摘要
目的探讨干扰素调节因子IRF9基因沉默对雨蛙素诱导的大鼠胰腺腺泡细胞AR42J细胞凋亡的影响,阐述IRF9在急性胰腺炎(AP)发生发展中的作用。方法 AR42J细胞培养至对数期,分为空白对照组、阴性对照组及IRF9沉默组。成功转染siRNA后应用雨蛙素诱导AR42J细胞构建体外急性胰腺炎模型。采用qRT-PCR和Western blot方法检测AR42J细胞中IRF9基因、Caspase 3及Caspase 9的mRNA及蛋白的表达水平;流式细胞术检测AR42J细胞凋亡率。结果①雨蛙素诱导24 h后,各组中的IRF9基因的mRNA及蛋白水平均较雨蛙素诱导前升高,Caspase 3及Caspase 9的mRNA及蛋白的表达水平亦明显升高,说明发生AP时IRF9及凋亡相关基因的表达水平均上调。②与空白对照组相比,雨蛙素诱导后IRF9沉默组AR42J细胞的凋亡率有明显降低,差异有统计学意义(P<0.05)。结论发生AP时IRF9、Caspase 3及Caspase 9的表达水平均上调,细胞凋亡率增加,IRF9基因沉默降低了雨蛙素诱导的体外AP细胞模型的细胞凋亡。
Objective To explore the effect of interferon regulatory factor IRF9 gene silencing on apoptosis of cerulein induced exocrine pancreas AR42 J cells and elucidate the role of IRF9 gene in the development of acute pancreatitis.Methods The AR42 J cells at logarithmic growth stage were divided into blank control group, negative control group and IRF9 silencing group. After successful transfection of siRNA, the model of acute pancreatitis in vitro was constructed by cerulean treatment. The mRNA and protein expression levels of IRF9 gene, Caspase 3 and Caspase 9 in AR42 J cells were detected by qRT-PCR and Western blot. The apoptosis rate of AR42 J cells in each group was detected by Annexin V-FITC/PI double staining assay.Results ① The mRNA and protein levels of IRF9 gene in each group increased after cerulean treated 24 hours, and the mRNA and protein levels of Caspase 3 and Caspase 9 also significantly increased, indicating that the expression levels of IRF9 and apoptosis-related genes were up-regulated when acute pancreatitis occurred.② Compared with the blank control group, the apoptosis rate of AR42 J cells in IRF9 silencing group significantly reduced after cerulean treatment(P<0.05).Conclusions The expression levels of IRF9, Caspase 3 and Caspase 9 were up-regulated and the apoptosis rate increased when acute pancreatitis occured. The silencing of IRF9 gene reduced the apoptosis of cerulean induced acute pancreatitis in vitro.
作者
薛彬华
余维丽
王小蝶
刘奕
Xue Binhua;Yu Weili;Wang Xiaodie(Dept of Inteniee Cars Unit,the Second Afiliated Hospital f Anhui Merical Unnersite,Hefei 230601)
出处
《安徽医科大学学报》
CAS
北大核心
2020年第4期508-512,共5页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金(编号:1508085QC49)
安徽医科大学第二附属医院博士科研启动基金(编号:2014bkj034)。
