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目的:观察经中药复方制剂处理后损伤的人血管内皮细胞表达细胞间黏附分子水平的变化,分析其抗动脉粥样硬化的可能途径。方法:实验于2004-01/2005-12在龙华医院科研实验中心完成。①选择SD大鼠70只,按随机数字表法分为7组,即正常对照组、模型组、西药组、中药复方0.4g/mL,0.8g/mL,1.6g/mL及3.2g/mL组,每组10只。以上各组大鼠分别灌胃生理盐水、生理盐水、1g/L氟伐他汀、0.4g/mL,0.8g/mL,1.6g/mL及3.2g/mL中药复方(以黄芪为君,栝楼、薤白为臣)生药,2mL/(次·只),分别腹主动脉采血,分离血清。②人血管内皮细胞HMEC-1与大鼠分组情况相同。除正常对照组外,其余各组HMEC-1细胞均通过高脂血清处理细胞建立细胞损伤模型。各组细胞分别应用相应组别的大鼠血清处理72h,应用酶联免疫吸附法测定培养上清液中可溶性细胞间黏附分子1表达水平,反转录-聚合酶链反应检测各组HMEC-1细胞可溶性细胞间黏附分子1mRNA表达水平。结果:各组HMEC-1细胞可溶性细胞间黏附分子1及其mRNA表达水平比较:中药复方0.4g/mL,0.8g/mL,1.6g/mL及3.2g/mL组细胞培养上清可溶性细胞间黏附分子1水平显著低于模型组[(4.33±0.19,4.44±0.34,3.46±0.17,4.33±0.27,6.27±0.25)μg/L(P<0.01)],其中中药复方1.6g/mL组作用最为明显,而且显著低于西药组(4.55±0.20)μg/L(P<0.01);以上各剂量中药同时能明显下调损伤的血管内皮细胞可溶性细胞间黏附分子1mRNA表达水平,其中中药复方3.2g/mL组作用最为明显,而且显著低于西药组[0.63±0.12,1.03±0.12(P<0.01)]。结论:以黄芪为君,栝楼、薤白为臣的中药复方制剂可明显下调损伤的血管内皮细胞培养上清液中可溶性细胞间黏附分子1及其mRNA表达水平,从而减少细胞间黏附及血管内皮损伤,其抗动脉粥样硬化的作用可能通过这一途径实现。
OBJECTIVE: To observe the changes of expression of intercellular adhesion molecules in human vascular endothelial cells damaged by Chinese herbal compound preparations, and to analyze their possible atherosclerosis pathways. METHODS: The experiment was completed at the Scientific Research Experiment Center of Longhua Hospital from January 2004 to December 2005. 1Select 70 SD rats and divide them into 7 groups according to the random number table method, namely normal control group, model group, western medicine group, Chinese herbal compound 0.4g/mL, 0.8g/mL, 1.6g/mL and 3.2g/mL Groups, 10 per group. Each group of rats were given normal saline, physiological saline, 1g/L fluvastatin, 0.4g/mL, 0.8g/mL, 1.6g/mL and 3.2g/mL Chinese herbal compound respectively. , White for the minister) raw medicine, 2mL / (secondary only), respectively, abdominal aorta blood collection, serum separation. The human vascular endothelial cell HMEC-1 was identical to the rat grouping. In addition to the normal control group, all other groups of HMEC-1 cells were treated with high-fat serum to establish a cell injury model. Each group of cells were treated with the corresponding group of rat serum for 72 hours. The level of soluble intercellular adhesion molecule 1 in the culture supernatant was determined by enzyme-linked immunosorbent assay. Reverse transcription-polymerase chain reaction was used to detect each group of HMEC-1. Cellular soluble intercellular adhesion molecule 1 mRNA expression levels. RESULTS: The levels of soluble intercellular adhesion molecule-1 and its mRNA expression in HMEC-1 cells in each group were compared: the cell culture supernatants of the Chinese herbal compound 0.4 g/mL, 0.8 g/mL, 1.6 g/mL and 3.2 g/mL groups were soluble The level of adhesion molecule 1 was significantly lower than that of model group [(4.33±0.19, 4.44±0.34, 3.46±0.17, 4.33±0.27, 6.27±0.25) μg/L (P<0.01)], of which the effect of Chinese herbal compound 1.6 g/mL group The most obvious and significantly lower than the Western medicine group (4.55 ± 0.20) μg / L (P <0.01); the above doses of traditional Chinese medicine can significantly reduce the vascular endothelial cell soluble intercellular adhesion molecule 1 mRNA expression levels, including traditional Chinese medicine compound 3.2g The effect of the /mL group was the most obvious, and it was significantly lower than that of the western medicine group [0.63±0.12, 1.03±0.12 (P<0.01)]. Conclusion: The traditional Chinese medicine compound preparations with Huangqirujun, Xiaolou and Qibai can significantly reduce the expression level of soluble intercellular adhesion molecule-1 and its mRNA in culture supernatants of injured vascular endothelial cells, thereby reducing intercellular adhesion and vascular endothelial cells. Damage, its antiatherosclerotic effects may be achieved through this approach.