siRNA沉默Smad3对肝星状细胞增殖与凋亡的影响及其机制研究  

Effect of siRNA-mediated Smad3 silence on proliferation and apoptosis in activated hepatic stellate cells

作　　者：刘晓兵[1,2] 郭晓红[1,2] 刘立新[1,2] 张骞骞[1,2] 

机构地区：[1]山西医科大学第一医院科研实验中心,山西太原030001 [2]山西医科大学肝病研究所,山西太原030001

出　　处：《中国病理生理杂志》2011年第7期1376-1381,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30871146);山西省留学人员科研基金资助项目(No.2009-44)

摘　　要：目的:观察Smad3小干扰RNA(siRNA-Smad3)沉默肝星状细胞(HSCs)Smad3基因对肝星状细胞增殖、凋亡的影响,并探讨其可能机制。方法:肝星状细胞株HSCs-T6分为3组:空白对照组、阴性对照组、siR-NA-Smad3转染组。siRNA-Smad3转染HSCs细胞株,转染不同时间后,CCK-8检测HSCs增殖的变化,流式细胞术检测HSCs凋亡的变化,免疫细胞化学法检测HSCs凋亡相关蛋白P53和Bcl-2表达的变化。结果:(1)24 h、48 h、72 h siRNA-Smad3转染组HSCs增殖受到显著抑制且凋亡明显增多(P<0.01)。(2)转染48 h后,siRNA-Smad3转染组P53蛋白表达显著增多,Bcl-2蛋白表达显著减少(P<0.01)。结论:siRNA-Smad3沉默Smad3基因可以在一定时间内显著抑制HSCs的增殖并诱导其凋亡,其可能通过上调凋亡相关蛋白P53表达、下调Bcl-2的表达来发挥诱导凋亡的作用;Smad3沉默有望成为抗肝纤维化治疗的一个新途径。AIM： To investigate the effects of siRNA-mediated Smad3 silence on proliferation and apoptosis in activated hepatic stellate cells（HSCs）.METHODS： HSCs-T6 cells were divided into 3 groups： blank group,negative control group and siRNA-Smad3 transfection group.The siRNA-Smad3 was transfected into HSCs-T6 cells.At different time points after transfection,cell proliferation was measured by CCK-8,cell apoptosis was detected by flow cytometry,and protein levels of P53 and Bcl-2 were determined by immunocytochemistry.RESULTS： HSCs proliferation was significantly inhibited at the time points of 24 h,48 h and 72 h after transfection.Meanwhile,the apoptosis of HSCs was significantly increased in siRNA-Smad3 transfection group（P0.01）.Compared to the control cells,the protein expression of P53 was significantly increased while Bcl-2 protein was significantly decreased 48 h after transfection in siRNA-Smad3 transfection group（P0.01）.CONCLUSION： The siRNA-mediated Smad3 silence significantly inhibits HSCs proliferation and induces apoptosis by up-regulating the P53 expression and down-regulating the Bcl-2 expression in HSCs.

关 键 词：RNA干扰 SMAD3 肝星状细胞 肝硬化 

分 类 号：R363[医药卫生—病理学]