抑制p70S6K对HSC-T6增殖活化的作用研究  

The study on the effect of silencing p70S6K on the proliferation and activation of HSC-T6

作　　者：占书箱 黄成[1,2] 马陶陶[1,2] 陈培杰[1,2] 李俊[1,2] 

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学肝病研究所,合肥230032

出　　处：《安徽医科大学学报》2014年第8期1062-1067,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81072686;81202978);安徽省高等学校省级自然科学研究项目(编号:KJ2010A162);安徽省自然科学基金重点项目(编号:KJ2012A156)

摘　　要：目的研究抑制核糖体40S小亚基S6蛋白激酶(p70S6K)的表达对大鼠肝星状细胞株(HSC-T6)增殖活化的影响。方法建立CCl4诱导大鼠肝纤维化模型,观察磷酸化p70S6K在纤维化肝组织中的表达变化;利用雷帕霉素抑制p70S6K蛋白表达,及靶向p70S6K基因的siRNA预处理HSC-T6,再添加重组小鼠血小板衍生生长因子(PDGFBB)诱导,观察HSC-T6的增殖活化情况;利用Western blot、RT-PCR检测p70S6K、P-p70S6K、α-SMA、Collagen I的蛋白和基因表达水平。结果 CCl4诱导大鼠肝纤维化模型组磷酸化p70S6K蛋白明显升高;雷帕霉素预处理HSC-T6,明显降低了p70S6K的磷酸化水平,并抑制了PDGF-BB诱导的细胞增殖及α-SMA、Collagen I的表达;siRNA转染HSC-T6,在基因和蛋白水平抑制了p70S6K的表达,同时减少了PDGF-BB诱导的α-SMA、Collagen I、P-p70S6K的表达。结论用siRNA和雷帕霉素阻断p70S6K的表达,在一定程度上可以抑制HSC-T6的增殖和活化,为肝纤维化的防治研究提供新的思路和靶点。Objective To investigate the effect of suppressing p70S6K（ribosomal protein S6 kinase） expression on the proliferation and activation of hepatic stellate cell strain （HSC-T6） in vitro.Methods CCl4 induced liver fibrosis model in rats and then the phosphorylated p70S6K（P-p70S6K） level was determined in fibrotic liver tissue.After pretreated with Rapamycin or siRNA targeting p70S6K,HSC-T6 was stimulated with PDGF and then the proliferation and activation of it were observed.Western blot and RT-PCR were used to detect the protein and mRNA expression of p70S6K,P-p70S6K,α-SMA and Collagen Ⅰ.Results The P-p70S6K level obviously increased in rat fibrotic liver tissue.The Rapamycin pretreatment significantly decreased the P-p70S6K level and inhibited PDGFmediated proliferation and the expression of α-SMA,Collagen Ⅰ in HSC-T6.p70S6K-siRNA down-regulated the expression of p70S6K in mRNA and protein level and meanwhile decreased the expression of α-SMA,Collagen Ⅰ and P-p70S6K induced by PDGF in HSC.Conclusion Blocking p70S6K expression by Rapamycin and p70S6K-siRNA can suppress the activation and proliferation of HSC to a certain extent,maybe providing new ideas and targets for the prevention and treatment of liver fibrosis.

关 键 词：肝星状细胞株 P70S6K SIRNA 雷帕霉素 肝纤维化 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]