【摘要】 目的探讨黏着斑激酶相关非激酶(FRNK)诱导肝星状细胞(HSC)凋亡后基质金属蛋白酶-2及其抑制剂的变化。方法在体外,以纤维连接蛋白(FN)刺激HSC增殖,在脂质体介导下用FRNK表达质粒瞬时转染HSC,应用膜联蛋白/碘化丙啶双标记流式细胞术和透射电镜检测细胞的凋亡,Western blot检测FRNK、FAK、p-FAK(Tyr397)、MMP-2、TIMP-2蛋白表达。结果FRNK表达质粒成功转染HSC,在翻译后水平抑制FAK磷酸化。与空质粒组比较,FRNK表达质粒转染HSC 48 h后,HSC凋亡率由(9.28%±1.05%)增加至(25.37%±1.92%)(P<0.01);同时,FRNK在翻译水平上调MMP-2表达、抑制TIMP-2表达。结论在脂质体介导下瞬时转染FRNK表达质粒可诱导HSC发生凋亡,MMP-2/TIMP-2比值上调可能参与了该调节过程。更多还原

【Abstract】 Objective To investigate the effect of breaking phosphorylation of FAK by FRNK on apoptosis and the expression of MMP-2/TIMP-2 in HSC.Methods After FN stimulated HSC,FRNK plasmid mediated by cationic liposome was transfected into HSC.The apoptosis of FRNK-induced HSC was examined by Annexin-V/Propidium Iodide double-labeled flow cytometry(FCM),gel electrophoresis and transmission electron microscopy.FRNK,FAK,p-FAK(Tyr397),MMP-2 and TIMP-2 in HSC were assayed by Western blot on protein level,and by RT-PCR on mRNA level,respectively.Results The expression of FRNK was enhanced after FRNK transfection into HSC in vitro.The apoptotic rate in HSC exposed to FRNK plasmid for 48 h was higher than that in the non-FRNK plasmid group[(25.37%±1.92%) vs(9.28%±1.05%),P<0.01].After exposure of HSC to FRNK plasmid,compared with the non-FRNK plasmid group,the expression of TIMP-2 in protein and mRNA level reduced;while the expression of MMP-2 increased.Conclusion FRNK can induce the HSC apoptosis and MMP-2/TIMP-2 was potentially involved in the process.更多还原