RhoA/Rho激酶在氟伐他汀对组织因子表达影响中的调控作用

The Regulatory Effect of RhoA/Rho Kinase in Tissue Factor Expression Influenced by Fluvastatin

目的研究RhoA/Rho激酶在氟伐他汀(fluvastatin,Flu)影响组织因子蛋白表达过程中的调控作用,探讨氟伐他汀在抗动脉粥样硬化血栓形成作用的新靶点。方法将同一簇人脐静脉内皮细胞(HUVEC)株进行培养、传代后,分设对照组、肿瘤坏死因子α(TNF-α)组、Flu组及干预组(TNF-α+Flu组),采用RT-PCR方法测定组织因子(tissue factor,TF)mRNA表达水平,用Western blot方法测定TF蛋白表达和RhoA的活化水平;随后,以血管紧张素Ⅱ(AngⅡ)、Y27632为干预因素,再分设空白对照组、TNF-α+AngⅡ组、AngⅡ组、TNF-α组、TNF-α+Y27632组,采用Western blot方法测定TF蛋白表达水平。结果 TNF-α在诱导HUVEC TF表达的同时,可使RhoA活化,氟伐他汀可抑制TF表达与RhoA活化;AngⅡ可促进TNF-α诱导的HUVEC TF蛋白表达水平,而Y27632可抑制TNF-α诱导的HUVEC TF蛋白表达水平。结论 RhoA/Rho激酶通路在G蛋白水平上参与了氟伐他汀抑制组织因子表达的调控机制。

Aim To observe the regulatory effect of RhoA/Rho kinase in expression of tissure factor（TF） protein influenced by fluvastatin,so as to explore the noval target point of fluvastatin in anti-artherosclerosis and anti-thrombosis.Methods The same human umbilical vein endothelial cells（HUVEC） was cultured and passage cultured,and was then divided into control group,TNF-α group,Flu group,and TNF-α＋Flu group,TNF-α＋AngⅡ group,AngⅡ group and TNF-α＋Y27632 group.RT-PCR method was used to examine the expression of TF mRNA.Western blot method was used to examine the TF protein expression and RhoA activity. TF protein expression of HUVECs in control group,TNF-α＋AngⅡ group,AngⅡ group,TNF-α group and TNF-α＋Y27632 group was examined using Western blot method.Results In the meanwhile of inducing the HUVECS TF expression,TNF-α also activated RhoA.Fluvastatin inhibited the activation of RhoA.AngⅡ could enhance the TF expression in HUVECS which induced by TNF-α,while Y27632 could inhibit the TF expression in HUVECS which induced by TNF-α. Conclusions The inhibitive effect of fluvastatin on the TF expression might be achieved through the RhoA/ Rho signal regulatory pathway at small G protein level.