补肾活血化痰方对动脉粥样硬化细胞黏附分子表达的研究
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摘要
动脉粥样硬化(atherosclerosis, AS)是严重危害人类健康的常见病,常累及心脑血管等重要部位,有极高发病率和死亡率。因此防治动脉粥样硬化,改善患者的生活质量,减少心脑血管事件的发生率和病死率,成为亟待解决的医学难题之一
本课题从文献研究、临床研究和实验研究三方面,通过对临床颈动脉粥样硬化患者观察及人脐静脉内皮细胞培养,对AS的发病机制进行分析,并对补肾活血化痰方抗AS的作用及炎症方面的作用机制进行探讨。
一、文献研究
中国古代医籍中没有动脉粥样硬化的病名,多将其归属于“胸痹”、“真心痛”、“中风”、“眩晕”、“头痛”、“痴呆”、“痰饮”、“瘀证”等病症。若机体脏腑功能失调,脾胃运化功能减弱,肾气渐衰,加之饮食肥甘厚味,气机失畅,易化生为痰浊,痰浊流滞于血脉日久则成痰瘀交结之证,正虚邪实相互影响,致使病变不断发展,多位学者多认为“痰浊”、“瘀血”为本病发病之关键。罗陆一教授认为,肾为人体之根本,主一身气血运行,肾虚气化无力则津聚成痰,温煦运化不足,血脉失于温运,血运无力,留而为瘀,故认为动脉粥样硬化属本虚标实之证,本虚则之于肾虚,标实在于痰瘀互结。
动脉粥样硬化是一个慢性炎症过程,细胞黏附分子在动脉粥样硬化的形成和进展中起关键作用。在动脉粥样硬化形成过程中,首先是血管内皮在各种致炎刺激因子下损伤,细胞间黏附分子-1(Intercellular adhesion molecule, ICAM-1)、血管细胞黏附分子-1(Vascular cell adhesion molecule, VC AM-1)等表达增加,调节白细胞的粘附,单核细胞穿越内皮摄取氧化型低密度脂蛋白(ox-LDL),变为激活的巨噬细胞,吞噬ox-LDL成为泡沫细胞,引起巨噬细胞的清除反应和血管平滑肌细胞的增殖,从而形成动脉粥样斑块。白细胞与血管内皮细胞黏附是炎症反应的起始步骤,介导白细胞与内皮细胞相互作用的分子基础就是细胞黏附分子。
二、临床研究
目的：观察补肾活血化痰方对颈动脉粥样硬化患者症状积分,颈动脉内膜中层厚度、斑块Crouse积分、血脂、血清可溶性细胞黏附分子水平的影响,探讨补肾活血化痰方抗AS的作用及其作用机制。
方法：选择颈动脉粥样硬化(carotid atherosclerosis, CAS)患者100例,随机分为2组,补肾活血化痰方(Bushenhuoxuehuatan Formula, BF)组50例和阿托伐他汀(Atorvastatin Calcium, AC)组50例,分别用补肾活血化痰方和阿托伐他汀钙进行治疗6个月,对两组CAS患者治疗前后的症状、颈动脉IMT、斑块Crouse积分、血脂水平、血清sIC AM-1和sVCAM-1进行检测。
结果：(1)治疗后BF组与AC组均可以明显降低症状积分(P<0.01,P<0.05),两组相比差异有显著性(P<0.05),BF组在改善症状方面优于AC组；(2)治疗后BF组与AC组均能明显降低双侧颈动脉内膜中层厚度(P<0.01,P<0.01),两组差异无显著性；(3)治疗后BF组与AC组均能显著降低颈动脉粥样硬化斑块Crouse积分(P<0.01,P<0.05)；(4)治疗后BF组与AC组均可以明显降低CAS血清中的TC(P<0.05,P<0.01)、TG(P<0.05,P<0.05)水平,BF组可以明显降低LDL-C(P<0.05),升高HDL-C(P<0.01),BF组与AC组相比在升高HDL-C方面差异有显著性(P<0.05)；(5)治疗后BF组与AC组可以明显降低CAS血清中sICAM-1 (P<0.01, P<0.01)、sVCAM-1 (P<0.01, P<0.01)水平。
结论：补肾活血化痰方可以改善颈动脉粥样硬化患者症状,使症状积分明显降低,降低颈动脉内膜中层厚度,降低斑块Crouse积分,明显降低CAS患者血清中的TC、TG、LDL-C水平,升高HDL-C,明显降低CAS患者血清中sIC AM-1、sVCAM-1,补肾活血化痰方可能通过调节血脂水平、减少血清可溶性细胞黏附分子水平来发挥抗AS的作用。
三、实验研究
目的：观察补肾活血化痰方对TNF-α诱导人脐静脉内皮细胞(HUVEC)存活率、ICAM-1、VCAM-1、NF-κB表达的影响。
方法：通过SD大鼠制备补肾活血化痰方血清,0.25%胰蛋白酶分离原代HUVEC,传代后将HUVEC分为五组：①对照组：HUVEC+DMEM培养基+10%对照组血清；②TNF-α组：HUVEC+DMEM培养基+10%对照组血清+10ng/mL TNF-a;③补肾活血化痰方高剂量组：HUVEC+DMEM培养基+10%补肾活血化痰方高剂量组血清+10ng/mL TNF-a;④补肾活血化痰方中剂量组：HUVEC+DMEM培养基+10%补肾活血化痰方中剂量组血清+10ng/mL TNF-a;⑤补肾活血化痰方低剂量组：HUVEC+DMEM培养基+10%补肾活血化痰方低剂量组血清+10ng/mL TNF-a。采用CCK-8法检测HUVEC的存活率,ELISA法检测细胞ICAM-1和VCAM-1的表达,免疫细胞化学方法观察细胞NF-κB表达,半定量逆转录聚合酶链反应(RT-PCR)法检测细胞ICAM-1 mRNA和VCAM-1 mRNA表达。
结果：(1)补肾活血化痰方高、中、低剂量组细胞存活率分别为87.2%、82%、80%,TNF-α组细胞存活率为74.2%,各组细胞存活率与对照相比均降低,TNF-a组的细胞存活率最低；(2)与对照组相比TNF-α组细胞ICAM-1和VCAM-1的表达明显增高(P<0.01,P<0.01),补肾活血化痰方不同剂量组均能抑制TNF-α诱导的内皮细胞ICAM-1和VCAM-1的表达,其中BF高剂量组与TNF-α组相比ICAM-1和VCAM-1的表达明显减少(P<0.01,P<0.01),BF中剂量组与TNF-a组相比ICAM-1的表达明显减少(P<0.05)；(3)TNF-α组可见细胞胞浆中有大量棕黄色颗粒,部分胞核深染,NF-κB表达阳性,补肾活血化痰方各剂量组NF-κB阳性表达信号均弱于TNF-a组,尤其以高、中剂量组明显；(4)与对照组相比TNF-a组中内皮细胞ICAM-1mRNA、VCAM-1 mRNA表达明显增高(P<0.01,P<0.01),补肾活血化痰方高、中剂量组均能下调TNF-α诱导的内皮细胞ICAM-1 mRNA表达(P<0.01,P<0.01),补肾活血化痰方高剂量组能下调TNF-a诱导的内皮细胞VCAM-1 mRNA表达(P<0.05)。
结论：补肾活血化痰方对TNF-a诱导人脐静脉内皮细胞的炎症反应有抑制作用,能抑制TNF-a诱导的内皮细胞ICAM-1、VCAM-1及NF-κB的表达,以高剂量组明显。这可能是补肾活血化痰方抗AS的作用机制之一。
Atherosclerosis (AS) is a commen disease which causing serious harm to human health, often involving the major organ, such as heart, brain and blood vessel. It has very high morbidity and mortality, therefore prevention and treatment of atherosclerosis and improving the quality of life of patients and reducing the incidence of cardiovascular events and mortality become one of medical problems urgently needed to be solved.
This dissertation includes three parts:literature research, clinical and experiment research. According to the observation to carotid atherosclerosis patients and human umbilical vein endothelial cells, the pathogenesis of AS is analysed, furthermore the effect of bushenhuoxuehuatan formula against AS and the inflammatory mechanism are discussed.
Part one: literature research
There have not the name of atherosclerosis in the ancient Chinese medical record, and generally atherosclerosis was called thoracic obstruction, angina pectoris, stroke, dizziness, headache, dementia, phlegm-fluid retention, blood stasis syndrome, etc. Zang-fu organ dysfunction, hypofunction of transportation and transformation of spleen and stomach, kidney-qi deficiency and excessive consumption of fatty and sweet foods may lead to disorder of qi activity and phlegm stagnation which gradually coming into being blood stasis. It is called syndrome of binding of phlegm and blood stasis. Many scholars believe the principal pathogenesis is phlegm stagnation and blood stasis. Professor Luo luoyi thinks that the kidney is considered as the foundation of human body and dominates the movement of qi and blood all over the body. If phlegm stagnation resulted from deficiency of kidney and disorder of qi activity, lack of warmth, transportation and transformation, blood flow slowly, stay and for blood stasis. So Professor Luo believe atherosclerosis is the syndrome of root deficiency and branch excess, the root is deficiency of kidney while the branch is binding of phlegm and blood stasis.
Atherosclerosis is a chronic inflammatory disease in which cell adhesion molecules play a key role. In the early process of atherogenesis, first the endothelial cell is injured by many inflammatory factors and expression of cell adhesion molecules such as intercellular adhesion molecule-1, vascular cell adhesion molecule-1 lead to increased adherence of leukocyte. Monocytes may migrate through the endothelium and situate themselves within the subendothelial layer, engulf oxidized low-density lipoprotein, then transform in activated macrophages which continuously accumulate xidized low-density lipoprotein and become foam cells. At the same time, necrotic macrophages and proliferative smooth muscle cells result in atherosclerotic plaque. Adherence of leukocyte with endothelial cells is the initiation of atherosclerosis, whose molecular foundation is cell adhesion molecules.
Part two:clinical research
Objective:To investigate the effect of Bushenhuoxuehuatan Formula on the clinical symptom score, carotid intima-media thickness, Crouse score in atherosclerotic plaque, blood lipids and the level of soluble cell adhesion molecules and explore the effect and its mechanism of Bushenhuoxuehuatan Formula on atherosclerosis.
Method: 100 carotid atherosclerosis (CAS) patients were ramdomly indived into 2 groups:Bushenhuoxuehuatan Formula (BF) group and Atorvastatin Calcium (AC) group, 50 patients in each group. After two-month treatment with Bushenhuoxuehuatan Formula and Atorvastatin Calcium respectively, the clinical symptom score, carotid intima-media thickness, Crouse score in atherosclerotic plaque, blood lipids and the level of soluble cell adhesion molecules were observed before and after treatment.
Results:(1) After treatment the clinical symptom score was obviously decreased in BF group and AC group (P<0.01, P<0.05). There was a significant difference between two groups (P<0.05), and compaired with AC group, BF group has better effect in improving symptom. (2) Bilateral carotid intima-media thickness was obviously decreased in BF group and AC group (P<0.01, P<0.01), and there was no significant difference between the two groups. (3) Crouse score in atherosclerotic plaque was markedly reduced in BF group and AC group (P<0.01, P<0.05). (4) The serum level of TC (P<0.05, P<0.01) and TG (P<0.05, P<0.05) in CAS patients was obviously decreased in BF group and AC group. The level of LDL-C (P<0.05) was decreased while the level of HDL-C (P<0.01) was increased in the BF group. There was significant difference in increasing HDL-C between two groups (P<0.05). (5) The serum level of soluble intercellular adhesion molecule-1 (sICAM-1) (P<0.01, P<0.01) and soluble vascular cell adhesion molecule-1 (sVCAM-1) (P<0.01, P<0.01) in CAS patients was obviously decreased in BF group and AC group.
Conclusions:Bushenhuoxuehuatan Formula can be improve symptom, decrease the clinical symptom score, decrease carotid intima-media thickness, reduce Crouse score in atherosclerotic plaque, decrease the level of TC, TG, LDL-C, increase the level of HDL-C, decrease the level of sICAM-1 and sVCAM-1 in CAS patients. The effect of Bushenhuoxuehuatan Formula against AS might be related with its regulating the blood lipids and decreasing the serum level of soluble cell adhesion molecules.
Part three: experimental research
Objective:To observe the effect of Bushenhuoxuehuatan Formula on the cell livability, the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and nuclear factor-kappa B (NF-κB) in human umbilical vein endothelial cells induced by tumor necrosis factor-a (TNF-a).
Method: The blank serum and Bushenhuoxuehuatan Formula serum was prepared respectively by using SD rats. The primarily HUVEC isolated by 0.25% Trypsin-EDTA, then passaged, was divided into 5 groups:①Control group:HUVEC+DMEM culture medium+10% blank serum,②TNF-a group:HUVEC+DMEM culture medium +10% blank serum+10ng/mL TNF-a,③High dose BF group:HUVEC+DMEM culture medium +10% high dose BF serum+10ng/mL TNF-a,④Medium dose BF group:HUVEC+DMEM culture medium+10% medium dose BF serum+10ng/mL TNF-a,⑤Low dose BF group: HUVEC+DMEM culture medium+10% low dose BF serum+10ng/mL TNF-a. The cell livability was detected by using cell counting (CCK-8) colorimetric analysis. ICAM-1 and VCAM-1 on cell surfaces were quantified by enzyme linked immuonosorbent assays (ELISA). The expression of NF-κB was detected by immunocytochemistry analysis. The expression of ICAM-1 mRNA and VCAM-1 mRNA were examined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).
Results:(1) The cell livability was 87.2%,82% and 80% in High, medium, low dose BF group respectively,74.2% in the TNF-a group. Compared with the control group, the cell livability was decreased in other four groups, especially in the TNF-a group. (2) The expression of ICAM-1 and VCAM-1 on cell surfaces were obviously increased in the TNF-a group (P<0.01, P<0.01), compared with the control group. All doses' Bushenhuoxuehuatan Formula could inhibit the increasing expression of ICAM-1 and VCAM-1 induced by TNF-a. Compared with TNF-a group, the expression of ICAM-1 and VCAM-1 were obviously decreased in high dose BF group (P<0.01, P<0.01), and the expression of ICAM-1 was obviously decreased in medium dose BF group (P<0.05). (3) A great deal of brown-yellow particles in the cytolymph and a small quantity of deep brown particles in the cytoplasm were found in endothelial cells of TNF-a group which showed NF-κB expression was positive. Campared with TNF-a group, the positive expression of NF-κB was significantly decreased in all doses'BF group, especially high and medium dose BF group. (4) Campared with TNF-a group, the mRNA level of ICAM-1 and VCAM-1 in HUVEC was obviously up-regulated (P<0.01, P<0.01). However, high and medium dose' Bushenhuoxuehuatan Formula could down-regulate the expression of ICAM-1 mRNA (P<0.01, P<0.01), while only high dose'Bushenhuoxuehuatan Formula could down-regulate the expression of VCAM-1 mRNA (P<0.05).
Conclusions:Bushenhuoxuehuatan Formula could mediate the inflammatory response of HUVEC induced by TNF-a and inhibit the expression of ICAM-1, VCAM-1 and NF-κB in endothelial cells, more significant effect in the high dose BF group. It might be the one of mechanism of Bushenhuoxuehuatan Formula against AS.
本课题从文献研究、临床研究和实验研究三方面,通过对临床颈动脉粥样硬化患者观察及人脐静脉内皮细胞培养,对AS的发病机制进行分析,并对补肾活血化痰方抗AS的作用及炎症方面的作用机制进行探讨。
一、文献研究
中国古代医籍中没有动脉粥样硬化的病名,多将其归属于“胸痹”、“真心痛”、“中风”、“眩晕”、“头痛”、“痴呆”、“痰饮”、“瘀证”等病症。若机体脏腑功能失调,脾胃运化功能减弱,肾气渐衰,加之饮食肥甘厚味,气机失畅,易化生为痰浊,痰浊流滞于血脉日久则成痰瘀交结之证,正虚邪实相互影响,致使病变不断发展,多位学者多认为“痰浊”、“瘀血”为本病发病之关键。罗陆一教授认为,肾为人体之根本,主一身气血运行,肾虚气化无力则津聚成痰,温煦运化不足,血脉失于温运,血运无力,留而为瘀,故认为动脉粥样硬化属本虚标实之证,本虚则之于肾虚,标实在于痰瘀互结。
动脉粥样硬化是一个慢性炎症过程,细胞黏附分子在动脉粥样硬化的形成和进展中起关键作用。在动脉粥样硬化形成过程中,首先是血管内皮在各种致炎刺激因子下损伤,细胞间黏附分子-1(Intercellular adhesion molecule, ICAM-1)、血管细胞黏附分子-1(Vascular cell adhesion molecule, VC AM-1)等表达增加,调节白细胞的粘附,单核细胞穿越内皮摄取氧化型低密度脂蛋白(ox-LDL),变为激活的巨噬细胞,吞噬ox-LDL成为泡沫细胞,引起巨噬细胞的清除反应和血管平滑肌细胞的增殖,从而形成动脉粥样斑块。白细胞与血管内皮细胞黏附是炎症反应的起始步骤,介导白细胞与内皮细胞相互作用的分子基础就是细胞黏附分子。
二、临床研究
目的：观察补肾活血化痰方对颈动脉粥样硬化患者症状积分,颈动脉内膜中层厚度、斑块Crouse积分、血脂、血清可溶性细胞黏附分子水平的影响,探讨补肾活血化痰方抗AS的作用及其作用机制。
方法：选择颈动脉粥样硬化(carotid atherosclerosis, CAS)患者100例,随机分为2组,补肾活血化痰方(Bushenhuoxuehuatan Formula, BF)组50例和阿托伐他汀(Atorvastatin Calcium, AC)组50例,分别用补肾活血化痰方和阿托伐他汀钙进行治疗6个月,对两组CAS患者治疗前后的症状、颈动脉IMT、斑块Crouse积分、血脂水平、血清sIC AM-1和sVCAM-1进行检测。
结果：(1)治疗后BF组与AC组均可以明显降低症状积分(P<0.01,P<0.05),两组相比差异有显著性(P<0.05),BF组在改善症状方面优于AC组；(2)治疗后BF组与AC组均能明显降低双侧颈动脉内膜中层厚度(P<0.01,P<0.01),两组差异无显著性；(3)治疗后BF组与AC组均能显著降低颈动脉粥样硬化斑块Crouse积分(P<0.01,P<0.05)；(4)治疗后BF组与AC组均可以明显降低CAS血清中的TC(P<0.05,P<0.01)、TG(P<0.05,P<0.05)水平,BF组可以明显降低LDL-C(P<0.05),升高HDL-C(P<0.01),BF组与AC组相比在升高HDL-C方面差异有显著性(P<0.05)；(5)治疗后BF组与AC组可以明显降低CAS血清中sICAM-1 (P<0.01, P<0.01)、sVCAM-1 (P<0.01, P<0.01)水平。
结论：补肾活血化痰方可以改善颈动脉粥样硬化患者症状,使症状积分明显降低,降低颈动脉内膜中层厚度,降低斑块Crouse积分,明显降低CAS患者血清中的TC、TG、LDL-C水平,升高HDL-C,明显降低CAS患者血清中sIC AM-1、sVCAM-1,补肾活血化痰方可能通过调节血脂水平、减少血清可溶性细胞黏附分子水平来发挥抗AS的作用。
三、实验研究
目的：观察补肾活血化痰方对TNF-α诱导人脐静脉内皮细胞(HUVEC)存活率、ICAM-1、VCAM-1、NF-κB表达的影响。
方法：通过SD大鼠制备补肾活血化痰方血清,0.25%胰蛋白酶分离原代HUVEC,传代后将HUVEC分为五组：①对照组：HUVEC+DMEM培养基+10%对照组血清；②TNF-α组：HUVEC+DMEM培养基+10%对照组血清+10ng/mL TNF-a;③补肾活血化痰方高剂量组：HUVEC+DMEM培养基+10%补肾活血化痰方高剂量组血清+10ng/mL TNF-a;④补肾活血化痰方中剂量组：HUVEC+DMEM培养基+10%补肾活血化痰方中剂量组血清+10ng/mL TNF-a;⑤补肾活血化痰方低剂量组：HUVEC+DMEM培养基+10%补肾活血化痰方低剂量组血清+10ng/mL TNF-a。采用CCK-8法检测HUVEC的存活率,ELISA法检测细胞ICAM-1和VCAM-1的表达,免疫细胞化学方法观察细胞NF-κB表达,半定量逆转录聚合酶链反应(RT-PCR)法检测细胞ICAM-1 mRNA和VCAM-1 mRNA表达。
结果：(1)补肾活血化痰方高、中、低剂量组细胞存活率分别为87.2%、82%、80%,TNF-α组细胞存活率为74.2%,各组细胞存活率与对照相比均降低,TNF-a组的细胞存活率最低；(2)与对照组相比TNF-α组细胞ICAM-1和VCAM-1的表达明显增高(P<0.01,P<0.01),补肾活血化痰方不同剂量组均能抑制TNF-α诱导的内皮细胞ICAM-1和VCAM-1的表达,其中BF高剂量组与TNF-α组相比ICAM-1和VCAM-1的表达明显减少(P<0.01,P<0.01),BF中剂量组与TNF-a组相比ICAM-1的表达明显减少(P<0.05)；(3)TNF-α组可见细胞胞浆中有大量棕黄色颗粒,部分胞核深染,NF-κB表达阳性,补肾活血化痰方各剂量组NF-κB阳性表达信号均弱于TNF-a组,尤其以高、中剂量组明显；(4)与对照组相比TNF-a组中内皮细胞ICAM-1mRNA、VCAM-1 mRNA表达明显增高(P<0.01,P<0.01),补肾活血化痰方高、中剂量组均能下调TNF-α诱导的内皮细胞ICAM-1 mRNA表达(P<0.01,P<0.01),补肾活血化痰方高剂量组能下调TNF-a诱导的内皮细胞VCAM-1 mRNA表达(P<0.05)。
结论：补肾活血化痰方对TNF-a诱导人脐静脉内皮细胞的炎症反应有抑制作用,能抑制TNF-a诱导的内皮细胞ICAM-1、VCAM-1及NF-κB的表达,以高剂量组明显。这可能是补肾活血化痰方抗AS的作用机制之一。
Atherosclerosis (AS) is a commen disease which causing serious harm to human health, often involving the major organ, such as heart, brain and blood vessel. It has very high morbidity and mortality, therefore prevention and treatment of atherosclerosis and improving the quality of life of patients and reducing the incidence of cardiovascular events and mortality become one of medical problems urgently needed to be solved.
This dissertation includes three parts:literature research, clinical and experiment research. According to the observation to carotid atherosclerosis patients and human umbilical vein endothelial cells, the pathogenesis of AS is analysed, furthermore the effect of bushenhuoxuehuatan formula against AS and the inflammatory mechanism are discussed.
Part one: literature research
There have not the name of atherosclerosis in the ancient Chinese medical record, and generally atherosclerosis was called thoracic obstruction, angina pectoris, stroke, dizziness, headache, dementia, phlegm-fluid retention, blood stasis syndrome, etc. Zang-fu organ dysfunction, hypofunction of transportation and transformation of spleen and stomach, kidney-qi deficiency and excessive consumption of fatty and sweet foods may lead to disorder of qi activity and phlegm stagnation which gradually coming into being blood stasis. It is called syndrome of binding of phlegm and blood stasis. Many scholars believe the principal pathogenesis is phlegm stagnation and blood stasis. Professor Luo luoyi thinks that the kidney is considered as the foundation of human body and dominates the movement of qi and blood all over the body. If phlegm stagnation resulted from deficiency of kidney and disorder of qi activity, lack of warmth, transportation and transformation, blood flow slowly, stay and for blood stasis. So Professor Luo believe atherosclerosis is the syndrome of root deficiency and branch excess, the root is deficiency of kidney while the branch is binding of phlegm and blood stasis.
Atherosclerosis is a chronic inflammatory disease in which cell adhesion molecules play a key role. In the early process of atherogenesis, first the endothelial cell is injured by many inflammatory factors and expression of cell adhesion molecules such as intercellular adhesion molecule-1, vascular cell adhesion molecule-1 lead to increased adherence of leukocyte. Monocytes may migrate through the endothelium and situate themselves within the subendothelial layer, engulf oxidized low-density lipoprotein, then transform in activated macrophages which continuously accumulate xidized low-density lipoprotein and become foam cells. At the same time, necrotic macrophages and proliferative smooth muscle cells result in atherosclerotic plaque. Adherence of leukocyte with endothelial cells is the initiation of atherosclerosis, whose molecular foundation is cell adhesion molecules.
Part two:clinical research
Objective:To investigate the effect of Bushenhuoxuehuatan Formula on the clinical symptom score, carotid intima-media thickness, Crouse score in atherosclerotic plaque, blood lipids and the level of soluble cell adhesion molecules and explore the effect and its mechanism of Bushenhuoxuehuatan Formula on atherosclerosis.
Method: 100 carotid atherosclerosis (CAS) patients were ramdomly indived into 2 groups:Bushenhuoxuehuatan Formula (BF) group and Atorvastatin Calcium (AC) group, 50 patients in each group. After two-month treatment with Bushenhuoxuehuatan Formula and Atorvastatin Calcium respectively, the clinical symptom score, carotid intima-media thickness, Crouse score in atherosclerotic plaque, blood lipids and the level of soluble cell adhesion molecules were observed before and after treatment.
Results:(1) After treatment the clinical symptom score was obviously decreased in BF group and AC group (P<0.01, P<0.05). There was a significant difference between two groups (P<0.05), and compaired with AC group, BF group has better effect in improving symptom. (2) Bilateral carotid intima-media thickness was obviously decreased in BF group and AC group (P<0.01, P<0.01), and there was no significant difference between the two groups. (3) Crouse score in atherosclerotic plaque was markedly reduced in BF group and AC group (P<0.01, P<0.05). (4) The serum level of TC (P<0.05, P<0.01) and TG (P<0.05, P<0.05) in CAS patients was obviously decreased in BF group and AC group. The level of LDL-C (P<0.05) was decreased while the level of HDL-C (P<0.01) was increased in the BF group. There was significant difference in increasing HDL-C between two groups (P<0.05). (5) The serum level of soluble intercellular adhesion molecule-1 (sICAM-1) (P<0.01, P<0.01) and soluble vascular cell adhesion molecule-1 (sVCAM-1) (P<0.01, P<0.01) in CAS patients was obviously decreased in BF group and AC group.
Conclusions:Bushenhuoxuehuatan Formula can be improve symptom, decrease the clinical symptom score, decrease carotid intima-media thickness, reduce Crouse score in atherosclerotic plaque, decrease the level of TC, TG, LDL-C, increase the level of HDL-C, decrease the level of sICAM-1 and sVCAM-1 in CAS patients. The effect of Bushenhuoxuehuatan Formula against AS might be related with its regulating the blood lipids and decreasing the serum level of soluble cell adhesion molecules.
Part three: experimental research
Objective:To observe the effect of Bushenhuoxuehuatan Formula on the cell livability, the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and nuclear factor-kappa B (NF-κB) in human umbilical vein endothelial cells induced by tumor necrosis factor-a (TNF-a).
Method: The blank serum and Bushenhuoxuehuatan Formula serum was prepared respectively by using SD rats. The primarily HUVEC isolated by 0.25% Trypsin-EDTA, then passaged, was divided into 5 groups:①Control group:HUVEC+DMEM culture medium+10% blank serum,②TNF-a group:HUVEC+DMEM culture medium +10% blank serum+10ng/mL TNF-a,③High dose BF group:HUVEC+DMEM culture medium +10% high dose BF serum+10ng/mL TNF-a,④Medium dose BF group:HUVEC+DMEM culture medium+10% medium dose BF serum+10ng/mL TNF-a,⑤Low dose BF group: HUVEC+DMEM culture medium+10% low dose BF serum+10ng/mL TNF-a. The cell livability was detected by using cell counting (CCK-8) colorimetric analysis. ICAM-1 and VCAM-1 on cell surfaces were quantified by enzyme linked immuonosorbent assays (ELISA). The expression of NF-κB was detected by immunocytochemistry analysis. The expression of ICAM-1 mRNA and VCAM-1 mRNA were examined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).
Results:(1) The cell livability was 87.2%,82% and 80% in High, medium, low dose BF group respectively,74.2% in the TNF-a group. Compared with the control group, the cell livability was decreased in other four groups, especially in the TNF-a group. (2) The expression of ICAM-1 and VCAM-1 on cell surfaces were obviously increased in the TNF-a group (P<0.01, P<0.01), compared with the control group. All doses' Bushenhuoxuehuatan Formula could inhibit the increasing expression of ICAM-1 and VCAM-1 induced by TNF-a. Compared with TNF-a group, the expression of ICAM-1 and VCAM-1 were obviously decreased in high dose BF group (P<0.01, P<0.01), and the expression of ICAM-1 was obviously decreased in medium dose BF group (P<0.05). (3) A great deal of brown-yellow particles in the cytolymph and a small quantity of deep brown particles in the cytoplasm were found in endothelial cells of TNF-a group which showed NF-κB expression was positive. Campared with TNF-a group, the positive expression of NF-κB was significantly decreased in all doses'BF group, especially high and medium dose BF group. (4) Campared with TNF-a group, the mRNA level of ICAM-1 and VCAM-1 in HUVEC was obviously up-regulated (P<0.01, P<0.01). However, high and medium dose' Bushenhuoxuehuatan Formula could down-regulate the expression of ICAM-1 mRNA (P<0.01, P<0.01), while only high dose'Bushenhuoxuehuatan Formula could down-regulate the expression of VCAM-1 mRNA (P<0.05).
Conclusions:Bushenhuoxuehuatan Formula could mediate the inflammatory response of HUVEC induced by TNF-a and inhibit the expression of ICAM-1, VCAM-1 and NF-κB in endothelial cells, more significant effect in the high dose BF group. It might be the one of mechanism of Bushenhuoxuehuatan Formula against AS.
引文
[1]段纪俊,常海燕.社会经济发展与全球疾病死亡率预测研究.中国社会医学杂志,2007：24(4)：254-256
[2]徐浩,陈可冀.从对动脉粥样硬化认识转变看中西医结合的优势与切入点.中国中西医结合杂志,2007；27(1)：5-7
[3]赵玲,魏海峰,张丽,等.中医痰浊血瘀证候的生物学基础研究.中华中医药杂志,2008：23(8)：680-683
[4]孟国凡.动脉粥样硬化的中医病因病机初探.云南中医中药杂志,2004；25(2)：55-56
[5]周学文,李曦明,陈民,等.以脾论治内清外柔治疗动脉粥样硬化.辽宁中医杂志,2007；34(2)：146
[6]罗陆一.从肾论治冠心病机理例析.中医药学刊,2005；23(4)：588-589
[7]顾耘,蔡霄月.冠心病中医可从肾论治.辽宁中医药大学学报,2007；9(6)：41-42
[8]许平,于睿.血脂异常当从肝论治.中医药学刊,2005；23(12)：2200-2201
[9]黄河清.谈肝气虚衰与动脉粥样硬化.四川中医,1997；15(2)：7-8
[10]宁珺.冠心病当心肝同治.中华现代中西医结合,2004；1(1)：2-3
[11]周仲瑛,金妙文,吴敏,等.滋肾养肝化痰消瘀法治疗动脉粥样硬化42例的临床研究.江苏中医药,2003；24(2)：13-15
[12]王丽英,张红敏,谢春光,等.中医释义动脉粥样硬化及2型糖尿病低度炎症的发生条件.时珍国医国药,2005；16(7)：662-663
[13]孙学刚,靖林林,赵益业,等.从气虚血瘀论治动脉粥样硬化潜证.中华中医药学刊,2007；25(8)：1637-1638
[14]杨宝元,张洁,史俊玲,等.冠状动脉粥样硬化形成与中医气虚血瘀病相关性研究.中西医结合心脑血管病杂志,2009；7(4)：385-387
[15]于艳丽,宋鲁成.中医“毒邪”在动脉粥样硬化中的地位和作用.河北中医,2009;31(2)：211-212
[16]林培政,杨开清.动脉粥样硬化性疾病与中医湿热证的关系.中药新药与临床药理,2006;17(2)：147-149
[17]赵明,孙立新,徐素芝.活血化瘀软胶囊治疗脑梗死伴发颈动脉粥样硬化斑块的临床观察.河北中医,2009；31(2)：191-192
[18]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.中国医院药学杂志,2006；26(10)：1275
[19]李辉,邱仕君.邓铁涛教授对“痰瘀相关”理论的阐释和发挥.湖北民族学院学报 (医学版),2005；22(1)：45-47
[20]王晓峰,李鹏,赵永东.沈宝藩教授运用痰瘀同治法经验摭拾.实用中医内科杂志,2004：18(4)：298-299
[21]韩素芳.益气活血汤治疗气虚血瘀型冠状动脉粥样硬化性心脏病心绞痛30例.河北中医,2009；31(5)：690
[22]晏庆德,张学颖.补阳还五汤片剂治疗早期动脉硬化性闭塞症临床研究.山东中医杂志,2005；24(3)：140-142
[23]王长垠,殷春萍,刘红霞.补肾法对颈动脉粥样硬化患者血清TNF-α、IL-6的影响.中国中医急症,2009；18(10)：1569-1570
[24]王少贤,杨晓锋.补肾活血化痰法对血管性痴呆患者血脂和血液流变学的影响.中华实用中西医杂志,2006；19(14)：1672-1673
[25]张芬梅,张慧岭,陈要玲,等.健脾降浊通脉方治疗动脉粥样硬化的临床观察.四川中医,2007；25(4)：45-46
[26]第五永长,刘修庄,陈亚文,等.健脾化痰活血利湿方药抗动脉粥样硬化作用的实验研究.陕西中医,2004；25(2)：181-182
[27]陈冰,宋剑南,牛晓红,等.健脾祛痰化瘀方对氧化型低密度脂蛋白诱导血管细胞信号分子钙离子和蛋白激酶C表达的影响.中国动脉硬化杂志,2004；12(2)：143-146
[28]王嵩,刘煜德,李荣,等.清热解毒法对50例动脉粥样硬化患者血脂、纤维蛋白原、D-二聚体的干预作用.山西中医,2009；24(6)：46-48
[29]王少英.清热解毒药物在治疗冠心病中的运用.北京中医,2004；23(1)：14-15
[30]杨俊卿,周岐新,李远宗.姜黄提取物对实验性动物高脂血症的防治作用研究.中国药房,2004；15(12)：598-600
[31]邱赛红,李福元,高雁,等.降脂消斑片对兔高脂血症合并动脉粥样硬化血脂的影响.湖南中医药大学学报,2009；29(3)：21-23
[32]朱明军,罗琰.调脂胶囊对动脉粥样硬化大鼠MDA、SOD的影响.中医药学刊,2006：24(10)：1821-1822
[33]罗陆一,冯润芬.地黄饮子治疗颈动脉粥样硬化斑块的临床研究.中华医学研究杂志,2004；4(5)：417-418
[34]甄艳军,牛丽颖,赵兰英,等.木贼提取物对早期AS大鼠血管平滑肌凋亡及Fas、FasL表达的干预.北京中医药大学学报,2007；30(1)：45-47
[35]程红,罗陆一.益肾通脉汤对兔动脉粥样硬化形成过程中平滑肌细胞增殖和凋亡的影响.江西中医学院学报,2009；21(4)：62-64
[36]Kervinen H, Palosuo T, Manninen V, et al. Joint effect of C-reactive protein and other risk factors on acute coronary events. Am Heart Journal,2001; 141:580-585 [37] Singh SK, Suresh MV, Voleti B, et al. The connection between C-reactive protein and atherosclerosis. Ann Med,2008; 40(2):110-120
[38]殷建明,罗陆一,华青.通心贴对冠心病血清Hs-CRP、M-CSF、IL-18的影响.江西中医药,2008；34(4)：28-29
[39]王晓杨,张志琴,张媛,等.CRP与在体兔动脉粥样粥样硬化相关性的研究及桑葚提取物的影响.中西医结合心脑血管病杂志,2009；7(9)：1053-1055
[40]Graham IM, Daly LE, Refsum HM, et al. Plasma homocysteine as a risk factor for vascular disease:The European Concerted Action Project. JAMA,1997; 277:1775-1781
[41]刘粉叶,张继东,胡连海,等.益肾活血胶囊对高同型半胱氨酸血症兔平滑肌细胞增殖及MMP-9基因表达的影响.南京中医药大学学报,2009；25(1)：35-37
[42]Blankenberg S, Rupprecht HJ, Bickel C, et al. Circulating cell adhesion molecules and death in patients with coronary artery disease. Circulation,2001; 104:1336-1342
[43]Sandra Tribolo, Federica Lodi, Carol Connor, et al. Comparative effect of quercetin and its predominant human metabolites on adhesion molecule expression in activated human vascular endothelial cells. Atherosclerosis,2008; 197:50-56
[44]张梅,温进坤,孙辉臣,等.丹参注射液对动脉粥样硬化大鼠血脂及细胞间黏附分子-1表达的影响.中国病理生理杂志,2004；20(10)：1871-1873
[45]Hojo Y, Ikeda U, Katsuki T, et al. Matrix metalloproteinase expression in the coronary circulation induced by coronary angioplasty. Atherosclerosis,2002; 161:185-192
[46]Parks WC, Wilson CI, Lopez-Boado YS, et al. Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol,2004; 4(8):617-629
[47]郑翠娥,郭伟星,李士涛,等.芪蛭合剂对大鼠实验性动脉粥样硬化主动脉MMP-9表达的影响.中华中医药学刊,2008；26(6)：1153-1155
[48]刘龙涛,吴敏,张文高,等.虎杖苷对颈动脉粥样硬化斑块稳定性的干预研究.北京中医药,2009；28(3)：172-175
[49]才向军.化浊降脂汤治疗高脂血症80例.陕西中医,2004；25(2)：135-136
[50]方微,张慧信,王绿娅,等.何首乌总甙抑制动脉粥样硬化病变形成.中国动脉硬化杂志,2005；13(2)：175-178
[51]Liu QL, Xiao JH, Ma R, et al. Effect of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside on lipoprotein oxidation and proliferation of coronary arterial smooth cells. J Asian Nat Prod Res,2007; 9(6-8):689-697
[52]李香珍.调脂饮治疗老年高脂血症86例.四川中医,2004；22(10)：55
[53]邓平珍,徐敏.补骨脂对去卵巢大鼠骨转换及血脂代谢影响的实验研究.新中医,2005：37(7)：94-96
[54]童琦燕.牛膝治疗高尿酸血症高脂血症.辽宁中医杂志,2003；30(11)：943
[55]崔瑛,侯士良.怀牛膝预防动脉粥样硬化的实验研究.基层中药杂志,1998；12 (1)：30-31
[56]左迎.三七皂苷注射液治疗冠状动脉粥样硬化前后患者血液流变学变化.黑龙江医学,2006；30(10)：748
[57]殷智,彭再生,杨长丰.复方血三七胶囊治疗脑动脉粥样硬化症42例疗效观察.中国民族民间医药杂志,2005：73(2)：90-92
[58]张翼冠,李晓辉,樊继山,等.三七皂甙通过抗炎和调血脂作用抑制大鼠动脉粥样硬化形成.现代生物医学进展,2007；7(11)：1601-1603,1607
[59]闫彦芳,张壮,孙塑伦,等.三七皂甙及其主要成分对血管内皮细胞缺氧损伤的保护作用.中国实验方剂学杂志,2002：8(1)：34-37
[60]赖光强,林中,韩正雪,等.地龙胶囊对缺血性脑梗死患者颈动脉粥样斑块的影响研究.中国中医急症,2009；18(7)：1034-1035
[61]白雪,杨思进,王丽萍,等.蚓激酶对不稳定型心绞痛患者C反应蛋白的影响.中国基层医药,2007；14(8)：1346-1347
[62]许超千,焦军东,孙宏丽,等.蚓激酶对实验动物血栓及相关血液学指标的影响.中草药,2005；36(9)：1367-1369
[63]陈奕芝,方若鸣,魏刚,等.石菖蒲挥发油、p-细辛醚对高脂血症大鼠血管舒张与抗血小板聚集的作用.中国中西医结合杂志,2004；24(基础理论研究特集)：16-18
[64]吴启端,吴清和,王绮雯,等.石菖蒲挥发油及p-细辛醚的抗血栓作用.中药新药与临床药理,2008；19(1)：29-31
[65]Hemanez A, M L, Chamorro C, et al. Inhibition of lipid synthesis and secretion in long-term cultures of adult rat bepatocytes by alpha-asarone. Planta Med,1993; 59 (2):121
[66]杨永宗.动脉粥样硬化性心血管病基础与临床,北京：科学出版社,2009,第二版：53
[67]Ross R. The pathogenesis of atherosclerosis. New Engl J Med,1976,295 (8):376-420
[68]Ross R. Atherosclerosis-an inflammatory disease, New Engl J Med,1999; 340 (2): 115-126
[69]Erling Falk. Pathogenesis of atherosclerosis. Journal of the American College of Cardiology.2006; 47(8):7-12
[70]Elena Galkina, Klaus Ley. Vascular adhesion molecules in atherosclerosis. Arteriosclerosis, Thrombosis, and Vascular Biology.2007; 27(11):2292-2301
[71]唐可京,李幼姬,谢灿茂.ICAM-1和VCAM-1的结构与表达调控.国外医学分子生物学分册,2002；24(3)：173-177
[72]Rothlein R, Dustin ML, Marlin SD, et al. A human intercellular adhesion molecule-1 (ICAM-1) distinct from LFA-1. J Immunol,1986; 137 (2):1270
[73]Jordi Bella, Prasanna R. Kolatkar, Christopher W. Marlor, et al. The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. Proc. Natl. Acad. Sci.,1998; 95:4140-4145
[74]Pietruczuk M, Pietruczuk A, Pancewicz S, et al. ICAM-1:structure, biological role and clinical significance. Pol Merkur Lekarski,2004; 17 (101):507-511
[75]Osborn L, Hession C, Tizard R, et al. Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes. Cell,2004; 59:1203-1211
[76]Rice GE, Bevilacqua MP.An inducible endothelial cell surface glycoprotein mediates melanoma adhesion. Science,1989; 246:1303-1306
[77]Jia-huai Wang, R Blake Pepinsky, Thilo Stehle, et al. The crystal structure of an N-terminal two-domain fragment of vascular cell adhesion molecule 1 (VCAM-1):a [75] cyclic peptide based on the domain 1 C-D loop can inhibit VCAM-1-a4 integrin interaction. Proc. Natl. Acad. Sci.,1995; 92:5714-5718
[78]Nureddin Cengiz, Ender Erdogan, Hanefi Ozbek, et al. Adhesion molecules in cerebral ischemia and atherosclerosis. Eur J Gen Med,2009; 6(4):249-256
[79]Blankenberg S, Barbaux S, Tiret L. Adhesion molecules and atherosclerosis. Atherosclerosis,2003; 170(2):191-203
[80]Poston RN, Haskard DO, Coucher JR, et al. Expression of intercellular adhesion molecule-1 in atherosclerotic plaques. Am J Pathol,1992; 140:665-673
[81]Printsera OY, Peclo MM, Gown AM. Various cell types in human atherosclerotic lesion express ICAM-1:Futher immuocytechemical and immuno chemical studies employing monoclonal antibody 10F3. Am J Pathol,1992; 140:889-896
[82]Daniel S. The neuropathology and neurochemistry of multiple system atrophy. In Mathias C, Bannister R, eds. Autonomic failure:a textbook of clinical disorders of the autonomic nervous system, Oxford:Oxford University Press,1999,3rd ed:321-328
[83]Asanori Aikawa, Peter Libby. The vulnerable atherosclerotic plaque:Pathogenesis and therapeutic approach. Cardio Vascular Pathology,2004; 13(3):125-138
[84]Thome SA, Abbot SE, Stevens CR, et al. Modified low density lipoprotein and cytokines mediate monocyte adhesion to smooth muscle cell. Atherosclerosis,1996; 95: 1263
[85]Cybulsky Myron L, Gimbrone Michael A, Jr. Endothelial expression of mononuclear leukocyte adhesion molecule during atherogenesis. Science,1991; 251(4995):788-791
[86]Kitagawa K, Matsumoto M, Sasaki T, et al. Involvement of ICAM-1 in the progression of atherosclerosis in APOE-knockout mice. Atherosclerosis,2002; 160(2):305-310
[87]Scalia R, Appel JZ 3rd, Lefer AM. 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,1998; 18(7):1093-1100
[88]Sen R, Baltimore D. Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism. Cell,1986; 47(6):921-928
[89]Paul P Tak, Gary S. Firestein. NF-κB:a key role in inflammatory disease. J Clin Invest, 2001; 107(1):7-11
[90]John Kuriyan, Dinitris Thanos. Structure of the NF-κB transcription factor: a holistic interaction with DNA. Structure,1995; 3:135-141
[91]Marc D Jacobs, Stephen C Harrison. Structure of an IκBα/NF-κB complex. Cell,1998; 95:749-758
[92]CollinsT. Endothelial nuclear factor-Kappa B and the unitation of the atherosclerotic lesion. lab Invest,1993; 69:499
[93]O'Connor S, Markovina S, Miyamoto S. Evidence for a phosphorylation-independent role for Ser 32 and 36 in proteasome inhibitor-resistant (PIR) IkappaBalpha degradation in B cells. Exp Cell Res,2005; 307(1):15-25
[94]Qing G, Qu Z, Xiao G. Stabilization of basally translated NF-kappaB-inducing kinase (NIK) protein functions as a molecular switch of processing of NF-kappaB2 p100. J Biol Chem,2005; 280(49):40578-40582
[95]Leena Hajra, Andrew I. Evans, Mian Chen, et al. The NF-κB singnal transduction pathway in aortic endothelial cells is primes for activation in regions predisposed to atherosclerosis lesion formation. Cell Biology,2000; 97(16):9052-9057
[96]K Brand, S Page, G Rogler, et al. Activated transcription factor nuclear-kappa B is present in the atherosclerotic lesion. J Clin Invest,1996; 97(7):1715-1722
[97]Burleigh ME, Babaev VR, Oates JA, et al. Cyclooxygenase-2 promotes early atherosclerotic lesion formation in LDL receptor-dificient mice. Circulation.2002; 105: 1816-1823
[98]Gu L, Okada Y, Clinton SK, et al. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low-density lipoprotein receptor-dificent mice. Mol Cell,1998; 2: 275-281
[99]Cybulsky MI, Liyama K, Li H, et al. A major role for VCAM-1, but not ICAM-1, in early atherosclerosis. J Clin Invest,2001; 107:1255-1262
[100]Park CW, Kim JH, Lee JH, et al. High glucose-induced intercellular adhesion molecule-1 (ICAM-1) expression through an osmotic effect in rat mesangial cells is PKC-NF-KB-dependent. Diabetologia,2000; 58(5):2166-2177
[101]Lawrence T, Gilroy DW, Colville-Nash PR, et al. Possible new role for NF-kappa B in the resolution of inflammation. Nat Med,2001; 7:1291
[102]方微,张慧信,王绿娅,等.何首乌总苷抗氧化与实验性小鼠动脉粥样硬化病变的形成.中国中药杂志,2007；32(13)：1320-1323
[103]Gerald Rimbach, Giuseppe Valacchi, Raffaella Canali, et al. Macrophages stimulated with IFN-y activate NF-κB and induce MCP-1 gene expression in primary human endothelial cells. Molecular Cell Biology Research Communication,2000; 3(4):238-242
[104]黄铮,龚兰生,施仲伟,等.颈动脉超声结合危险因素调查冠心病高危患者,中国超声医学杂志,1998；14(3)：16-18
[105]Crouse JR, Harpold GH, Kahl FR, et al. Evaluation of a scoring system of extracranial carotid atherosclerosis extent with B-mode ultrasound. Stroke,1986; 17(2): 270-275
[106]Lemne C, Jogestrand T, de Faire U. Carotid intima-media thickness and plaque in borderline hypertension. Stroke,1995; 26(1):34-39
[107]Kawasaki J. Diagnostic accuracy of carotid ultrasonography in screening for coronary artery disease. J Cardiol,2000; 36(5):295-302
[108]Sonia S Anand, Salim Yusuf, Vladmir Vuksan, et al. Differences in risk factors, atherosclerosis, and cardiovascular disease between ethnic groups in Canada: the Study of Health Assessment and Risk in Ethnic groups (SHARE). Lancet,2000; 356(9226):279-284
[109]Pognoli P, Tremoli E, Poli A, et al. Intimal plus medial thichness of the arterial wall:a direct measurement with ultrasound imaging. Circulation, 1986; 74(6):1399-1406
[110]Salonen JT, Korpela K, Salonen R, et al. Autoantibody against oxidized LDL and progression of carotid atherosclerosis. Lancet,1992; 339(8798):883-887
[111]Libby P, Aikawa M, Schonbeck U. Cholesterol and atherosclerosis. Biochim Biophys Acta,2000; 1529(1-3):299-309
[112]Rader DJ. High-density lipoprotein and atherosclerosis. Am J Cardiol, 2002; 90(8A): 62i-70i
[113]John S, Jacobi J, Delles C, et al. Plasma soluble adhesion molecules and endothelium-dependent vasodilation in early human atherosclerosis. Clin Sci (Lond),2000; 98(5):521-529
[114]Jie J Cao, Chau Thach, Teri A Manolio, et al. C-reactive protein, carotid intima-media thickness, and incidence of ischemic stroke in the elderly. Circulation,2003; 108:166-167
[115]Stefan Blankenberg, Rupprecht HJ, Christoph Bickel, et al. Circulating cell adhesion molecules and death in patients with coronary artery disease. Circulation,2001; 104: 1336-1342
[116]Yu CM, Zhang Q, Lam L, et al. Comparison of intensive and low-dose atovastatin therapy in the reduction of carotid intimal-medial thickness in patients with coronary heart disease. Heart,2007; 93(8):933-939
[117]殷建明,罗陆一.通心贴对冠状动脉粥样硬化性心脏病患者血清Hs-CRP、IL-18、GMP-140的影响.上海中医药杂志,2008；42(4)：26-27
[118]程红,罗陆一.益肾通络汤对兔动脉粥样硬化模型平滑肌细胞凋亡的干预作用.中西医结合心脑血管病杂志,2009；7(5)：551-552
[119]周凌,伍津津,鲁元刚,等.人脐静脉内皮细胞的分离培养和冻存.重庆医学,2006：35(3)：226-230
[120]Fan Zhang, Wei Yu, James L. Hargrove,et al. Inhibition of TNF-α induced ICAM-1, VCAM-1 and E-selectin expression by selenium, Atherosclerosis,2002; 161:381-386
[121]Carswell EA, Old LJ, Kassel RL, et al. An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci,1975; 72(9):3666-3670
[122]Shalaby MR, Aggarwal BB, Rinderknecht E, et al. Activation of human polymorphonuclear neutrophil functions by interferon-gamma and tumor necrosis factors. J Immunol,1985; 135(3):2069-2073
[123]Stannard AK, Riddell DR, Bradley NJ, et al. Apolipoprotein E and regulation of cytokine-induced cell adhesion molecule expression in endothelial cells. Atherosclerosis. 1998; 139:57-64
[124]Barath P, Fishbein MC, Cao J, et al. Detection and localization of tumor necrosis factor in human athroma. Am J Cardiol,1990; 65(5):297-302
[125]张寄南,曹克将,杨志健.心脏标志物学,南京：江苏科学技术出版社,2007,第一版：80
[126]Price DT, Loscalzo J. Cellular adhesion molecules and atherogenesis. Am J Med, 1999; 107(1):85-97
[127]Calixto JB, Campos MM, Otuki MF, et al. Anti-inflammatory compounds of plant origin. Planta Med,2004; 70 (2):9
[128]Okadas, Shikatak, Matsudam, et al. Intercellular adhesion molecule-1-deficient mice a resistant against renal injury after induction of diabetes. Diabetes,2003; 52 (10):2586-2593
[129]Pirisi M, Vitulli D, Falleti E, et al. Increased soluble ICAM-1 concentration and impaired delay-type hypersensitivity skin tests in patients with chronic liver disease. J Clin Pathol,1997; 50(1):50-53
[130]Collins T. Endothelial nuclear factor-Kappa B and the unitation of the atherosclerotic lesion. Lab Invest,1993; 69:499
[131]Kim CD, Kim YK, Lee SH, et al. Rebamipide inhibits neutrophil adhesion to hypoxia/reoxygenation-stimulated endothelial cell via nuclear factor kappaB-dependent pathway. J Pharmacol Exp Ther,2000; 294 (3): 864-869
[132]朱陵群,王硕仁,赵明镜,等.活血注射液对ox-LDL诱导人脐静脉内皮细胞活化的影响.中国中药杂志,2008；33(13)：1617-1621
[2]徐浩,陈可冀.从对动脉粥样硬化认识转变看中西医结合的优势与切入点.中国中西医结合杂志,2007；27(1)：5-7
[3]赵玲,魏海峰,张丽,等.中医痰浊血瘀证候的生物学基础研究.中华中医药杂志,2008：23(8)：680-683
[4]孟国凡.动脉粥样硬化的中医病因病机初探.云南中医中药杂志,2004；25(2)：55-56
[5]周学文,李曦明,陈民,等.以脾论治内清外柔治疗动脉粥样硬化.辽宁中医杂志,2007；34(2)：146
[6]罗陆一.从肾论治冠心病机理例析.中医药学刊,2005；23(4)：588-589
[7]顾耘,蔡霄月.冠心病中医可从肾论治.辽宁中医药大学学报,2007；9(6)：41-42
[8]许平,于睿.血脂异常当从肝论治.中医药学刊,2005；23(12)：2200-2201
[9]黄河清.谈肝气虚衰与动脉粥样硬化.四川中医,1997；15(2)：7-8
[10]宁珺.冠心病当心肝同治.中华现代中西医结合,2004；1(1)：2-3
[11]周仲瑛,金妙文,吴敏,等.滋肾养肝化痰消瘀法治疗动脉粥样硬化42例的临床研究.江苏中医药,2003；24(2)：13-15
[12]王丽英,张红敏,谢春光,等.中医释义动脉粥样硬化及2型糖尿病低度炎症的发生条件.时珍国医国药,2005；16(7)：662-663
[13]孙学刚,靖林林,赵益业,等.从气虚血瘀论治动脉粥样硬化潜证.中华中医药学刊,2007；25(8)：1637-1638
[14]杨宝元,张洁,史俊玲,等.冠状动脉粥样硬化形成与中医气虚血瘀病相关性研究.中西医结合心脑血管病杂志,2009；7(4)：385-387
[15]于艳丽,宋鲁成.中医“毒邪”在动脉粥样硬化中的地位和作用.河北中医,2009;31(2)：211-212
[16]林培政,杨开清.动脉粥样硬化性疾病与中医湿热证的关系.中药新药与临床药理,2006;17(2)：147-149
[17]赵明,孙立新,徐素芝.活血化瘀软胶囊治疗脑梗死伴发颈动脉粥样硬化斑块的临床观察.河北中医,2009；31(2)：191-192
[18]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.中国医院药学杂志,2006；26(10)：1275
[19]李辉,邱仕君.邓铁涛教授对“痰瘀相关”理论的阐释和发挥.湖北民族学院学报 (医学版),2005；22(1)：45-47
[20]王晓峰,李鹏,赵永东.沈宝藩教授运用痰瘀同治法经验摭拾.实用中医内科杂志,2004：18(4)：298-299
[21]韩素芳.益气活血汤治疗气虚血瘀型冠状动脉粥样硬化性心脏病心绞痛30例.河北中医,2009；31(5)：690
[22]晏庆德,张学颖.补阳还五汤片剂治疗早期动脉硬化性闭塞症临床研究.山东中医杂志,2005；24(3)：140-142
[23]王长垠,殷春萍,刘红霞.补肾法对颈动脉粥样硬化患者血清TNF-α、IL-6的影响.中国中医急症,2009；18(10)：1569-1570
[24]王少贤,杨晓锋.补肾活血化痰法对血管性痴呆患者血脂和血液流变学的影响.中华实用中西医杂志,2006；19(14)：1672-1673
[25]张芬梅,张慧岭,陈要玲,等.健脾降浊通脉方治疗动脉粥样硬化的临床观察.四川中医,2007；25(4)：45-46
[26]第五永长,刘修庄,陈亚文,等.健脾化痰活血利湿方药抗动脉粥样硬化作用的实验研究.陕西中医,2004；25(2)：181-182
[27]陈冰,宋剑南,牛晓红,等.健脾祛痰化瘀方对氧化型低密度脂蛋白诱导血管细胞信号分子钙离子和蛋白激酶C表达的影响.中国动脉硬化杂志,2004；12(2)：143-146
[28]王嵩,刘煜德,李荣,等.清热解毒法对50例动脉粥样硬化患者血脂、纤维蛋白原、D-二聚体的干预作用.山西中医,2009；24(6)：46-48
[29]王少英.清热解毒药物在治疗冠心病中的运用.北京中医,2004；23(1)：14-15
[30]杨俊卿,周岐新,李远宗.姜黄提取物对实验性动物高脂血症的防治作用研究.中国药房,2004；15(12)：598-600
[31]邱赛红,李福元,高雁,等.降脂消斑片对兔高脂血症合并动脉粥样硬化血脂的影响.湖南中医药大学学报,2009；29(3)：21-23
[32]朱明军,罗琰.调脂胶囊对动脉粥样硬化大鼠MDA、SOD的影响.中医药学刊,2006：24(10)：1821-1822
[33]罗陆一,冯润芬.地黄饮子治疗颈动脉粥样硬化斑块的临床研究.中华医学研究杂志,2004；4(5)：417-418
[34]甄艳军,牛丽颖,赵兰英,等.木贼提取物对早期AS大鼠血管平滑肌凋亡及Fas、FasL表达的干预.北京中医药大学学报,2007；30(1)：45-47
[35]程红,罗陆一.益肾通脉汤对兔动脉粥样硬化形成过程中平滑肌细胞增殖和凋亡的影响.江西中医学院学报,2009；21(4)：62-64
[36]Kervinen H, Palosuo T, Manninen V, et al. Joint effect of C-reactive protein and other risk factors on acute coronary events. Am Heart Journal,2001; 141:580-585 [37] Singh SK, Suresh MV, Voleti B, et al. The connection between C-reactive protein and atherosclerosis. Ann Med,2008; 40(2):110-120
[38]殷建明,罗陆一,华青.通心贴对冠心病血清Hs-CRP、M-CSF、IL-18的影响.江西中医药,2008；34(4)：28-29
[39]王晓杨,张志琴,张媛,等.CRP与在体兔动脉粥样粥样硬化相关性的研究及桑葚提取物的影响.中西医结合心脑血管病杂志,2009；7(9)：1053-1055
[40]Graham IM, Daly LE, Refsum HM, et al. Plasma homocysteine as a risk factor for vascular disease:The European Concerted Action Project. JAMA,1997; 277:1775-1781
[41]刘粉叶,张继东,胡连海,等.益肾活血胶囊对高同型半胱氨酸血症兔平滑肌细胞增殖及MMP-9基因表达的影响.南京中医药大学学报,2009；25(1)：35-37
[42]Blankenberg S, Rupprecht HJ, Bickel C, et al. Circulating cell adhesion molecules and death in patients with coronary artery disease. Circulation,2001; 104:1336-1342
[43]Sandra Tribolo, Federica Lodi, Carol Connor, et al. Comparative effect of quercetin and its predominant human metabolites on adhesion molecule expression in activated human vascular endothelial cells. Atherosclerosis,2008; 197:50-56
[44]张梅,温进坤,孙辉臣,等.丹参注射液对动脉粥样硬化大鼠血脂及细胞间黏附分子-1表达的影响.中国病理生理杂志,2004；20(10)：1871-1873
[45]Hojo Y, Ikeda U, Katsuki T, et al. Matrix metalloproteinase expression in the coronary circulation induced by coronary angioplasty. Atherosclerosis,2002; 161:185-192
[46]Parks WC, Wilson CI, Lopez-Boado YS, et al. Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol,2004; 4(8):617-629
[47]郑翠娥,郭伟星,李士涛,等.芪蛭合剂对大鼠实验性动脉粥样硬化主动脉MMP-9表达的影响.中华中医药学刊,2008；26(6)：1153-1155
[48]刘龙涛,吴敏,张文高,等.虎杖苷对颈动脉粥样硬化斑块稳定性的干预研究.北京中医药,2009；28(3)：172-175
[49]才向军.化浊降脂汤治疗高脂血症80例.陕西中医,2004；25(2)：135-136
[50]方微,张慧信,王绿娅,等.何首乌总甙抑制动脉粥样硬化病变形成.中国动脉硬化杂志,2005；13(2)：175-178
[51]Liu QL, Xiao JH, Ma R, et al. Effect of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside on lipoprotein oxidation and proliferation of coronary arterial smooth cells. J Asian Nat Prod Res,2007; 9(6-8):689-697
[52]李香珍.调脂饮治疗老年高脂血症86例.四川中医,2004；22(10)：55
[53]邓平珍,徐敏.补骨脂对去卵巢大鼠骨转换及血脂代谢影响的实验研究.新中医,2005：37(7)：94-96
[54]童琦燕.牛膝治疗高尿酸血症高脂血症.辽宁中医杂志,2003；30(11)：943
[55]崔瑛,侯士良.怀牛膝预防动脉粥样硬化的实验研究.基层中药杂志,1998；12 (1)：30-31
[56]左迎.三七皂苷注射液治疗冠状动脉粥样硬化前后患者血液流变学变化.黑龙江医学,2006；30(10)：748
[57]殷智,彭再生,杨长丰.复方血三七胶囊治疗脑动脉粥样硬化症42例疗效观察.中国民族民间医药杂志,2005：73(2)：90-92
[58]张翼冠,李晓辉,樊继山,等.三七皂甙通过抗炎和调血脂作用抑制大鼠动脉粥样硬化形成.现代生物医学进展,2007；7(11)：1601-1603,1607
[59]闫彦芳,张壮,孙塑伦,等.三七皂甙及其主要成分对血管内皮细胞缺氧损伤的保护作用.中国实验方剂学杂志,2002：8(1)：34-37
[60]赖光强,林中,韩正雪,等.地龙胶囊对缺血性脑梗死患者颈动脉粥样斑块的影响研究.中国中医急症,2009；18(7)：1034-1035
[61]白雪,杨思进,王丽萍,等.蚓激酶对不稳定型心绞痛患者C反应蛋白的影响.中国基层医药,2007；14(8)：1346-1347
[62]许超千,焦军东,孙宏丽,等.蚓激酶对实验动物血栓及相关血液学指标的影响.中草药,2005；36(9)：1367-1369
[63]陈奕芝,方若鸣,魏刚,等.石菖蒲挥发油、p-细辛醚对高脂血症大鼠血管舒张与抗血小板聚集的作用.中国中西医结合杂志,2004；24(基础理论研究特集)：16-18
[64]吴启端,吴清和,王绮雯,等.石菖蒲挥发油及p-细辛醚的抗血栓作用.中药新药与临床药理,2008；19(1)：29-31
[65]Hemanez A, M L, Chamorro C, et al. Inhibition of lipid synthesis and secretion in long-term cultures of adult rat bepatocytes by alpha-asarone. Planta Med,1993; 59 (2):121
[66]杨永宗.动脉粥样硬化性心血管病基础与临床,北京：科学出版社,2009,第二版：53
[67]Ross R. The pathogenesis of atherosclerosis. New Engl J Med,1976,295 (8):376-420
[68]Ross R. Atherosclerosis-an inflammatory disease, New Engl J Med,1999; 340 (2): 115-126
[69]Erling Falk. Pathogenesis of atherosclerosis. Journal of the American College of Cardiology.2006; 47(8):7-12
[70]Elena Galkina, Klaus Ley. Vascular adhesion molecules in atherosclerosis. Arteriosclerosis, Thrombosis, and Vascular Biology.2007; 27(11):2292-2301
[71]唐可京,李幼姬,谢灿茂.ICAM-1和VCAM-1的结构与表达调控.国外医学分子生物学分册,2002；24(3)：173-177
[72]Rothlein R, Dustin ML, Marlin SD, et al. A human intercellular adhesion molecule-1 (ICAM-1) distinct from LFA-1. J Immunol,1986; 137 (2):1270
[73]Jordi Bella, Prasanna R. Kolatkar, Christopher W. Marlor, et al. The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. Proc. Natl. Acad. Sci.,1998; 95:4140-4145
[74]Pietruczuk M, Pietruczuk A, Pancewicz S, et al. ICAM-1:structure, biological role and clinical significance. Pol Merkur Lekarski,2004; 17 (101):507-511
[75]Osborn L, Hession C, Tizard R, et al. Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes. Cell,2004; 59:1203-1211
[76]Rice GE, Bevilacqua MP.An inducible endothelial cell surface glycoprotein mediates melanoma adhesion. Science,1989; 246:1303-1306
[77]Jia-huai Wang, R Blake Pepinsky, Thilo Stehle, et al. The crystal structure of an N-terminal two-domain fragment of vascular cell adhesion molecule 1 (VCAM-1):a [75] cyclic peptide based on the domain 1 C-D loop can inhibit VCAM-1-a4 integrin interaction. Proc. Natl. Acad. Sci.,1995; 92:5714-5718
[78]Nureddin Cengiz, Ender Erdogan, Hanefi Ozbek, et al. Adhesion molecules in cerebral ischemia and atherosclerosis. Eur J Gen Med,2009; 6(4):249-256
[79]Blankenberg S, Barbaux S, Tiret L. Adhesion molecules and atherosclerosis. Atherosclerosis,2003; 170(2):191-203
[80]Poston RN, Haskard DO, Coucher JR, et al. Expression of intercellular adhesion molecule-1 in atherosclerotic plaques. Am J Pathol,1992; 140:665-673
[81]Printsera OY, Peclo MM, Gown AM. Various cell types in human atherosclerotic lesion express ICAM-1:Futher immuocytechemical and immuno chemical studies employing monoclonal antibody 10F3. Am J Pathol,1992; 140:889-896
[82]Daniel S. The neuropathology and neurochemistry of multiple system atrophy. In Mathias C, Bannister R, eds. Autonomic failure:a textbook of clinical disorders of the autonomic nervous system, Oxford:Oxford University Press,1999,3rd ed:321-328
[83]Asanori Aikawa, Peter Libby. The vulnerable atherosclerotic plaque:Pathogenesis and therapeutic approach. Cardio Vascular Pathology,2004; 13(3):125-138
[84]Thome SA, Abbot SE, Stevens CR, et al. Modified low density lipoprotein and cytokines mediate monocyte adhesion to smooth muscle cell. Atherosclerosis,1996; 95: 1263
[85]Cybulsky Myron L, Gimbrone Michael A, Jr. Endothelial expression of mononuclear leukocyte adhesion molecule during atherogenesis. Science,1991; 251(4995):788-791
[86]Kitagawa K, Matsumoto M, Sasaki T, et al. Involvement of ICAM-1 in the progression of atherosclerosis in APOE-knockout mice. Atherosclerosis,2002; 160(2):305-310
[87]Scalia R, Appel JZ 3rd, Lefer AM. 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,1998; 18(7):1093-1100
[88]Sen R, Baltimore D. Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism. Cell,1986; 47(6):921-928
[89]Paul P Tak, Gary S. Firestein. NF-κB:a key role in inflammatory disease. J Clin Invest, 2001; 107(1):7-11
[90]John Kuriyan, Dinitris Thanos. Structure of the NF-κB transcription factor: a holistic interaction with DNA. Structure,1995; 3:135-141
[91]Marc D Jacobs, Stephen C Harrison. Structure of an IκBα/NF-κB complex. Cell,1998; 95:749-758
[92]CollinsT. Endothelial nuclear factor-Kappa B and the unitation of the atherosclerotic lesion. lab Invest,1993; 69:499
[93]O'Connor S, Markovina S, Miyamoto S. Evidence for a phosphorylation-independent role for Ser 32 and 36 in proteasome inhibitor-resistant (PIR) IkappaBalpha degradation in B cells. Exp Cell Res,2005; 307(1):15-25
[94]Qing G, Qu Z, Xiao G. Stabilization of basally translated NF-kappaB-inducing kinase (NIK) protein functions as a molecular switch of processing of NF-kappaB2 p100. J Biol Chem,2005; 280(49):40578-40582
[95]Leena Hajra, Andrew I. Evans, Mian Chen, et al. The NF-κB singnal transduction pathway in aortic endothelial cells is primes for activation in regions predisposed to atherosclerosis lesion formation. Cell Biology,2000; 97(16):9052-9057
[96]K Brand, S Page, G Rogler, et al. Activated transcription factor nuclear-kappa B is present in the atherosclerotic lesion. J Clin Invest,1996; 97(7):1715-1722
[97]Burleigh ME, Babaev VR, Oates JA, et al. Cyclooxygenase-2 promotes early atherosclerotic lesion formation in LDL receptor-dificient mice. Circulation.2002; 105: 1816-1823
[98]Gu L, Okada Y, Clinton SK, et al. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low-density lipoprotein receptor-dificent mice. Mol Cell,1998; 2: 275-281
[99]Cybulsky MI, Liyama K, Li H, et al. A major role for VCAM-1, but not ICAM-1, in early atherosclerosis. J Clin Invest,2001; 107:1255-1262
[100]Park CW, Kim JH, Lee JH, et al. High glucose-induced intercellular adhesion molecule-1 (ICAM-1) expression through an osmotic effect in rat mesangial cells is PKC-NF-KB-dependent. Diabetologia,2000; 58(5):2166-2177
[101]Lawrence T, Gilroy DW, Colville-Nash PR, et al. Possible new role for NF-kappa B in the resolution of inflammation. Nat Med,2001; 7:1291
[102]方微,张慧信,王绿娅,等.何首乌总苷抗氧化与实验性小鼠动脉粥样硬化病变的形成.中国中药杂志,2007；32(13)：1320-1323
[103]Gerald Rimbach, Giuseppe Valacchi, Raffaella Canali, et al. Macrophages stimulated with IFN-y activate NF-κB and induce MCP-1 gene expression in primary human endothelial cells. Molecular Cell Biology Research Communication,2000; 3(4):238-242
[104]黄铮,龚兰生,施仲伟,等.颈动脉超声结合危险因素调查冠心病高危患者,中国超声医学杂志,1998；14(3)：16-18
[105]Crouse JR, Harpold GH, Kahl FR, et al. Evaluation of a scoring system of extracranial carotid atherosclerosis extent with B-mode ultrasound. Stroke,1986; 17(2): 270-275
[106]Lemne C, Jogestrand T, de Faire U. Carotid intima-media thickness and plaque in borderline hypertension. Stroke,1995; 26(1):34-39
[107]Kawasaki J. Diagnostic accuracy of carotid ultrasonography in screening for coronary artery disease. J Cardiol,2000; 36(5):295-302
[108]Sonia S Anand, Salim Yusuf, Vladmir Vuksan, et al. Differences in risk factors, atherosclerosis, and cardiovascular disease between ethnic groups in Canada: the Study of Health Assessment and Risk in Ethnic groups (SHARE). Lancet,2000; 356(9226):279-284
[109]Pognoli P, Tremoli E, Poli A, et al. Intimal plus medial thichness of the arterial wall:a direct measurement with ultrasound imaging. Circulation, 1986; 74(6):1399-1406
[110]Salonen JT, Korpela K, Salonen R, et al. Autoantibody against oxidized LDL and progression of carotid atherosclerosis. Lancet,1992; 339(8798):883-887
[111]Libby P, Aikawa M, Schonbeck U. Cholesterol and atherosclerosis. Biochim Biophys Acta,2000; 1529(1-3):299-309
[112]Rader DJ. High-density lipoprotein and atherosclerosis. Am J Cardiol, 2002; 90(8A): 62i-70i
[113]John S, Jacobi J, Delles C, et al. Plasma soluble adhesion molecules and endothelium-dependent vasodilation in early human atherosclerosis. Clin Sci (Lond),2000; 98(5):521-529
[114]Jie J Cao, Chau Thach, Teri A Manolio, et al. C-reactive protein, carotid intima-media thickness, and incidence of ischemic stroke in the elderly. Circulation,2003; 108:166-167
[115]Stefan Blankenberg, Rupprecht HJ, Christoph Bickel, et al. Circulating cell adhesion molecules and death in patients with coronary artery disease. Circulation,2001; 104: 1336-1342
[116]Yu CM, Zhang Q, Lam L, et al. Comparison of intensive and low-dose atovastatin therapy in the reduction of carotid intimal-medial thickness in patients with coronary heart disease. Heart,2007; 93(8):933-939
[117]殷建明,罗陆一.通心贴对冠状动脉粥样硬化性心脏病患者血清Hs-CRP、IL-18、GMP-140的影响.上海中医药杂志,2008；42(4)：26-27
[118]程红,罗陆一.益肾通络汤对兔动脉粥样硬化模型平滑肌细胞凋亡的干预作用.中西医结合心脑血管病杂志,2009；7(5)：551-552
[119]周凌,伍津津,鲁元刚,等.人脐静脉内皮细胞的分离培养和冻存.重庆医学,2006：35(3)：226-230
[120]Fan Zhang, Wei Yu, James L. Hargrove,et al. Inhibition of TNF-α induced ICAM-1, VCAM-1 and E-selectin expression by selenium, Atherosclerosis,2002; 161:381-386
[121]Carswell EA, Old LJ, Kassel RL, et al. An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci,1975; 72(9):3666-3670
[122]Shalaby MR, Aggarwal BB, Rinderknecht E, et al. Activation of human polymorphonuclear neutrophil functions by interferon-gamma and tumor necrosis factors. J Immunol,1985; 135(3):2069-2073
[123]Stannard AK, Riddell DR, Bradley NJ, et al. Apolipoprotein E and regulation of cytokine-induced cell adhesion molecule expression in endothelial cells. Atherosclerosis. 1998; 139:57-64
[124]Barath P, Fishbein MC, Cao J, et al. Detection and localization of tumor necrosis factor in human athroma. Am J Cardiol,1990; 65(5):297-302
[125]张寄南,曹克将,杨志健.心脏标志物学,南京：江苏科学技术出版社,2007,第一版：80
[126]Price DT, Loscalzo J. Cellular adhesion molecules and atherogenesis. Am J Med, 1999; 107(1):85-97
[127]Calixto JB, Campos MM, Otuki MF, et al. Anti-inflammatory compounds of plant origin. Planta Med,2004; 70 (2):9
[128]Okadas, Shikatak, Matsudam, et al. Intercellular adhesion molecule-1-deficient mice a resistant against renal injury after induction of diabetes. Diabetes,2003; 52 (10):2586-2593
[129]Pirisi M, Vitulli D, Falleti E, et al. Increased soluble ICAM-1 concentration and impaired delay-type hypersensitivity skin tests in patients with chronic liver disease. J Clin Pathol,1997; 50(1):50-53
[130]Collins T. Endothelial nuclear factor-Kappa B and the unitation of the atherosclerotic lesion. Lab Invest,1993; 69:499
[131]Kim CD, Kim YK, Lee SH, et al. Rebamipide inhibits neutrophil adhesion to hypoxia/reoxygenation-stimulated endothelial cell via nuclear factor kappaB-dependent pathway. J Pharmacol Exp Ther,2000; 294 (3): 864-869
[132]朱陵群,王硕仁,赵明镜,等.活血注射液对ox-LDL诱导人脐静脉内皮细胞活化的影响.中国中药杂志,2008；33(13)：1617-1621