摘要
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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第11期2155-2158,共4页
Chinese Journal of Pathophysiology
基金
河北省自然科学基金资助项目(No.C2008001133)
河北省普通高校强势特色学科专项经费资助项目
关键词
肝星状细胞
黏着斑激酶
黏着斑激酶相关非激酶
膜型基质金属蛋白酶-1
胶原
肝硬化
Hepatic stellate cells
Focal adhesion kinase
Focal adhesion kinase -related non -kinase
Membrane - type matrix metalloproteinase - 1
Collagen
Liver cirrhosis