RhoGDI1在TNF-α诱导内皮细胞损伤中的作用研究

The role of RhoGDI1 in endothelial cell injury induced by TNF-α

目的:阐明Rho特异鸟苷酸解离抑制因子1(Rho GDI1)和Rho/ROCK信号通路在肿瘤坏死因子α(TNF-α)诱导内皮细胞损伤中的作用机制。方法:采用siRNA技术干扰人脐静脉内皮细胞Rho GDI1表达,实验分为正常对照组,10μg·L^(-1) TNF-α处理组和Rho GDI1干扰组。MTT法检测细胞存活率,TUNEL法检测细胞凋亡,Western blot检测Rho GDI1、Rho A、ROCK表达及活化。结果:TNF-α处理和Rho GDI1干扰均显著降低内皮细胞存活率,提高内皮细胞的凋亡。TNF-α处理后Rho GDI1表达显著下降,TNF-α处理和Rho GDI1干扰均可显著提高Rho A表达以及ROCK活化。结论:TNF-α可能通过抑制Rho GDI1促进Rho/ROCK信号通路的活化,最终引起内皮细胞凋亡。

Objective:To elucidate the mechanism of Rho-specific guanine nucleotide dissociation inhibitor 1(RhoGDI1)and Rho/ROCK signaling pathway in endothelial cell injury induced by tumor necrosis factor-α(TNF-α).Methods:RhoGDI1 expression was interfered in human umbilical vein endothelial cells by siRNA technique.The experiment was divided into 3 groups:the normal control group,the 10μg·L-1 TNF-αtreatment group,and the RhoGDI1 interference group.MTT method was used to detect cell viability.TUNEL assay was used to detect cell apoptosis.The expression and activation of RhoGDI1,RhoA and ROCK were detected by Western blot analysis.Results:TNF-αtreatment or RhoGDI1 suppression could significantly decrease the survival rate of endothelial cells and increase the endothelial apoptosis.Western blot showed that TNF-αtreatment significantly decrease the expression of RhoGDI1.Both TNF-αtreatment and RhoGDI1 suppression could significantly enhance the expression of RhoA and activation of ROCK.Conclusion:TNF-αmay promote the activation of Rho/ROCK signaling pathway by inhibiting RhoGDI1,and finally induce apoptosis in endothelial cells.