中药复方对损伤人血管内皮细胞表达细胞间黏附分子的影响

Effects of compound prescription on soluble intercellular adhesion molecule-1 expression of injured human vascular endothelial cells

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。

AIM： To observe the protective effect of Chinese compound prescription on expression of soluble intercellular adhesion molecule-1 （sICAM-1） of injured human vascular endothelial ceils （cell line： HMEC-1）, and study its possible anti-atherosclerotic mechanism. METHODS： The experiment was conducted in the Experimental Center of Science and Technology of Hualong Hospital from January 2004 to December 2005. ①Seventy SD rats were selected and randomly divided into 7 groups： normal control group, model group, western medicine group and 0.4 g/mL, 0.8 g/mL, 1.6 g/mL , 3.2 g/mL Chinese compound groups with 10 rats in each group. Rats of each group were given gastric perfusion of normal saline, normal saline, 1 g/L fluvastatin, 0.4 g/mL, 0.8 g/mL, 1.6 g/mL, 3.2 g/mL Chinese compound gruffs （based on radix astragali and mainly were trichosanthis and allii bulbus） respectively, and 2 mL for each rat in each time. Blood was drawn from abdominal aorta, and the serum was separated. ② The HMEC-1 of human vascular endothelial cells were the same as grouping of rats. Except the normal control group, the injured cell models of HMEC-1 cells of each group were established by cultured with hyperlipidemic serum. Cells in each group were treated with the serum of the same group for 72 hours respectively, and enzyme linked immunosorbent assay was adopted to detect the expression of sICAM-1 cultured in supernatant. The expressions of mRNA in sICAM-1 of HMEC-1 of each group were tested by RT-PCR. RESULTS： Comparison of sICAM-1 of HMEC-1 and its mRNA levels among all groups： the levels of sICAM-1 cultured in supernatant in 0.4 g/mL, 0.8 g/mL, 1.6 g/mL and 3.2 g/mL Chinese prescription groups were lower than the 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）], and the effects of 1.6 g/mL Chinese prescription group was the most significant, which was obviously lower than the western medicine group （4.55 ±0.20） μg/L, （P 〈 0.01 ）. Chinese prescription could remarkably down-regulate the levels of sICAM-1 of injured vascular endothelial cells and its mRNA level at abovementioned doses, and the effects of 3.2 g/mL Chinese prescription group was the most marked, which was obviously lower than the western medicine group[0.63±0.12,1.03±0.12, （P 〈 0.01 ）]. CONCLUSION： TCM prescription can improve the function of endothelial cells possibly by down-regulating the sICAM-1 cultured in supernatant and it is mRNA level, accordingly reduce the adhesion between cells and injury of vascular endothelial cells, so as to exert the function of anti-atherosclerosis.