血脉宁颗粒对动脉粥样硬化模型家兔胆固醇逆转运作用机制的研究
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摘要
动脉粥样硬化（atherosclerosis，AS）是指发生在动脉的非炎症性、退行性、增生性的病变，最终导致动脉血管壁增厚变硬、失去弹性及管腔缩小。
AS常累及大、中型动脉，病变从内膜开始，局部有脂质和复合糖类积聚、出血和血栓形成、纤维组织增生和钙质沉着，并有动脉中层的逐渐退变和钙化。AS的病程漫长，是在生命过程中缓慢形成和发展的，病机十分复杂，病变在发展过程中可出现活动、进展、静止和消退等不同时期的相应变化。临床将AS分为早期和晚期两大类病变，早期病变的显著特点就是脂质条纹，即脂纹，也有人提出微血栓和胶样病变也是AS的早期病变，称之为横纹；晚期病变的显著特点就是粥样斑块或纤维斑块的形成，并由此引发各种疾病包括脑血管意外或心血管疾病。
动脉粥样硬化属于现代医学的病名，中医学与该病相关的论述散见于“痰浊”、“胸痹”、“中风”、“眩晕”、“健忘”、“风痒”、“痴呆”、“真心痛”、“消渴”、“肾痹”、“脱疽”等疾病的记载当中。AS是导致心、脑及其他重要器官血管疾病的主要原因，临床以心脑血管发生病变为主，也可见于部分周围血管病。病人临床表现多为眩晕，肢麻，心悸，气短，头疼，胸闷，语言不利，半身活动障碍，颈部不适，阵发性疼痛，心前区疼痛，下肢跛行，舌质红、苔多白腻边有癖点，脉多沉细涩、弦紧等症状。临床表现形式多种多样，各个阶段证候不同，其症状也不尽相同。其病因病机颇为复杂，中医学认为，AS的发生与机体年老体衰，嗜食膏粱厚味，内伤七情，寒邪侵袭等因素有关，气虚血疲，脾虚湿盛，痰癖互结，痰癖化毒，肝肾阴虚是本病的主要病机。
现代医学对动脉粥样硬化疾病的研究较为详尽，胆固醇逆转运在动脉粥样硬化的发生发展中起着至关重要的作用，因为研究发现，胆固醇逆转运是胆固醇排出体外的唯一有效途径，因此在对于该病发病机理及以此为靶点的新药研制过程中，多涉及胆固醇逆转运的研究。
多年来，导师陈民教授以中医理论为指导，结合现代医学前沿的研究成果，综合治疗动脉粥样硬化疾病的经验，探索出了中药复方血脉宁颗粒，该方针对动脉粥样硬化的治疗和预防均取得了良好的临床效果。陈民教授认为，动脉粥样硬化主要是因为年老体弱，正气不足，饮食失调，嗜食肥甘，从而导致脾失健运，气虚血瘀，痰瘀互结，郁而化热，热毒伤脉，由此导致痰、血、热毒瘀结于脉中。因此，该病病机为气虚血瘀，脉络痹阻。故以“补气活血，降脂通络”为根本治疗大法。方中以黄芪、绞股蓝为君药，行补气降脂之功；辅以丹参、川芎、三七，以助活血化瘀之力；配以山楂、茯苓、泽泻，祛脂除湿；诸药相辅相成，共奏补气活血、降脂通络之功。
本研究在动脉粥样硬化中医病因病机理论的指导下，结合现代医学检测手段，以家兔为实验对象，拟探明中药复方血脉宁颗粒治疗动脉粥样硬化疾病的部分作用机理，为临床AS的治疗以及血脉宁颗粒的应用提供科学依据。
第一部分血脉宁颗粒对AS模型家兔血脂影响的实验研究
目的：
观察血脉宁颗粒对动脉粥样硬化模型家兔血脂的调节作用，验证血脉宁颗粒的临床疗效。
材料与方法：
本研究以家兔为实验对象，将40只家兔按照随机数字法，随机分为四组，分别为：空白对照组，模型组，血脉宁颗粒组，西药对照组。家兔适应性饲养一周后，对模型组、血脉宁颗粒组和西药对照组进行模型复制，即采用高脂饲料喂饲的方法进行动脉粥样硬化模型的复制：模型组、血脉宁颗粒组和西药对照组家兔均给予高脂饲料喂养，并于实验第一天一次性的腹腔注射维生素D3（60万IU/kg）。空白对照组给予正常颗粒饲料喂养，并于实验第一天给予等量生理盐水腹腔注射。连续以高脂饲料喂养4周后，开始对血脉宁颗粒组和西药对照组进行治疗，分别经口灌饲血脉宁颗粒和阿托伐他汀，每天一次，连续灌饲4周。至实验第八周末，经耳缘静脉采集各组家兔静脉血4ml，分离血清，采用酶法检测各组家兔血清中甘油三酯、胆固醇、高密度脂蛋白、低密度脂蛋白的含量。截取各组家兔1.5～2cm的腹主动脉组织，浸泡于4%多聚甲醛内，常规石蜡包埋、切片、HE染色，观察腹主动脉的病理变化。采用SPSS10.0软件包对各组实验数据进行统计分析，计量资料以均数±标准差（x±s）表示，多组均数比较采用单因素方差分析，以p＜0.05为有统计学意义。
结果：
空白对照组家兔腹主动脉平滑肌细胞排列整齐，未见脂质沉积及动脉粥样硬化斑块。模型组家兔腹主动脉可见明显的动脉粥样硬化斑块形成，而血脉宁颗粒组和西药对照组家兔腹主动脉病理改变明显轻于模型组。
与空白对照组比较，模型组，血脉宁颗粒组，西药对照组家兔血清中胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白含量显著升高，有统计学意义（p＜0.01）。
与模型组比较，血脉宁颗粒组，西药对照组家兔血清中胆固醇、甘油三酯、低密度脂蛋白含量显著降低，而高密度脂蛋白含量显著增高，有统计学意义（p＜0.01）。
两治疗组家兔血清中胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白含量比较，无统计学意义（p＞0.05）。
结论：
1.单纯高脂饲料连续喂饲家兔8周即可在腹主动脉等大动脉处形成明显的动脉粥样硬化斑块。
2.血脉宁颗粒可有效降低高脂血症或动脉粥样硬化模型家兔血清中的甘油三酯、总胆固醇、低密度脂蛋白；上调高密度脂蛋白水平。
3.血脉宁颗粒对血脂具有双向调节作用。
第二部分血脉宁颗粒对AS模型家兔巨噬细胞胆固醇含量及ABCA1表达影响的实验研究
目的：
观察血脉宁颗粒对动脉粥样硬化模型家兔腹腔巨噬细胞内脂质的清除作用，观察AS模型家兔腹腔巨噬细胞表面ABCA1基因及蛋白表达的调节作用，探索中药复方血脉宁颗粒治疗动脉粥样硬化的作用机制之一。
材料与方法：
动物实验的分组及药物干预措施同第一部分，于实验结束前的第三天，每只家兔经腹腔注射入无菌淀粉溶液2ml。实验结束后，处死各组家兔，并向腹腔中注射PBS缓冲液40ml，轻揉腹部3分钟，然后剖开腹部，收集腹腔液，分离各组家兔腹腔巨噬细胞，采用油红O染色法观察各组家兔腹腔巨噬细胞荷脂情况，再用4%NP-40异丙醇溶液洗脱染色的巨噬细胞中的染料，492nm波长处检测各洗脱液的吸光度值，以反应各组家兔腹腔巨噬细胞荷脂量；免疫细胞化学法检测腹腔巨噬细胞表面ABCA1蛋白的表达；RT-PCR方法检测腹腔巨噬细胞表面ABCA1mRNA的表达情况。采用SPSS10.0软件包对各组实验数据进行统计分析，计量资料以均数±标准差（x±s）表示，多组均数比较采用单因素方差分析，以p＜0.05为有统计学意义。
结果：
模型组家兔腹腔巨噬细胞荷脂量显著高于其余三组，两治疗组之间无显著性差异。
免疫细胞化学检测结果发现，AS模型组家兔巨噬细胞ABCA1蛋白表达水平显著上调，与空白对照组比较有统计学意义（p＜0.01）。通过阳性细胞百分率的统计比较可知，与模型组比较，血脉宁颗粒能够显著上调ABCA1蛋白的表达，有统计学意义（p＜0.01）。
RT-PCR检测结果发现，AS模型家兔ABCA1mRNA表达均显著上调，药物干预后，上调更加显著，说明中药血脉宁颗粒对于ABCA1mRNA的表达有一定的上调作用。
两治疗组比较，家兔腹腔巨噬细胞荷脂量、细胞表面ABCA1基因及蛋白的表达水平均无统计学意义（p＞0.05）。
结论：
1.血脉宁颗粒能够有效清除泡沫细胞内脂质含量，从而有效抑制动脉粥样硬化疾病的发生。
2.血脉宁颗粒通过上调ABCA1基因及蛋白的表达水平来实现其清除泡沫细胞内脂质的作用。
3.血脉宁颗粒上调ABCA1基因及蛋白的表达，清除泡沫细胞内脂质可能是血脉宁颗粒防治动脉粥样硬化的机制之一。
第三部分血脉宁颗粒对AS模型家兔肝脏组织中SR-BI表达影响的实验研究
目的：
观察血脉宁颗粒对动脉粥样硬化模型家兔肝脏组织中SR-BI mRNA及SR-BI蛋白表达的调节作用，探索中药复方血脉宁颗粒治疗动脉粥样硬化的又一作用机制。
材料与方法：
动物实验的分组及药物干预措施同第一部分，待实验结束后，经耳缘静脉注入空气，导致空气栓塞，处死各组家兔，剖开腹部，摘取肝脏尾状叶，采用RT-PCR方法检测各组家兔肝脏组织中SR-BI mRNA的表达；采用Western-blot方法检测各组家兔肝脏组织中SR-BI蛋白的表达水平。采用SPSS10.0软件包对各组实验数据进行统计分析，计量资料以均数±标准差（x±s）表示，多组均数比较采用单因素方差分析，以p＜0.05为有统计学意义。
结果：
Western-blot结果显示，目的条带位于大约180kDa处，积分光密度值统计结果表明，与空白对照组比较，AS模型各组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）；与模型组比较，两治疗组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）。
RT-PCR结果显示，目的条带位于大约485bp处，积分光密度值统计结果表明，与空白对照组比较，AS模型各组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）；与模型组比较，两治疗组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）。
两治疗组比较，家兔肝脏组织中SR-BI基因及蛋白的表达水平均无统计学意义（p＞0.05）。
结论：
1.血脉宁颗粒可显著上调动脉粥样硬化模型家兔肝细胞中SR-BI mRNA的表达。
2.血脉宁颗粒可显著上调动脉粥样硬化模型家兔肝细胞中SR-BI蛋白的表达。
3.血脉宁颗粒可通过上调肝细胞中SR-BI基因及蛋白的表达而实现其对胆固醇的逆向转运的促进作用。
4.上调肝细胞中SR-BI基因及蛋白的表达，从而促进胆固醇逆转运可能是血脉宁颗粒防治动脉粥样硬化的又一机制。
Atherosclerosis (atherosclerosis，AS) refers to an event in the arteriesof the non inflammatory，degenerative，hyperplastic lesions，eventually leadingto arterial wall thickening and hardening，loss of flexibility and luminalnarrowing．
AS often involving the big，medium-sized arteries，intimal lesions from thestart，local lipid accumulation and complex carbohydrates，bleeding andthrombosis，fibrous tissue hyperplasia and calcinosis，and arterial medialgradual degeneration and calcification．AS is a long course, life in slowformation and development，pathogenesis is very complex，lesions in thedevelopment process can be seen in the activity，progress，static and subsidesin different periods corresponding changes．Clinical AS divided into early andlate two categories of lesions．Early lesion is characterized by a significantlipid stripe，namely fatty streaks，also somebody puts forward micro thrombusand gelatinous lesions is AS early lesions，also called striated．Late lesionis characterized by a significant atheromatous plaque or fibrous plaqueformation，and thus cause a variety of diseases including cardiovascular diseaseor cerebrovascular accident．
Atherosclerosis disease belongs to the modern medicine，Chinese medicineand disease related records scattered in“phlegm”，“chest”，“stroke”，“vertigo”，“forget”，“the itch”，“dementia”，“true heart”，“diabetes”，“kidney paralysis”，“gangrene” diseases such as therecord．AS is the leading cause of heart，brain and other important organs ofvascular disease is the major cause for the incidence of cardiovascular andcerebrovascular disease，clinical disease，can also be found in the section ofperipheral vascular diseases．The patient manifested as dizziness，palpitations，shortness of breath，limb numbness， headache， chest tightness， negativelanguage， body movement disorder，neck discomfort，paroxysmal pain，precordial pain，lower limb claudication，red tongue，fur more greasy side stereotypies，Mai Shen fine tart，tension and other symptoms. Clinical manifestations ofvarious forms，various stages of syndrome，the symptoms are not the same．Itspathogenesis is complex， in medicine，AS occurs and the infirm，eat rich foodis delicious， internal causes，pathogenic cold invasion and other factors，Qi deficiency and blood stasis syndrome of spleen deficiency and dampness，Sheng，phlegm，phlegm and blood stasis of detoxification，liver and kidneydeficiency is the major pathogenesis of this disease．
Modern medicine for atherosclerotic disease research in detail，reversecholesterol transport in atherosclerosis development plays a vital role，because the study found，reverse cholesterol transport cholesterol excretionis the only effective way，therefore, for the disease pathogenesis and as a targetfor the development of new drug research process，many involved in reversecholesterol transportation research．
Over the years，tutor to traditional Chinese medicine theory，combined withmodern medical cutting-edge research results，as well as the rich treatment ofatherosclerotic disease experience， explored a compound Chinese herbal Xue MaiNing granule，the compound preparation for the treatment and prevention ofatherosclerosis have achieved good clinical curative effect. We think，atherosclerosis is mainly because of frail，the deficiency of vital qi，eatingdisorders，eat fatness，which leads to the spleen fails，stagnation of Qi andblood，phlegm and blood stasis syndrome，depression and of heat，heat injuryof vein，resulting in the sputum， blood stasis， poison， heat， etc. in theveins. Therefore，the disease pathogenesis is deficiency of Qi and blood stasis，vein blockage. The “Buqi Huoxue Tongluo，lipid-lowering treatment” as thefundamental law. Party in Astragalus， Gynostemma pentaphyllum as monarch drug，line Qi lipid-lowering of reactive power， supplemented with Danshen， RhizomaChuanxiong， three seven， to help blood circulation power． With hawthorn，tuckahoe， Alisma， Quzhi desiccant. The drug supplement each other， playeda total of Buqi Huoxue Tongluo， lipid-lowering．
The study of atherosclerosis in the pathogenesis of Chinese medicine underthe guidance，combined with modern medical detection means，in atheroscleroticrabbits as experimental object，through to the rabbit serum cholesterol，triglyceride， high density lipoprotein，low density lipoprotein content in foamcells， lipid content， ABCA1gene and protein expression levels，the liver cellSR-BI gene and protein expression level monitoring， of Xue Mai Ning Granuleon blood lipid regulation role in atherosclerotic rabbits atherosclerosis modelof rabbit peritoneal macrophages， the surface of ABCA1gene and proteinexpression of Xue Mai Ning Granule on regulation and atherosclerosis model rabbitliver tissue in SR-BI mRNA and SR-BI protein expression influence. Thus fromthe blood rheology, gene expression，protein expression in different anglefurther proved the compound Chinese herbal Xue Mai Ning granule in the treatmentof atherosclerotic disease mechanisms， clinical atherosclerosis in patientswith treatment of Xue Mai Ning particles and clinical application and promotionto provide scientific experimental basis．
PART ONEXue Mai Ning Granule on the experimental study of the effectsof AS rabbit lipid
Purpose：
Observation of Xue Mai Ning Granule on blood lipid regulation role inatherosclerotic rabbits，verify the clinical curative effect of Xue Mai Ninggranule．
Materials and methods：
This study in rabbits as experimental objects，40rabbits were in accordancewith the random number method， were randomly divided into four groups， respectively： blank control group， model group， Xue Mai Ning granule group，the control group of Western medicine．Adaptive feeding rabbits after a week，the model group， Xue Mai Ning granule group and Western medicine group weremodel replication. Using high fat diet feeding methods atherosclerosis modelreplication，model group， Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet， and in experiment first day one-timeintraperitoneal injection of vitamin D3（600000IU/kg）. Blank control groupgiven normal granular feed， and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous with high fat diet for4weeks,beginning of Xue Mai Ning granule group and Western medicine group were treatedwith, by mouth filling feeding Xue Mai Ning granule and atorvastatin， once aday，after4weeks of continuous irrigation．To experiment eighth weekend，through ear vein collecting all rabbits venous blood4ml，isolated from theserum by enzyme assay，all rabbits serum triglyceride，cholesterol，high densitylipoprotein，low density lipoprotein in. The interception of the abdominal aortain rabbits of each group1.5～2CM, soaked in4%paraformaldehyde, embedded inparaffin, sections, HE staining was observed in the abdominal aorta,pathological changes．Using SPSS10.0software package for each of the groupsof experimental data for statistical analysis，measurement data to mean±SD(x±s)，multiple group were compared with single factor analysis of variance，with P＜0.05was considered statistically significant．
Result：
Blank control group rabbit abdominal aorta smooth muscle cells arrangedregularly, no lipid deposition and atherosclerotic plaque. Model group rabbitabdominal aorta showed obvious atherosclerotic plaque formation, and Xue MaiNing granule group and Western medicine group rabbit abdominal aortapathological change was significantly milder than that in the model group。
Compared with the blank control group, model group，Xue Mai Ning granule group， the control group of Western medicine in rabbit serum cholesterol，triglyceride，high density lipoprotein，low density lipoprotein contentincreased significantly，with statistical significance (P＜0.01)．
Compared with the model group， Xue Mai Ning granule group，the control groupof Western medicine in rabbit serum cholesterol，triglyceride，low densitylipoprotein content decreased，and high density lipoprotein content decreased，with statistical significance (P＜0.01)．
The two treatment groups in rabbit serum cholesterol，triglyceride，highdensity lipoprotein，low density lipoprotein content， without statisticalsignificance (P＞0.05)．
Conclusion：
1．Simple high-fat feed continuous feeding rabbits after8weeks in suchlarge abdominal aortic artery formation was evident in the atheroscleroticplaque．
2．Xue Mai Ning granule can effectively reduce hyperlipidemia andatherosclerosis model in rabbits serum triglyceride，total cholesterol，lowdensity lipoprotein；increase high density lipoprotein level．
3．Xue Mai Ning Granule on blood lipids has bidirectional regulation effect．
PART TWOXue Mai Ning Granule on the experimental study of the effectsof AS rabbit macrophage cholesterol content and ABCA1expression
Purpose：
Observation of Xue Mai Ning Granule on peritoneal macrophages inatherosclerotic rabbits lipid clearance effect of AS model； observation of rabbit peritoneal macrophages surface ABCA1gene and protein expressionregulating effect of compound traditional Chinese medicine，exploration of XueMai Ning granule in treating atherosclerosis mechanism．Materials and methods：
This study in rabbits as experimental objects，40rabbits were in accordancewith the random number method，were randomly divided into four groups，respectively：blank control group， model group，Xue Mai Ning granule group，the control group of Western medicine. Adaptive feeding rabbits after a week，the model group，Xue Mai Ning granule group and Western medicine group were modelreplication. Using high fat diet feeding methods atherosclerosis modelreplication，model group， Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet，and in experiment first day one-timeintraperitoneal injection of vitamin D3(600000IU/kg). Blank control groupgiven normal granular feed，and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous to high fat diet for4weeks，all rabbits through ear vein collecting venous blood3ml，isolated from the serumwere detected，the level of blood lipid of rabbits. By statistical analysis，compared with the blank control group，model group three rabbits blood lipidswere significantly higher than control group (P＜0.01)，that successfully mould，and the start of Xue Mai Ning granule group and the control group of Westernmedicine treatment，respectively，by mouth filling feeding Xue Mai Ning granuleand atorvastatin，once a day，after4weeks of continuous irrigation．In theexperiment until the end of the third day，every rabbits by intraperitonealinjection into the sterile starch solution2ml/only. After the end of theexperiment，put to death all rabbits，and to the peritoneal injection of PBSbuffer40ml，gently massage the abdomen for3minutes，then cut open the belly，collection of peritoneal fluid，separation of rabbit peritoneal macrophages wereobserved in each group，rabbit peritoneal macrophage lipid loaded condition， then the4%NP-40isopropanol solution elution staining of macrophages in the dye，492nm wavelength detection the eluate of the absorbance value，in response toall rabbits peritoneal macrophage lipid loaded weight；immunocytochemicaldetection of peritoneal macrophage surface expression of ABCA1protein；RT-PCRmethod for the detection of peritoneal macrophage surface ABCA1mRNA expression.Using SPSS10.0software package for each of the groups of experimental data forstatistical analysis，measurement data to mean±SD(x±s)，multiple group werecompared with single factor analysis of variance，with P＜0.05was consideredstatistically significant．Result：
Model rabbits peritoneal macrophage lipid loaded weight was significantlyhigher than the other three groups，between the two treatment groups with nosignificant difference．
Immunocytochemical detection results，AS model rabbits of group ABCA1ofmacrophages protein expression level was significantly higher，compared withthe blank control group was statistically significant (P＜0.01). Through thepositive cell percentage statistics study shows that，compared with the modelgroup，Xue Mai Ning granule could significantly increase the ABCA1proteinexpression，with statistical significance (P＜0.01)．
The results of RT-PCR，AS ABCA1mRNA expression in rabbits were significantlyup-regulated，drug intervention，increase more significantly，indicating thatChinese medicine Xue Mai Ning granule for the ABCA1mRNA expression has someupregulate．
The two treatment groups were comparable，rabbit peritoneal macrophageslipid loaded weight，cell surface ABCA1gene and protein expression levels werenot statistically significant (P＞0.05)．
Conclusion：
1．Xue Mai Ning granule can effectively remove foam intracellular lipidcontent，thereby effectively inhibit atherosclerosis disease．
2．Xue Mai Ning granule through the up-regulation of ABCA1gene and proteinexpression level to achieve the removal of foam cells role of lipid．
3．Xue Mai Ning granule upregulate ABCA1gene and protein expression，intracellular lipid removal of foam may be Xue Mai Ning granule in preventingand treating atherosclerosis is one of the mechanisms．
PART THREEXue Mai Ning Granule on the experimental study of the effectsof AS rabbit liver tissue SR-BI expression
Purpose：
Observation of Xue Mai Ning Granule on liver tissue in rabbit atherosclerosismodel of SR-BI mRNA and protein expression of SR-BI regulation．Exploration ofcompound Chinese herbal Xue Mai Ning granule in treating atherosclerosismechanism．
Materials and methods：
This study in rabbits as experimental objects,40rabbits were in accordancewith the random number method，were randomly divided into four groups，respectively：blank control group，model group，Xue Mai Ning granule group，the control group of Western medicine．Adaptive feeding rabbits after a week，the model group，Xue Mai Ning granule group and Western medicine group were modelreplication．Using high fat diet feeding methods atherosclerosis modelreplication，model group，Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet，and in experiment first day one-timeintraperitoneal injection of vitamin D3(600000IU/kg). Blank control group given normal granular feed，and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous to high fat diet for4weeks，all rabbits through ear vein collecting venous blood3ml，isolated from the serumwere detected，the level of blood lipid of rabbits. By statistical analysis，compared with the blank control group，model group three rabbits blood lipidswere significantly higher than control group（P＜0.01），that successfully mould，and the start of Xue Mai Ning granule group and the control group of Westernmedicine treatment，respectively，by mouth filling feeding Xue Mai Ning granuleand atorvastatin，once a day，after4weeks of continuous irrigation．In theend of the experiment，through ear vein injection of air，cause an air embolism，put to death all rabbits，open abdominal，removal of liver caudate lobe，usingRT-PCR method to detect all rabbits liver tissues of SR-BI mRNA expression；using Western-blot for detecting clusters of rabbit liver tissue SR-BI proteinexpression level. Using SPSS10.0software package for each of the groups ofexperimental data for statistical analysis，measurement data to mean±SD(x±s)，multiple group were compared with single factor analysis of variance，with P＜0.05was considered statistically significant．
Result：
Western-blot results show，objective strap located approximately180kDa，integral optical density value of statistical results indicate that，comparedwith the blank control group，model group rabbit AS expression level of SR-BIwas significantly higher，with statistical significance (P＜0.01)；comparedwith the model group，treatment group two in rabbit SR-BI expression level wassignificantly higher，with statistical significance (P＜0.01)．
RT-PCR results show，objective strap located approximately485bp，integraloptical density value of statistical results indicate that，compared with theblank control group，model group rabbit AS expression level of SR-BI wassignificantly higher，with statistical significance (P＜0.01)；compared with the model group，treatment group two in rabbit SR-BI expression level wassignificantly higher，with statistical significance (P＜0.01)．
The two treatment groups were comparable，the rabbit liver tissue SR-BI geneand protein expression levels were not statistically significant (P＞0.05)．
Conclusion：
1．Xue Mai Ning granule can obviously increase in atherosclerotic rabbitsand SR-BI mRNA expression in liver cells．
2．Xue Mai Ning granule can obviously increase in atherosclerotic rabbitsand expression of SR-BI protein in liver cells．
3．Xue Mai Ning granule through upregulation of liver cell SR-BI gene andprotein expression and its effect on cholesterol reverse transport role．
4．upregulation of liver cell SR-BI gene and protein expression，whichpromotes reverse cholesterol transport may be Xue Mai Ning granule in preventingand treating atherosclerosis and mechanism．
AS常累及大、中型动脉，病变从内膜开始，局部有脂质和复合糖类积聚、出血和血栓形成、纤维组织增生和钙质沉着，并有动脉中层的逐渐退变和钙化。AS的病程漫长，是在生命过程中缓慢形成和发展的，病机十分复杂，病变在发展过程中可出现活动、进展、静止和消退等不同时期的相应变化。临床将AS分为早期和晚期两大类病变，早期病变的显著特点就是脂质条纹，即脂纹，也有人提出微血栓和胶样病变也是AS的早期病变，称之为横纹；晚期病变的显著特点就是粥样斑块或纤维斑块的形成，并由此引发各种疾病包括脑血管意外或心血管疾病。
动脉粥样硬化属于现代医学的病名，中医学与该病相关的论述散见于“痰浊”、“胸痹”、“中风”、“眩晕”、“健忘”、“风痒”、“痴呆”、“真心痛”、“消渴”、“肾痹”、“脱疽”等疾病的记载当中。AS是导致心、脑及其他重要器官血管疾病的主要原因，临床以心脑血管发生病变为主，也可见于部分周围血管病。病人临床表现多为眩晕，肢麻，心悸，气短，头疼，胸闷，语言不利，半身活动障碍，颈部不适，阵发性疼痛，心前区疼痛，下肢跛行，舌质红、苔多白腻边有癖点，脉多沉细涩、弦紧等症状。临床表现形式多种多样，各个阶段证候不同，其症状也不尽相同。其病因病机颇为复杂，中医学认为，AS的发生与机体年老体衰，嗜食膏粱厚味，内伤七情，寒邪侵袭等因素有关，气虚血疲，脾虚湿盛，痰癖互结，痰癖化毒，肝肾阴虚是本病的主要病机。
现代医学对动脉粥样硬化疾病的研究较为详尽，胆固醇逆转运在动脉粥样硬化的发生发展中起着至关重要的作用，因为研究发现，胆固醇逆转运是胆固醇排出体外的唯一有效途径，因此在对于该病发病机理及以此为靶点的新药研制过程中，多涉及胆固醇逆转运的研究。
多年来，导师陈民教授以中医理论为指导，结合现代医学前沿的研究成果，综合治疗动脉粥样硬化疾病的经验，探索出了中药复方血脉宁颗粒，该方针对动脉粥样硬化的治疗和预防均取得了良好的临床效果。陈民教授认为，动脉粥样硬化主要是因为年老体弱，正气不足，饮食失调，嗜食肥甘，从而导致脾失健运，气虚血瘀，痰瘀互结，郁而化热，热毒伤脉，由此导致痰、血、热毒瘀结于脉中。因此，该病病机为气虚血瘀，脉络痹阻。故以“补气活血，降脂通络”为根本治疗大法。方中以黄芪、绞股蓝为君药，行补气降脂之功；辅以丹参、川芎、三七，以助活血化瘀之力；配以山楂、茯苓、泽泻，祛脂除湿；诸药相辅相成，共奏补气活血、降脂通络之功。
本研究在动脉粥样硬化中医病因病机理论的指导下，结合现代医学检测手段，以家兔为实验对象，拟探明中药复方血脉宁颗粒治疗动脉粥样硬化疾病的部分作用机理，为临床AS的治疗以及血脉宁颗粒的应用提供科学依据。
第一部分血脉宁颗粒对AS模型家兔血脂影响的实验研究
目的：
观察血脉宁颗粒对动脉粥样硬化模型家兔血脂的调节作用，验证血脉宁颗粒的临床疗效。
材料与方法：
本研究以家兔为实验对象，将40只家兔按照随机数字法，随机分为四组，分别为：空白对照组，模型组，血脉宁颗粒组，西药对照组。家兔适应性饲养一周后，对模型组、血脉宁颗粒组和西药对照组进行模型复制，即采用高脂饲料喂饲的方法进行动脉粥样硬化模型的复制：模型组、血脉宁颗粒组和西药对照组家兔均给予高脂饲料喂养，并于实验第一天一次性的腹腔注射维生素D3（60万IU/kg）。空白对照组给予正常颗粒饲料喂养，并于实验第一天给予等量生理盐水腹腔注射。连续以高脂饲料喂养4周后，开始对血脉宁颗粒组和西药对照组进行治疗，分别经口灌饲血脉宁颗粒和阿托伐他汀，每天一次，连续灌饲4周。至实验第八周末，经耳缘静脉采集各组家兔静脉血4ml，分离血清，采用酶法检测各组家兔血清中甘油三酯、胆固醇、高密度脂蛋白、低密度脂蛋白的含量。截取各组家兔1.5～2cm的腹主动脉组织，浸泡于4%多聚甲醛内，常规石蜡包埋、切片、HE染色，观察腹主动脉的病理变化。采用SPSS10.0软件包对各组实验数据进行统计分析，计量资料以均数±标准差（x±s）表示，多组均数比较采用单因素方差分析，以p＜0.05为有统计学意义。
结果：
空白对照组家兔腹主动脉平滑肌细胞排列整齐，未见脂质沉积及动脉粥样硬化斑块。模型组家兔腹主动脉可见明显的动脉粥样硬化斑块形成，而血脉宁颗粒组和西药对照组家兔腹主动脉病理改变明显轻于模型组。
与空白对照组比较，模型组，血脉宁颗粒组，西药对照组家兔血清中胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白含量显著升高，有统计学意义（p＜0.01）。
与模型组比较，血脉宁颗粒组，西药对照组家兔血清中胆固醇、甘油三酯、低密度脂蛋白含量显著降低，而高密度脂蛋白含量显著增高，有统计学意义（p＜0.01）。
两治疗组家兔血清中胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白含量比较，无统计学意义（p＞0.05）。
结论：
1.单纯高脂饲料连续喂饲家兔8周即可在腹主动脉等大动脉处形成明显的动脉粥样硬化斑块。
2.血脉宁颗粒可有效降低高脂血症或动脉粥样硬化模型家兔血清中的甘油三酯、总胆固醇、低密度脂蛋白；上调高密度脂蛋白水平。
3.血脉宁颗粒对血脂具有双向调节作用。
第二部分血脉宁颗粒对AS模型家兔巨噬细胞胆固醇含量及ABCA1表达影响的实验研究
目的：
观察血脉宁颗粒对动脉粥样硬化模型家兔腹腔巨噬细胞内脂质的清除作用，观察AS模型家兔腹腔巨噬细胞表面ABCA1基因及蛋白表达的调节作用，探索中药复方血脉宁颗粒治疗动脉粥样硬化的作用机制之一。
材料与方法：
动物实验的分组及药物干预措施同第一部分，于实验结束前的第三天，每只家兔经腹腔注射入无菌淀粉溶液2ml。实验结束后，处死各组家兔，并向腹腔中注射PBS缓冲液40ml，轻揉腹部3分钟，然后剖开腹部，收集腹腔液，分离各组家兔腹腔巨噬细胞，采用油红O染色法观察各组家兔腹腔巨噬细胞荷脂情况，再用4%NP-40异丙醇溶液洗脱染色的巨噬细胞中的染料，492nm波长处检测各洗脱液的吸光度值，以反应各组家兔腹腔巨噬细胞荷脂量；免疫细胞化学法检测腹腔巨噬细胞表面ABCA1蛋白的表达；RT-PCR方法检测腹腔巨噬细胞表面ABCA1mRNA的表达情况。采用SPSS10.0软件包对各组实验数据进行统计分析，计量资料以均数±标准差（x±s）表示，多组均数比较采用单因素方差分析，以p＜0.05为有统计学意义。
结果：
模型组家兔腹腔巨噬细胞荷脂量显著高于其余三组，两治疗组之间无显著性差异。
免疫细胞化学检测结果发现，AS模型组家兔巨噬细胞ABCA1蛋白表达水平显著上调，与空白对照组比较有统计学意义（p＜0.01）。通过阳性细胞百分率的统计比较可知，与模型组比较，血脉宁颗粒能够显著上调ABCA1蛋白的表达，有统计学意义（p＜0.01）。
RT-PCR检测结果发现，AS模型家兔ABCA1mRNA表达均显著上调，药物干预后，上调更加显著，说明中药血脉宁颗粒对于ABCA1mRNA的表达有一定的上调作用。
两治疗组比较，家兔腹腔巨噬细胞荷脂量、细胞表面ABCA1基因及蛋白的表达水平均无统计学意义（p＞0.05）。
结论：
1.血脉宁颗粒能够有效清除泡沫细胞内脂质含量，从而有效抑制动脉粥样硬化疾病的发生。
2.血脉宁颗粒通过上调ABCA1基因及蛋白的表达水平来实现其清除泡沫细胞内脂质的作用。
3.血脉宁颗粒上调ABCA1基因及蛋白的表达，清除泡沫细胞内脂质可能是血脉宁颗粒防治动脉粥样硬化的机制之一。
第三部分血脉宁颗粒对AS模型家兔肝脏组织中SR-BI表达影响的实验研究
目的：
观察血脉宁颗粒对动脉粥样硬化模型家兔肝脏组织中SR-BI mRNA及SR-BI蛋白表达的调节作用，探索中药复方血脉宁颗粒治疗动脉粥样硬化的又一作用机制。
材料与方法：
动物实验的分组及药物干预措施同第一部分，待实验结束后，经耳缘静脉注入空气，导致空气栓塞，处死各组家兔，剖开腹部，摘取肝脏尾状叶，采用RT-PCR方法检测各组家兔肝脏组织中SR-BI mRNA的表达；采用Western-blot方法检测各组家兔肝脏组织中SR-BI蛋白的表达水平。采用SPSS10.0软件包对各组实验数据进行统计分析，计量资料以均数±标准差（x±s）表示，多组均数比较采用单因素方差分析，以p＜0.05为有统计学意义。
结果：
Western-blot结果显示，目的条带位于大约180kDa处，积分光密度值统计结果表明，与空白对照组比较，AS模型各组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）；与模型组比较，两治疗组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）。
RT-PCR结果显示，目的条带位于大约485bp处，积分光密度值统计结果表明，与空白对照组比较，AS模型各组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）；与模型组比较，两治疗组家兔SR-BI表达水平显著上调，有统计学意义（p＜0.01）。
两治疗组比较，家兔肝脏组织中SR-BI基因及蛋白的表达水平均无统计学意义（p＞0.05）。
结论：
1.血脉宁颗粒可显著上调动脉粥样硬化模型家兔肝细胞中SR-BI mRNA的表达。
2.血脉宁颗粒可显著上调动脉粥样硬化模型家兔肝细胞中SR-BI蛋白的表达。
3.血脉宁颗粒可通过上调肝细胞中SR-BI基因及蛋白的表达而实现其对胆固醇的逆向转运的促进作用。
4.上调肝细胞中SR-BI基因及蛋白的表达，从而促进胆固醇逆转运可能是血脉宁颗粒防治动脉粥样硬化的又一机制。
Atherosclerosis (atherosclerosis，AS) refers to an event in the arteriesof the non inflammatory，degenerative，hyperplastic lesions，eventually leadingto arterial wall thickening and hardening，loss of flexibility and luminalnarrowing．
AS often involving the big，medium-sized arteries，intimal lesions from thestart，local lipid accumulation and complex carbohydrates，bleeding andthrombosis，fibrous tissue hyperplasia and calcinosis，and arterial medialgradual degeneration and calcification．AS is a long course, life in slowformation and development，pathogenesis is very complex，lesions in thedevelopment process can be seen in the activity，progress，static and subsidesin different periods corresponding changes．Clinical AS divided into early andlate two categories of lesions．Early lesion is characterized by a significantlipid stripe，namely fatty streaks，also somebody puts forward micro thrombusand gelatinous lesions is AS early lesions，also called striated．Late lesionis characterized by a significant atheromatous plaque or fibrous plaqueformation，and thus cause a variety of diseases including cardiovascular diseaseor cerebrovascular accident．
Atherosclerosis disease belongs to the modern medicine，Chinese medicineand disease related records scattered in“phlegm”，“chest”，“stroke”，“vertigo”，“forget”，“the itch”，“dementia”，“true heart”，“diabetes”，“kidney paralysis”，“gangrene” diseases such as therecord．AS is the leading cause of heart，brain and other important organs ofvascular disease is the major cause for the incidence of cardiovascular andcerebrovascular disease，clinical disease，can also be found in the section ofperipheral vascular diseases．The patient manifested as dizziness，palpitations，shortness of breath，limb numbness， headache， chest tightness， negativelanguage， body movement disorder，neck discomfort，paroxysmal pain，precordial pain，lower limb claudication，red tongue，fur more greasy side stereotypies，Mai Shen fine tart，tension and other symptoms. Clinical manifestations ofvarious forms，various stages of syndrome，the symptoms are not the same．Itspathogenesis is complex， in medicine，AS occurs and the infirm，eat rich foodis delicious， internal causes，pathogenic cold invasion and other factors，Qi deficiency and blood stasis syndrome of spleen deficiency and dampness，Sheng，phlegm，phlegm and blood stasis of detoxification，liver and kidneydeficiency is the major pathogenesis of this disease．
Modern medicine for atherosclerotic disease research in detail，reversecholesterol transport in atherosclerosis development plays a vital role，because the study found，reverse cholesterol transport cholesterol excretionis the only effective way，therefore, for the disease pathogenesis and as a targetfor the development of new drug research process，many involved in reversecholesterol transportation research．
Over the years，tutor to traditional Chinese medicine theory，combined withmodern medical cutting-edge research results，as well as the rich treatment ofatherosclerotic disease experience， explored a compound Chinese herbal Xue MaiNing granule，the compound preparation for the treatment and prevention ofatherosclerosis have achieved good clinical curative effect. We think，atherosclerosis is mainly because of frail，the deficiency of vital qi，eatingdisorders，eat fatness，which leads to the spleen fails，stagnation of Qi andblood，phlegm and blood stasis syndrome，depression and of heat，heat injuryof vein，resulting in the sputum， blood stasis， poison， heat， etc. in theveins. Therefore，the disease pathogenesis is deficiency of Qi and blood stasis，vein blockage. The “Buqi Huoxue Tongluo，lipid-lowering treatment” as thefundamental law. Party in Astragalus， Gynostemma pentaphyllum as monarch drug，line Qi lipid-lowering of reactive power， supplemented with Danshen， RhizomaChuanxiong， three seven， to help blood circulation power． With hawthorn，tuckahoe， Alisma， Quzhi desiccant. The drug supplement each other， playeda total of Buqi Huoxue Tongluo， lipid-lowering．
The study of atherosclerosis in the pathogenesis of Chinese medicine underthe guidance，combined with modern medical detection means，in atheroscleroticrabbits as experimental object，through to the rabbit serum cholesterol，triglyceride， high density lipoprotein，low density lipoprotein content in foamcells， lipid content， ABCA1gene and protein expression levels，the liver cellSR-BI gene and protein expression level monitoring， of Xue Mai Ning Granuleon blood lipid regulation role in atherosclerotic rabbits atherosclerosis modelof rabbit peritoneal macrophages， the surface of ABCA1gene and proteinexpression of Xue Mai Ning Granule on regulation and atherosclerosis model rabbitliver tissue in SR-BI mRNA and SR-BI protein expression influence. Thus fromthe blood rheology, gene expression，protein expression in different anglefurther proved the compound Chinese herbal Xue Mai Ning granule in the treatmentof atherosclerotic disease mechanisms， clinical atherosclerosis in patientswith treatment of Xue Mai Ning particles and clinical application and promotionto provide scientific experimental basis．
PART ONEXue Mai Ning Granule on the experimental study of the effectsof AS rabbit lipid
Purpose：
Observation of Xue Mai Ning Granule on blood lipid regulation role inatherosclerotic rabbits，verify the clinical curative effect of Xue Mai Ninggranule．
Materials and methods：
This study in rabbits as experimental objects，40rabbits were in accordancewith the random number method， were randomly divided into four groups， respectively： blank control group， model group， Xue Mai Ning granule group，the control group of Western medicine．Adaptive feeding rabbits after a week，the model group， Xue Mai Ning granule group and Western medicine group weremodel replication. Using high fat diet feeding methods atherosclerosis modelreplication，model group， Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet， and in experiment first day one-timeintraperitoneal injection of vitamin D3（600000IU/kg）. Blank control groupgiven normal granular feed， and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous with high fat diet for4weeks,beginning of Xue Mai Ning granule group and Western medicine group were treatedwith, by mouth filling feeding Xue Mai Ning granule and atorvastatin， once aday，after4weeks of continuous irrigation．To experiment eighth weekend，through ear vein collecting all rabbits venous blood4ml，isolated from theserum by enzyme assay，all rabbits serum triglyceride，cholesterol，high densitylipoprotein，low density lipoprotein in. The interception of the abdominal aortain rabbits of each group1.5～2CM, soaked in4%paraformaldehyde, embedded inparaffin, sections, HE staining was observed in the abdominal aorta,pathological changes．Using SPSS10.0software package for each of the groupsof experimental data for statistical analysis，measurement data to mean±SD(x±s)，multiple group were compared with single factor analysis of variance，with P＜0.05was considered statistically significant．
Result：
Blank control group rabbit abdominal aorta smooth muscle cells arrangedregularly, no lipid deposition and atherosclerotic plaque. Model group rabbitabdominal aorta showed obvious atherosclerotic plaque formation, and Xue MaiNing granule group and Western medicine group rabbit abdominal aortapathological change was significantly milder than that in the model group。
Compared with the blank control group, model group，Xue Mai Ning granule group， the control group of Western medicine in rabbit serum cholesterol，triglyceride，high density lipoprotein，low density lipoprotein contentincreased significantly，with statistical significance (P＜0.01)．
Compared with the model group， Xue Mai Ning granule group，the control groupof Western medicine in rabbit serum cholesterol，triglyceride，low densitylipoprotein content decreased，and high density lipoprotein content decreased，with statistical significance (P＜0.01)．
The two treatment groups in rabbit serum cholesterol，triglyceride，highdensity lipoprotein，low density lipoprotein content， without statisticalsignificance (P＞0.05)．
Conclusion：
1．Simple high-fat feed continuous feeding rabbits after8weeks in suchlarge abdominal aortic artery formation was evident in the atheroscleroticplaque．
2．Xue Mai Ning granule can effectively reduce hyperlipidemia andatherosclerosis model in rabbits serum triglyceride，total cholesterol，lowdensity lipoprotein；increase high density lipoprotein level．
3．Xue Mai Ning Granule on blood lipids has bidirectional regulation effect．
PART TWOXue Mai Ning Granule on the experimental study of the effectsof AS rabbit macrophage cholesterol content and ABCA1expression
Purpose：
Observation of Xue Mai Ning Granule on peritoneal macrophages inatherosclerotic rabbits lipid clearance effect of AS model； observation of rabbit peritoneal macrophages surface ABCA1gene and protein expressionregulating effect of compound traditional Chinese medicine，exploration of XueMai Ning granule in treating atherosclerosis mechanism．Materials and methods：
This study in rabbits as experimental objects，40rabbits were in accordancewith the random number method，were randomly divided into four groups，respectively：blank control group， model group，Xue Mai Ning granule group，the control group of Western medicine. Adaptive feeding rabbits after a week，the model group，Xue Mai Ning granule group and Western medicine group were modelreplication. Using high fat diet feeding methods atherosclerosis modelreplication，model group， Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet，and in experiment first day one-timeintraperitoneal injection of vitamin D3(600000IU/kg). Blank control groupgiven normal granular feed，and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous to high fat diet for4weeks，all rabbits through ear vein collecting venous blood3ml，isolated from the serumwere detected，the level of blood lipid of rabbits. By statistical analysis，compared with the blank control group，model group three rabbits blood lipidswere significantly higher than control group (P＜0.01)，that successfully mould，and the start of Xue Mai Ning granule group and the control group of Westernmedicine treatment，respectively，by mouth filling feeding Xue Mai Ning granuleand atorvastatin，once a day，after4weeks of continuous irrigation．In theexperiment until the end of the third day，every rabbits by intraperitonealinjection into the sterile starch solution2ml/only. After the end of theexperiment，put to death all rabbits，and to the peritoneal injection of PBSbuffer40ml，gently massage the abdomen for3minutes，then cut open the belly，collection of peritoneal fluid，separation of rabbit peritoneal macrophages wereobserved in each group，rabbit peritoneal macrophage lipid loaded condition， then the4%NP-40isopropanol solution elution staining of macrophages in the dye，492nm wavelength detection the eluate of the absorbance value，in response toall rabbits peritoneal macrophage lipid loaded weight；immunocytochemicaldetection of peritoneal macrophage surface expression of ABCA1protein；RT-PCRmethod for the detection of peritoneal macrophage surface ABCA1mRNA expression.Using SPSS10.0software package for each of the groups of experimental data forstatistical analysis，measurement data to mean±SD(x±s)，multiple group werecompared with single factor analysis of variance，with P＜0.05was consideredstatistically significant．Result：
Model rabbits peritoneal macrophage lipid loaded weight was significantlyhigher than the other three groups，between the two treatment groups with nosignificant difference．
Immunocytochemical detection results，AS model rabbits of group ABCA1ofmacrophages protein expression level was significantly higher，compared withthe blank control group was statistically significant (P＜0.01). Through thepositive cell percentage statistics study shows that，compared with the modelgroup，Xue Mai Ning granule could significantly increase the ABCA1proteinexpression，with statistical significance (P＜0.01)．
The results of RT-PCR，AS ABCA1mRNA expression in rabbits were significantlyup-regulated，drug intervention，increase more significantly，indicating thatChinese medicine Xue Mai Ning granule for the ABCA1mRNA expression has someupregulate．
The two treatment groups were comparable，rabbit peritoneal macrophageslipid loaded weight，cell surface ABCA1gene and protein expression levels werenot statistically significant (P＞0.05)．
Conclusion：
1．Xue Mai Ning granule can effectively remove foam intracellular lipidcontent，thereby effectively inhibit atherosclerosis disease．
2．Xue Mai Ning granule through the up-regulation of ABCA1gene and proteinexpression level to achieve the removal of foam cells role of lipid．
3．Xue Mai Ning granule upregulate ABCA1gene and protein expression，intracellular lipid removal of foam may be Xue Mai Ning granule in preventingand treating atherosclerosis is one of the mechanisms．
PART THREEXue Mai Ning Granule on the experimental study of the effectsof AS rabbit liver tissue SR-BI expression
Purpose：
Observation of Xue Mai Ning Granule on liver tissue in rabbit atherosclerosismodel of SR-BI mRNA and protein expression of SR-BI regulation．Exploration ofcompound Chinese herbal Xue Mai Ning granule in treating atherosclerosismechanism．
Materials and methods：
This study in rabbits as experimental objects,40rabbits were in accordancewith the random number method，were randomly divided into four groups，respectively：blank control group，model group，Xue Mai Ning granule group，the control group of Western medicine．Adaptive feeding rabbits after a week，the model group，Xue Mai Ning granule group and Western medicine group were modelreplication．Using high fat diet feeding methods atherosclerosis modelreplication，model group，Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet，and in experiment first day one-timeintraperitoneal injection of vitamin D3(600000IU/kg). Blank control group given normal granular feed，and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous to high fat diet for4weeks，all rabbits through ear vein collecting venous blood3ml，isolated from the serumwere detected，the level of blood lipid of rabbits. By statistical analysis，compared with the blank control group，model group three rabbits blood lipidswere significantly higher than control group（P＜0.01），that successfully mould，and the start of Xue Mai Ning granule group and the control group of Westernmedicine treatment，respectively，by mouth filling feeding Xue Mai Ning granuleand atorvastatin，once a day，after4weeks of continuous irrigation．In theend of the experiment，through ear vein injection of air，cause an air embolism，put to death all rabbits，open abdominal，removal of liver caudate lobe，usingRT-PCR method to detect all rabbits liver tissues of SR-BI mRNA expression；using Western-blot for detecting clusters of rabbit liver tissue SR-BI proteinexpression level. Using SPSS10.0software package for each of the groups ofexperimental data for statistical analysis，measurement data to mean±SD(x±s)，multiple group were compared with single factor analysis of variance，with P＜0.05was considered statistically significant．
Result：
Western-blot results show，objective strap located approximately180kDa，integral optical density value of statistical results indicate that，comparedwith the blank control group，model group rabbit AS expression level of SR-BIwas significantly higher，with statistical significance (P＜0.01)；comparedwith the model group，treatment group two in rabbit SR-BI expression level wassignificantly higher，with statistical significance (P＜0.01)．
RT-PCR results show，objective strap located approximately485bp，integraloptical density value of statistical results indicate that，compared with theblank control group，model group rabbit AS expression level of SR-BI wassignificantly higher，with statistical significance (P＜0.01)；compared with the model group，treatment group two in rabbit SR-BI expression level wassignificantly higher，with statistical significance (P＜0.01)．
The two treatment groups were comparable，the rabbit liver tissue SR-BI geneand protein expression levels were not statistically significant (P＞0.05)．
Conclusion：
1．Xue Mai Ning granule can obviously increase in atherosclerotic rabbitsand SR-BI mRNA expression in liver cells．
2．Xue Mai Ning granule can obviously increase in atherosclerotic rabbitsand expression of SR-BI protein in liver cells．
3．Xue Mai Ning granule through upregulation of liver cell SR-BI gene andprotein expression and its effect on cholesterol reverse transport role．
4．upregulation of liver cell SR-BI gene and protein expression，whichpromotes reverse cholesterol transport may be Xue Mai Ning granule in preventingand treating atherosclerosis and mechanism．
引文
[1]范永臻，郭志刚，王振坤，等.罗格列酮对动脉粥样硬化兔高密度脂蛋白的影响[J].实用医学杂志，2010，26（9）：1508-1512。
[2]骆杨平.苯扎贝特促进小鼠巨噬细胞胆固醇逆转运体内和体外研究[D].中南大学2009届博士研究生.
[3]赵培.刺芒柄花素上调胆固醇逆转运途径关键蛋白SR-BI和ABCA1及抗动脉粥样硬化作用研究[D].北京协和医学院硕士学位论文，2009.
[4]邵晓军，王海萍，张晨，等.免疫损伤-高脂喂养法建立兔动脉粥样硬化模型评价[J].齐鲁医学杂志，2009，24（3）：264-266.
[5]Green T J，Mohgadasian M H．Species-related variations in lipoproteinmetabolism：The impact of FER（HDL）on susceptibility to atherogenesis[J]．LifeSci，2004，74（19）：2441－2449．
[6]李贵才，刘占军，徐德锌，等.家兔动脉粥样硬化模型稳定性的观察[J].中国比较医学杂志，2004，14(5):294-297.
[7]张勇，娄桂予，钱民章，等.建立兔高脂血症模型及动脉粥样硬化斑块的方法[J].遵义医学院学报，2001，24(4):304-306.
[8]李跃华，陆建平，张亚明.兔动脉粥样硬化模型的建立及其HR检测[J].中国CT和MRI杂志，2006，4(1):34-35.
[9]覃伟，钱妍.番茄红素对动脉粥样硬化家兔血脂及低密度脂蛋白受体mRNA表达的影响[J].中国医院药学杂志，2010，30（3）：191-194.
[10] RekhterMD， Hicks GW， Brammer DW，et al. Animal model that mimicsatherosclerotic plaque rupture[J]. Circulation research，1998，83(7):705.
[11] AbelaGS，Picon PD，FriedlSE，etal. Triggering of plaque disruption andarterial thrombosis in an atherosclerotic rabbitmode[J].l Circulation，1995，91(3):776.
[12] Breslow JL. Mouse models of atherosclerosis[J]. Science，1996，272(5262):685.
[13]赵欣，赵全明.动脉粥样硬化动物模型研究进展[J].医学研究杂志，2010，39（10）：15-20.
[14]武凤翔，李耀卿，耿新生，等.益心康片降脂作用研究[J].河南中医药学刊，1999，14（6）：18-19.
[15]孙文娟，刘洁，杨世杰，等.不同产地长梗薤白提取物对高脂血症大鼠脂代谢的影响及其抗氧化作用.白求恩医科大学学报[J]，1999，25（3）：259-261.
[16]宋小鸽，唐照亮，侯正明，等.茶多酚对高脂血症动物的预防作用[J].中国中药杂志，1999，24（9）：562-564．
[17]张平，魏喜保，常明项.降脂啤酒预防血脂升高及抗氧化的实验研究[J].湖北中医杂志，1999，21（7）：332-335．
[18]储益平，殷书梅.甲壳聚多糖降脂降胆固醇实验研究.时珍国医国药[J]，1999，10（3）：178-180．
[19]余大鹏，方肇年.降脂灵胶囊对高脂血症模型动物血脂的影响[J].中国中医药科技，1999，6（4）：257-259．
[20]Angelin B，Parini P Eriksson M.Reverse cholesterol transport in man：promotion of fecal steroid excretion by infusion of reconstituted HDL[J].Atherosclerosis.Supplements，2002-3(4)．
[21]张愍.冠心病痰浊辨证分型与血清脂蛋白动态平衡关系的研究[J].中西医结合杂志，1995，15(1):9.
[22]龙庆余.从痰瘀辨治高血脂症60例[J].安徽中医临床杂志，1998，10(1):20.
[23]姚克裘，周曾绮，陈教先.血通胶囊对老年高脂血症影响的临床研究[J].上海中医药杂志，1995，1:37.
[24]刘洪.气虚血瘀证与血管内皮细胞相关因子的研究进展[J].中国中医基础医学杂志，2005，11(7)：553-556.
[25]周学文，陈民，李曦明，等.血脂异常中医病因病机探讨[J].中华中医药学刊，2007,25（2）：197-198.
[26]徐浩，鹿小燕，陈可冀，等.血瘀证及其兼证与冠脉造影所示病变及介入治疗后再狭窄的相关性研究[J].中国中西医结合杂志，2007，27(1)：8-13.
[27]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机相关性研究[J].中西医结合心脑血管病杂志，2009，7（4）：385-387.
[28]麻春杰，丁连平.健脾化湿法治疗高脂血症78例[J].现代中西医结合杂志，2002，11(4)：321.
[29]熊文生，曾益宏.理脾化痰降脂片对高脂血症影响的临床研究[J].中国医药学报，2000，(2)：35.
[30]孙建芝，牛晓亚.痰浊证微观辩证指标的实验研究[J]．河南中医，1996，16(2):21-22.
[31]于艳丽，宋鲁成．中医“毒邪”在动脉粥样硬化中的地位和作用[J].河北中医，2009，31(2):211－212．
[32]许迎春，王化良，丁晶．动脉粥样硬化从毒论治探讨[J].中医杂志，2004，45(6)：405－407．
[33]王永炎.关于提高脑血管疾病疗效难点的思考[J].中国中西医结合杂志，1997，17(4)：195-196．
[34]章小平．益气养阴活血法治疗糖尿病合并冠心病研究进展[J].安徽中医学院学报，2001，20（1）：60。
[35]樊力．补阳还五汤治疗糖尿病周围神经病变30例[J]．四川中医，2000，18（3）：24．
[36]王筠，张军平．冠心病之络脉虚滞论[J]．中医药学刊，2006，24(4)：629-630．
[37]陈浩，胡业彬．冠心病某些危险因素与“瘀毒”病因关系探讨[J].安徽中医学院学报，2009，28（4）：2-4．
[38]陈可翼，史载祥．实用瘀证学[M]．人民卫生出版社，1999，6．
[39]周强，冯健兰．地黄饮子治疗颈动脉粥样硬化斑块的疗效及对血内皮素、丙二醛和C反应蛋白的影响[J]．中国全科医学，2006，9(4)：335-337.
[40]王志强，张学平，张伟，等.化痰通络汤治疗颈动脉内膜粥样硬化斑块临床研究［J］.中国医药指南，2008，6(23)：259-261.
[41]刘煜德，吴辉，吴伟，等．黄连解毒汤对高胆固醇兔血液流变学和动脉粥样硬化干预作用［J］.中国中药杂志，2009，34(5)：600-604.
[42]谢彬．ATP结合盒转运蛋白A1研究进展[J]．国际内科学杂志，2009，36（1）：37-40.
[43]ProbstMC，Thum ann H，A slanidisa C，et a1. Screerfing for functional sequencevariations and mutations in ABCA1[J].A therosclerosis，2004，175(2):269-279．
[44]苟连平，吕湛，秦俭，等．ATP结合盒转运蛋白A1与动脉粥样硬化[J].心血管病学进展．2007，28（6）：951-954．
[45]唐朝克，杨永宗．ABCA1在动脉粥样硬化发生与发展中的作用[J].生命的化学，2003，23(2)：138-140．
[46]李红玲，朱敏佳，党怀欣，等．ATF6调节ABCA1的表达[J]．中国生物化学与分子生物学报，2006，22（10）：799-805.
[47]Santamaina-fojo S，Pterson K，Knapper C，et al.Complete genomic sequenceof the human and mouse ATP blinding cassette promoter A[J].Proc Natl Acad Sci，2000，97（14）：7987-7992.
[48]Schmitz G，Langmann T.Structure function and regulation of the ABCA1geneproduct[J].Curr Opin Lipidol，2001，12（2）：129-140．
[49] Kielar D，Dietmaier W，Langmann T，et al.Rapid quantification of human ABCA1mRNA in various cell types and tissues by real-time reverse transcription-PCR．ClinChem，2001，47：2089-2097．
[50]罗丹.ATP结合框转运分子A1的结构、功能及调控的研究进展[J].国外医学临床生物化学与检验学分册，2003，24(5)：252-253．
[51]Lee JY And Parks JS. ATP-binding Cassette Transporter Al And Its Roel InHDL formation. Curr Opin Lipidol，2005，16：19-25．
[52]Denis M，Haidar B，Marcil M，et al. Characterization Of Oligomeric HumanATP Binding Cassette Transporter Al. Potential Implications For Determing Thestructure of nascent high density lipoprotein Particles. J Biol Chem，2004，279：41529-41536．
[53]李颖庆，唐朝克，文红艳．载脂蛋白A-I与ATP结合盒转运体A1[J]．医学分子生物学杂志，2005，2(5):385-388．
[54] Adorni MP，Zimetti F，Billheimer JT，et al.The roles of different pathwaysin the release of cholesterol from macrophages.J Lipid Res，2007，48:2453-2462.
[55] Haghpassand M，Bourassa P-AK，Francone OL，et al.Monocyte/macrophageexpression of ABCA1has minimal contribution to plasma HDL levels.J Clin Invest，2001，108：1315-1320.
[56] Wang，X.，Collins，H.L.，Ranalletta，M.，et al．Macrophage ABCA1and ABCGI，but not SR-Bl，promote macrophage reverse cholesterol transport in vivo.J ClinInvest，2007，117:2216-2224.
[57] Ming-DongWang，Vivian Franklin，Yves L.Marcel.In vivo reverse cholesteroltransport from macrophages lacking ABCA1expression is impaired arteriosclerThromb Vase Biol，2007，27:1837-1842．
[58] Wendy Jessupa，IngridC，Gelissena，Katharina Gausa，et al.Roles of ATP bindingcassette transporters A1and G1，scavenger receptor B1and membrane lipid domainsin cholesterol export from macrophages.Curr Opin Lipidol，2006，17:247-257.
[59]Van Eck M，Singaraja RR，Ye D，et al.Macrophage ATP-binding cassettetransPorter A1overexpression inhibits atherosclerotic lesion progression inlow-density lipoprotein receptor knockout mice.Arterioscler Thromb Vasc Biol，2006，26：929-934.
[60]Wang N，Lan D，Chen W，Matsuura F，Tall AR.ATP-binding cassette transportersG1and G4mediate cellular cholesterol efflux to high-density lipoproteins.ProcNatl Acad Sci USA，2004，101：9774-9779．
[61]Kennedy MA，Barrera GC，Nakamura K，Baldan A，Tarr P，Fishbein MC，FrankJ，Francone OL，Edwards PA，ABCG1has a critical role in mediating cholesterolefflux to HDL and preventing cellular lipid accumulation．Cell Metab，2005，1：121-131．
[62]张晓菲，谭晓华，杨磊，等．ABCA1在胆固醇逆转运中作用的研究进展[J]．健康基础理论，2010，30（3）：214-217．
[63]WadeDP，Owen JS.Regulation of the cholesterol efflux gene，ABCA1[J]. Lancet，2001，357(9251)：161-163．
[64]Huuskonen J，Vishnum M，PullingerC R，et al. Regulation of ATP-BindingCassette Transporter A1Transcription by Thyroid Hormone Receptor[J].Biochemistry，2004，43(6)：1626-1632．
[65]PanousisC G，Zuckerman S H.Interferon-gamma induce downregulation ofTangiger diease gene(ATP-binding-cassette transporter) in macrophage-derivedfoam cells[J].A rterioscler Thromb Vasc Biol，2000，20(6)：1565-1571.
[66]Pullinger CR，H akamata H，Duchateau PN， et a1.Analysis of hABC1gene5，end: additional peptide sequence， promoter region， and fourpolymorphisms[J].Biochem Biophys Res Commun，2000，271(2):451-455.
[67]祁莉萍，严晓伟，叶平，等．ABCA1-565C/T基因多态性对心梗患者来源的巨噬细胞胆固醇外流功能影响及其机制研究[J]．中国全科医学，2010，13（13）：1427-1430．
[68]Zhao SP，Xiao ZJ，Nie S，et al．The study on ABCA1R219K gentic variationin patients with coronary heart disease[J]．Chin J Cardiol，2004，32(8)：712-716.
[69]ASZTALOS B F.HDL Atherosclerosis Treatment Study. High-density lipoproteinmetabolism and progression of atherosclerosis: new insights from the HDLAtherosclerosis Treatment Study[J]. Curr Opin Cardiol，2004，19(4)：385-391.
[70]Vedhchalam C， Liu L， NicketM，et al. Influence of apoA-I structure onthe ABCA1-mediated efflux of cellular lipids [J]. Biol Chem，2004，278(48)：49931-49939.
[71]AzumaY，TakadaM，ShinH.W et al.Retroendocytosis pathway of ABCA1/apoA-Icontributes to HDL formation[J].Genes Cells，2009，14(2):191-204.
[72]ZHANGQing-Hai，YI Guang-Hui，LI Yuan-Bin，et al.Expression Regulation andSignal Transduction of Scavenger Receptor Class B Type1[J]. Chinese Journalof Biochemistry and Molecular Biology，2009，25（11）：1003-1009．
[73]常冠楠，徐新，张社兵，等．B类I型清道夫受体的研究进展[J]．中西医结合心脑血管病杂志，2010，8（9）：1126-1129．
[74]秦媛媛，凌文华．B族I型清道夫受体与高密度脂蛋白代谢[J]．国际内科学杂志，2009，36（5）：286-289．
[75]Ahars M，Naing T， M c Pherson R．Scavenger receptor-BI localizes to a lateendosom al compartment．J Lipid Res，2008，10(4)：50．
[76]Babitt J，Trigatti B，Rigotti A，et al．SR-BI，a high density lipoprote inreceptor that mediates selective lipid uptake，is N-glycosy lated and fattyacylated and colocalizes with plasm a membrane caveolae．J Biol Chem，1997，272:13242-13249．
[77]Vifials M，Xu S，Vasile E，et al. Identification of the N-1inked glycosylationsites on the high density lipoprotein(HDL) receptor SR-BI and assessment of theireffects on HDL binding and selective lipid uptake [J].J Biol Chem，2003，278(7):5325-5332．
[78]KriegerM．Charting the fate of the“good cholesterol”: identification andcharacterization of the high-density lipoprote in receptorSR-BI．Annu RevBiochem，1999，68:5232-5581．
[79] Acton SL， Rigotti A， Landschulz KT， et al．Identification of scavengerreceptor SR-BI as a high density lipoprote in receptor．Science，1996，271，518-520．
[80]Mineo C，Yuhanna IS，QuonMJ，et al.High density lipoprotein inducedendothelial nitric-oxide synthase activation is mediated by Akt and MAPkinases[J].J Biol Chem，2003，278(11)：9142-9149．
[81]Grove J，Huby T，Stamataki Z，et al.Scavenger receptor B I and BⅡexpressionlevels modulate hepatitis C virus infectivity[J].JViro，2007，81(7):3162-3169.
[82]Thuahnai ST，Lund-Katz S，Dhanasekaran P，et al.Scavenger receptor classB type I mediated cholestery lester-selective up take and efflux of unesterifedcholesterol.Influence of high density lipoprotein size and structure[J].J BiolChem，2004，279（13）：448-455．
[83]刘卉．SR-BI在胆固醇代谢和动脉粥样硬化中的作用．基层医学论坛，2009，13（12）：1143-1144．
[84]孙蓓．辛伐他汀对高脂血症大鼠肝细胞B族I型清道夫受体表达的影响．社区医学杂志，2009，7（1）：31-32．
[85]Covey SD，Krieger M，Wang W，et al.Scavenger receptor class B typeI-mediated protection against atherosclerosis in LDL receptor-negative miceinvolves its expression in bone marrow derived cells[J].Arterioscler Thromb VascBiol，2003，23（9）：1589-1594．
[86]曾颖，谭玉林，冯惊涛，等．阿托伐他汀对THP-1巨噬细胞CD36表达的影响[J].中国心血管病研究杂志，2007，5(2)：132-134．
[87]朱贵月，朱兴雷，耿庆信，等．阿托伐他汀对冠心病患者外周血单核细胞/巨噬细胞清道夫受体的影响[J].中国病理生理杂志，2006，22(2)：292-294．
[88]王志宏，常云华，刘玉环，等．氟伐他汀对阿霉素肾病大鼠肾脏清道夫受体A基因表达的影响及肾保护作用[J].中国中西医结合肾病杂志，2007，8(10)：576-579．
[89]尹小波，莫中成，叶延平，等．丙丁酚可干预小鼠动脉粥样硬化病并调节B类I型清道夫受体的表达[J]．中国动脉硬化杂志，2007，15(7)：569．
[90]范春雷，高丽萍，沃兴德，等．川芎嗪对爪蟾卵母细胞外源清道夫受体基因表达的影响[J].中国药理学通报，2005，21(6)：746-751．
[91]郑伟，周洁，陈娟，等．甲基莲心碱对RAW264.7巨噬细胞泡沫化的影响[J]．医学分子生物学杂志，2005，2(3)：170-174．
[92]童孟立，王宇晖，鲁盈，等．复方积雪草对人肾系膜细胞清道夫受体表达的影响[J]．中国中医药科技，2005，12(2)：89-91．
[93]林培政，杨开清，等．清热化湿消瘀法对湿热证模型兔肝脏清道夫受体基因表达的影响[J]．中药新药与临床药理，2007，18(1)：15-17．
[94]姜浩．脉心康干预载脂蛋白E基因敲除小鼠动脉粥样硬化及其巨噬细胞泡沫化的分子机制研究[M].山东中医药大学博士学位论文.
[95]樊继山，李晓辉，李淑慧，等．三七皂甙对泡沫细胞形成的影响极其信号机制[J]．中草药，2007，38（6）：893-895．
[96]倪占玲.罗布考促进小鼠巨噬细胞胆固醇逆转运的体内外实验研究[D].中南大学，2007届博士研究生.
[97] Schmilz G，Langman T.Structure， function and regulation of the ABCA1geneproduct[J].Curr opin lipidol，2001，12(2):129-140.
[2]骆杨平.苯扎贝特促进小鼠巨噬细胞胆固醇逆转运体内和体外研究[D].中南大学2009届博士研究生.
[3]赵培.刺芒柄花素上调胆固醇逆转运途径关键蛋白SR-BI和ABCA1及抗动脉粥样硬化作用研究[D].北京协和医学院硕士学位论文，2009.
[4]邵晓军，王海萍，张晨，等.免疫损伤-高脂喂养法建立兔动脉粥样硬化模型评价[J].齐鲁医学杂志，2009，24（3）：264-266.
[5]Green T J，Mohgadasian M H．Species-related variations in lipoproteinmetabolism：The impact of FER（HDL）on susceptibility to atherogenesis[J]．LifeSci，2004，74（19）：2441－2449．
[6]李贵才，刘占军，徐德锌，等.家兔动脉粥样硬化模型稳定性的观察[J].中国比较医学杂志，2004，14(5):294-297.
[7]张勇，娄桂予，钱民章，等.建立兔高脂血症模型及动脉粥样硬化斑块的方法[J].遵义医学院学报，2001，24(4):304-306.
[8]李跃华，陆建平，张亚明.兔动脉粥样硬化模型的建立及其HR检测[J].中国CT和MRI杂志，2006，4(1):34-35.
[9]覃伟，钱妍.番茄红素对动脉粥样硬化家兔血脂及低密度脂蛋白受体mRNA表达的影响[J].中国医院药学杂志，2010，30（3）：191-194.
[10] RekhterMD， Hicks GW， Brammer DW，et al. Animal model that mimicsatherosclerotic plaque rupture[J]. Circulation research，1998，83(7):705.
[11] AbelaGS，Picon PD，FriedlSE，etal. Triggering of plaque disruption andarterial thrombosis in an atherosclerotic rabbitmode[J].l Circulation，1995，91(3):776.
[12] Breslow JL. Mouse models of atherosclerosis[J]. Science，1996，272(5262):685.
[13]赵欣，赵全明.动脉粥样硬化动物模型研究进展[J].医学研究杂志，2010，39（10）：15-20.
[14]武凤翔，李耀卿，耿新生，等.益心康片降脂作用研究[J].河南中医药学刊，1999，14（6）：18-19.
[15]孙文娟，刘洁，杨世杰，等.不同产地长梗薤白提取物对高脂血症大鼠脂代谢的影响及其抗氧化作用.白求恩医科大学学报[J]，1999，25（3）：259-261.
[16]宋小鸽，唐照亮，侯正明，等.茶多酚对高脂血症动物的预防作用[J].中国中药杂志，1999，24（9）：562-564．
[17]张平，魏喜保，常明项.降脂啤酒预防血脂升高及抗氧化的实验研究[J].湖北中医杂志，1999，21（7）：332-335．
[18]储益平，殷书梅.甲壳聚多糖降脂降胆固醇实验研究.时珍国医国药[J]，1999，10（3）：178-180．
[19]余大鹏，方肇年.降脂灵胶囊对高脂血症模型动物血脂的影响[J].中国中医药科技，1999，6（4）：257-259．
[20]Angelin B，Parini P Eriksson M.Reverse cholesterol transport in man：promotion of fecal steroid excretion by infusion of reconstituted HDL[J].Atherosclerosis.Supplements，2002-3(4)．
[21]张愍.冠心病痰浊辨证分型与血清脂蛋白动态平衡关系的研究[J].中西医结合杂志，1995，15(1):9.
[22]龙庆余.从痰瘀辨治高血脂症60例[J].安徽中医临床杂志，1998，10(1):20.
[23]姚克裘，周曾绮，陈教先.血通胶囊对老年高脂血症影响的临床研究[J].上海中医药杂志，1995，1:37.
[24]刘洪.气虚血瘀证与血管内皮细胞相关因子的研究进展[J].中国中医基础医学杂志，2005，11(7)：553-556.
[25]周学文，陈民，李曦明，等.血脂异常中医病因病机探讨[J].中华中医药学刊，2007,25（2）：197-198.
[26]徐浩，鹿小燕，陈可冀，等.血瘀证及其兼证与冠脉造影所示病变及介入治疗后再狭窄的相关性研究[J].中国中西医结合杂志，2007，27(1)：8-13.
[27]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机相关性研究[J].中西医结合心脑血管病杂志，2009，7（4）：385-387.
[28]麻春杰，丁连平.健脾化湿法治疗高脂血症78例[J].现代中西医结合杂志，2002，11(4)：321.
[29]熊文生，曾益宏.理脾化痰降脂片对高脂血症影响的临床研究[J].中国医药学报，2000，(2)：35.
[30]孙建芝，牛晓亚.痰浊证微观辩证指标的实验研究[J]．河南中医，1996，16(2):21-22.
[31]于艳丽，宋鲁成．中医“毒邪”在动脉粥样硬化中的地位和作用[J].河北中医，2009，31(2):211－212．
[32]许迎春，王化良，丁晶．动脉粥样硬化从毒论治探讨[J].中医杂志，2004，45(6)：405－407．
[33]王永炎.关于提高脑血管疾病疗效难点的思考[J].中国中西医结合杂志，1997，17(4)：195-196．
[34]章小平．益气养阴活血法治疗糖尿病合并冠心病研究进展[J].安徽中医学院学报，2001，20（1）：60。
[35]樊力．补阳还五汤治疗糖尿病周围神经病变30例[J]．四川中医，2000，18（3）：24．
[36]王筠，张军平．冠心病之络脉虚滞论[J]．中医药学刊，2006，24(4)：629-630．
[37]陈浩，胡业彬．冠心病某些危险因素与“瘀毒”病因关系探讨[J].安徽中医学院学报，2009，28（4）：2-4．
[38]陈可翼，史载祥．实用瘀证学[M]．人民卫生出版社，1999，6．
[39]周强，冯健兰．地黄饮子治疗颈动脉粥样硬化斑块的疗效及对血内皮素、丙二醛和C反应蛋白的影响[J]．中国全科医学，2006，9(4)：335-337.
[40]王志强，张学平，张伟，等.化痰通络汤治疗颈动脉内膜粥样硬化斑块临床研究［J］.中国医药指南，2008，6(23)：259-261.
[41]刘煜德，吴辉，吴伟，等．黄连解毒汤对高胆固醇兔血液流变学和动脉粥样硬化干预作用［J］.中国中药杂志，2009，34(5)：600-604.
[42]谢彬．ATP结合盒转运蛋白A1研究进展[J]．国际内科学杂志，2009，36（1）：37-40.
[43]ProbstMC，Thum ann H，A slanidisa C，et a1. Screerfing for functional sequencevariations and mutations in ABCA1[J].A therosclerosis，2004，175(2):269-279．
[44]苟连平，吕湛，秦俭，等．ATP结合盒转运蛋白A1与动脉粥样硬化[J].心血管病学进展．2007，28（6）：951-954．
[45]唐朝克，杨永宗．ABCA1在动脉粥样硬化发生与发展中的作用[J].生命的化学，2003，23(2)：138-140．
[46]李红玲，朱敏佳，党怀欣，等．ATF6调节ABCA1的表达[J]．中国生物化学与分子生物学报，2006，22（10）：799-805.
[47]Santamaina-fojo S，Pterson K，Knapper C，et al.Complete genomic sequenceof the human and mouse ATP blinding cassette promoter A[J].Proc Natl Acad Sci，2000，97（14）：7987-7992.
[48]Schmitz G，Langmann T.Structure function and regulation of the ABCA1geneproduct[J].Curr Opin Lipidol，2001，12（2）：129-140．
[49] Kielar D，Dietmaier W，Langmann T，et al.Rapid quantification of human ABCA1mRNA in various cell types and tissues by real-time reverse transcription-PCR．ClinChem，2001，47：2089-2097．
[50]罗丹.ATP结合框转运分子A1的结构、功能及调控的研究进展[J].国外医学临床生物化学与检验学分册，2003，24(5)：252-253．
[51]Lee JY And Parks JS. ATP-binding Cassette Transporter Al And Its Roel InHDL formation. Curr Opin Lipidol，2005，16：19-25．
[52]Denis M，Haidar B，Marcil M，et al. Characterization Of Oligomeric HumanATP Binding Cassette Transporter Al. Potential Implications For Determing Thestructure of nascent high density lipoprotein Particles. J Biol Chem，2004，279：41529-41536．
[53]李颖庆，唐朝克，文红艳．载脂蛋白A-I与ATP结合盒转运体A1[J]．医学分子生物学杂志，2005，2(5):385-388．
[54] Adorni MP，Zimetti F，Billheimer JT，et al.The roles of different pathwaysin the release of cholesterol from macrophages.J Lipid Res，2007，48:2453-2462.
[55] Haghpassand M，Bourassa P-AK，Francone OL，et al.Monocyte/macrophageexpression of ABCA1has minimal contribution to plasma HDL levels.J Clin Invest，2001，108：1315-1320.
[56] Wang，X.，Collins，H.L.，Ranalletta，M.，et al．Macrophage ABCA1and ABCGI，but not SR-Bl，promote macrophage reverse cholesterol transport in vivo.J ClinInvest，2007，117:2216-2224.
[57] Ming-DongWang，Vivian Franklin，Yves L.Marcel.In vivo reverse cholesteroltransport from macrophages lacking ABCA1expression is impaired arteriosclerThromb Vase Biol，2007，27:1837-1842．
[58] Wendy Jessupa，IngridC，Gelissena，Katharina Gausa，et al.Roles of ATP bindingcassette transporters A1and G1，scavenger receptor B1and membrane lipid domainsin cholesterol export from macrophages.Curr Opin Lipidol，2006，17:247-257.
[59]Van Eck M，Singaraja RR，Ye D，et al.Macrophage ATP-binding cassettetransPorter A1overexpression inhibits atherosclerotic lesion progression inlow-density lipoprotein receptor knockout mice.Arterioscler Thromb Vasc Biol，2006，26：929-934.
[60]Wang N，Lan D，Chen W，Matsuura F，Tall AR.ATP-binding cassette transportersG1and G4mediate cellular cholesterol efflux to high-density lipoproteins.ProcNatl Acad Sci USA，2004，101：9774-9779．
[61]Kennedy MA，Barrera GC，Nakamura K，Baldan A，Tarr P，Fishbein MC，FrankJ，Francone OL，Edwards PA，ABCG1has a critical role in mediating cholesterolefflux to HDL and preventing cellular lipid accumulation．Cell Metab，2005，1：121-131．
[62]张晓菲，谭晓华，杨磊，等．ABCA1在胆固醇逆转运中作用的研究进展[J]．健康基础理论，2010，30（3）：214-217．
[63]WadeDP，Owen JS.Regulation of the cholesterol efflux gene，ABCA1[J]. Lancet，2001，357(9251)：161-163．
[64]Huuskonen J，Vishnum M，PullingerC R，et al. Regulation of ATP-BindingCassette Transporter A1Transcription by Thyroid Hormone Receptor[J].Biochemistry，2004，43(6)：1626-1632．
[65]PanousisC G，Zuckerman S H.Interferon-gamma induce downregulation ofTangiger diease gene(ATP-binding-cassette transporter) in macrophage-derivedfoam cells[J].A rterioscler Thromb Vasc Biol，2000，20(6)：1565-1571.
[66]Pullinger CR，H akamata H，Duchateau PN， et a1.Analysis of hABC1gene5，end: additional peptide sequence， promoter region， and fourpolymorphisms[J].Biochem Biophys Res Commun，2000，271(2):451-455.
[67]祁莉萍，严晓伟，叶平，等．ABCA1-565C/T基因多态性对心梗患者来源的巨噬细胞胆固醇外流功能影响及其机制研究[J]．中国全科医学，2010，13（13）：1427-1430．
[68]Zhao SP，Xiao ZJ，Nie S，et al．The study on ABCA1R219K gentic variationin patients with coronary heart disease[J]．Chin J Cardiol，2004，32(8)：712-716.
[69]ASZTALOS B F.HDL Atherosclerosis Treatment Study. High-density lipoproteinmetabolism and progression of atherosclerosis: new insights from the HDLAtherosclerosis Treatment Study[J]. Curr Opin Cardiol，2004，19(4)：385-391.
[70]Vedhchalam C， Liu L， NicketM，et al. Influence of apoA-I structure onthe ABCA1-mediated efflux of cellular lipids [J]. Biol Chem，2004，278(48)：49931-49939.
[71]AzumaY，TakadaM，ShinH.W et al.Retroendocytosis pathway of ABCA1/apoA-Icontributes to HDL formation[J].Genes Cells，2009，14(2):191-204.
[72]ZHANGQing-Hai，YI Guang-Hui，LI Yuan-Bin，et al.Expression Regulation andSignal Transduction of Scavenger Receptor Class B Type1[J]. Chinese Journalof Biochemistry and Molecular Biology，2009，25（11）：1003-1009．
[73]常冠楠，徐新，张社兵，等．B类I型清道夫受体的研究进展[J]．中西医结合心脑血管病杂志，2010，8（9）：1126-1129．
[74]秦媛媛，凌文华．B族I型清道夫受体与高密度脂蛋白代谢[J]．国际内科学杂志，2009，36（5）：286-289．
[75]Ahars M，Naing T， M c Pherson R．Scavenger receptor-BI localizes to a lateendosom al compartment．J Lipid Res，2008，10(4)：50．
[76]Babitt J，Trigatti B，Rigotti A，et al．SR-BI，a high density lipoprote inreceptor that mediates selective lipid uptake，is N-glycosy lated and fattyacylated and colocalizes with plasm a membrane caveolae．J Biol Chem，1997，272:13242-13249．
[77]Vifials M，Xu S，Vasile E，et al. Identification of the N-1inked glycosylationsites on the high density lipoprotein(HDL) receptor SR-BI and assessment of theireffects on HDL binding and selective lipid uptake [J].J Biol Chem，2003，278(7):5325-5332．
[78]KriegerM．Charting the fate of the“good cholesterol”: identification andcharacterization of the high-density lipoprote in receptorSR-BI．Annu RevBiochem，1999，68:5232-5581．
[79] Acton SL， Rigotti A， Landschulz KT， et al．Identification of scavengerreceptor SR-BI as a high density lipoprote in receptor．Science，1996，271，518-520．
[80]Mineo C，Yuhanna IS，QuonMJ，et al.High density lipoprotein inducedendothelial nitric-oxide synthase activation is mediated by Akt and MAPkinases[J].J Biol Chem，2003，278(11)：9142-9149．
[81]Grove J，Huby T，Stamataki Z，et al.Scavenger receptor B I and BⅡexpressionlevels modulate hepatitis C virus infectivity[J].JViro，2007，81(7):3162-3169.
[82]Thuahnai ST，Lund-Katz S，Dhanasekaran P，et al.Scavenger receptor classB type I mediated cholestery lester-selective up take and efflux of unesterifedcholesterol.Influence of high density lipoprotein size and structure[J].J BiolChem，2004，279（13）：448-455．
[83]刘卉．SR-BI在胆固醇代谢和动脉粥样硬化中的作用．基层医学论坛，2009，13（12）：1143-1144．
[84]孙蓓．辛伐他汀对高脂血症大鼠肝细胞B族I型清道夫受体表达的影响．社区医学杂志，2009，7（1）：31-32．
[85]Covey SD，Krieger M，Wang W，et al.Scavenger receptor class B typeI-mediated protection against atherosclerosis in LDL receptor-negative miceinvolves its expression in bone marrow derived cells[J].Arterioscler Thromb VascBiol，2003，23（9）：1589-1594．
[86]曾颖，谭玉林，冯惊涛，等．阿托伐他汀对THP-1巨噬细胞CD36表达的影响[J].中国心血管病研究杂志，2007，5(2)：132-134．
[87]朱贵月，朱兴雷，耿庆信，等．阿托伐他汀对冠心病患者外周血单核细胞/巨噬细胞清道夫受体的影响[J].中国病理生理杂志，2006，22(2)：292-294．
[88]王志宏，常云华，刘玉环，等．氟伐他汀对阿霉素肾病大鼠肾脏清道夫受体A基因表达的影响及肾保护作用[J].中国中西医结合肾病杂志，2007，8(10)：576-579．
[89]尹小波，莫中成，叶延平，等．丙丁酚可干预小鼠动脉粥样硬化病并调节B类I型清道夫受体的表达[J]．中国动脉硬化杂志，2007，15(7)：569．
[90]范春雷，高丽萍，沃兴德，等．川芎嗪对爪蟾卵母细胞外源清道夫受体基因表达的影响[J].中国药理学通报，2005，21(6)：746-751．
[91]郑伟，周洁，陈娟，等．甲基莲心碱对RAW264.7巨噬细胞泡沫化的影响[J]．医学分子生物学杂志，2005，2(3)：170-174．
[92]童孟立，王宇晖，鲁盈，等．复方积雪草对人肾系膜细胞清道夫受体表达的影响[J]．中国中医药科技，2005，12(2)：89-91．
[93]林培政，杨开清，等．清热化湿消瘀法对湿热证模型兔肝脏清道夫受体基因表达的影响[J]．中药新药与临床药理，2007，18(1)：15-17．
[94]姜浩．脉心康干预载脂蛋白E基因敲除小鼠动脉粥样硬化及其巨噬细胞泡沫化的分子机制研究[M].山东中医药大学博士学位论文.
[95]樊继山，李晓辉，李淑慧，等．三七皂甙对泡沫细胞形成的影响极其信号机制[J]．中草药，2007，38（6）：893-895．
[96]倪占玲.罗布考促进小鼠巨噬细胞胆固醇逆转运的体内外实验研究[D].中南大学，2007届博士研究生.
[97] Schmilz G，Langman T.Structure， function and regulation of the ABCA1geneproduct[J].Curr opin lipidol，2001，12(2):129-140.