【作者】 唐绪刚；

【导师】 黄文权；

【作者基本信息】 第三军医大学 ， 中西医结合临床， 2009， 硕士

【摘要】 背景动脉粥样硬化(atherosclerosis,AS)所致冠心病严重威胁着人类健康,也给家庭和社会带来了沉重负担。研究显示,AS是一种慢性炎症过程,细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)与此过程关系密切。ICAM-1属于免疫球蛋白超家族成员,主要表达于血管内皮细胞、单核细胞、淋巴细胞等,但正常情况下仅为低水平或不表达。在AS炎症发展过程中单核细胞与内皮细胞间的黏附主要靠ICAM-1介导。细胞外的黏附分子能被酶分解并在血清中检测到,称为可溶性黏附分子。中医学“胸痹心痛”相当于现代医学的冠心病心绞痛。活血化瘀中药具有很好的抗AS作用,但其机制尚不明了。本课题旨在观察五灵脂水提取物对实验性AS大鼠ICAM-1表达的影响,探讨其抗AS作用的机制,丰富中医药治疗冠心病的理论基础。方法将健康、清洁、雄性6 w龄Wistar大鼠60只随机分为空白对照组、模型组、五灵脂低剂量组、五灵脂高剂量组,采用高脂饮食结合腹腔注射维生素D3注射液复制动脉粥样硬化模型,经病理检测证实模型复制成功后五灵脂低、高剂量组大鼠分别给予五灵脂水提取物不同剂量(2.5g/kg体重、10g/kg体重)灌胃干预。给药6 w后采用酶联免疫吸附法测大鼠血清可溶性细胞间黏附分子-1(soluble intercellular adhesion molecule-1,sICAM-1)水平,免疫组织化学法观察主动脉ICAM-1表达,透射电镜观察主动脉内膜病变情况。结果大鼠动脉粥样硬化模型复制成功。ELISA结果显示,空白对照组大鼠血清含微量sICAM-1(为3.10±0.42 ng/ml),明显低于其它组(P<0.01)。药物干预后五灵脂低剂量组、五灵脂高剂量组大鼠血清sICAM-1(分别为42.73±6.97、21.29±4.27 ng/ml)较干预前(分别为82.26±6.72、82.41±7.06 ng/ml)显著降低,差异有统计学意义(P<0.01),也显著低于模型组(P<0.01)。免疫组化结果显示,模型组大鼠主动脉内皮细胞ICAM-1表达强阳性,药物干预后五灵脂低剂量组、五灵脂高剂量组大鼠ICAM-1表达明显减少(P<0.01),五灵脂高剂量组轻于五灵脂低剂量组(P<0.05),但仍明显高于空白对照组(P<0.05)。透射电镜结果显示,空白对照组大鼠主动脉透射电镜观察可见内皮细胞间连接紧密,无细胞肿胀、脱落等,内皮下间隙正常;模型组见内皮细胞明显肿胀,部分脱落,细胞间连接中断,表面有单核细胞黏附,内皮下泡沫细胞堆积,内皮下间隙变窄;五灵脂低剂量、五灵脂高剂量组大鼠主动脉病变明显轻于模型组,五灵脂高剂量组病变轻于五灵脂低剂量组。结论1.高脂饲料(配方为3%胆固醇,0.5%胆酸钠,0.2%丙基硫氧嘧啶,5%白糖,10%猪油,81.3%基础饲料)配合腹腔注射维生素D3注射液能够形成大鼠AS动物模型,但该配方仍有待改进。2.AS大鼠主动脉内皮细胞ICAM-1表达强阳性,血清sICAM-1水平明显升高,其与AS炎症关系密切。3.中药五灵脂能抑制实验性动脉粥样硬化大鼠ICAM-1表达,减轻血管内膜病变程度,10g/kg体重剂量优于2.5g/kg体重剂量,这可能是其抗AS炎症的分子机制之一。更多还原

【Abstract】 Background: That atherosclerosis (AS) result in coronary heart disease(CHD) acutely threatens human being’s health, and bring heavy burden to the family and society. Research display that AS is an especially kind chronic inflammation process, and intercellular adhesion molecule-1(ICAM-1)associates with this process closely. ICAM-1 is a member of the immunoglobulin superfamily, and express in vascular endothelial cell(VEC)、monocaryon(MN)、leukomonocyte(LY) etc. ICAM-1 commonly expresses lowly or doesn’t express even. In the development process of AS chronic inflammation, MNs adhere to ECs mostly by ICAM-1. Adhesion molecules out of cell can be resolved by especial enzymes and detected in blood serum that are called dissolubility adhesion molecule. In Chinese traditional medicine“XIONGBI”corresponds to CHD of Western medicine. The traditional Chinese medicine is good for the patients of CHD, but the mechanisms isn’t clear .This study is to explore the effect of Faeces Trogopteri (FT) aqueous extract on the expression of intercellular adhesion molecule (ICAM-1) and endarteropathy and its anti artherosclerosis (AS) in the experimental AS rats, and enhance the rationale of TCM for CHD.Methods: Sixty healthy male clean six weeks Wistar rats were randomly divided into 4 groups of 15 rats per group: blank, model, FT low dose(FTL, 2.5 g/kg weight) and FT high dose (FTH, 10 g/kg weight) groups. The blank group was fed standard feedstuff, the other three groups were duplicated by feeding high cholesterol diet and peritoneal injecting vitamin D3 6 weeks later, the serum soluble intercellular adhesion molecule-1 (sICAM-1) level in the rats was measured by enzyme liked immunosorbent assay kit; the expression of ICAM-1 proteinum in the endarterium of AS rats was detected by immunohistochemical technique; the pathological changes in the endarterium of AS rats was observed by transmission electron microscope.Results: The rat AS model was successfully duplicated. Compared with the other groups, the serum sICAM-1 level of blank group was markedly lower〔( 3.10±0.42 ) vs (81.62±7.30),(82.26±6.72) ,(82.41±7.06) ng/ml,P<0.01〕; after the FT interfering, the serum sICAM-1 level of FTL and FTH group rats was markedly lower〔(42.73±6.97) vs (82.26±6.72), (21.29±4.27) vs (82.41±7.06) ng/ml,P<0.01〕, and lower than that of model group〔(42.73±6.97) vs (85.63±11.43), (21.29±4.27) vs (85.63±11.43) ng/ml, P<0.01〕. The expression of ICAM-1 proteinum at the endarterium of FTL and FTH group rats was obviously fewer than those of model group, and those of FTL group was fewer than those of FTH group. Transmission electron microscope result showed hat the endothelial damage of the aortas in FTL and FTH groups was less severe than that in model group.Conclusion: 1.The AS model of rat can be made by feeding HFD and injecting vitamin D3,but the prescription of HFD may need to be improved.2. The serum soluble intercellular adhesion molecule-1 (sICAM-1) level in the AS rats is conspicuously higher than that of the blank group, which indicates that sICAM-1 correlates with AS inflammation process closely.3. FT can downregulate the expression of ICAM-1 in the artery of experimental AS rats, and lighten the endothelial damage of the aortas, higher doe(s10 g/kg weight) is better than lower dose(2.5 g/kg weight), which may account for the anti AS effects of FT.更多还原