siRNA沉默Smad3对肝星状细胞增殖与凋亡的影响及其机制研究

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、siRNA-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。

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

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 immunocytochemist...