小干扰RNA沉默IKK/NF-κB信号通路对HSC-T6细胞凋亡的影响  

Effects of small interfering RNA silencing IKK/NF-κB on the apoptosis of HSC-T6

作　　者：张彦亮[1] 吴会玲[1] 岳巧艳[1] 宋爽[1] 宋希[1] 陶臻[1] 

机构地区：[1]南京医科大学附属南京医院感染科(南京市第一医院),江苏南京210006

出　　处：《胃肠病学和肝病学杂志》2016年第11期1258-1261,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：南京市卫生科技发展项目(QYK11173)

摘　　要：目的研究小干扰RNA(siRNA)沉默IKK/NF-κB信号通路对于肝星状细胞株HSC-T6凋亡的影响。方法根据Gen Bank数据库IKKβ的c DNA序列,设计构建合成IKKβsiRNA,采用阳性脂质体转染,设立空白对照组、阴性对照组及siRNA实验组,采用TUNEL法和流式细胞仪Annexin-V/PI双染法测定IKKβsiRNA转染后24 h、48 h和72 h细胞凋亡率,同时采用Western blotting检测NF-κB P65、Bcl-2、Bax和Caspase3的蛋白表达情况。结果与空白对照组和阴性对照组相比,实验组24 h、48 h和72 h HSC-T6细胞凋亡率均显著增加(P<0.05),且具有时间依赖性;实验组的NF-κB P65及Bcl-2蛋白的表达均明显升高,而Caspase3和Bax则显著增加,与空白对照和阴性对照组相比差异有统计学意义(P<0.05)。结论 SiRNA沉默IKK/NF-κB信号通路能够下调NF-κB P65的表达,同时提高Bax和Caspase3的表达,促进HSC-T6凋亡,这种肝星状细胞凋亡诱导效应具有潜在的逆转肝纤维化的治疗作用。Objective To investigate the effects of small interfering RNA （siRNA） silencing IKK/NF-κB on the ap- optosis of hepate stellate cell line T6 （HSC-T6）. Methods Chemically synthesized siRNA directed against IKKβ was transfected into HSC-T6 by cationic liposome Lipofectamine. The cells were divided into blank control group, negative control group and experimental group. TUNEL staining and AnnexinV/PI double staining with Flow cytometry （FCM） were used to detect the apoptosis rate of the cells at 24 hours,48 hours, 72 hours. The protein levels of NF-κB P65, Bcl-2, Bax and Caspase3 were analyzed by Western blotting. Results Compared with the negative control group and blank control group, the apoptosis rate in experimental group was increased significantly with time （P 〈 0.05）. In the experimental group, the expressions of NF-κB P65 and Bcl-2 were decreased significantly, while the expressions of Caspase and Bax were increased compared with the negative control group and blank control group （P 〈 0.05）. Con- clusion SiRNA silencing IKK/NF-κB can down-regulate the expression of NF-κB P65, while can increase the expres- sions of Bax and Caspase3, by which can promote apoptosis of HSC-T6. This may be a potential strategy for the reverse effect of hepatic fibrosis.

关 键 词：小干扰RNA 肝纤维化 肝星状细胞 IKK/NF-κB信号通路 凋亡 

分 类 号：R575[医药卫生—消化系统]