益气活血复方含药血清对人脐静脉内皮细胞TLR4/NF-ΚB信号通路及TNF-a ICAM-1 mRNA表达的影响

Effect of Yiqi Huoxue Compound Drug Serum on the Expression of Toll Like Receptor 4 /NF-ΚB Signals and TNF-a ICAM-1 in Huvecs

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)/NF-KB及肿瘤坏死因子-a(TNF-a)、细胞间黏附分子(ICAM-1)mRNA表达的影响,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方法测定TLR4、NF-KB、TNF-a及ICAM-1 mRNA的表达。结果:用LPS刺激人脐静脉内皮细胞后,引起TLR4、NF-KB、TNF-a及ICAM-1 mRNA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、NF-KB、TNF-a及ICAM-1 mRNA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/NF-KB信号通路的高表达,同时抑制TNF-a及ICAM-1的表达,这可能是其发挥抗动脉粥样硬化作用的机制之一。

Objective： To investigate the influence of Yiqihuoxue compound drug serum on expression of TLR4/NF-KB signals and TNF-a, ICAM-1 in HUVECs, and to study its possible anti-atheroselerotic mechanism. Methods： （1） 20 New Zealand male rabbits were divided into 4 groups in random： the normal control group, the high density Chinese compound group, the middle density Chinese compound group, and low density Chinese compound group, with 5 rabbits in each group. Each group was given gastric perfusion of normal saline and Chinese compound respectively everyday. In the 7th day, Blood was drawn from heart 2h after gastric perfusion, and then the serum was separated. （2） HUVECs was cultured in vitro and stimulated with LPS, then treated separately with normal, high, middle and low density of the Yiqihuoxue drug serum compound for 24h, measured the expression of TLR4, NF-KB, TNF-a and ICAM-1 mRNA with real- time PCR. Results： LPS enhanced the expression of TLR4, NF-KB, TNF-a and ICAM-1 mRNA（vs normal control group, P〈0.01 ）, Yiqihuoxue compound drug serum can decreased significantly the high expression of TLR4, NF-KB, TNF-a and ICAM-1 mRNA （vs model group, P〈0.01 or P〈0.05 ）which were stimulated by LPS. Conclusion： Yiqihuoxue compound can block the high expression of TLR4/NF-KB signals, and decreased the high expression of TNF-a and ICAM-1. Maybe it＇ s the mechanism of action of Yiqihuoxue compound exert the function of anti-artherosclerosis.