CAJ | 학술논문

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48 h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HSC 48 h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.
목적:탐토점착반격매상관비격매(FRNK)대섬유련접단백(FN)자격적간성상세포(HSC)막형기질금속단백매-1(MT1-MMP)표체적영향병탐토MT1-MMP재FRNK조공HSC효원대사중적작용.방법:응용체외세포배양기술,지질체개도법진행FRNK질립순시전염;채용Western blotting법측정FRNK단백재순시전염시상응적표체,감정전염효과;분별응용Western blotting화RT-PCR방법측정MT1-MMP재HSC중적상응표체.결과:FRNK질립성공전염HSC,우전염48 h시,FRNK단백표체최강,P<0.05;FRNK전염후재기인수평상:MT1-MMP mRNA표체명현증가;번역단백수평상:FRNK질립전염HSC 48 h후,MT1-MMP단백표체량현저승고.결론:지질체개도하FRNK질립전염,가사외원성FRNK재HSC내대량표체,FRNK가능통과상조MT1-MMP치래억제HSC효원합성.
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 trans-fected into HSCs by cationic liposome method. The protein levels of FRNK in HSC were assayed by Western blotting. The levels of MT1 - 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 MT1 - 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.