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目的:研究益气活血复方含药血清对脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)Toll样受体4(TLR4)表达的影响,从mRNA和蛋白水平探讨益气活血复方含药血清防治动脉粥样硬化(AS)的机制。方法:(1)选择新西兰大耳白兔20只,随机分为4组,即正常组、中药高浓度组、中药中浓度组、中药低浓度组,每组5只。以上各组白兔分别以生理盐水和高、中、低浓度益气活血复方连续灌胃7天。末次灌胃给药2h后,心脏采血,离心后分离血清。(2)体外培养人脐静脉内皮细胞,用LPS刺激后,分别加入高、中、低浓度益气活血复方含药血清干预24h,收集细胞,用荧光定量PCR方法和Western blotting法分别测定TLR4 mRNA和蛋白的表达。结果:用LPS刺激人脐静脉内皮细胞后,引起TLR4 mRNA和蛋白的高表达(与空白对照组比较P<0.01),用益气活血复方含药血清干预以后显著抑制TLR4 mRNA和蛋白的高表达(与模型组比较P<0.01或P<0.05)。结论:益气活血复方可阻断TLR4高表达,有TLR4拮抗作用,这可能是其抗动脉粥样硬化的作用机制之一。
Objective: To investigate the effect of Yiqihuoxue Compound-containing serum on lipopolysaccharide (LPS) -induced Toll-like receptor 4 (TLR4) expression in human umbilical vein endothelial cells (HUVECs), to explore the effects of Yiqi Huoxue Compound Serum prevention and treatment of atherosclerosis (AS) mechanism. Methods: (1) Twenty New Zealand white rabbits were randomly divided into 4 groups: normal group, high concentration group, medium concentration group and low concentration group. There were 5 rats in each group. The above groups of rabbits were given saline and high, medium and low concentration of Qi and blood for 7 days. After the last gavage administration 2h, the heart blood, serum after centrifugation. (2) Human umbilical vein endothelial cells were cultured in vitro, stimulated with LPS, respectively, high, medium and low concentrations of Yiqi Huoxue compound serum containing intervention 24h, collected cells, fluorescence quantitative PCR and Western blotting were measured TLR4 mRNA And protein expression. Results: The stimulation of LPS stimulated the expression of TLR4 mRNA and protein (compared with the blank control group, P <0.01). After the intervention of Yiqi Huoxue Compound-containing serum, the high expression of TLR4 mRNA and protein was significantly inhibited P <0.01 or P <0.05 compared with model group). Conclusion: Yiqi Huoxue Compound can block TLR4 overexpression and TLR4 antagonism, which may be one of its anti-atherosclerosis mechanisms.