摘要
目的:探讨SB431542对肝星状细胞T6(HSC T6)Smad4蛋白细胞内转位及表达的影响.方法:HSCT6细胞贴壁后,在细胞培养瓶中加入SB43154210mol/L培养2h,用免疫荧光法结合激光共聚焦显微镜观察细胞内Smad4蛋白细胞内转位.将HSCT6细胞分成3组:空白对照组、SB4315421mol/L组、SB43154210mol/L组,培养24h后分别提取细胞浆蛋白及核蛋白,以Western blot法检测Smad4蛋白表达水平变化.结果:SB43154210mol/L作用2h后,未出现Smad4蛋白向细胞核内转位.HSC T6细胞贴壁后被SB4315421mol/L、SB43154210mol/L作用24h后,细胞浆内Smad4蛋白表达量较空白对照组减少,呈剂量依赖型;而细胞核内Smad4蛋白表达量较空白对照组增多.结论:SB431542通过抑制大鼠HSC活化过程中Smad4蛋白在细胞内的核转位,可起到阻断TGF-1/Smad通路的细胞内信号转导的作用.
AIM： To investigate the expression and intracellular translocation of Smad4 protein in HSC-T6 cells treated with SB431542. METHODS： Adherent HSC-T6 cells were incubated with SB431542 （10 mol/L） for two hours, and the intracellular translocation of Smad4 protein was observed by immunofluorescence using a Laser scanning confocal microscope. Another part of HSC-T6 cells were randomly divided into three groups： a normal control group and two SB431542 treatment groups （treated with 1 and 10 mol/L of SB431542, respectively）. After incubation with SB431542 for 24 hours, cytoplasmic protein and nuclear protein were extracted and detected by Western blot. RESULTS： Intracellular translocation of Smad4 protein did not occur when the cells were incubated with 10 mol/L of SB431542 for two hours. Compared to the control group, the expression of Smad4 protein in the cytoplasm increased and that in the nucleus decreased after treatment with SB431542 for 24 h, and the effects were does-dependent. CONCLUSION： SB431542 blocks the TGFβ1/ Smads signaling pathway by inhibiting Smad4 nuclear translocation in HSC-T6 cells.
出处
《世界华人消化杂志》
CAS
北大核心
2012年第18期1653-1656,共4页
World Chinese Journal of Digestology
