基础医学与临床 ›› 2009, Vol. 29 ›› Issue (9): 943-947.

• 研究论文 • 上一篇    下一篇

FRNK高表达诱导HSC凋亡后MMP-2及TIMP-2的变化

申建刚 张晓岚 魏娟 霍晓霞 桑荣霞 安君艳   

  1. 深圳市人民医院 河北医科大学第二医院 河北医科大学第二医院 河北医科大学第二医院 石家庄市第一医院 河北医科大学第二医院消化内科
  • 收稿日期:2008-12-16 修回日期:2009-04-10 出版日期:2009-09-20 发布日期:2009-09-20
  • 通讯作者: 张晓岚

Change of MMP-2/TIMP-2 after apoptosis of Hepatic Stellate Cell induced by FRNK overexpression

Jian-gang SHEN, Xiao-lan ZHANG, Juan WEI, Xiao-xia HUO, Rong-xia SANG, Jun-yan AN   

  1. Shenzhen People's Hospital the Second Hospital of Hebei Medical University the First Hospital of Shijiazhuang Dept. Gastroenterology, the Second Hospital, Hebei Medical University
  • Received:2008-12-16 Revised:2009-04-10 Online:2009-09-20 Published:2009-09-20
  • Contact: Xiao-lan ZHANG,

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。

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

Abstract: Objective To investigate the effect of breaking the phosphorylation of FAK by FRNK on apoptosis and the expression of MMP-2/TIMP-2 in HSC in vitro. 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 microscope. And the levels of 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 has been transiently transfected 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; On the contrary, the expression of MMP-2 increased. Conclusions FRNK can induce the HSC apoptosis and MMP-2/TIMP-2 was perhaps involved in the process.

Key words: hepatic stellate cell, apoptosis, FAK related non-kinase, matrix metalloproteinase-2, tissue inhibitors of matrix metalloproteinasea-2