oxLDL刺激,VCAM-1的mRNA表达降低,CD40、MMP-9的蛋白表达下降。
3.随着益肾活血提取液浓度的增加,VCAM-1、CD40和MMP-9的表达呈下降趋势。提示益肾活血提取液对oxLDL诱导的免疫炎症因子的调节作用在一定范围内具有剂量依赖性。
Part 1 Effect of Yishenhuoxue capsule on regulating immune response and plaque stability in apolipoprotein E-knockout mice
Background
Atherosclerosis(AS) is a common disease which done great harm to human health.Multitude studies had indicated that both humoral and cellular immunity participate the processes of AS.So regulating the immune response will become an important method in treatment of AS.
Cell differentiation antigen 40(CD40) and CD40 ligand(CD40L) is a pair of transmembrane glycoprotein which belongs to tumor necrosis factor receptor(TNFR) and tumor necrosis factor(TNF) superfamilies.It has been shown that the costimulatory signal delivered by CD40-CD40L interactions participate the immune inflammatory response caused by oxLDL.CD40-CD40L interactions can induce endothelial cells(ECs),vascular smooth muscle cells(VSMCs) and macrophage(MΦ) expressing adhesion molecules such as vascular adhesion molecule-1(VCAM-1), intercellular adhesion molecule-1(ICAM-1),E selectin and chemotatic factors such as interleukin-8(IL-8),monocyte inflammatory protein-1(MIP-1) and monecyte chemoattractant protein-1(MCP-1),et al.These adhesion molecules and chemotatic factors can promote the leukocyte dipping into the vascular wall and the migration and proliferation of VSMCs in intima.In addition,CD40-CD40L interactions can promote ECs,VSMCs and MΦexpressing matrix metalloproteinase and tissue factor, thus can cause plaque unstability,rupture and thrombopoiesis.
Yishenhuoxue Capsule(YC),an extract of traditional Chinese medicine,has been used as an antiatherosclerotic agent for years in clinic.Previous studies have shown that YC can lower atherosclerotic index,down-regulate the expression of inflammatory factors caused by high hyperhomocysteinemia,but the effect of YC on modulating the immune response and plaque stability has not been clarified.In this study,we observed the effects of YC on serum levels of lipid and oxLDL and the expression of CD40,matrix metalloproteinase-9(MMP-9) and VCAM-1 in atherosclerotic plaques and on the plaque stability in vivo.The study coule provide scientific evidence for the YC on treatment of AS in clinic.
Aim
To investigate the possible mechanisms of YC on modulating immune response in atherosclerosis and on plaque stability through observing the plasma oxLDL level, changes of the morphology and the ultrastructure and detecting the variation of CD40, VCAM-1 and MMP-9 mRNA and protein expression and the composition of plaques.
Methods
Mail 45 apolipoprotein E-knockout(apoE~(-/-)) mice at 8 weeks of age were fed a high fat high cholesterol diet including 15%fat and 0.25%cholesterol.Mail 9 C57BL/6J mice at 8 weeks of age were fed a normal chow diet.At 20 weeks of age, apoE~(-/-) mice were randomly divided into five groups,the ApoE-KO group,the apoE~(-/-) +YSHXH group(YSHXH group),the ApoE~(-/-)+ YSHXL group(YSHXL group),the apoE~(-/-) + total panax notoginsenoside group(TPNS group) and the apoE~(-/-)+ simvastain group(Simvastain group)(n=9 for each group).C57BL/6J mice were the normal control group(Control group).Mice in apoE~(-/-)+YSHXH and apoE~(-/-)+YSHXL group were given YC at 2500 mg/kg,500 mg/kg,respectively.Mice in TPNS group were given TPNS at 60 mg/kg.And mice in Simvastain group were given Simvastain at 3.3 mg/kg.All drugs were given orally,once a day for 12 weeks.Mice in ApoE-KO and control groups were given 0.2 ml distilled water,once a day for 12 weeks.After administered for 12 weeks,all mice were sacrificed.1 ml blood was obtained from each mouse by removing eyeball.Serum levels of total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) and trigliyceride(TG) was detected by oxidation enzyme method.Serum oxLDL level was mas determined by enzyme linked immunosorbent assay(ELISA).The aorta was perfused with sterile phosphatebuffered saline(PBS) by cardiac puncture.The aortic arch about 1mm was fixed in 3% glutaraldehyde to make transmission electron microscope sample,and the surplus aortic arch and the root of aorta was fixed in 4%paraformaldehyde overnight for histology and immunohistochemistry.The thoracic aorta was rapidly frozen in liquid nitrogen for later detection of CD40,VCAM-1,MMP-9 mRNA expression.
Results
1.Comparison of serum lipid concentration
The serum TC,TG and LDL-C concentration was significantly higher in apoE~(-/-) group than that in Comal group(P<0.001).Lipid levels in YSHXH,Simvastain and TPNS group were significantly lower compared with that in apoE~(-/-) group(P<0.001). The TC,LDL-C levels were no difference between YSHXL and apoE~(-/-) group mice(P >0.05),while the TG concentration in YSHXL group was lower than that in apoE~(-/-) group(P<0.05).High dose YC was superior to low dose YC in lowering serum lipid level(P<0.001).
2.Comparison of serum oxLDL concentration
The serum oxLDL concentration in apoE~(-/-) group was significantly higher than that in Control group(P<0.001).OxLDL levels in drugs treated groups were lower than that in apoE~(-/-) group(P<0.001 or P<0.01).There were significant difference between YSHXH and YSHXL group(P<0.001).
3.Effect of YC on areas of atherosclerotic lesion in apoE~(-/-) mice
A normal aorta wall was observed in Control group mice.Atherosclerotic lesions were observed in all apoE~(-/-) mice and the lumens were narrowed in a different degree. The ratio of lesion area to vessel area in apoE~(-/-) group was higher than that in YSHXH, TPNS and Simvastain group(32.54±5.13%vs 14.16±5.18%,12.04±5.02%,14.43±4.29%,P<0.05 or P<0.01).No difference between YSHXL and apoE~(-/-) group (32.54±5.13%vs 26.22±4.74%,P>0.05).
4.Effect of YC on plaque composition in apoE~(-/-) mice
Compared with the apoE~(-/-) group,MΦand lipid infiltration in plaques were lower and VSMCs in plaques were higher in the YSHXH,Simvastain and TPNS group(P<0.001).Lipid infiltration in the YSHXL group was also lower than the apoE~(-/-) group(P<0.05),but the level of MΦand VSMCs in plaques of the two groups was no significant difference(P>0.05).These indicated that YC could promote plaque stability,and high dose YC was superior to low dose YC.
5.Ultrastructural observation of the ECs in aorta In the Control group,the ECs exhibited characteristics of monolayer pavement cells with a good succession,the cell membrane was intacted and kytoplasm was uniformity.Morphous of mitochondria is normal.The membrana elastica interna was succession and uniformity.In the apoE~(-/-) group,the ECs had obviously engorgement and the succession between ECs was interruption.ECs amotic phenomenon was observed and nuclei heterochromatin obviously increased.The mitochondria obviously swelled with the ridge swelling and dissolved.The membrana elastica interna was uneven and interrupted and VSMCs migrated into intima from the gap. However,these changes were ameliorated when treated with YC,Simvastain and TPNS.The nuclei had slight heterochromatin.The numbers of mitochondria with slight increasing,a consecutive membrana elastica intema was observed.
6.Comparison of expression of CD40,VCAM-1 and MMP-9 protein in aorta wall
(1) Comparison of expression of CD40 protein:The expression of CD40 protein in the apoE~(-/-) group was higher than that in the Control group(P<0.001).The expression of CD40 protein was significantly decreased after treatment with YC, simvastain and TPNS(P<0.001).The expression of CD40 protein was decreased significantly in the Simvastain group,the high dose YC group compared with that in the low dose group(P<0.05).
(2) Comparison of expression of VCAM-1 protein:The expression of VCAM-1 protein in the apoE~(-/-) group was higher than that in the Control group(P<0.001). After treatment with YC,simvastain and TPNS,the expression of VCAM-1 protein in each group was decreased in a different degree compared with the apoE~(-/-) group(P< 0.01 or P<0.05).The expression of VCAM-1 protein was decreased significantly in the YSHXH group compared with that in the YSHXL group(P<0.05).
(3) Comparison of expression of MMP-9 protein:The expression of MMP-9 protein in the apoE~(-/-) group was higher than that in the Control group(P<0.001).The expression of VCAM-1 protein in drugs treated groups was significantly decreased compared with the apoE~(-/-) group(P<0.001 or P<0.01 or P<0.05).The expression of VCAM-1 protein was decreased significantly in the YSHXH group,the Simvastain group and the TPNS group compared with that in the YSHXL group(P<0.01 or P< 0.05).
7.Comparison of expression of CD40,VCAM-1 and MMP-9 mRNA in aorta wall
(1) Comparison of expression of CD40 mRNA:The expression of CD40 mRNA in the apoE~(-/-) group was higher than that in the normal group(P<0.001). After treatment with YC,simvastain and TPNS,CD40 mRNA was decreased in a different degree compared with the apoE~(-/-) group,and among those treatment groups, the Simvastain group had the lowest CD40 mRNA expression(P<0.001),next was the TPNS group and the YSHXH group.The expression of CD40 mRNA in the YSHXH group,the Simvastain group and the TPNS group were lower than that in the YSHXL group(all P<0.001).
(2) Comparison expression of VCAM-1 mRNA:The expression of VCAM-1 mRNA in the apoE~(-/-) group was higher than that in the normal group(P<0.001). VCAM-1 mRNA was decreased in a different degree in drugs treatment groups compared with the apoE~(-/-) group.The expression of VCAM-1 mRNA in the YSHXH group and the Simvastain group were lower than that in the TPNS group and YSHXL group(P<0.001).
(3) Comparison expression of MMP-9 mRNA:The expression of MMP-9 mRNA in the apoE~(-/-) group was higher than that in the normal group(P<0.001).VCAM-1 mRNA was decreased significantly in drugs treatment groups compared with the apoE~(-/-) group(P<0.001).The expression of VCAM-1 mRNA in the YSHXH group, the TPNS group and the Simvastain group were lower than that in the YSHXL group (P<0.001).
Conclusion
1.YC could decrease blood lipid level in apoE~(-/-) mice.
2.YC could decrease serum oxLDL concentration in apoE~(-/-) mice.
3.YC could down-regulated the mRNA and protein expression of CD40,MMP-9 and VCAM-1.
4.YC could regulate immune response in AS,inhibit the development of AS and prevent transformation from stable plaque to unstable plaque.The mechanisms were possibly associated with the suppressive effect of YC on blood levels of lipid and oxLDL and CD40、MMP-9 and VCAM-1 expression in aortas of apoE~(-/-) mice.
Part 2 Effect of Yishenhuoxue extraction on CD40,MMP-9 and VCAM-1 expression in human umbilical vein endothelial cells induced by oxLDL
Aim
To observe the Yishenhuoxue extraction on the CD40、MMP-9 and VCAM-1 expression in human umbilical vein endothelial cells(HUVECs) induced by oxLDL. Investigate the effect of Yishenhuoxue extraction on atherosclerosis in vitro.
Methods
The Yishenhuoxue extraction was prepared by decocting with distilled water and precipitating with ethanol.The HUVECs were obtained by perfusing trypsinase liquid in umbilical cord and cultured in vitro.The 3-5 generation cells were used in the study. Cells were divided into 7 groups,the normal group,the oxLDL stimulated group (stimulate group),the oxLDL+low dose Yishenhuoxue extraction group(low dose group),the oxLDL+middle dose Yishenhuoxue extraction group(middle dose group), the oxLDL+high dose Yishenhuoxue extraction group(high dose group),the oxLDL +simvastain group(simvastain group) and the oxLDL+total panax notoginsenosidum group(TPNS group).Cells in stimulate group were admoved oxLDL at 50μg/ml into cell culture fluid and cultured for 24 h.Cells in low dose group、middle dose group and high dose group were pretreated with Yishenhuoxue extraction at 100μg/ml,500μg/ml and 1 mg/ml,respectively for 24 h,after 24 h, cells were given oxLDL at 50μg/ml and cultured for another 24 h.Cells in simvastain group and TPNS group were pretreated with simvastain at 10μmol/l and TPNS at 100μg/ml for 24 h,after 24 h,cells were given oxLDL at 50μg/ml and cultured for another 24 h.The expression of VCAM-1 mRNA was detected by real time RT-PCR, the expression of CD40,MMP-9 protein was detected by western blot.All experimental data analyzed by statistics.
Results
1.Comparison of VCAM-1 mRNA:The expression of VCAM-1 mRNA in stimulate group was significantly higher than that in the normal group(P<0.001). The VCAM-1 mRNA expression was significantly lower in lose dose group,middle dose group,high dose group,simvastain group and TPNS group compared with that in stimulate group(P<0.05 or P<0.001).Among those five groups,the simvastain group had the lowest expression of VCAM-1 mRNA,next was the high dose group and TPNS group.The VCAM-1 mRNA was significantly lower in simvastain group and high dose group compared with low dose group(P<0.05).
2.Comparison of CD40 protein:The expression of CD40 protein in stimulate group was significantly higher than that in the normal group(P<0.001).HUVECs treated with Yishenhuoxue extraction,simvastain and TPNS for 24 hours indicated a decrease of CD40 protein expression.CD40 protein was the lowest in simvastain and high dose group,high dose Yishenhuoxue extraction and middle dose Yishenhuoxue extraction was superior to low dose Yishenhuoxue extraction(P<0.05 or P<0.01).
3.Comparison of MMP-9 protein:The expression of MMP-9 protein in stimulate group was significantly higher than that in the normal group(P<0.001).The MMP-9 protein expression was significantly lower in lose dose group,middle dose group, high dose group,simvastain group and TPNS group compared with that in stimulate group(P<0.01 or P<0.001).The MMP-9 protein was significantly lower in high dose group and simvastain group compared with that in middle dose group,TPNS group and low dose group(P<0.05,P<0.01 or P<0.001).
Conclusion
1.oxLDL could induce the expression of CD40、VCAM-1 and MMP-9 in HUVECs at a high level.
2.Pretreatment with Yishenhuoxue extraction,simvastain and TPNS could down-regnlate the expression of CD40、VCAM-1 and MMP-9 in HUVECs induced by oxLDL.
3.The expression of CD40、VCAM-1 and MMP-9 in HUVECs had a descending tendency along with the increasing concentration of Yishenhuoxue extraction.It indicated that the effect of Yishenhuoxue extraction on modulating immune inflammatory factors in HUVECs induced by oxLDL was in a dose dependent manner.
引文
1. Libby P. Inflammation in atherosclerosis[J]. Nature, 2002, 420 (6917): 868- 874.
2. Palinski W, Tangirala RK, Miller E, et al . Increased autoantibody titers against epitopes of oxidized LDL in LDL receptor-deficient mice with increased atherosclerosis[J].Arterioscler Thromb Vasc Biol, 1995,15: 1569-1576.
3. Hasson GK. Regulation of immune mechanisms in atheroscleresis[J]. Ann NY Acad Sci, 2001, 947: 157-165.
4. Suzuki H, Kurihara Y, Takeya M, et al. A role for macrophages cavenger receptors in atherescleresis and susceptibility to infection[J]. Nature, 1997, 386 (6622): 292—296.
5. Sliverstein RL, Febbraio M. CD36 and atheroscleresis[J]. Curt Opin Lipid, 2000, 11:483—489.
6. Febbraio M, Podmz EA, Smith JD, et al. Targeted disruption of the class B scarenger receptor CD36 protects against atherosclerotic lesion development in mice[J]. J Clin Invest, 2000,105: 1049 —1056.
7. Shmaoka T, Kume N, Minami M, et al. Molecular cloning of a novel scavenger receptorfor oxidized LDL, SR—PSOX on macrophages[J]. J Biol Chem, 2000, 275: 40663—40666.
8. Hakkinen T, Karkola K, Yla-Herttuala S. Macrophages, smooth muscle cells, endothelial cells, and T-cells express CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions: colocalization with epitopes of oxidized low- density lipoprotein, scavenger receptor, and CD16 (FcγRIII) [J]. Virchows Archiv:an international journal of pathology, 2000,437: 396-405.
9. Li D, Liu L, Chen H, et al. LOX-1, an oxidized LDL endothelial receptor, induces CD40/CD40L signaling in human coronary artery endothelial cells[J]. Arteriosclerosis,Thrombosis, and Vascular biology, 2003, 23: 816-821.
10. Schonbeck U, Gerdes N, Varo N, et al. Oxidized low-density lipoprotein augments and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors limit CD40 and CD40L expression in human vascular cells[J]. Circulation, 2002, 106: 2888-2893.
11.Shen LH,Zhou L,Wang BY,et al.Oxidized low-density lipoprotein induces differentiation of RAW264.7 murine macrophage cell line into dendritic-like cells[J].Atherosclerosis,2008,199:257-264.
12.Cybulsky MI,Iiyama K,Li H,et al.A major role for VCAM-1,but not ICAM-1,in early atherosclerosis[J].J Clin Invest,2001,107:1255-1262.
13.Kotowicz K,Dixon GL,Klein NJ,et al.Biological function of CD40 on human endothelial cells:costimulation with CD40 ligand and interleukin-4 selectively induces expression of vascular cell adhesion molecule-1 and P-selectin resulting in preferential adhesion of lymphocytes[J],Immunology,2000,100:441-448.
14.Reinders ME,Sho M,Robertson SW,et al.Proangiogenic function of CD40ligand-CD40 interactions[J].J Immunol,2003,171:1534-1541.
15.Sch(o|¨)nbeck U,Libby P.CD40 signaling and plaque instability[J].Circulation research,2001,89:1092-1103.
16.Lutgens E,Cleutjens KB,Heeneman S,et al.Both early and delayed anti-CD40L antibody treatment induces a stable plaque phenotype[J].PNAS,2000,97:7464-7469.
17.张继东,罗玮敏,杨瑞雪,等.益肾活血胶囊治疗冠心病高同型半胱氨酸血症的临床研究[J].南京中医药大学学报,2005,21(5):277-280.
18.刘粉叶,张继东,胡连海,等.益肾活血胶囊对同型半胱氨酸致兔动脉粥样硬化MCP-1 mRNA表达及NF-κB活化的影响[J].山东大学学报(医学版,2008,46(4):353-356.
19.来丽萍,张继东,李丽珍,等.益肾活血胶囊对高同型半胱氨酸致兔血管平滑肌细胞核因子活化及血管细胞黏附分子表达的影响[J].上海中医药大学学报,2008,22(1):45-48.
20.Livak KJ,Schmittqen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J].Methods,2001,25(4):402-408.
21.Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Res,2001,29(9):e45.
22.杨永宗.动脉粥样硬化实验研究模型百年回溯[J].中国动脉硬化杂志,2008,16(12):925-927.
23.Nakashima Y,Plump AS,Raines EW,et al.ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree[J].Arteriosclerosis and Thrombosis,1994,14:133-140.
24.Piedrahita JA,Zhang SH,Hagaman JR,et al.Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E[J].Proc Natl Acad Sci USA,1992,89:4471-4475.
25.Plump AS,Smith JD,Hayek T,et al.Severe hypercholesterolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells[J].Cell,1992,71:343-353.
26.胡琴,张运.基因工程小鼠动脉粥样硬化模型的研究进展[J].中国动脉硬化杂志.2006,14(8):725-727.
27.Ross R.Atherosclerosis -an inflammatory disease[J].N Eng J Med,1999,340(2):115-126.
28.Hansson GK.Immune Mechanisms in Atherosclerosis[J].Arterioscler Thromb Vasc Biol.2001,21:1876-1890.
29.Steinberg D.Low density lipoprotein oxidation and its pathobiological significance[J].J Biol Chem,1997,272(34):20963-20966.
30.Palinski W,H(o|¨)rkk(o|¨) S,Miller E,et al.Cloning monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E deficient mice.Demonstration of epitopes of oxidized low density lipoprotein in human plasma[J].The Journal of Clinical Investigation,1996,98:800-814.
31.Stemme S,Faber B,Holm J,et al.T lymphocytes from human atherosclerotic plaques recognize oxidized low density lipoprotein[J].Proc Natl Acad Sci USA,1995,92:3893-3897.
32.Wu R,Huang YH,Elinder LS,et al.Lysophosphatidylcholine is involved in the antigenicity of oxidized LDL[J].Arterioscler Thromb Vasc Biol,1998,18(4):626-630.
33.Friedman P,Horkko S,Steinberg D,et al.Correlation of antiphospholipid antibody recognition with the structure of synthetic oxidized phospholipids. Importance of Schiff base formation and aldol concentration [J]. J Biol Chem, 2002, 277(9): 7010—7020.
34. Lee TS, Yen HC, Pan CC, et al. The role of interleukin 12 in the development of atherosclerosis in apoE-deficient mice[J]. Arteriosclerosis, Thrombosis, and Vascular biology, 1999, 19: 734-742.
35. Vickers KC, Maguire CT, Wolfert R, et al. Relationship of lipoprotein-associated phospholipase A2 and oxidized low-density lipoprotein in carotid atherosclerosis[J]. J Lipid Res. 2009, Apr 9.
36. Verreth W, De Keyzer D, Davey PC, et al. Rosuvastatin restores superoxide dismutase expression and inhibits accumulation of oxidized LDL in the aortic arch of obese dyslipidemic mice[J]. Br J Pharmacol, 2007, 151 (3):347-55.
37. Virella G, Koskinen S, Krings G, et al. Immunochemical characterization of purified human oxidized low-density lipoprotein antibodies[J]. Clin Immunol, 2000, 95(2): 135—144.
38. Shaw PX, Horkko S, Tsimikas S, et al. Human-derived anti-oxidized LDL autoantibody blocks uptake of oxidized LDL by macrophages and localizes to atherosclerotic lesions in vivo[J]. Arterioscler Thromb Vasc Biol, 2001, 8: 1333-1339.
39. Tsimikas S, Bergmark C, Reinaldo W, et al. Temporal increases in plasma markers of oxidized low density lipoprotein strongly reflect the presence of acute coronary syndrome[J]. J Am Coll Cardio, 2003, 3: 360—370.
40. Liang KW, Huang JL, Kao CH, et al. Significantly higher levels of oxidized LDL autoantibody in coronary artery disease patients [J]. Chin Med J (Tai pei), 2000, 63(2): 101 — 106.
41. van Puijvelde GH, Hauer AD, de Vos P, et al. Induction of oral tolerance to oxidized low-density lipoprotein ameliorates atherosclerosis [J]. Circulation, 2006, 114(18): 1968-76.
42. Locksley R, Killeen N, Lenardo M. The TNF and TNF receptor super2 families: integrating mammalian biology [J]. Cell, 2001, 104: 487 - 501.
43.Gerritse K,Laman JD,Noelle RJ,et al.CD40-CD40 ligand interactions in experimental allergic encephalomyelitis andmultip le sclerosis[J].Proc Natl Acad Sci USA,1996,93:2499-2504.
44.Mach F,Schonbeck U,Sukhova GK,et al.Functional CD40 ligand is expressed on human vascular endothelial cells,smooth muscle cells,and macrophages:Implicationfor CD40-CD40 ligand signaling in atherosclerosis[J].J Proc Natl Acad Sci USA,1997,94:1931-936.
45.Garlichs CD,John S,Eskafi S,et al Upregulation of CD40 and CD40ligand(CD154) in patients with moderate hypercholesterolemia[J].Circulation,2001,104:2395-2400.
46.Melter M,Reinders MEJ,Masayuki S,et al.Ligation of CD40 induces the expression of vascular endothelial growth factor by endothelial cells and monocytes and promotes angiogenesis in vivo[J].Blood,2000,96:3801-3808.
47.Hansson GK,Lippy P,Sch(o|¨)nbeck U,et al.Innate and adaptive immunity in the pathogenesis of atherosclerosis[J].Circ Res,2002,91(4):281-291.
48.Peng X,Kasaran A,Warmerdam PAM,et al.Accessory signaling by CD40 for T cell activation:inductionof Th1 and Th2 cytokines and synergy with interleukin-12 for interferon- gamma production[J].J Eur J Immunol,1996,26:1621-1627.
49.Ruedl C,Bachmann MF,Kopf M.The antigen dose determines T helper subset development by regulation of CD40 ligand[J].European Journal of Immunology,2000,30:2056-2064.
50.Hansson GK,Zhou X,Tornquist E,et al.The role of adaptive immunity in atherosclerosis[J].Ann NY Acad Sci.2000,902(5):53-62.
51.Math F,Schonbeck U,Sukhova GK,et al.Reduction of atherosclerosis in mice by inhibition of CD40 signalling[J].Nature,1998,394(6 689):200-203.
52.杨瑞雪,张继东,罗玮敏,等.益肾活血胶囊对不稳定型心绞痛患者Hcy及ET、TNF-α、NO的影响[J].上海中医药杂志,2006,40(1):22-24.
53.Swerlick RA,Lee KH,Sepp NT,et al.Regulation of vascular cell adhesion molecule-1 in human dermal microvascular endothelial cells[J].J Immunol,1992, 149(2):698-705.
54.Cybulsky MI,Gimbrone MA Jr.Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis[J].Science,1991,251:788-791.
55.Sakai A,Kume N,Nishi E,et al.P-selectin and vascular cell adhesion molecule-1are focally expression in aortas of hypercholesterolemic rabbits before intimal accumulation of macrophages and T lymphocytes[J].Arterioscler Thromb and Vase Biol,1997,17(2):310-316.
56.Bocher BS,Luscinskas FW,Gimbrone MA,et al.Adhesion of human basophils,eosinophils,and neutrophils to interleukin-12 activated human vascular endothelial cells:contributions of endothelial cell adhesion molecules[J].Exp Med,1991,173(6):1553-1557.
57.Barks JL,Mc Quillan JJ,Iademareo MF.TNF-alpha and IL-4 synergistically increase vascular cell adhesion molecule-1 expression in cultured vascular smooth muscle cells[J].J Immunol,1997,159(9):4532-4538.
58.Thorne SA,Abbot SE,Stevens CR,et al.Modified low density lipoprotein and cytokines mediate monocyte adhesion to smooth muscle cells[J].Atherosclerosis,1996,127(2):167-176.
59.Blankenberg S,Barbaux S,Tiret L.Adhesion molecules and atherosclerosis[J].Atherosclerosis,2003,170:191-203.
60.Davies MJ,Gordon JL,Gearing AJ,et al.The expression of the adhesion molecules ICAM-1,VCAM-1,PECAM,and E-selectin in human atherosclerosis[J].J Pathol,1993,171:223-229.
61.Liuba P,Pesonen E,Paakkari I,et al.Co-infection with Chlamydia pneumoniae and Helicobacter pylori results in vascular endothelial dysfunction and enhanced VCAM-1 expression in apoE-knockout mice[J].J Vasc Res,2003,40:115-122.
62.饶丹,姜红,曾秋棠.黏附分子细胞间黏附分子-1/E-选择素与冠心病[J].心血管病学进展,2005,26(3):278-281.
63.Singh RJ,Mason JC,Lidington EA,et al.Cytokine stimulated vascular cell adhesion molecule-1(VCAM-1) eetodomain release is regulated by TMP-3[J]. Cardiovasc Res,2005,67(1):39-49.
64.Li H,Cybulsky MI,Gimbrone MA Jr,etal.Inducible expression of vascular cell adhesion molecule-1 by vascula smooth muscle cells in vitro and within rabbit atheroma[J].Am J Pathol,1993,143:1551-1559.
65.Dansky HM,Barlow CB,Lominska C,et al.Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage[J].Arterioscler Thromb Vasc Biol,2001,21:1662-1667.
66.Barringhaus KG,Phillips JW,Thatte JS,et al.Alpha4 beta1 integrin(VLA-4)blockade attenuates both early and late leukocyte recruitment and neointimal growth following carotid injury in apolipoprotein E~(-/-) mice[J].J Vasc Res,2004,41:252-260.
67.王梅芳,杨林花.MMP-9与冠状动脉粥样硬化[J].临床心血管病杂志,2004,20(11):699-701.
68.Vine N,Powell JT.Metalloproteinases in degenerative aortic disease[J].Clin Sci,1991,81(2):233-239.
69.Xu,XP,Meisel SR,Ong JM,et al.Oxidized low-density lipoprotein regulates matrix metalloproteinase-9 and its tissue inhibitor in human monocyte-derived macrophages[J].Circulation,1999,99:993-998.
70.Sch(o|¨)nbeck U,Mach F,Sukhova G K,et al.Regulation of matrix metalloproteinase expression in human vascular smooth muscle cells by T lymphocytes:a role for CD40 signaling in plaque rupture?[J].Circ Res,1997,81:448-454.
71.郭爱桃,韦立新.基质金属蛋白酶1与冠状动脉粥样硬化斑块破裂的关系[J].中华病理学杂志,2000,29(3):263-266.
72.Sullivan JL.Macrophage iron,hepcidin,and atherosclerotic plaque stability[J].Exp Biol Med(Maywood).2007,232(8):1014-1020.
73.Boren J,Olin K,Lee I,et al.Identification of the principal proteoglycan-binding site in LDL.A single-point mutation in apo-B100 severely affects proteoglycan interaction without affecting LDL receptor binding[J].J Clin Invest,1998,101: 2658-2664.
74. Shiomi M, Ito T, Tsukada T, et al. Reduction of serum cholesterol levels alters lesional composition of atherosclerotic plaques. Effect of pravastatin sodium on atherosclerosis in mature WHHL rabbits[J]. Arterioscler Thromb Vasc Biol, 1995, 15: 1938-1944.
75. Aikawa M, Rabkin E, Okada Y, et al. Lipid lowering by diet reduces matrix metalloproteinase activity and increases collagen content of rabbit atheroma: a potential mechanism of lesion stabilization[J]. Circulation, 1998, 97: 2433-2444.
76. Schachinger V, Zeiher AM. Atherosclerosis-associated endothelial dysfunction[J]. Z Kardiol, 2000, 89 Suppl 9: IX/70-74.
77. Riazy M, Chen JH, Steinbrecher UP. VEGF secretion by macrophages is stimulated by lipid and protein components of OxLDL via PI3-kinase and PKCzeta activation and is independent of OxLDL uptake[J]. Atherosclerosis, 2009, 204(1): 47-54.
78. Lippy P, Geng YJ, Sukhova GK, et al. Molecular determinants of atherosclerotic plaque vulnerability[J]. Ann N Y Acad Sci, 1997, 811:134-145.
79. Ikeda U, Shimada K. Matrix metalloproteinases and coronary artery diseases[J]. Clinical Cardiology, 2003,26: 55-59.
80. Sukhova GK, Schonbeck U, Rabkin E, et al. Evidence for increased collagenolysis by interstitial collagenases-1 and -3 in vulnerable human atheromatous plaques[J]. Circulation, 1999, 99: 2503-2509.
81. Aikawa M, Voglic SJ, Sugiyama S, et al. Dietary lipid lowering reduces tissue factor expression in rabbit atheroma[J]. Circulation, 1999,100:1215-1222.
82. Zhou L, Stordeur P, de Lavareille A, et al. CD40 engagement on endothelial cells promotes tissue factor-dependent procoagulant activity[J]. Thromb Haemost, 1998, 79: 1025-1028.
83. Miller DL, Yaron R, Yellin MJ. CD40L-CD40 interactions regulate endothelial cell surface tissue factor and thrombomodulin expression[J]. J Leukoc Biol, 1998, 63: 373-379.
84. Slupsky JR, Kalbas M, Willuweit A, et al. Activated platelets induce tissue factor expression on human umbilical vein endothelial cells by ligation of CD40[J].Thromb Haemost,1998,80:1008-1014.
85.Sch(o|¨)nbeck U,Mach F,Sukhova GK,Herman M,Graber P,Kehry MR,Libby P.CD40 ligation induces tissue factor expression in human vascular smooth muscle cells[J].Am J Pathol,2000,156:7-14.
86.Blankenberg S,Rupprecht HJ,Bickel C,et al.Circulating cell adhesion molecules and death in patients with coronary artery disease[J].Circulation,2001,104:1336-1342.
87.Koga M,Kai H,Yasukawa H,et al.Postnatal blocking of interferon-gamma function prevented atherosclerotic plaque formation in apolipoprotein E-knockout mice[J].Hypertens Res,2007,30:259-267.
88.Dansky HM,Barlow CB,Lominska C,et al.Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage[J].Arterioscler Thromb Vase Biol,2001,21:1662-1667.
89.Zhang B,Ye S,Herrmann SM,et al.Functional polymorphism in the regulatory region of gelatinase B gene in relation to severity of coronary atherosclerosis[J].Circulation,1999,99(14):1788-1794.
90.Bennett MR.Apoptosis of vascular smooth muscle cells in vascular remodeling and atherosclerotic plaque rupture[J].Cardiovasc Res,1999,41(2):361-368.
91.Brown DL,Hibbs MS,Kearney M,et al.Identification of 92-kD gelatinase in human coronary atherosclerotic lesions.Association of active enzyme synthesis with unstable angina[J].Circulation,1995,91(8):2125-2131.
92.詹小萍,楼建国,金晓莹,等.益肾活血方对家兔血瘀模型纤溶系统作用的实验研究[J].中国中药杂志,2004,29(5):440-443.
93.孙可丰,胡晓春,胡晓晨,等.抗凝降压胶囊治疗肾虚血瘀型高血压病的临床研究[J].中医药信息,1998,(2):27-28.
94.王旭,陈金锭.益肾化瘀法对2型糖尿病血液流变学、SOD、LPO水平及血管病变的影响[J].南京中医药大学学报(自然科学版),2001,17(1):20-22.
95.黄妙珍.益糖宁颗粒剂改善2型糖尿病胰岛素抵抗的临床研究[J].中华实用中西医杂志,2003,3(16):626-628.
96.李敏.补肾活血方的调脂作用探要[J].中医药学刊,2003,21(5):667-668.
97.陆小青.益肾活血通脉汤治疗颈动脉粥样硬化的临床观察.吉林中医药,2004,24(5):12-13.
98.赵玉霞,刘运芳,刘艳,燕芳芳,于会明.祛瘀消斑胶囊对易损斑块的稳定作用.中医杂志,2006,47(11):856-858.
99.赵玉霞,刘运芳,刘艳.脑心通胶囊对不稳定斑块兔TLR-2和TLR-4炎性信号转导因子的影响.中西医结合心脑血管病杂志,2006,4(12):1071-1073.
100.钱彦丛,宇文萍.枸杞子的化学成分及药理研究新进展[J].中医药学报,2000,(4):33-35
101.刘秀英,胡怡秀,臧雪冰,等.枸杞油胶丸对高脂大鼠血脂水平的影响.中国自然医学杂志,2004,6(3):134-136.
102.姜怡邓,曹军,董泉洲,等.PKC和MMPs在枸杞籽油抗动脉粥样硬化中的作用和机制[J].华西药学杂志,2007,22(1):009-012.
103.马灵筠,陈群力,杨五彪,等.枸杞多糖对动脉粥样硬化模型兔血脂、脂质过氧化、NO和C-反应蛋白的影响[J].郑州大学学报(医学版),2005,40(2):328-330.
104.马灵筠,孙海双,陈群力,等.枸杞多糖对动脉粥样硬化内皮细胞功能及炎症反应的影响[J].江苏中医药,2006,27(5):61-63.
105.杜光.枸杞子免疫药理研究及临床应用.湖北中医杂志,2005,27(2):54-55.
106.陶大勇.枸杞多糖的免疫调节作用.安徽农业科学,2007,35(22):6816-6818.
107.赵娜,郭治昕,赵雪,等.丹参的化学成分与药理作用[J].国外医药·植物药分册,2007,22(4):155-159.
108.田季雨,陈建宗,顾宜,等.复方丹参滴丸对家兔血脂水平和颈动脉粥样硬化斑块的影响[J].中国临床康复,2004,8(9):1708-1709.
109.王南,蔡海江,朱宇,等.丹参素对牛主动脉平滑肌细胞氧化修饰LDL的抑制作用[J].南京医科大学学报,1994,14(4):529-531.
110.成之福,王毅华,王家安,等.复方丹参注射液对冠心病患者一氧化氮、内 皮素-1的影响[J].中国医院药学杂志,2006,26(10):1275.
111.许涛,喻莉,郑智,等.丹参对实验性动脉粥样硬化形成的预防[J].临床心血管病杂志,2005,21(1):54-55.
112.姜开余,顾振纶,阮长耿.丹参素对CD11b、P-Selectin、ICAM-1、VCAM-1、E-Selectin表达的影响[J].中国药理学通报,2000,16(6):682-685.
113.刘虹彬,温进坤.丹参对血管平滑肌细胞基质金属蛋白酶和骨桥蛋白基因表达及细胞增殖的影响[J].中国中西医结合杂志,2002,22(10):764-766.
114.孙学刚,贾钰华,陈育鹩.定心方及丹参酮对血小板膜粘附分子表达的影响[J].山东中医药大学学报,2001,25(1):61-63.
115.姜开余,阮长耿,陈振纶,等.丹参酮Ⅱ-A磺酸钠对内皮细胞和血小板体外表达粘附分子的作用[J].中国药理学报,1998,19(1):47-50.
116.费丽萍,李进禧,郑道声,等.丹参酮Ⅱ-A磺酸钠对红细胞内游离钙浓度的影响[J].微循环杂志,1997,7(4):17-19.
117.崔伊梅.黄芪及其有效成分黄芪苷Ⅳ在心血管药理作用的研究进展[J].天津医科大学学报,2005,11(4):656-659.
118.汪德清,丁保国,Tomas G Neil,等.黄芪总黄酮对动脉粥样硬化早期形成的影响[J].中国药理学通报,2003,19(6):637-640.
119.吴勇,石显水,王石顺,等.黄芪多糖对动脉粥样硬化内皮细胞的保护作用[J].中国临床康复,2005,9(23):238-240.
120.黄进宇,单江,程刚,等.黄芪注射液体外对大鼠血管平滑肌细胞基质金属蛋白酶-2活性的抑制作用[J].中医杂志,2003,44(2):140-142.
121.刘宽芝,李静波,吕海莉,等.黄芪、三七总皂苷对2型糖尿病大血管病变患者MMP-9的影响[J].中国中药杂志,2004,29(3):264-266.
122.杨志红,赵晓龙,陈凤玲,等.黄芪多糖对THP-1巨噬细胞源性泡沫细胞功能的影响[J].中国分子心脏病学杂志,2007,7(3):138-142.
123.王冬梅,朱玮,张存莉,等.黄精化学成分及其生物活性[J].西北林学院学报,2006,21(2):142-145.
124.李友元,邓洪波,向大雄,等.黄精多糖的降血脂及抗动脉粥样硬化作用[J].中国动脉硬化杂志,2005,13(4):429-431.
125.吉中强,高晓昕,宋鲁卿,等.63种调脂中药抗人血小板聚集的实验研究[J].山东中医杂志,1998,17(8):365-367.
126.张萍,刘丹,李友元.黄精多糖对动脉粥样硬化家兔血清IL-6及CRP的影响[J].医学临床研究,2006,23(7):1100-1101.
127.李友元,张萍,邓洪波,等.动脉粥样硬化家兔VCAM-1表达及黄精多糖对其表达的影响[J].医学临床研究,2005,22(9):1287-1288.
128.陈金水,陈松苍,王耀华.黄精口服液对剧烈运动小鼠氧自由基代谢及肌酸激酶影响[J].中医研究,1996,9(4):6-8.
129.李秋怡,干国平,刘焱文.川芎的化学成分及药理研究进展[J].时珍国医国药,2006;17(7):1298-1299.
130.梅家俊,蔡大勇,赵婷秀,等.川芎防治动脉粥样硬化症机制的实验研究[J].湖北中医学院学报,2004,6(2):5-7.
131.阮秋蓉,瞿智玲,朱敏,等.川芎嗪对血管壁细胞血管细胞粘附分子1和单核细胞趋化蛋白1表达的作用及可能机制[J].中国动脉硬化杂志,2006,14(7):560-564.
132.刘红.川芎嗪对动脉粥样硬化家兔血清一氧化氮、血浆内皮素和脂质过氧化物的影响[J].湖北民族学院学报(医学版),2000,17(4):4-5.
133.吴文元.川芎嗪治疗周围动脉粥样硬化闭塞症疗效观察[J].实用中医药杂志,2002,18(8):5-6.
134.Sheu JR,Kan YC,Hung WC,et al.The antiplatelet activity of tetramethylpyrazine is mediated through activation of NO synthase[J].Life Sci,2000,67(8):937-947.
135.潘贺,刚宏林,苏云明.中药水蛭的活性成分及药理作用研究概况[J].中医药信息,2006,23(1):20-22.
136.李凤文,刘晓颖,张立石,等.水蛭及其复方对实验性动脉粥样硬化家兔主动脉内皮素基因表达的影响[J].中医杂志,1999,40(1):50-51
137.张玉娟,田景振,郭之平.水蛭研究概述[J].食品与药品,2005,7(6):9-11.
138.朱智明,刘伊丽,万华印,等.血小板膜GPⅡb/Ⅲa单抗及水蛭素对损伤动脉内膜血小板沉积量的影响[J].心肺血管病杂志,1998,17(1):39-41.
139.王敏,崔连群,张承俊,等.凝血酶诱导血管内皮细胞生长因子的表达及水蛭素的抑制作用[J].中国新药杂志,2004,13(3):226-230.
140.刘京生,苗智慧,董力,等.水蛭抗肿瘤作用的实验研究[J].时珍国医国药,2001,12(10):884-885.
141.屠婕红,余菁,陈伟光.瓜蒌的化学成分和药理作用研究概况[J].中国药师,2004,7(7):562-563.
142.贺立勍.瓜蒌、薤白降脂作用的析因研究[J].湖南中医药导报,2002,8(4):205-207.
143.凌宏,鲁翔,董传仁,等.瓜蒌、心得安对血小板体外聚集和TXA2合成的影响[J].湖北医学院学报,1988,9(2):138.
144.陈燕,赵辉,谢锐.大黄的药理及临床研究近况[J].西南国防医药,2005,15(2):178-180.
145.徐在品,卢占军,陈眷华,等.大黄醇提液抗家兔实验性高脂血症及脂肪肝的实验研究[J].中国应用生理学杂志,2007,23(3):375-379.
146.Abe I,Seki T,Noguchi H,et al.Galloylesters from rhubarb are potent inhibitors of squalene epoxidase,a key enzyme in cholesterol biosynthesis[J].Planta Med,2000,66(8):753-756.
147.Liu Y,Yan F,Liu Y,Zhang C,Yu H,Zhang Y,Zhao Y.Aqueous extract of rhubarb stabilizes vulnerable atherosclerotic plaques due to depression of inflammation and lipid accumulation.Phytother Res.2008,22(7):935-942.
148.Kumar A,Dhawan S,Aggarwal BB.Emodin(3-methyl-1,6,8-trihydroxyanthraquinone)inhibits TNF-induced NF-κB activation,IkB degradation,and expression of cell surface adhesion proteins in human vascular endothelial cells[J].Oncogene,1998,17(7):913-918.
149.Li HL,Chen HL,Li H,et al.Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW 264.7macrophages[J].Int J Mol Med,2005,16(1):41-47.
150.文川,徐浩,黄启福,等.活血中药对ApoE基因缺陷小鼠血脂及动脉粥样硬化斑块炎症反应的影响[J].中国中西医结合杂志,2005,25(4):345-349.
151.黄政德,蒋孟良,易延逵,等.酒制丹参、大黄对大鼠血小板功能及抗凝血作用的研究[J].中成药,2001,23(5):341-342.
152.熊辉岩,谭大风.大黄属三种植物不同部分提取物清除羟基自由基的体外实验研究[J].食品科学,2003,24(1):128-130.
153.兰进,杨世林,郑玉权,等.黄连的研究进展[J].中草药,2001,32(12):1139-1141.
154.勾祥辉.黄连素治疗高血脂59例[J].人民军医,1995,(2):35.
155.王秀杰.黄连的药理研究及现代应用[J].China Pharmacist,2003,6(6):370.
156.何国厚,艾志兵,刘勇,等.小檗碱对兔颈动脉粥样硬化中内膜增生和巨噬细胞趋化作用的影响[J].中风与神经疾病杂志,2006,23(1):94-96.
157.段浩,张宏考,刘继军,等.小檗碱对兔颈动脉损伤后白介素-6等的影响[J].医药论坛杂志,2006,27(10):6-9.
158.周岚,何国厚,李承晏,等.小檗碱对颈动脉粥样硬化家兔血管紧张素的影响[J].卒中与神经疾病,2006,13(4):212-214,218.
159.陈其明,谢明智.小檗碱对正常小鼠血糖调节的影响[J].药学学报,1987,22(3):161-165.
1.Ross R.Atherosclerosis -an inflammatory disease[J].N Eng J Med,1999,340(2):115-126.
2.DeM meyer GR,Herman AG.Vascular endothelial dysfunction[J].Prog Cardiovasc Dis,1997,39(4):325-342.
3.Lusis AJ.Atherosclerosis.Nature,2000,407(2):233-241.
4.刘志忠,陈绍良,刘玲玲,等氧化低密度脂蛋白抗体水平与冠状动脉再狭窄的相关性:独立与年龄、性别、体质量指数、高血压、高血脂及吸烟史的危险因子.中国临床康复[J].2004,8(3):446-447.
5.Sch(o|¨)nbeck U,Gerdes N,Varo N,et al.Oxidized low-density lipoprotein augments and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors limit CD40 and CD40L expression in human vascular cells[J].Circulation,2002,106:2888-2893.
6.Li D,Liu L,Chen H,et al.LOX-1,an oxidized LDL endothelial receptor,induces CD40/CD40L signaling in human coronary artery endothelial cells[J].Arteriosclerosis,Thrombosis,and Vascular biology,2003,23:816-821.
7.姜立清,胡大一,庞文跃.阿托伐他汀对氧化低密度脂蛋白诱导的内皮基质金属蛋白酶-1表达的影响[J].中国老年学杂志.2005,25(11):1358-1360.
8.Romanic AM,Harrison SM,Bao W,et al.Myocardial protection from ischemia/reperfusion injury by targeted deletion of matrix metalloproteinase-9[J].Cardiovas Res,2002,54(3):549-558.
9.朱陵群,庞鹤,王硕仁,牛福玲,崔巍。活血注射液对oxLDL诱导的单核-内皮细胞黏附作用的影响[J].北京中医药大学学报,2007,30(3):173-176.
10.顾耘.软脉煎对血管内皮细胞动脉硬化模型MCP-1的影响[J].实用老年医学,2005,19(5):265-267.
11.Livak KJ,Schmittqen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J].Methods,2001,25(4):402-408.
12.Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Res,2001,29(9):e45.
13. Ehara S, Ueda M, Naruko T, et al. Elevated levels of oxidized low density lipoprotein show a positive relationship with the severity of acute coronary syndromes. circulation[J], 2001,103:1995-1960.
14. Toshima S, Hasegama A, Kurabayashi M, et al.Circulating oxidized lowdensity lipoprotein levels: a biochemical risk marker for coronary heart disease[J]. Arterioscler Thromb Vasc Biol, 2000,20: 2243-2247.
15. Holvoet P, Van Cleemput J, Collen D, et al. oxidized low dendity lipoprotein is aprognostic marker of transplant associated coronary disease[J]. Arterioscler Thromb Vasc Biol, 2000,20: 698-702.
16. Carr A, Mccall MR, Frei B. Oxidation of LDL by myeloperoxidase and reactive nitrogen species: reation pathways and antioxidation protection[J]. Arterioscler Thromb Vasc Biol, 2000,20: 1716-1723.
17. Jay W Heinedce. Oxidants and antioxidants in the pathogenesis for oxidized low density lipoprotein hypothesis[J]. Atherosclerosis, 1998, 141: 11-15.
18. Mehta JL, Li DY, Chen HJ, et al. Inhibition of LOX-1 by statins may relate to up regulation of eNOS[J]. Biochem Biophys Res Conunu, 2001,289: 857 - 861.
19. Akira T, Yan H, Maria F, et al. Expression of adhesion molecules by human endothelial cells exposed to oxidized low density lipoprotein influences of degree of oxidation and location of oxidized LDL[J]. Atherosclerosis, 2001, (154): 79-86.
20. Amberger A, Maczek C, Jürgens G, et al. Co-expression of ICAM-1, VCAM-1, ELAM-1 and Hsp60 in human arterial and venous endothelial cells in response to cytokines and oxidized low-density lipoproteins[J]. Cell Stress Chaperones, 1997, 2 (2): 94-103.
21. Li D, Liu L, Chen H, et al. Lox-1 mediates oxidized low density lipoprotein -induced expression of matrix metalloproteinases in human coronary artery endothelial cells[J]. Circulation, 2003,107: 612-617.
22. Sawanura T, Kume N, Aoyama T, et al. A novel receptor for oxidized low density lipoprotein[J]. Nature, 1997,386(6): 73-77.
23. Okak K, Sawamnra T, Kikuta S, et al. Lectin-like oxidized low-density lipoprotein receptor 1 mediates phagocytosis of aged/apoptotic cells in endothelial cells[J].Proc Natl Acad,Sci USA,1998,95:9535-9540.
24.Li D,Chen H,Sawamura T,et al.Statins modulate oxjdized low-density lipoprotein-mediated adhesion molecule expression in human coronary artery endothelial cells:role of Lox-1[J].J Pharmacol Exp Ther,2002,302:601-605.
25.杨瑞雪,张继东,罗玮敏,等.益肾活血胶囊对不稳定型心绞痛患者Hcy及ET、TNF-α、NO的影响[J].上海中医药杂志2006,40(1):22-24.
26.Hakkinen T,Karkola K,Yla-Herttuala S.Macrophages,smooth muscle cells,endothelial cells,and T-cells express CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions:colocalization with epitopes of oxidized low-density lipoprotein,scavenger receptor,and CD16(FcγRⅢ)[J].Virchows Archiv:an international journal of pathology,2000,437:396-405.
27.Aukrust P,Fredrik M,Ueland T,et al Enhanced level of soluble and membrane-bound CD40 ligand in patients with unstable angina[J].Circulation,1999,100:614-620.
28.Hakonarson H,Kim C,Whelan R,et al.Bidirectional activation between human airway smooth muscle cells and T lymphocytes:role in induction of altered airway responsiveness[J].J Immunol,2001,166:293.
29.Garlichs CD,Eskafi S,Raaz D,et al.Patients with acute coronary syndromes express enhanced CD40 ligand/CD154 on platelets[J].Heart,2001,86(6):649-655.
30.Mach F,Sch(o|¨)nbeck U,Rosalind P,et al.T lymphocytes induce endothelial cell matrix metalloproteinase expression by a CD40L-dependent mechanism[J].AmJ Pathol,1999,154:229-238.
31.Viallard JF,Solanilla A,Gauthier B,et al.Increased soluble and platelet-associated CD40 ligand in essential thrombocythemia and reactive thrombocytosis[J].Blood,2002,99(7):2612-2614.
32.Mach F,Sch(o|¨)nbeck U,Sukhova GK,et al.Reduction of atherosclerosis in mice by inhibition of CD40 signalling[J].Nature,1998,394(6689):200-203.
33.Sch(o|¨)nbeck U,Sukhova GK,Shimizu K,et al.Inhibition of CD40 signaling limits evolution of established atherosclerosis in mice[J].Proc Natl Acad Sci USA,2000,97(13):7458-7463.
34.饶丹,姜红,曾秋棠.黏附分子细胞间黏附分子-1/E-选择素与冠心病[J].心血管病学进展,2005,26(3):278-281.
35.Singh RJ,Mason JC,Lidington EA,et al.Cytokine stimulated vascular cell adhesion molecule-1(VCAM-1) ectodomain release is regulated by TMP-3[J].Cardiovasc Res,2005,67(1):39-49.
36.Davies MJ,Gordon JL,Gearing AJ,et al.The expression of the adhesion molecules ICAM-1,VCAM-1,PECAM,and E-selectin in human atherosclerosis[J].J Pathol,1993,171(3):223-229.
37.Wu YT,Gao YC,Zhou SM.Dysfunction of vascular endothelium and coronary disease[J].China Journal of Modern Medicin.2003,5(13):47-54.
38.Li H,Cybulsky MI,Gimbrone MA Jr,etal.Inducible expression of vascular cell adhesion molecule-1 by vascula smooth muscle cells in vitro and within rabbit atheroma[J].Am J Pathol,1993,143:1551-1559.
39.Blankenberg S,Rupprecht HJ,Bickel C,et al.Circulating cell adhesion molecules and death in patients with coronary artery disease[J].Circulation,2001,104:1336-1342.
40.Mulvihill NT,Foley JB,Murphy R,et al.Evidence of prolonged inflammation in unstable angina and non-Q wave myocardial infarction[J].J Am Coll Cardiol,2000,36(4):1210-1216.
41.Dansky HM,Barlow CB,Lominska C,et al.Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage[J].Arterioscler Thromb Vasc Biol,2001,21:1662-1667.
42.Barringhaus KG,Phillips JW,Thatte JS,et al.Alpha4beta1 integrin(VLA-4)blockade attenuates both early and late leukocyte recruitment and neointimal growth following carotid injury in apolipoprotein E-/- mice[J].J Vasc Res,2004,41:252-260.
43.Brown DL,Hibbs MS,Kearney M,et al.Identification of 92-kD gelatinase in human coronary atherosclerotic lesions:association of active enzyme synthesis with unstable angina[J].Circulation,1995,91:2125-2131.
44.Alvarez B,Ruiz C,Chacon P,et al.Serum values of metalloproteinaseo2 and metalloproteinase-9 as related to unstable plaque and inflammatory cells in patients with greater than 70%carotid artery stenosis[J].Journal of Vascular Surgery,2004,40:469-475.
45.Loftus IM,Naylor AR,Goodall S,et al.Increased matrix metalloproteinase-9activity in unstable carotid plaques.A potential role in acute plaque disruption[J].Stroke,2000,31:40-47.
46.Longo GM,Xiong W,Greiner TC,et al.Matrix metalloproteinases 2 and 9 work in concert to produce aortic aneurysms[J].The Journal of Clinical Investigation,2002,110:613-614.
47.Romanic AM,Harrison SM,Bao W,et al.Myocardial protection from ischemia/reperfusion injury by targeted deletion of matrix metalloproteinase-9[J].Cardiovas Res,2002,54(3):549-558.
48.姜怡邓,曹军,董泉洲,等.PKC和MMPs在枸杞籽油抗动脉粥样硬化中的作用和机制[J].华西药学杂志,2007,22(1):009-012.
49.马灵筠,陈群力,杨五彪,等.枸杞多糖对动脉粥样硬化模型兔血脂、脂质过氧化、NO和C-反应蛋白的影响[J].郑州大学学报(医学版),2005,40(2):328-330.
50.马灵筠,孙海双,陈群力,等.枸杞多糖对动脉粥样硬化内皮细胞功能及炎症反应的影响[J].江苏中医药,2006,27(5):61-63.
51.王南,蔡海江,朱宇,等.丹参素对牛主动脉平滑肌细胞氧化修饰LDL的抑制作用[J].南京医科大学学报,1994,14(4):529-531.
52.成之福,王毅华,王家安,等.复方丹参注射液对冠心病患者一氧化氮、内皮素-1的影响[J].中国医院药学杂志,2006,26(10):1275.
53.许冬青,张旭,蔡文峰,等.丹参药物血清保护血管内皮细胞分子机制研究[J].中成药,2005,27(2):229-230.
54.崔广智,金树梅,赵桂峰,等.丹参酮ⅡA对TNFα诱导的ECV304细胞 NF-k B、kB-α表达及粘附分子ICAM-1、VCAM-1mRNA表达的影响[J].中国药理学通报,2007,23(12):1671-1675.
55.李永胜,王进,王照华,等.丹参酮ⅡA对主动脉内皮细胞功能损伤的保护机制[J].中国急救医学,2007,27(8):720-723.
56.陈海明,叶攀.丹参酮ⅡA对血管内皮细胞氧化应激损伤的保护作用[J].中药材,2008,31(4):569-572.
57.修春,宓穗卿,王宁生.隐丹参酮对TNF-α诱导的人脐静脉内皮细胞粘附分子ICAM-1表达的作用[J].中药药理与临床,2009,25(2):16-19.
58.杨顺清,洪小苏.丹参多酚酸盐对电离辐射损伤血管内皮细胞影响的初步研究[J].中国误诊学杂志2008,8(36):8829-8831.
59.何胜虎,燕建锋,袁彬,等.丹参多酚酸盐对高糖诱导的人脐静脉内皮细胞损伤的保护作用[J].中国动脉硬化杂志,2008,,16(12):948-952.
60.杨亚安,唐丽华,蒋小岗,等.丹参醇提取物对培养内皮细胞T-AOC、LPO、NO、SOD变化的影响[J].苏州大学学报,2005,25(6):963-965.
61.叶泽兵,汪凡军,张刚庆,等.黄芪注射液对TNF-α诱导人脐静脉内皮细胞炎症因子的影响及其分子机制[J].南方医科大学学报,2009,29(6):1137-1140.
62.叶泽兵,汪凡军,张刚庆,等.黄芪注射液对脂多糖诱导内皮细胞损伤保护作用的实验研究[J].实用医学杂志,2009,29(5):1368-1370.
63.范可,陈培红,吴青.黄芪注射液对高糖所致血管内皮细胞损伤的保护作用[J].中国中西医结合临床,2008,20(4):295-298.
64.陈立新,朱海燕,朱陵群,等.黄芪多糖对人心脏微血管内皮细胞增殖及再灌注后内皮细胞与中性粒细胞黏附的影响[J].中国组织工程研究与临床康复,2007,11(27):5382-5386.
65.杨五彪,陈群力,马灵筠,等.黄芪多糖对兔动脉粥样硬化血管内皮细胞功能的影响[J].陕西医学杂志,2005,34(8):914-918.
66.樊懿,吴大正,胡之璧.黄芪甲甙对组胺引起的血管内皮细胞单层通透性增高的作用[J].中国中医基础医学杂志,2001,7(2):36-38.
67.陈金水,陈松苍,王耀华.黄精口服液对剧烈运动小鼠氧自由基代谢及肌酸激酶影响[J].中医研究,1996,9(4):6-8.
68.李友元,邓洪波,向大雄,等.黄精多糖的降血脂及抗动脉粥样硬化作用[J].中国动脉硬化杂志,2005,13(4):429-431.
69.张萍,刘丹,李友元.黄精多糖对动脉粥样硬化家兔血清IL-6及CRP的影响[J].医学临床研究,2006,23(7):1100-1101.
70.李友元,张萍,邓洪波,等.动脉粥样硬化家兔VCAM-1表达及黄精多糖对其表达的影响[J].医学临床研究,2005,22(9):1287-1288.
71.崔磊,王守训,刘新泳.川芎嗪在抗动脉粥样硬化中的作用和机制[J].医学综述,2008,14(22):3491-3493.
72.梁德,姚珍松,杨志东.川芎嗪对家兔血管内皮细胞修复的影响[J].广州中医药大学学报,2001,18(2):155-158.
73.高雪,杨洁,元英进.盐酸川芎嗪对血管紧张素Ⅱ损伤内皮细胞的保护作用[J].细胞生物学杂志,2007,29:895-900.
74.朱际平,袁冬平.川芎嗪对缺氧损伤的人脐静脉内皮细胞分泌一氧化氮的影响[J].实用临床医药杂志,2009,13(7):39-41.
75.李翠乔.天然药物川芎嗪对脂质过氧化内皮细胞的保护作用研究[J].科学技术与工程,2008,8(17):4789-4792.
76.熊石龙,志斌,王前,等.川芎嗪对肿瘤坏死因子致血管内皮细胞组织因子表达的影响[J].中国现代医学杂志,2007,17(19):2330-2333.
77.李凤文,刘晓颖,张立石,等.水蛭及其复方对实验性动脉粥样硬化家兔主动脉内皮素基因表达的影响[J].中医杂志,1999,40(1):50-51.
78.李宁,赵霞,张文高.水蛭微粉对不稳定性心绞痛血管内皮保护作用的观察[J].中国误诊学杂志,2009,9(7):1539.
79.朱智明,刘伊丽,万华印,等.血小板膜GPⅡb/Ⅲa单抗及水蛭素对损伤动脉内膜血小板沉积量的影响[J].心肺血管病杂志,1998,17(1):39-41.
80.高春林,贾俊亚,范生尧.水蛭素对凝血酶所致脑微血管内皮细胞损伤的保护作用[J].中国临床药学杂志,2007,16(2):73-77.
81.王敏,崔连群,张承俊,等.凝血酶诱导血管内皮细胞生长因子的表达及水蛭素的抑制作用[J].中国新药杂志,2004,13(3):226-230.
82.王敏,崔连群,王晓军,韩秋霞,刘鲁宁.水蛭素对凝血酶诱导血管内皮细胞 与中性粒细胞表达黏附分子的影响[J].中国新药杂志,2005,14(8):989-992.
83.芦丽莉,王冬梅.瓜蒌皮提取液对实验性高脂血症大鼠血清NO,SOD,MDA 的影响[J].北华大学学报(自然科学版).2008,9(5):423-425.
84.熊辉岩,谭大风.大黄属三种植物不同部分提取物清除羟基自由基的体外实验研究[J].食品科学,2003,24(1):128-130.
85.Kumar A,Dhawan S,Aggarwal BB.Emodin(3-methyl-1,6,8-trihydroxyanthraquinone)inhibits TNF-induced NF-κB activation,IkB degradation,and expression of cell surface adhesion proteins in human vascular endothelial cells[J].Oncogene,1998,17(7):913-918.
86.Li HL,Chen HL,Li H,et al.Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW 264.7 macrophages[J].Int J Mol Med,2005,16(1):41-47.
87.王为民,虞燕琴,钱令波,舒强,刘广义,林茹,杨军。大黄素对大鼠体外血管内皮一氧化氮cGMP信号途径的影响[J].中国中西医结合杂志,2006,26(7):636.639.
88.何国厚,艾志兵,刘勇,等.小檗碱对兔颈动脉粥样硬化中内膜增生和巨噬细胞趋化作用的影响[J].中风与神经疾病杂志,2006,23(1):94-96.
89.段浩,张宏考,刘继军,等.小檗碱对兔颈动脉损伤后白介素-6等的影响[J].医药论坛杂志,2006,27(10):6-9.
90.周岚,何国厚,李承晏,等.小檗碱对颈动脉粥样硬化家兔血管紧张素的影响[J].卒中与神经疾病,2006,13(4):212-214,218.
91.尹文清,陈琪,佘志刚.盐酸小蘖碱对LPS诱导的人脐静脉内皮细胞与粒性白细胞粘附的影响[J].广东药学院学报,2002,18(2):122-124.
Alvarez B., Ruiz C., Chacon P., Alvarez-Sabin J., Matas M., 2004. Serum values of metalloproteinase-2 and metalloproteinase-9 as related to unstable plaque and inflammatory cells in patients with greater than 70% carotid artery stenosis. Journal of Vascular Surgery 40, 469-475.
Brown D.L., Hibbs M.S., Kearney M., Loushin C., Isner J.M., 1995. Identification of 92-kD gelatinase in human coronary atherosclerotic lesions: association of active enzyme synthesis with unstable angina. Circulation 91, 2125-2131.
Hakkinen T., Karkola K., Yla-Herttuala S., 2000. Macrophages, smooth muscle cells, endothelial cells, and T-cells express CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions: colocalization with epitopes of oxidized low- density lipoprotein, scavenger receptor, and CD16 (FcγRIII). Virchows Archiv: an international journal of pathology 437, 396-405.
He X., Li X., Li S., He C., 2007. Preventive effect of total saponins of panax notoginseng on experimental atherosclerosis in rabbits. China Pharmacy 18,408-410.
He X., Zhang L., Chen P., Li S., 2004. Effect of TPNS on hyperlipoidemia rats. Joural of Yunnan Traditional Chinese Medicine 25, 32-33.
Huang K., Deng X., Luo J., 1997. TPNS enhances relaxation of pulmonary artery in chronic hypoxia rats induced by NO. Guangzhou Medicine 18,54-56.
Ikeda U., Shimada K., 2003. Matrix metalloproteinases and coronary artery diseases. Clinical Cardiology 26, 55-59.
Ji W., Gong BQ., 2007. Hypolipidemic effects and mechanisms of Panax notoginseng on lipid profile in hyperlipidemic rats. Journal of Ethnopharmacology 113, 318-324.
Koyuncu FM., Ozbilgin K., Kuscu NK., Inan S., Vatansever S., Ceylan E., 2006. The effect of oestradiol and neta on immunohistochemical staining of iNOS and eNOS in coronary arteries of ovariectomized rats. Histol Histopathol 21,367-371.
Lee T.S., Yen H.C., Pan C.C., Chau L.Y., 1999. The role of interleukin 12 in the development of atherosclerosis in apoE-deficient mice. Arteriosclerosis, Thrombosis, and Vascular biology 19, 734-742.
Li D., Liu L., Chen H., Sawamura T., Mehta J.L., 2003. LOX-1, an oxidized LDL endothelial receptor, induces CD40/CD40L signaling in human coronary artery endothelial cells. Arteriosclerosis, Thrombosis, and Vascular biology 23, 816-821.
Li X., Wang G., Sun J., Hao H., Xiong Y., Yan B., 2007. Pharmacokinetic and absolute bioavailability study of total Panax notoginsenoside, a typical multiple constituent traditional Chinese medicine (TCM) in rats. Biological and Pharmaceutical Bulletin 30, 847-851.
Ling Y., 1984. Science of Chinese Materia Medica, Fifth ed. Science and Technology Publishing Company, Shanghai, China, pp. 144-145.
Liu Y., Li X., 2005. Effect of pannax notoginseng total saponin on inflammatory immune factors in atherosclerosis. Chinese Traditional and Herbal Drags 36,728-730.
Livak K.J., Schmittgen, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-ΔΔCT) method. Methods 25,402-408.
Loftus I.M., Naylor A.R., Goodall S., Crowther M., Jones L., Bell P.R., Thompson M.M., 2000. Increased matrix metalloproteinase-9 activity in unstable carotid plaques. A potential role in acute plaque disruption. Stroke 31,40-47.
Longo G.M., Xiong W., Greiner T.C., Zhao Y., Fiotti N., Baxter B.T., 2002. Matrix metalloproteinases 2 and 9 work in concert to produce aortic aneurysms. The Journal of Clinical Investigation 110, 613-614.
Lu P., Chen H., 2004. Effects of notoginseng folium saponins on regulating blood lipid. Chinese Journal of Biochemical Pharmaceuti 25,235-236.
Mach F., Schonbeck U., Sukhova G.K., Atkinson E., Libby P., 1998. Reduction of atherosclerosis in mice by inhibition of CD40 signaling. Nature 394,200-203.
Mach F., Schonbeck U., Sukhova G.K., Bourcier T., Bonnefoy J.Y., Pober J.S., Libby P., 1997. Functional CD40 ligand is expressed on human vascular endothelial cells, smooth muscle cells, and macrophages: implications for CD40-CD40 ligand signaling in atherosclerosis. Proceedings of the National Academy of Sciences of the United States of America 94, 1931-1936.
Miida T., Nakamura Y., Mezaki T., Hanyu O., Maruyama S., Horikawa Y., Izawa S., Yamada Y., Matsui H., Okada M., 2002. LDL-cholesterol and HDL-cholesterol concentrations decrease during the day, Annals of Clinical Biochemistry 39, 241-249.
Nakashima Y., Plump A.S., Raines E.W., Breslow J.L., Ross R., 1994. ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree. Arteriosclerosis and Thrombosis 14, 133-140.
Ross R., 1999. Atherosclerosis-an inflammation diease. The New England Journal of Medicine 340, 115-126.
Ruedl C., Bachmann M.F., Kopf M., 2000. The antigen dose determines T helper subset development by regulation of CD40 ligand. European Journal of Immunology 30, 2056-2064.
Schonbeck U., Gerdes N., Varo N., Reynolds R.S., Horton D.B., Bavendiek U., Robbie L., Ganz P., Kinlay S., Libby P., 2002. Oxidized low-density lipoprotein augments and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors limit CD40 and CD40L expression in human vascular cells. Circulation 106,2888-2893.
Schonbeck U., Libby P., 2001. CD40 signaling and plaque instability. Circulation Research 89, 1092-1103.
Schonbeck U., Mach F., Sukhova G.K., Murphy C., Bonnefoy J.Y., Fabunmi R.P., Libby P., 1997. Regulation of matrix metalloproteinase expression in human vascular smooth muscle cells by T lymphocytes: a role for CD40 signaling in plaque rupture? Circulation Research 81,48-454.
Shen L.H., Zhou L., Wang B.Y., Pu J., Hu L.H., Chai D.J., Wang L., Zeng J.Z., He B., 2008. Oxidized low-density lipoprotein induces differentiation of RAW264.7 murine macrophage cell line into dendritic-like cells. Atherosclerosis 199,257-264.
Su N., He M., Zhong B., Zhang S., 2002. Effects of Xuesaitong on blood lipid and plasma viscosity in hyperlipemia patients. Journal of Hunan College of Traditional Chinese Medicine 22,65-66.
Tarchalski J., Guzik P., Wysocki H., 2003. Correlation between the extent of coronary atherosclerosis and lipid profile. Molecular and Cellular Biochemistry 246,25-30.
Xu X.P., Meisel S.R., Ong J.M., Kaul S., Cercek B., Rajavashisth T.B., Sharifi B., Shah P.K., 1999. Oxidized low-density lipoprotein regulates matrix metalloproteinase-9 and its tissue inhibitor in human monocyte-derived macrophages. Circulation 99, 993-998.
Yang J., Qin C., 2003. Effects of Complex Danshen formula, Danshen and Sanqi on the function of platelet. China Journal of Experimental Traditional Medical Formulae 9, 59-62.
Yuan Z., Li X., Li S., Fan J., 2006. Effects of PNS on stability of atherosclerotic plaque and initial investigation for its mechanism. Chinese Journal of Nature Medicine 4,62-64.
Yuan Z., Li X., Li S., Zhang H., Liu Y., 2006. Mechanism approach on TPNS improving stability of atherosclerotic plaque in rabbits. Chinese Traditional and Herbal Drugs 37,741-743.
Zhang S.H., Reddick R.L., Piedrahita J.A., Maeda N., 1992. Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E. Science 258,468-471.
Zhou X., Chen M., Li X., Bai G., Zhou Q., Jiang S., 2007. An approach to etiological factors and pathogenesis of dyslipidemia in TCM. Chinese Archives of Traditional Chinese Medicine 25, 197-198.
Aikawa M, Rabkin E, Okada Y, Voglic SJ, Clinton SK, Brinckerhoff CE, Sukhova GK, Libby P. 1998. Lipid lowering by diet reduces matrix metalloproteinase activity and increase collagen content of rabbit atheroma: a potential mechanism of lesion stabilization. Circulation 97: 2433-2444.
Barringhaus KG, Phillips JW, Thatte JS, Sanders JM, Czarnik AC, Bennett DK, Ley KF, Sarembock IJ. 2004. Alpha4betal integrin (VLA-4) blockade attenuates both early and late leukocyte recruitment and neointimal growth following carotid injury in apolipoprotein E-/- mice. J Vasc Res 41: 252-260.
Blankenberg S, Barbaux S, Tiret L. 2003. Adhesion molecules and atherosclerosis. Atherosclerosis 170: 191-203.
Blankenberg S, Rupprecht HJ, Bickel C, Peetz D, Hafner G, Tiret L, Meyer J. 2001. Circulating cell adhesion molecules and death in patients with coronary artery disease. Circulation 104: 1336-1342.
Couffinhal T, Duplaa C, Moreau C, Lamaziere JM, Bonnet J. 1994. Regulation of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 in human vascular smooth muscle cells. Circ Res 74: 225-234.
Cybulsky MI, Gimbrone MA Jr. 1991. Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis. Science 251: 788 -791.
Dansky HM, Barlow CB, Lominska C, Sikes JL, Kao C, Weinsaft J, Cybulsky MI, Smith JD. 2001. Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage. Arterioscler Thromb Vasc Biol 21: 1662-1667.
Davies MJ, Gordon JL, Gearing AJ, Pigott R, Woolf N, Katz D, Kyriakopoulos A. 1993. The expression of the adhesion molecules ICAM-1, VCAM-1, PECAM, and E-selectin in human atherosclerosis. J Pathol 171: 223-229.
Dong GJ, Liu JG, Shi DZ, Wang YY, Luan LJ, Cheng YY. 2007. Effects of water extractives of a qi and blood regulating prescription on early atherosclerosis of apolipoprotein E-deficient mice. Zhong Xi Yi Jie He Xue Bao 5:45-49.
Falk E. 1992. Why do plaques rupture? Circulation 86: 30-42.
Gutstein DE, Fuster V. 1999. Pathophysiology and clinical significance of atherosclerotic plaque rupture. Cardiovasc Res 41: 323-33.
Jawien J, Nastalek P, Korbut R. 2004. Mouse models of experimental atherosclerosis. J Physiol Pharmacol 55: 503-517.
Khan BV, Parthasarathy SS, Alexander RW, Medford RM. 1995. Modified low density lipoprotein and its constituents augment cytokine-activated vascular cell adhesion molecule-1 gene expression in human vascular endothelial cells. J Clin Invest 95: 1262-1270.
Koga M, Kai H, Yasukawa H, Kato S, Yamamoto T, Kawai Y, Kusaba K, Seki Y, Kai M, Egashira K, Kataoka Y, Imaizumi T. 2007. Postnatal blocking of interferon-gamma function prevented atherosclerotic plaque formation in apolipoprotein E-knockout mice. Hypertens Res 30: 259-267.
Li H, Cybulsky MI, Gimbrone MA Jr, Libby P. 1993. An atherogenic diet rapidly induces VCAM-1, a cytokine-regulatable mononuclear leukocyte adhesion molecule, in rabbit aortic endothelium. Arterioscler Thromb 13: 197-204.
Li X, Liu Q, Zhou T, Zhao S, Zhou S. 2008. PAPP-A: a possible pathogenic link to the instability of atherosclerotic plaque. Med Hypotheses 70: 597-599.
Libby P. 2001. Current concepts of the pathogenesis of the acute coronary syndromes. Circulation 104: 365-372.
Libby P, Sukhova G, Lee RT, Galis ZS. 1995. Cytokines regulate vascular functions related to stability of the atherosclerotic plaque. J Cardiovasc Pharmacol 25: S9-12.
Liuba P, Pesonen E, Paakkari I, Batra S, Andersen L, Forslid A, Yla-Herttuala S, Persson K, Wadstrom T, Wang X, Laurini R. 2003. Co-infection with Chlamydia pneumoniae and Helicobacter pylori results in vascular endothelial dysfunction and enhanced VCAM-1 expression in apoE-knockout mice. J Vasc Res 40: 115-122.
Lutters BC, Leeuwenburgh MA, Appeldoorn CC, Molenaar TJ, Van Berkel TJ, Biessen EA. 2004. Blocking endothelial adhesion molecules: a potential therapeutic strategy to combat atherogenesis. Curr Opin Lipidol 15: 545-552.
Mulvihill NT, Foley JB, Murphy R, Crean P, Walsh M. 2000. Evidence of prolonged inflammation in unstable angina and non-Q wave myocardial infarction. J Am Coll Cardiol 36: 1210-1216.
Piedrahita JA, Zhang SH, Hagaman JR, Oliver PM, Maeda N. 1992. Generation of mice carrying a mutant apolipoprotein E gene inactivated by gene targeting in embryonic stem cells. PNAS 89: 4471-4475.
Ramana KV, Bhatnagar A, Srivastava SK. 2004. Inhibition of aldose reductase attenuates TNF-alpha-induced expression of adhesion molecules in endothelial cells. FASEB J 18: 1209 -1218.
Reiner Z, Tedeschi-Reiner E. 2001. New information on the pathophysiology of atherosclerosis. Lijec Vjesn 123: 26-31.
Ross R. 1999. Atherosclerosis: an inflammatory disease. N Engl J Med. 340: 115-126.
Scalia R, Appel JZ 3rd, Lefer AM. 1998. Leukocyte-endothelium interaction during the early stages of hypercholesterolemia in the rabbit: role of P-selectin, ICAM-1, and VCAM-1. Arterioscler Thromb Vasc Biol 18: 1093-1100.
Wang B, Zhang JD, Feng JB, Yin HQ, Liu FY, Wang Y. 2007. Effect of traditional Chinese medicine Qin-Dan-Jiang-Ya-Tang on remodeled vascular phenotype and osteopontin in spontaneous hypertensive rats. J Ethnopharmacol 110: 176-182.
Yang RuiXue, Zhang JiDong, Luo WeiMin, Guo RuiChen, Zhu YuanYuan, Wang ShiHua. 2006. Effects of "YiShen HuoXue Capsule" on Hcy, ET, TNF-α and NO in Patients with Unstable Angina. Shanghai Journal of Traditional Chinese Medicine 40: 22-24.
Yokozawa T, Cho EJ, Sasaki S, Satoh A, Okamoto T, Sei Y. 2006. The Protective Role of Chinese Prescription Kangen-karyu Extract on Diet-Induced Hypercholesterolemia in Rats. Biol. Pharm. Bull 29: 760-765.
2. Palinski W, Tangirala RK, Miller E, et al . Increased autoantibody titers against epitopes of oxidized LDL in LDL receptor-deficient mice with increased atherosclerosis[J].Arterioscler Thromb Vasc Biol, 1995,15: 1569-1576.
3. Hasson GK. Regulation of immune mechanisms in atheroscleresis[J]. Ann NY Acad Sci, 2001, 947: 157-165.
4. Suzuki H, Kurihara Y, Takeya M, et al. A role for macrophages cavenger receptors in atherescleresis and susceptibility to infection[J]. Nature, 1997, 386 (6622): 292—296.
5. Sliverstein RL, Febbraio M. CD36 and atheroscleresis[J]. Curt Opin Lipid, 2000, 11:483—489.
6. Febbraio M, Podmz EA, Smith JD, et al. Targeted disruption of the class B scarenger receptor CD36 protects against atherosclerotic lesion development in mice[J]. J Clin Invest, 2000,105: 1049 —1056.
7. Shmaoka T, Kume N, Minami M, et al. Molecular cloning of a novel scavenger receptorfor oxidized LDL, SR—PSOX on macrophages[J]. J Biol Chem, 2000, 275: 40663—40666.
8. Hakkinen T, Karkola K, Yla-Herttuala S. Macrophages, smooth muscle cells, endothelial cells, and T-cells express CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions: colocalization with epitopes of oxidized low- density lipoprotein, scavenger receptor, and CD16 (FcγRIII) [J]. Virchows Archiv:an international journal of pathology, 2000,437: 396-405.
9. Li D, Liu L, Chen H, et al. LOX-1, an oxidized LDL endothelial receptor, induces CD40/CD40L signaling in human coronary artery endothelial cells[J]. Arteriosclerosis,Thrombosis, and Vascular biology, 2003, 23: 816-821.
10. Schonbeck U, Gerdes N, Varo N, et al. Oxidized low-density lipoprotein augments and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors limit CD40 and CD40L expression in human vascular cells[J]. Circulation, 2002, 106: 2888-2893.
11.Shen LH,Zhou L,Wang BY,et al.Oxidized low-density lipoprotein induces differentiation of RAW264.7 murine macrophage cell line into dendritic-like cells[J].Atherosclerosis,2008,199:257-264.
12.Cybulsky MI,Iiyama K,Li H,et al.A major role for VCAM-1,but not ICAM-1,in early atherosclerosis[J].J Clin Invest,2001,107:1255-1262.
13.Kotowicz K,Dixon GL,Klein NJ,et al.Biological function of CD40 on human endothelial cells:costimulation with CD40 ligand and interleukin-4 selectively induces expression of vascular cell adhesion molecule-1 and P-selectin resulting in preferential adhesion of lymphocytes[J],Immunology,2000,100:441-448.
14.Reinders ME,Sho M,Robertson SW,et al.Proangiogenic function of CD40ligand-CD40 interactions[J].J Immunol,2003,171:1534-1541.
15.Sch(o|¨)nbeck U,Libby P.CD40 signaling and plaque instability[J].Circulation research,2001,89:1092-1103.
16.Lutgens E,Cleutjens KB,Heeneman S,et al.Both early and delayed anti-CD40L antibody treatment induces a stable plaque phenotype[J].PNAS,2000,97:7464-7469.
17.张继东,罗玮敏,杨瑞雪,等.益肾活血胶囊治疗冠心病高同型半胱氨酸血症的临床研究[J].南京中医药大学学报,2005,21(5):277-280.
18.刘粉叶,张继东,胡连海,等.益肾活血胶囊对同型半胱氨酸致兔动脉粥样硬化MCP-1 mRNA表达及NF-κB活化的影响[J].山东大学学报(医学版,2008,46(4):353-356.
19.来丽萍,张继东,李丽珍,等.益肾活血胶囊对高同型半胱氨酸致兔血管平滑肌细胞核因子活化及血管细胞黏附分子表达的影响[J].上海中医药大学学报,2008,22(1):45-48.
20.Livak KJ,Schmittqen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J].Methods,2001,25(4):402-408.
21.Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Res,2001,29(9):e45.
22.杨永宗.动脉粥样硬化实验研究模型百年回溯[J].中国动脉硬化杂志,2008,16(12):925-927.
23.Nakashima Y,Plump AS,Raines EW,et al.ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree[J].Arteriosclerosis and Thrombosis,1994,14:133-140.
24.Piedrahita JA,Zhang SH,Hagaman JR,et al.Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E[J].Proc Natl Acad Sci USA,1992,89:4471-4475.
25.Plump AS,Smith JD,Hayek T,et al.Severe hypercholesterolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells[J].Cell,1992,71:343-353.
26.胡琴,张运.基因工程小鼠动脉粥样硬化模型的研究进展[J].中国动脉硬化杂志.2006,14(8):725-727.
27.Ross R.Atherosclerosis -an inflammatory disease[J].N Eng J Med,1999,340(2):115-126.
28.Hansson GK.Immune Mechanisms in Atherosclerosis[J].Arterioscler Thromb Vasc Biol.2001,21:1876-1890.
29.Steinberg D.Low density lipoprotein oxidation and its pathobiological significance[J].J Biol Chem,1997,272(34):20963-20966.
30.Palinski W,H(o|¨)rkk(o|¨) S,Miller E,et al.Cloning monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E deficient mice.Demonstration of epitopes of oxidized low density lipoprotein in human plasma[J].The Journal of Clinical Investigation,1996,98:800-814.
31.Stemme S,Faber B,Holm J,et al.T lymphocytes from human atherosclerotic plaques recognize oxidized low density lipoprotein[J].Proc Natl Acad Sci USA,1995,92:3893-3897.
32.Wu R,Huang YH,Elinder LS,et al.Lysophosphatidylcholine is involved in the antigenicity of oxidized LDL[J].Arterioscler Thromb Vasc Biol,1998,18(4):626-630.
33.Friedman P,Horkko S,Steinberg D,et al.Correlation of antiphospholipid antibody recognition with the structure of synthetic oxidized phospholipids. Importance of Schiff base formation and aldol concentration [J]. J Biol Chem, 2002, 277(9): 7010—7020.
34. Lee TS, Yen HC, Pan CC, et al. The role of interleukin 12 in the development of atherosclerosis in apoE-deficient mice[J]. Arteriosclerosis, Thrombosis, and Vascular biology, 1999, 19: 734-742.
35. Vickers KC, Maguire CT, Wolfert R, et al. Relationship of lipoprotein-associated phospholipase A2 and oxidized low-density lipoprotein in carotid atherosclerosis[J]. J Lipid Res. 2009, Apr 9.
36. Verreth W, De Keyzer D, Davey PC, et al. Rosuvastatin restores superoxide dismutase expression and inhibits accumulation of oxidized LDL in the aortic arch of obese dyslipidemic mice[J]. Br J Pharmacol, 2007, 151 (3):347-55.
37. Virella G, Koskinen S, Krings G, et al. Immunochemical characterization of purified human oxidized low-density lipoprotein antibodies[J]. Clin Immunol, 2000, 95(2): 135—144.
38. Shaw PX, Horkko S, Tsimikas S, et al. Human-derived anti-oxidized LDL autoantibody blocks uptake of oxidized LDL by macrophages and localizes to atherosclerotic lesions in vivo[J]. Arterioscler Thromb Vasc Biol, 2001, 8: 1333-1339.
39. Tsimikas S, Bergmark C, Reinaldo W, et al. Temporal increases in plasma markers of oxidized low density lipoprotein strongly reflect the presence of acute coronary syndrome[J]. J Am Coll Cardio, 2003, 3: 360—370.
40. Liang KW, Huang JL, Kao CH, et al. Significantly higher levels of oxidized LDL autoantibody in coronary artery disease patients [J]. Chin Med J (Tai pei), 2000, 63(2): 101 — 106.
41. van Puijvelde GH, Hauer AD, de Vos P, et al. Induction of oral tolerance to oxidized low-density lipoprotein ameliorates atherosclerosis [J]. Circulation, 2006, 114(18): 1968-76.
42. Locksley R, Killeen N, Lenardo M. The TNF and TNF receptor super2 families: integrating mammalian biology [J]. Cell, 2001, 104: 487 - 501.
43.Gerritse K,Laman JD,Noelle RJ,et al.CD40-CD40 ligand interactions in experimental allergic encephalomyelitis andmultip le sclerosis[J].Proc Natl Acad Sci USA,1996,93:2499-2504.
44.Mach F,Schonbeck U,Sukhova GK,et al.Functional CD40 ligand is expressed on human vascular endothelial cells,smooth muscle cells,and macrophages:Implicationfor CD40-CD40 ligand signaling in atherosclerosis[J].J Proc Natl Acad Sci USA,1997,94:1931-936.
45.Garlichs CD,John S,Eskafi S,et al Upregulation of CD40 and CD40ligand(CD154) in patients with moderate hypercholesterolemia[J].Circulation,2001,104:2395-2400.
46.Melter M,Reinders MEJ,Masayuki S,et al.Ligation of CD40 induces the expression of vascular endothelial growth factor by endothelial cells and monocytes and promotes angiogenesis in vivo[J].Blood,2000,96:3801-3808.
47.Hansson GK,Lippy P,Sch(o|¨)nbeck U,et al.Innate and adaptive immunity in the pathogenesis of atherosclerosis[J].Circ Res,2002,91(4):281-291.
48.Peng X,Kasaran A,Warmerdam PAM,et al.Accessory signaling by CD40 for T cell activation:inductionof Th1 and Th2 cytokines and synergy with interleukin-12 for interferon- gamma production[J].J Eur J Immunol,1996,26:1621-1627.
49.Ruedl C,Bachmann MF,Kopf M.The antigen dose determines T helper subset development by regulation of CD40 ligand[J].European Journal of Immunology,2000,30:2056-2064.
50.Hansson GK,Zhou X,Tornquist E,et al.The role of adaptive immunity in atherosclerosis[J].Ann NY Acad Sci.2000,902(5):53-62.
51.Math F,Schonbeck U,Sukhova GK,et al.Reduction of atherosclerosis in mice by inhibition of CD40 signalling[J].Nature,1998,394(6 689):200-203.
52.杨瑞雪,张继东,罗玮敏,等.益肾活血胶囊对不稳定型心绞痛患者Hcy及ET、TNF-α、NO的影响[J].上海中医药杂志,2006,40(1):22-24.
53.Swerlick RA,Lee KH,Sepp NT,et al.Regulation of vascular cell adhesion molecule-1 in human dermal microvascular endothelial cells[J].J Immunol,1992, 149(2):698-705.
54.Cybulsky MI,Gimbrone MA Jr.Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis[J].Science,1991,251:788-791.
55.Sakai A,Kume N,Nishi E,et al.P-selectin and vascular cell adhesion molecule-1are focally expression in aortas of hypercholesterolemic rabbits before intimal accumulation of macrophages and T lymphocytes[J].Arterioscler Thromb and Vase Biol,1997,17(2):310-316.
56.Bocher BS,Luscinskas FW,Gimbrone MA,et al.Adhesion of human basophils,eosinophils,and neutrophils to interleukin-12 activated human vascular endothelial cells:contributions of endothelial cell adhesion molecules[J].Exp Med,1991,173(6):1553-1557.
57.Barks JL,Mc Quillan JJ,Iademareo MF.TNF-alpha and IL-4 synergistically increase vascular cell adhesion molecule-1 expression in cultured vascular smooth muscle cells[J].J Immunol,1997,159(9):4532-4538.
58.Thorne SA,Abbot SE,Stevens CR,et al.Modified low density lipoprotein and cytokines mediate monocyte adhesion to smooth muscle cells[J].Atherosclerosis,1996,127(2):167-176.
59.Blankenberg S,Barbaux S,Tiret L.Adhesion molecules and atherosclerosis[J].Atherosclerosis,2003,170:191-203.
60.Davies MJ,Gordon JL,Gearing AJ,et al.The expression of the adhesion molecules ICAM-1,VCAM-1,PECAM,and E-selectin in human atherosclerosis[J].J Pathol,1993,171:223-229.
61.Liuba P,Pesonen E,Paakkari I,et al.Co-infection with Chlamydia pneumoniae and Helicobacter pylori results in vascular endothelial dysfunction and enhanced VCAM-1 expression in apoE-knockout mice[J].J Vasc Res,2003,40:115-122.
62.饶丹,姜红,曾秋棠.黏附分子细胞间黏附分子-1/E-选择素与冠心病[J].心血管病学进展,2005,26(3):278-281.
63.Singh RJ,Mason JC,Lidington EA,et al.Cytokine stimulated vascular cell adhesion molecule-1(VCAM-1) eetodomain release is regulated by TMP-3[J]. Cardiovasc Res,2005,67(1):39-49.
64.Li H,Cybulsky MI,Gimbrone MA Jr,etal.Inducible expression of vascular cell adhesion molecule-1 by vascula smooth muscle cells in vitro and within rabbit atheroma[J].Am J Pathol,1993,143:1551-1559.
65.Dansky HM,Barlow CB,Lominska C,et al.Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage[J].Arterioscler Thromb Vasc Biol,2001,21:1662-1667.
66.Barringhaus KG,Phillips JW,Thatte JS,et al.Alpha4 beta1 integrin(VLA-4)blockade attenuates both early and late leukocyte recruitment and neointimal growth following carotid injury in apolipoprotein E~(-/-) mice[J].J Vasc Res,2004,41:252-260.
67.王梅芳,杨林花.MMP-9与冠状动脉粥样硬化[J].临床心血管病杂志,2004,20(11):699-701.
68.Vine N,Powell JT.Metalloproteinases in degenerative aortic disease[J].Clin Sci,1991,81(2):233-239.
69.Xu,XP,Meisel SR,Ong JM,et al.Oxidized low-density lipoprotein regulates matrix metalloproteinase-9 and its tissue inhibitor in human monocyte-derived macrophages[J].Circulation,1999,99:993-998.
70.Sch(o|¨)nbeck U,Mach F,Sukhova G K,et al.Regulation of matrix metalloproteinase expression in human vascular smooth muscle cells by T lymphocytes:a role for CD40 signaling in plaque rupture?[J].Circ Res,1997,81:448-454.
71.郭爱桃,韦立新.基质金属蛋白酶1与冠状动脉粥样硬化斑块破裂的关系[J].中华病理学杂志,2000,29(3):263-266.
72.Sullivan JL.Macrophage iron,hepcidin,and atherosclerotic plaque stability[J].Exp Biol Med(Maywood).2007,232(8):1014-1020.
73.Boren J,Olin K,Lee I,et al.Identification of the principal proteoglycan-binding site in LDL.A single-point mutation in apo-B100 severely affects proteoglycan interaction without affecting LDL receptor binding[J].J Clin Invest,1998,101: 2658-2664.
74. Shiomi M, Ito T, Tsukada T, et al. Reduction of serum cholesterol levels alters lesional composition of atherosclerotic plaques. Effect of pravastatin sodium on atherosclerosis in mature WHHL rabbits[J]. Arterioscler Thromb Vasc Biol, 1995, 15: 1938-1944.
75. Aikawa M, Rabkin E, Okada Y, et al. Lipid lowering by diet reduces matrix metalloproteinase activity and increases collagen content of rabbit atheroma: a potential mechanism of lesion stabilization[J]. Circulation, 1998, 97: 2433-2444.
76. Schachinger V, Zeiher AM. Atherosclerosis-associated endothelial dysfunction[J]. Z Kardiol, 2000, 89 Suppl 9: IX/70-74.
77. Riazy M, Chen JH, Steinbrecher UP. VEGF secretion by macrophages is stimulated by lipid and protein components of OxLDL via PI3-kinase and PKCzeta activation and is independent of OxLDL uptake[J]. Atherosclerosis, 2009, 204(1): 47-54.
78. Lippy P, Geng YJ, Sukhova GK, et al. Molecular determinants of atherosclerotic plaque vulnerability[J]. Ann N Y Acad Sci, 1997, 811:134-145.
79. Ikeda U, Shimada K. Matrix metalloproteinases and coronary artery diseases[J]. Clinical Cardiology, 2003,26: 55-59.
80. Sukhova GK, Schonbeck U, Rabkin E, et al. Evidence for increased collagenolysis by interstitial collagenases-1 and -3 in vulnerable human atheromatous plaques[J]. Circulation, 1999, 99: 2503-2509.
81. Aikawa M, Voglic SJ, Sugiyama S, et al. Dietary lipid lowering reduces tissue factor expression in rabbit atheroma[J]. Circulation, 1999,100:1215-1222.
82. Zhou L, Stordeur P, de Lavareille A, et al. CD40 engagement on endothelial cells promotes tissue factor-dependent procoagulant activity[J]. Thromb Haemost, 1998, 79: 1025-1028.
83. Miller DL, Yaron R, Yellin MJ. CD40L-CD40 interactions regulate endothelial cell surface tissue factor and thrombomodulin expression[J]. J Leukoc Biol, 1998, 63: 373-379.
84. Slupsky JR, Kalbas M, Willuweit A, et al. Activated platelets induce tissue factor expression on human umbilical vein endothelial cells by ligation of CD40[J].Thromb Haemost,1998,80:1008-1014.
85.Sch(o|¨)nbeck U,Mach F,Sukhova GK,Herman M,Graber P,Kehry MR,Libby P.CD40 ligation induces tissue factor expression in human vascular smooth muscle cells[J].Am J Pathol,2000,156:7-14.
86.Blankenberg S,Rupprecht HJ,Bickel C,et al.Circulating cell adhesion molecules and death in patients with coronary artery disease[J].Circulation,2001,104:1336-1342.
87.Koga M,Kai H,Yasukawa H,et al.Postnatal blocking of interferon-gamma function prevented atherosclerotic plaque formation in apolipoprotein E-knockout mice[J].Hypertens Res,2007,30:259-267.
88.Dansky HM,Barlow CB,Lominska C,et al.Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage[J].Arterioscler Thromb Vase Biol,2001,21:1662-1667.
89.Zhang B,Ye S,Herrmann SM,et al.Functional polymorphism in the regulatory region of gelatinase B gene in relation to severity of coronary atherosclerosis[J].Circulation,1999,99(14):1788-1794.
90.Bennett MR.Apoptosis of vascular smooth muscle cells in vascular remodeling and atherosclerotic plaque rupture[J].Cardiovasc Res,1999,41(2):361-368.
91.Brown DL,Hibbs MS,Kearney M,et al.Identification of 92-kD gelatinase in human coronary atherosclerotic lesions.Association of active enzyme synthesis with unstable angina[J].Circulation,1995,91(8):2125-2131.
92.詹小萍,楼建国,金晓莹,等.益肾活血方对家兔血瘀模型纤溶系统作用的实验研究[J].中国中药杂志,2004,29(5):440-443.
93.孙可丰,胡晓春,胡晓晨,等.抗凝降压胶囊治疗肾虚血瘀型高血压病的临床研究[J].中医药信息,1998,(2):27-28.
94.王旭,陈金锭.益肾化瘀法对2型糖尿病血液流变学、SOD、LPO水平及血管病变的影响[J].南京中医药大学学报(自然科学版),2001,17(1):20-22.
95.黄妙珍.益糖宁颗粒剂改善2型糖尿病胰岛素抵抗的临床研究[J].中华实用中西医杂志,2003,3(16):626-628.
96.李敏.补肾活血方的调脂作用探要[J].中医药学刊,2003,21(5):667-668.
97.陆小青.益肾活血通脉汤治疗颈动脉粥样硬化的临床观察.吉林中医药,2004,24(5):12-13.
98.赵玉霞,刘运芳,刘艳,燕芳芳,于会明.祛瘀消斑胶囊对易损斑块的稳定作用.中医杂志,2006,47(11):856-858.
99.赵玉霞,刘运芳,刘艳.脑心通胶囊对不稳定斑块兔TLR-2和TLR-4炎性信号转导因子的影响.中西医结合心脑血管病杂志,2006,4(12):1071-1073.
100.钱彦丛,宇文萍.枸杞子的化学成分及药理研究新进展[J].中医药学报,2000,(4):33-35
101.刘秀英,胡怡秀,臧雪冰,等.枸杞油胶丸对高脂大鼠血脂水平的影响.中国自然医学杂志,2004,6(3):134-136.
102.姜怡邓,曹军,董泉洲,等.PKC和MMPs在枸杞籽油抗动脉粥样硬化中的作用和机制[J].华西药学杂志,2007,22(1):009-012.
103.马灵筠,陈群力,杨五彪,等.枸杞多糖对动脉粥样硬化模型兔血脂、脂质过氧化、NO和C-反应蛋白的影响[J].郑州大学学报(医学版),2005,40(2):328-330.
104.马灵筠,孙海双,陈群力,等.枸杞多糖对动脉粥样硬化内皮细胞功能及炎症反应的影响[J].江苏中医药,2006,27(5):61-63.
105.杜光.枸杞子免疫药理研究及临床应用.湖北中医杂志,2005,27(2):54-55.
106.陶大勇.枸杞多糖的免疫调节作用.安徽农业科学,2007,35(22):6816-6818.
107.赵娜,郭治昕,赵雪,等.丹参的化学成分与药理作用[J].国外医药·植物药分册,2007,22(4):155-159.
108.田季雨,陈建宗,顾宜,等.复方丹参滴丸对家兔血脂水平和颈动脉粥样硬化斑块的影响[J].中国临床康复,2004,8(9):1708-1709.
109.王南,蔡海江,朱宇,等.丹参素对牛主动脉平滑肌细胞氧化修饰LDL的抑制作用[J].南京医科大学学报,1994,14(4):529-531.
110.成之福,王毅华,王家安,等.复方丹参注射液对冠心病患者一氧化氮、内 皮素-1的影响[J].中国医院药学杂志,2006,26(10):1275.
111.许涛,喻莉,郑智,等.丹参对实验性动脉粥样硬化形成的预防[J].临床心血管病杂志,2005,21(1):54-55.
112.姜开余,顾振纶,阮长耿.丹参素对CD11b、P-Selectin、ICAM-1、VCAM-1、E-Selectin表达的影响[J].中国药理学通报,2000,16(6):682-685.
113.刘虹彬,温进坤.丹参对血管平滑肌细胞基质金属蛋白酶和骨桥蛋白基因表达及细胞增殖的影响[J].中国中西医结合杂志,2002,22(10):764-766.
114.孙学刚,贾钰华,陈育鹩.定心方及丹参酮对血小板膜粘附分子表达的影响[J].山东中医药大学学报,2001,25(1):61-63.
115.姜开余,阮长耿,陈振纶,等.丹参酮Ⅱ-A磺酸钠对内皮细胞和血小板体外表达粘附分子的作用[J].中国药理学报,1998,19(1):47-50.
116.费丽萍,李进禧,郑道声,等.丹参酮Ⅱ-A磺酸钠对红细胞内游离钙浓度的影响[J].微循环杂志,1997,7(4):17-19.
117.崔伊梅.黄芪及其有效成分黄芪苷Ⅳ在心血管药理作用的研究进展[J].天津医科大学学报,2005,11(4):656-659.
118.汪德清,丁保国,Tomas G Neil,等.黄芪总黄酮对动脉粥样硬化早期形成的影响[J].中国药理学通报,2003,19(6):637-640.
119.吴勇,石显水,王石顺,等.黄芪多糖对动脉粥样硬化内皮细胞的保护作用[J].中国临床康复,2005,9(23):238-240.
120.黄进宇,单江,程刚,等.黄芪注射液体外对大鼠血管平滑肌细胞基质金属蛋白酶-2活性的抑制作用[J].中医杂志,2003,44(2):140-142.
121.刘宽芝,李静波,吕海莉,等.黄芪、三七总皂苷对2型糖尿病大血管病变患者MMP-9的影响[J].中国中药杂志,2004,29(3):264-266.
122.杨志红,赵晓龙,陈凤玲,等.黄芪多糖对THP-1巨噬细胞源性泡沫细胞功能的影响[J].中国分子心脏病学杂志,2007,7(3):138-142.
123.王冬梅,朱玮,张存莉,等.黄精化学成分及其生物活性[J].西北林学院学报,2006,21(2):142-145.
124.李友元,邓洪波,向大雄,等.黄精多糖的降血脂及抗动脉粥样硬化作用[J].中国动脉硬化杂志,2005,13(4):429-431.
125.吉中强,高晓昕,宋鲁卿,等.63种调脂中药抗人血小板聚集的实验研究[J].山东中医杂志,1998,17(8):365-367.
126.张萍,刘丹,李友元.黄精多糖对动脉粥样硬化家兔血清IL-6及CRP的影响[J].医学临床研究,2006,23(7):1100-1101.
127.李友元,张萍,邓洪波,等.动脉粥样硬化家兔VCAM-1表达及黄精多糖对其表达的影响[J].医学临床研究,2005,22(9):1287-1288.
128.陈金水,陈松苍,王耀华.黄精口服液对剧烈运动小鼠氧自由基代谢及肌酸激酶影响[J].中医研究,1996,9(4):6-8.
129.李秋怡,干国平,刘焱文.川芎的化学成分及药理研究进展[J].时珍国医国药,2006;17(7):1298-1299.
130.梅家俊,蔡大勇,赵婷秀,等.川芎防治动脉粥样硬化症机制的实验研究[J].湖北中医学院学报,2004,6(2):5-7.
131.阮秋蓉,瞿智玲,朱敏,等.川芎嗪对血管壁细胞血管细胞粘附分子1和单核细胞趋化蛋白1表达的作用及可能机制[J].中国动脉硬化杂志,2006,14(7):560-564.
132.刘红.川芎嗪对动脉粥样硬化家兔血清一氧化氮、血浆内皮素和脂质过氧化物的影响[J].湖北民族学院学报(医学版),2000,17(4):4-5.
133.吴文元.川芎嗪治疗周围动脉粥样硬化闭塞症疗效观察[J].实用中医药杂志,2002,18(8):5-6.
134.Sheu JR,Kan YC,Hung WC,et al.The antiplatelet activity of tetramethylpyrazine is mediated through activation of NO synthase[J].Life Sci,2000,67(8):937-947.
135.潘贺,刚宏林,苏云明.中药水蛭的活性成分及药理作用研究概况[J].中医药信息,2006,23(1):20-22.
136.李凤文,刘晓颖,张立石,等.水蛭及其复方对实验性动脉粥样硬化家兔主动脉内皮素基因表达的影响[J].中医杂志,1999,40(1):50-51
137.张玉娟,田景振,郭之平.水蛭研究概述[J].食品与药品,2005,7(6):9-11.
138.朱智明,刘伊丽,万华印,等.血小板膜GPⅡb/Ⅲa单抗及水蛭素对损伤动脉内膜血小板沉积量的影响[J].心肺血管病杂志,1998,17(1):39-41.
139.王敏,崔连群,张承俊,等.凝血酶诱导血管内皮细胞生长因子的表达及水蛭素的抑制作用[J].中国新药杂志,2004,13(3):226-230.
140.刘京生,苗智慧,董力,等.水蛭抗肿瘤作用的实验研究[J].时珍国医国药,2001,12(10):884-885.
141.屠婕红,余菁,陈伟光.瓜蒌的化学成分和药理作用研究概况[J].中国药师,2004,7(7):562-563.
142.贺立勍.瓜蒌、薤白降脂作用的析因研究[J].湖南中医药导报,2002,8(4):205-207.
143.凌宏,鲁翔,董传仁,等.瓜蒌、心得安对血小板体外聚集和TXA2合成的影响[J].湖北医学院学报,1988,9(2):138.
144.陈燕,赵辉,谢锐.大黄的药理及临床研究近况[J].西南国防医药,2005,15(2):178-180.
145.徐在品,卢占军,陈眷华,等.大黄醇提液抗家兔实验性高脂血症及脂肪肝的实验研究[J].中国应用生理学杂志,2007,23(3):375-379.
146.Abe I,Seki T,Noguchi H,et al.Galloylesters from rhubarb are potent inhibitors of squalene epoxidase,a key enzyme in cholesterol biosynthesis[J].Planta Med,2000,66(8):753-756.
147.Liu Y,Yan F,Liu Y,Zhang C,Yu H,Zhang Y,Zhao Y.Aqueous extract of rhubarb stabilizes vulnerable atherosclerotic plaques due to depression of inflammation and lipid accumulation.Phytother Res.2008,22(7):935-942.
148.Kumar A,Dhawan S,Aggarwal BB.Emodin(3-methyl-1,6,8-trihydroxyanthraquinone)inhibits TNF-induced NF-κB activation,IkB degradation,and expression of cell surface adhesion proteins in human vascular endothelial cells[J].Oncogene,1998,17(7):913-918.
149.Li HL,Chen HL,Li H,et al.Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW 264.7macrophages[J].Int J Mol Med,2005,16(1):41-47.
150.文川,徐浩,黄启福,等.活血中药对ApoE基因缺陷小鼠血脂及动脉粥样硬化斑块炎症反应的影响[J].中国中西医结合杂志,2005,25(4):345-349.
151.黄政德,蒋孟良,易延逵,等.酒制丹参、大黄对大鼠血小板功能及抗凝血作用的研究[J].中成药,2001,23(5):341-342.
152.熊辉岩,谭大风.大黄属三种植物不同部分提取物清除羟基自由基的体外实验研究[J].食品科学,2003,24(1):128-130.
153.兰进,杨世林,郑玉权,等.黄连的研究进展[J].中草药,2001,32(12):1139-1141.
154.勾祥辉.黄连素治疗高血脂59例[J].人民军医,1995,(2):35.
155.王秀杰.黄连的药理研究及现代应用[J].China Pharmacist,2003,6(6):370.
156.何国厚,艾志兵,刘勇,等.小檗碱对兔颈动脉粥样硬化中内膜增生和巨噬细胞趋化作用的影响[J].中风与神经疾病杂志,2006,23(1):94-96.
157.段浩,张宏考,刘继军,等.小檗碱对兔颈动脉损伤后白介素-6等的影响[J].医药论坛杂志,2006,27(10):6-9.
158.周岚,何国厚,李承晏,等.小檗碱对颈动脉粥样硬化家兔血管紧张素的影响[J].卒中与神经疾病,2006,13(4):212-214,218.
159.陈其明,谢明智.小檗碱对正常小鼠血糖调节的影响[J].药学学报,1987,22(3):161-165.
1.Ross R.Atherosclerosis -an inflammatory disease[J].N Eng J Med,1999,340(2):115-126.
2.DeM meyer GR,Herman AG.Vascular endothelial dysfunction[J].Prog Cardiovasc Dis,1997,39(4):325-342.
3.Lusis AJ.Atherosclerosis.Nature,2000,407(2):233-241.
4.刘志忠,陈绍良,刘玲玲,等氧化低密度脂蛋白抗体水平与冠状动脉再狭窄的相关性:独立与年龄、性别、体质量指数、高血压、高血脂及吸烟史的危险因子.中国临床康复[J].2004,8(3):446-447.
5.Sch(o|¨)nbeck U,Gerdes N,Varo N,et al.Oxidized low-density lipoprotein augments and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors limit CD40 and CD40L expression in human vascular cells[J].Circulation,2002,106:2888-2893.
6.Li D,Liu L,Chen H,et al.LOX-1,an oxidized LDL endothelial receptor,induces CD40/CD40L signaling in human coronary artery endothelial cells[J].Arteriosclerosis,Thrombosis,and Vascular biology,2003,23:816-821.
7.姜立清,胡大一,庞文跃.阿托伐他汀对氧化低密度脂蛋白诱导的内皮基质金属蛋白酶-1表达的影响[J].中国老年学杂志.2005,25(11):1358-1360.
8.Romanic AM,Harrison SM,Bao W,et al.Myocardial protection from ischemia/reperfusion injury by targeted deletion of matrix metalloproteinase-9[J].Cardiovas Res,2002,54(3):549-558.
9.朱陵群,庞鹤,王硕仁,牛福玲,崔巍。活血注射液对oxLDL诱导的单核-内皮细胞黏附作用的影响[J].北京中医药大学学报,2007,30(3):173-176.
10.顾耘.软脉煎对血管内皮细胞动脉硬化模型MCP-1的影响[J].实用老年医学,2005,19(5):265-267.
11.Livak KJ,Schmittqen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J].Methods,2001,25(4):402-408.
12.Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Res,2001,29(9):e45.
13. Ehara S, Ueda M, Naruko T, et al. Elevated levels of oxidized low density lipoprotein show a positive relationship with the severity of acute coronary syndromes. circulation[J], 2001,103:1995-1960.
14. Toshima S, Hasegama A, Kurabayashi M, et al.Circulating oxidized lowdensity lipoprotein levels: a biochemical risk marker for coronary heart disease[J]. Arterioscler Thromb Vasc Biol, 2000,20: 2243-2247.
15. Holvoet P, Van Cleemput J, Collen D, et al. oxidized low dendity lipoprotein is aprognostic marker of transplant associated coronary disease[J]. Arterioscler Thromb Vasc Biol, 2000,20: 698-702.
16. Carr A, Mccall MR, Frei B. Oxidation of LDL by myeloperoxidase and reactive nitrogen species: reation pathways and antioxidation protection[J]. Arterioscler Thromb Vasc Biol, 2000,20: 1716-1723.
17. Jay W Heinedce. Oxidants and antioxidants in the pathogenesis for oxidized low density lipoprotein hypothesis[J]. Atherosclerosis, 1998, 141: 11-15.
18. Mehta JL, Li DY, Chen HJ, et al. Inhibition of LOX-1 by statins may relate to up regulation of eNOS[J]. Biochem Biophys Res Conunu, 2001,289: 857 - 861.
19. Akira T, Yan H, Maria F, et al. Expression of adhesion molecules by human endothelial cells exposed to oxidized low density lipoprotein influences of degree of oxidation and location of oxidized LDL[J]. Atherosclerosis, 2001, (154): 79-86.
20. Amberger A, Maczek C, Jürgens G, et al. Co-expression of ICAM-1, VCAM-1, ELAM-1 and Hsp60 in human arterial and venous endothelial cells in response to cytokines and oxidized low-density lipoproteins[J]. Cell Stress Chaperones, 1997, 2 (2): 94-103.
21. Li D, Liu L, Chen H, et al. Lox-1 mediates oxidized low density lipoprotein -induced expression of matrix metalloproteinases in human coronary artery endothelial cells[J]. Circulation, 2003,107: 612-617.
22. Sawanura T, Kume N, Aoyama T, et al. A novel receptor for oxidized low density lipoprotein[J]. Nature, 1997,386(6): 73-77.
23. Okak K, Sawamnra T, Kikuta S, et al. Lectin-like oxidized low-density lipoprotein receptor 1 mediates phagocytosis of aged/apoptotic cells in endothelial cells[J].Proc Natl Acad,Sci USA,1998,95:9535-9540.
24.Li D,Chen H,Sawamura T,et al.Statins modulate oxjdized low-density lipoprotein-mediated adhesion molecule expression in human coronary artery endothelial cells:role of Lox-1[J].J Pharmacol Exp Ther,2002,302:601-605.
25.杨瑞雪,张继东,罗玮敏,等.益肾活血胶囊对不稳定型心绞痛患者Hcy及ET、TNF-α、NO的影响[J].上海中医药杂志2006,40(1):22-24.
26.Hakkinen T,Karkola K,Yla-Herttuala S.Macrophages,smooth muscle cells,endothelial cells,and T-cells express CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions:colocalization with epitopes of oxidized low-density lipoprotein,scavenger receptor,and CD16(FcγRⅢ)[J].Virchows Archiv:an international journal of pathology,2000,437:396-405.
27.Aukrust P,Fredrik M,Ueland T,et al Enhanced level of soluble and membrane-bound CD40 ligand in patients with unstable angina[J].Circulation,1999,100:614-620.
28.Hakonarson H,Kim C,Whelan R,et al.Bidirectional activation between human airway smooth muscle cells and T lymphocytes:role in induction of altered airway responsiveness[J].J Immunol,2001,166:293.
29.Garlichs CD,Eskafi S,Raaz D,et al.Patients with acute coronary syndromes express enhanced CD40 ligand/CD154 on platelets[J].Heart,2001,86(6):649-655.
30.Mach F,Sch(o|¨)nbeck U,Rosalind P,et al.T lymphocytes induce endothelial cell matrix metalloproteinase expression by a CD40L-dependent mechanism[J].AmJ Pathol,1999,154:229-238.
31.Viallard JF,Solanilla A,Gauthier B,et al.Increased soluble and platelet-associated CD40 ligand in essential thrombocythemia and reactive thrombocytosis[J].Blood,2002,99(7):2612-2614.
32.Mach F,Sch(o|¨)nbeck U,Sukhova GK,et al.Reduction of atherosclerosis in mice by inhibition of CD40 signalling[J].Nature,1998,394(6689):200-203.
33.Sch(o|¨)nbeck U,Sukhova GK,Shimizu K,et al.Inhibition of CD40 signaling limits evolution of established atherosclerosis in mice[J].Proc Natl Acad Sci USA,2000,97(13):7458-7463.
34.饶丹,姜红,曾秋棠.黏附分子细胞间黏附分子-1/E-选择素与冠心病[J].心血管病学进展,2005,26(3):278-281.
35.Singh RJ,Mason JC,Lidington EA,et al.Cytokine stimulated vascular cell adhesion molecule-1(VCAM-1) ectodomain release is regulated by TMP-3[J].Cardiovasc Res,2005,67(1):39-49.
36.Davies MJ,Gordon JL,Gearing AJ,et al.The expression of the adhesion molecules ICAM-1,VCAM-1,PECAM,and E-selectin in human atherosclerosis[J].J Pathol,1993,171(3):223-229.
37.Wu YT,Gao YC,Zhou SM.Dysfunction of vascular endothelium and coronary disease[J].China Journal of Modern Medicin.2003,5(13):47-54.
38.Li H,Cybulsky MI,Gimbrone MA Jr,etal.Inducible expression of vascular cell adhesion molecule-1 by vascula smooth muscle cells in vitro and within rabbit atheroma[J].Am J Pathol,1993,143:1551-1559.
39.Blankenberg S,Rupprecht HJ,Bickel C,et al.Circulating cell adhesion molecules and death in patients with coronary artery disease[J].Circulation,2001,104:1336-1342.
40.Mulvihill NT,Foley JB,Murphy R,et al.Evidence of prolonged inflammation in unstable angina and non-Q wave myocardial infarction[J].J Am Coll Cardiol,2000,36(4):1210-1216.
41.Dansky HM,Barlow CB,Lominska C,et al.Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage[J].Arterioscler Thromb Vasc Biol,2001,21:1662-1667.
42.Barringhaus KG,Phillips JW,Thatte JS,et al.Alpha4beta1 integrin(VLA-4)blockade attenuates both early and late leukocyte recruitment and neointimal growth following carotid injury in apolipoprotein E-/- mice[J].J Vasc Res,2004,41:252-260.
43.Brown DL,Hibbs MS,Kearney M,et al.Identification of 92-kD gelatinase in human coronary atherosclerotic lesions:association of active enzyme synthesis with unstable angina[J].Circulation,1995,91:2125-2131.
44.Alvarez B,Ruiz C,Chacon P,et al.Serum values of metalloproteinaseo2 and metalloproteinase-9 as related to unstable plaque and inflammatory cells in patients with greater than 70%carotid artery stenosis[J].Journal of Vascular Surgery,2004,40:469-475.
45.Loftus IM,Naylor AR,Goodall S,et al.Increased matrix metalloproteinase-9activity in unstable carotid plaques.A potential role in acute plaque disruption[J].Stroke,2000,31:40-47.
46.Longo GM,Xiong W,Greiner TC,et al.Matrix metalloproteinases 2 and 9 work in concert to produce aortic aneurysms[J].The Journal of Clinical Investigation,2002,110:613-614.
47.Romanic AM,Harrison SM,Bao W,et al.Myocardial protection from ischemia/reperfusion injury by targeted deletion of matrix metalloproteinase-9[J].Cardiovas Res,2002,54(3):549-558.
48.姜怡邓,曹军,董泉洲,等.PKC和MMPs在枸杞籽油抗动脉粥样硬化中的作用和机制[J].华西药学杂志,2007,22(1):009-012.
49.马灵筠,陈群力,杨五彪,等.枸杞多糖对动脉粥样硬化模型兔血脂、脂质过氧化、NO和C-反应蛋白的影响[J].郑州大学学报(医学版),2005,40(2):328-330.
50.马灵筠,孙海双,陈群力,等.枸杞多糖对动脉粥样硬化内皮细胞功能及炎症反应的影响[J].江苏中医药,2006,27(5):61-63.
51.王南,蔡海江,朱宇,等.丹参素对牛主动脉平滑肌细胞氧化修饰LDL的抑制作用[J].南京医科大学学报,1994,14(4):529-531.
52.成之福,王毅华,王家安,等.复方丹参注射液对冠心病患者一氧化氮、内皮素-1的影响[J].中国医院药学杂志,2006,26(10):1275.
53.许冬青,张旭,蔡文峰,等.丹参药物血清保护血管内皮细胞分子机制研究[J].中成药,2005,27(2):229-230.
54.崔广智,金树梅,赵桂峰,等.丹参酮ⅡA对TNFα诱导的ECV304细胞 NF-k B、kB-α表达及粘附分子ICAM-1、VCAM-1mRNA表达的影响[J].中国药理学通报,2007,23(12):1671-1675.
55.李永胜,王进,王照华,等.丹参酮ⅡA对主动脉内皮细胞功能损伤的保护机制[J].中国急救医学,2007,27(8):720-723.
56.陈海明,叶攀.丹参酮ⅡA对血管内皮细胞氧化应激损伤的保护作用[J].中药材,2008,31(4):569-572.
57.修春,宓穗卿,王宁生.隐丹参酮对TNF-α诱导的人脐静脉内皮细胞粘附分子ICAM-1表达的作用[J].中药药理与临床,2009,25(2):16-19.
58.杨顺清,洪小苏.丹参多酚酸盐对电离辐射损伤血管内皮细胞影响的初步研究[J].中国误诊学杂志2008,8(36):8829-8831.
59.何胜虎,燕建锋,袁彬,等.丹参多酚酸盐对高糖诱导的人脐静脉内皮细胞损伤的保护作用[J].中国动脉硬化杂志,2008,,16(12):948-952.
60.杨亚安,唐丽华,蒋小岗,等.丹参醇提取物对培养内皮细胞T-AOC、LPO、NO、SOD变化的影响[J].苏州大学学报,2005,25(6):963-965.
61.叶泽兵,汪凡军,张刚庆,等.黄芪注射液对TNF-α诱导人脐静脉内皮细胞炎症因子的影响及其分子机制[J].南方医科大学学报,2009,29(6):1137-1140.
62.叶泽兵,汪凡军,张刚庆,等.黄芪注射液对脂多糖诱导内皮细胞损伤保护作用的实验研究[J].实用医学杂志,2009,29(5):1368-1370.
63.范可,陈培红,吴青.黄芪注射液对高糖所致血管内皮细胞损伤的保护作用[J].中国中西医结合临床,2008,20(4):295-298.
64.陈立新,朱海燕,朱陵群,等.黄芪多糖对人心脏微血管内皮细胞增殖及再灌注后内皮细胞与中性粒细胞黏附的影响[J].中国组织工程研究与临床康复,2007,11(27):5382-5386.
65.杨五彪,陈群力,马灵筠,等.黄芪多糖对兔动脉粥样硬化血管内皮细胞功能的影响[J].陕西医学杂志,2005,34(8):914-918.
66.樊懿,吴大正,胡之璧.黄芪甲甙对组胺引起的血管内皮细胞单层通透性增高的作用[J].中国中医基础医学杂志,2001,7(2):36-38.
67.陈金水,陈松苍,王耀华.黄精口服液对剧烈运动小鼠氧自由基代谢及肌酸激酶影响[J].中医研究,1996,9(4):6-8.
68.李友元,邓洪波,向大雄,等.黄精多糖的降血脂及抗动脉粥样硬化作用[J].中国动脉硬化杂志,2005,13(4):429-431.
69.张萍,刘丹,李友元.黄精多糖对动脉粥样硬化家兔血清IL-6及CRP的影响[J].医学临床研究,2006,23(7):1100-1101.
70.李友元,张萍,邓洪波,等.动脉粥样硬化家兔VCAM-1表达及黄精多糖对其表达的影响[J].医学临床研究,2005,22(9):1287-1288.
71.崔磊,王守训,刘新泳.川芎嗪在抗动脉粥样硬化中的作用和机制[J].医学综述,2008,14(22):3491-3493.
72.梁德,姚珍松,杨志东.川芎嗪对家兔血管内皮细胞修复的影响[J].广州中医药大学学报,2001,18(2):155-158.
73.高雪,杨洁,元英进.盐酸川芎嗪对血管紧张素Ⅱ损伤内皮细胞的保护作用[J].细胞生物学杂志,2007,29:895-900.
74.朱际平,袁冬平.川芎嗪对缺氧损伤的人脐静脉内皮细胞分泌一氧化氮的影响[J].实用临床医药杂志,2009,13(7):39-41.
75.李翠乔.天然药物川芎嗪对脂质过氧化内皮细胞的保护作用研究[J].科学技术与工程,2008,8(17):4789-4792.
76.熊石龙,志斌,王前,等.川芎嗪对肿瘤坏死因子致血管内皮细胞组织因子表达的影响[J].中国现代医学杂志,2007,17(19):2330-2333.
77.李凤文,刘晓颖,张立石,等.水蛭及其复方对实验性动脉粥样硬化家兔主动脉内皮素基因表达的影响[J].中医杂志,1999,40(1):50-51.
78.李宁,赵霞,张文高.水蛭微粉对不稳定性心绞痛血管内皮保护作用的观察[J].中国误诊学杂志,2009,9(7):1539.
79.朱智明,刘伊丽,万华印,等.血小板膜GPⅡb/Ⅲa单抗及水蛭素对损伤动脉内膜血小板沉积量的影响[J].心肺血管病杂志,1998,17(1):39-41.
80.高春林,贾俊亚,范生尧.水蛭素对凝血酶所致脑微血管内皮细胞损伤的保护作用[J].中国临床药学杂志,2007,16(2):73-77.
81.王敏,崔连群,张承俊,等.凝血酶诱导血管内皮细胞生长因子的表达及水蛭素的抑制作用[J].中国新药杂志,2004,13(3):226-230.
82.王敏,崔连群,王晓军,韩秋霞,刘鲁宁.水蛭素对凝血酶诱导血管内皮细胞 与中性粒细胞表达黏附分子的影响[J].中国新药杂志,2005,14(8):989-992.
83.芦丽莉,王冬梅.瓜蒌皮提取液对实验性高脂血症大鼠血清NO,SOD,MDA 的影响[J].北华大学学报(自然科学版).2008,9(5):423-425.
84.熊辉岩,谭大风.大黄属三种植物不同部分提取物清除羟基自由基的体外实验研究[J].食品科学,2003,24(1):128-130.
85.Kumar A,Dhawan S,Aggarwal BB.Emodin(3-methyl-1,6,8-trihydroxyanthraquinone)inhibits TNF-induced NF-κB activation,IkB degradation,and expression of cell surface adhesion proteins in human vascular endothelial cells[J].Oncogene,1998,17(7):913-918.
86.Li HL,Chen HL,Li H,et al.Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW 264.7 macrophages[J].Int J Mol Med,2005,16(1):41-47.
87.王为民,虞燕琴,钱令波,舒强,刘广义,林茹,杨军。大黄素对大鼠体外血管内皮一氧化氮cGMP信号途径的影响[J].中国中西医结合杂志,2006,26(7):636.639.
88.何国厚,艾志兵,刘勇,等.小檗碱对兔颈动脉粥样硬化中内膜增生和巨噬细胞趋化作用的影响[J].中风与神经疾病杂志,2006,23(1):94-96.
89.段浩,张宏考,刘继军,等.小檗碱对兔颈动脉损伤后白介素-6等的影响[J].医药论坛杂志,2006,27(10):6-9.
90.周岚,何国厚,李承晏,等.小檗碱对颈动脉粥样硬化家兔血管紧张素的影响[J].卒中与神经疾病,2006,13(4):212-214,218.
91.尹文清,陈琪,佘志刚.盐酸小蘖碱对LPS诱导的人脐静脉内皮细胞与粒性白细胞粘附的影响[J].广东药学院学报,2002,18(2):122-124.
Alvarez B., Ruiz C., Chacon P., Alvarez-Sabin J., Matas M., 2004. Serum values of metalloproteinase-2 and metalloproteinase-9 as related to unstable plaque and inflammatory cells in patients with greater than 70% carotid artery stenosis. Journal of Vascular Surgery 40, 469-475.
Brown D.L., Hibbs M.S., Kearney M., Loushin C., Isner J.M., 1995. Identification of 92-kD gelatinase in human coronary atherosclerotic lesions: association of active enzyme synthesis with unstable angina. Circulation 91, 2125-2131.
Hakkinen T., Karkola K., Yla-Herttuala S., 2000. Macrophages, smooth muscle cells, endothelial cells, and T-cells express CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions: colocalization with epitopes of oxidized low- density lipoprotein, scavenger receptor, and CD16 (FcγRIII). Virchows Archiv: an international journal of pathology 437, 396-405.
He X., Li X., Li S., He C., 2007. Preventive effect of total saponins of panax notoginseng on experimental atherosclerosis in rabbits. China Pharmacy 18,408-410.
He X., Zhang L., Chen P., Li S., 2004. Effect of TPNS on hyperlipoidemia rats. Joural of Yunnan Traditional Chinese Medicine 25, 32-33.
Huang K., Deng X., Luo J., 1997. TPNS enhances relaxation of pulmonary artery in chronic hypoxia rats induced by NO. Guangzhou Medicine 18,54-56.
Ikeda U., Shimada K., 2003. Matrix metalloproteinases and coronary artery diseases. Clinical Cardiology 26, 55-59.
Ji W., Gong BQ., 2007. Hypolipidemic effects and mechanisms of Panax notoginseng on lipid profile in hyperlipidemic rats. Journal of Ethnopharmacology 113, 318-324.
Koyuncu FM., Ozbilgin K., Kuscu NK., Inan S., Vatansever S., Ceylan E., 2006. The effect of oestradiol and neta on immunohistochemical staining of iNOS and eNOS in coronary arteries of ovariectomized rats. Histol Histopathol 21,367-371.
Lee T.S., Yen H.C., Pan C.C., Chau L.Y., 1999. The role of interleukin 12 in the development of atherosclerosis in apoE-deficient mice. Arteriosclerosis, Thrombosis, and Vascular biology 19, 734-742.
Li D., Liu L., Chen H., Sawamura T., Mehta J.L., 2003. LOX-1, an oxidized LDL endothelial receptor, induces CD40/CD40L signaling in human coronary artery endothelial cells. Arteriosclerosis, Thrombosis, and Vascular biology 23, 816-821.
Li X., Wang G., Sun J., Hao H., Xiong Y., Yan B., 2007. Pharmacokinetic and absolute bioavailability study of total Panax notoginsenoside, a typical multiple constituent traditional Chinese medicine (TCM) in rats. Biological and Pharmaceutical Bulletin 30, 847-851.
Ling Y., 1984. Science of Chinese Materia Medica, Fifth ed. Science and Technology Publishing Company, Shanghai, China, pp. 144-145.
Liu Y., Li X., 2005. Effect of pannax notoginseng total saponin on inflammatory immune factors in atherosclerosis. Chinese Traditional and Herbal Drags 36,728-730.
Livak K.J., Schmittgen, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-ΔΔCT) method. Methods 25,402-408.
Loftus I.M., Naylor A.R., Goodall S., Crowther M., Jones L., Bell P.R., Thompson M.M., 2000. Increased matrix metalloproteinase-9 activity in unstable carotid plaques. A potential role in acute plaque disruption. Stroke 31,40-47.
Longo G.M., Xiong W., Greiner T.C., Zhao Y., Fiotti N., Baxter B.T., 2002. Matrix metalloproteinases 2 and 9 work in concert to produce aortic aneurysms. The Journal of Clinical Investigation 110, 613-614.
Lu P., Chen H., 2004. Effects of notoginseng folium saponins on regulating blood lipid. Chinese Journal of Biochemical Pharmaceuti 25,235-236.
Mach F., Schonbeck U., Sukhova G.K., Atkinson E., Libby P., 1998. Reduction of atherosclerosis in mice by inhibition of CD40 signaling. Nature 394,200-203.
Mach F., Schonbeck U., Sukhova G.K., Bourcier T., Bonnefoy J.Y., Pober J.S., Libby P., 1997. Functional CD40 ligand is expressed on human vascular endothelial cells, smooth muscle cells, and macrophages: implications for CD40-CD40 ligand signaling in atherosclerosis. Proceedings of the National Academy of Sciences of the United States of America 94, 1931-1936.
Miida T., Nakamura Y., Mezaki T., Hanyu O., Maruyama S., Horikawa Y., Izawa S., Yamada Y., Matsui H., Okada M., 2002. LDL-cholesterol and HDL-cholesterol concentrations decrease during the day, Annals of Clinical Biochemistry 39, 241-249.
Nakashima Y., Plump A.S., Raines E.W., Breslow J.L., Ross R., 1994. ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree. Arteriosclerosis and Thrombosis 14, 133-140.
Ross R., 1999. Atherosclerosis-an inflammation diease. The New England Journal of Medicine 340, 115-126.
Ruedl C., Bachmann M.F., Kopf M., 2000. The antigen dose determines T helper subset development by regulation of CD40 ligand. European Journal of Immunology 30, 2056-2064.
Schonbeck U., Gerdes N., Varo N., Reynolds R.S., Horton D.B., Bavendiek U., Robbie L., Ganz P., Kinlay S., Libby P., 2002. Oxidized low-density lipoprotein augments and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors limit CD40 and CD40L expression in human vascular cells. Circulation 106,2888-2893.
Schonbeck U., Libby P., 2001. CD40 signaling and plaque instability. Circulation Research 89, 1092-1103.
Schonbeck U., Mach F., Sukhova G.K., Murphy C., Bonnefoy J.Y., Fabunmi R.P., Libby P., 1997. Regulation of matrix metalloproteinase expression in human vascular smooth muscle cells by T lymphocytes: a role for CD40 signaling in plaque rupture? Circulation Research 81,48-454.
Shen L.H., Zhou L., Wang B.Y., Pu J., Hu L.H., Chai D.J., Wang L., Zeng J.Z., He B., 2008. Oxidized low-density lipoprotein induces differentiation of RAW264.7 murine macrophage cell line into dendritic-like cells. Atherosclerosis 199,257-264.
Su N., He M., Zhong B., Zhang S., 2002. Effects of Xuesaitong on blood lipid and plasma viscosity in hyperlipemia patients. Journal of Hunan College of Traditional Chinese Medicine 22,65-66.
Tarchalski J., Guzik P., Wysocki H., 2003. Correlation between the extent of coronary atherosclerosis and lipid profile. Molecular and Cellular Biochemistry 246,25-30.
Xu X.P., Meisel S.R., Ong J.M., Kaul S., Cercek B., Rajavashisth T.B., Sharifi B., Shah P.K., 1999. Oxidized low-density lipoprotein regulates matrix metalloproteinase-9 and its tissue inhibitor in human monocyte-derived macrophages. Circulation 99, 993-998.
Yang J., Qin C., 2003. Effects of Complex Danshen formula, Danshen and Sanqi on the function of platelet. China Journal of Experimental Traditional Medical Formulae 9, 59-62.
Yuan Z., Li X., Li S., Fan J., 2006. Effects of PNS on stability of atherosclerotic plaque and initial investigation for its mechanism. Chinese Journal of Nature Medicine 4,62-64.
Yuan Z., Li X., Li S., Zhang H., Liu Y., 2006. Mechanism approach on TPNS improving stability of atherosclerotic plaque in rabbits. Chinese Traditional and Herbal Drugs 37,741-743.
Zhang S.H., Reddick R.L., Piedrahita J.A., Maeda N., 1992. Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E. Science 258,468-471.
Zhou X., Chen M., Li X., Bai G., Zhou Q., Jiang S., 2007. An approach to etiological factors and pathogenesis of dyslipidemia in TCM. Chinese Archives of Traditional Chinese Medicine 25, 197-198.
Aikawa M, Rabkin E, Okada Y, Voglic SJ, Clinton SK, Brinckerhoff CE, Sukhova GK, Libby P. 1998. Lipid lowering by diet reduces matrix metalloproteinase activity and increase collagen content of rabbit atheroma: a potential mechanism of lesion stabilization. Circulation 97: 2433-2444.
Barringhaus KG, Phillips JW, Thatte JS, Sanders JM, Czarnik AC, Bennett DK, Ley KF, Sarembock IJ. 2004. Alpha4betal integrin (VLA-4) blockade attenuates both early and late leukocyte recruitment and neointimal growth following carotid injury in apolipoprotein E-/- mice. J Vasc Res 41: 252-260.
Blankenberg S, Barbaux S, Tiret L. 2003. Adhesion molecules and atherosclerosis. Atherosclerosis 170: 191-203.
Blankenberg S, Rupprecht HJ, Bickel C, Peetz D, Hafner G, Tiret L, Meyer J. 2001. Circulating cell adhesion molecules and death in patients with coronary artery disease. Circulation 104: 1336-1342.
Couffinhal T, Duplaa C, Moreau C, Lamaziere JM, Bonnet J. 1994. Regulation of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 in human vascular smooth muscle cells. Circ Res 74: 225-234.
Cybulsky MI, Gimbrone MA Jr. 1991. Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis. Science 251: 788 -791.
Dansky HM, Barlow CB, Lominska C, Sikes JL, Kao C, Weinsaft J, Cybulsky MI, Smith JD. 2001. Adhesion of monocytes to arterial endothelium and initiation of atherosclerosis are critically dependent on vascular cell adhesion molecule-1 gene dosage. Arterioscler Thromb Vasc Biol 21: 1662-1667.
Davies MJ, Gordon JL, Gearing AJ, Pigott R, Woolf N, Katz D, Kyriakopoulos A. 1993. The expression of the adhesion molecules ICAM-1, VCAM-1, PECAM, and E-selectin in human atherosclerosis. J Pathol 171: 223-229.
Dong GJ, Liu JG, Shi DZ, Wang YY, Luan LJ, Cheng YY. 2007. Effects of water extractives of a qi and blood regulating prescription on early atherosclerosis of apolipoprotein E-deficient mice. Zhong Xi Yi Jie He Xue Bao 5:45-49.
Falk E. 1992. Why do plaques rupture? Circulation 86: 30-42.
Gutstein DE, Fuster V. 1999. Pathophysiology and clinical significance of atherosclerotic plaque rupture. Cardiovasc Res 41: 323-33.
Jawien J, Nastalek P, Korbut R. 2004. Mouse models of experimental atherosclerosis. J Physiol Pharmacol 55: 503-517.
Khan BV, Parthasarathy SS, Alexander RW, Medford RM. 1995. Modified low density lipoprotein and its constituents augment cytokine-activated vascular cell adhesion molecule-1 gene expression in human vascular endothelial cells. J Clin Invest 95: 1262-1270.
Koga M, Kai H, Yasukawa H, Kato S, Yamamoto T, Kawai Y, Kusaba K, Seki Y, Kai M, Egashira K, Kataoka Y, Imaizumi T. 2007. Postnatal blocking of interferon-gamma function prevented atherosclerotic plaque formation in apolipoprotein E-knockout mice. Hypertens Res 30: 259-267.
Li H, Cybulsky MI, Gimbrone MA Jr, Libby P. 1993. An atherogenic diet rapidly induces VCAM-1, a cytokine-regulatable mononuclear leukocyte adhesion molecule, in rabbit aortic endothelium. Arterioscler Thromb 13: 197-204.
Li X, Liu Q, Zhou T, Zhao S, Zhou S. 2008. PAPP-A: a possible pathogenic link to the instability of atherosclerotic plaque. Med Hypotheses 70: 597-599.
Libby P. 2001. Current concepts of the pathogenesis of the acute coronary syndromes. Circulation 104: 365-372.
Libby P, Sukhova G, Lee RT, Galis ZS. 1995. Cytokines regulate vascular functions related to stability of the atherosclerotic plaque. J Cardiovasc Pharmacol 25: S9-12.
Liuba P, Pesonen E, Paakkari I, Batra S, Andersen L, Forslid A, Yla-Herttuala S, Persson K, Wadstrom T, Wang X, Laurini R. 2003. Co-infection with Chlamydia pneumoniae and Helicobacter pylori results in vascular endothelial dysfunction and enhanced VCAM-1 expression in apoE-knockout mice. J Vasc Res 40: 115-122.
Lutters BC, Leeuwenburgh MA, Appeldoorn CC, Molenaar TJ, Van Berkel TJ, Biessen EA. 2004. Blocking endothelial adhesion molecules: a potential therapeutic strategy to combat atherogenesis. Curr Opin Lipidol 15: 545-552.
Mulvihill NT, Foley JB, Murphy R, Crean P, Walsh M. 2000. Evidence of prolonged inflammation in unstable angina and non-Q wave myocardial infarction. J Am Coll Cardiol 36: 1210-1216.
Piedrahita JA, Zhang SH, Hagaman JR, Oliver PM, Maeda N. 1992. Generation of mice carrying a mutant apolipoprotein E gene inactivated by gene targeting in embryonic stem cells. PNAS 89: 4471-4475.
Ramana KV, Bhatnagar A, Srivastava SK. 2004. Inhibition of aldose reductase attenuates TNF-alpha-induced expression of adhesion molecules in endothelial cells. FASEB J 18: 1209 -1218.
Reiner Z, Tedeschi-Reiner E. 2001. New information on the pathophysiology of atherosclerosis. Lijec Vjesn 123: 26-31.
Ross R. 1999. Atherosclerosis: an inflammatory disease. N Engl J Med. 340: 115-126.
Scalia R, Appel JZ 3rd, Lefer AM. 1998. Leukocyte-endothelium interaction during the early stages of hypercholesterolemia in the rabbit: role of P-selectin, ICAM-1, and VCAM-1. Arterioscler Thromb Vasc Biol 18: 1093-1100.
Wang B, Zhang JD, Feng JB, Yin HQ, Liu FY, Wang Y. 2007. Effect of traditional Chinese medicine Qin-Dan-Jiang-Ya-Tang on remodeled vascular phenotype and osteopontin in spontaneous hypertensive rats. J Ethnopharmacol 110: 176-182.
Yang RuiXue, Zhang JiDong, Luo WeiMin, Guo RuiChen, Zhu YuanYuan, Wang ShiHua. 2006. Effects of "YiShen HuoXue Capsule" on Hcy, ET, TNF-α and NO in Patients with Unstable Angina. Shanghai Journal of Traditional Chinese Medicine 40: 22-24.
Yokozawa T, Cho EJ, Sasaki S, Satoh A, Okamoto T, Sei Y. 2006. The Protective Role of Chinese Prescription Kangen-karyu Extract on Diet-Induced Hypercholesterolemia in Rats. Biol. Pharm. Bull 29: 760-765.