膜型基质金属蛋白酶-1在黏着斑激酶相关非激酶调控肝星状细胞胶原代谢中的作用

Effects of MT1-MMP on collagen metabolism regulated by FRNK in hepatic stellate cells

目的:探讨黏着斑激酶相关非激酶(FRNK)对纤维连接蛋白(FN)刺激的肝星状细胞(HSC)膜型基质金属蛋白酶-1(MT1-MMP)表达的影响并探讨MT1-MMP在FRNK调控HSC胶原代谢中的作用。方法:应用体外细胞培养技术,脂质体介导法进行FRNK质粒瞬时转染;采用Western blotting法测定FRNK蛋白在瞬时转染时相应的表达,鉴定转染效果;分别应用Western blotting和RT-PCR方法测定MT1-MMP在HSC中的相应表达。结果:FRNK质粒成功转染HSC,于转染48h时,FRNK蛋白表达最强,P<0·05;FRNK转染后在基因水平上:MT1-MMP mRNA表达明显增加;翻译蛋白水平上:FRNK质粒转染HSC48h后,MT1-MMP蛋白表达量显著升高。结论:脂质体介导下FRNK质粒转染,可使外源性FRNK在HSC内大量表达,FRNK可能通过上调MT1-MMP值来抑制HSC胶原合成。

AIM ： To investigate the effect of FAK - related non - kinase （FRNK） on the expression of membrane - type matrix metalloproteinase - 1 （ MT1 - MMP） in hepatic stellate cells （HSC）. METHODS ： FRNK were transfected into HSCs by cationic liposome method. The protein levels of FRNK in HSC were assayed by Western blotting. The levels of MTI - MMP were determined by RT - PCR for mRNA and by Western blotting for protein, respectively. RESULTS ： The up - regulated expression of FRNK protein was observed and it was at 48 h after transfection that the FRNK protein content was the highest （ P 〈0. 05 ）. The expressions of MT1 - MMP mRNA and protein were also up - regulated by the transfection of FRNK, and it was at 48 h after transfection that the MTI - MMP protein content was significantly increased. CONCLUSION： The mRNA and protein of FRNK were over - expressed in HSC transfected with the gene of FRNK. The inhibitory, effect of FRNK on the collagen synthesis in HSC may be through the up - regulation of MT1 - MMP.