FRNK高表达诱导HSC凋亡后MMP-2及TIMP-2的变化  

Change of MMP-2/TIMP-2 after apoptosis of hepatic stellate cell induced by FRNK overexpression

作　　者：申建刚 张晓岚[2] 魏娟[2] 霍晓霞[2] 桑荣霞[3] 安君艳[2] 

机构地区：[1]深圳市人民医院,广东深圳518020 [2]河北医科大学第二医院消化科,河北石家庄050000 [3]石家庄市第一医院消化科,河北石家庄050011

出　　处：《基础医学与临床》2009年第9期943-947,共5页Basic and Clinical Medicine

基　　金：河北省自然科学基金(C2008001133);河北省卫生厅资助项目(2003055)

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。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 plas- mid, compared with the laon-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.

关 键 词：肝星状细胞 凋亡 黏着斑激酶相关非激酶 基质金属蛋白酶-2 基质金属蛋白酶组织抑制剂-2 

分 类 号：R363[医药卫生—病理学]