摘要
活化STAT蛋白抑制物（PIAS）1基因沉默可增强雨蛙肽诱导的胰腺腺泡细胞炎症反应.目的：探讨PIAS1基因沉默对雨蛙肽刺激胰腺腺泡细胞凋亡的影响.方法：AR42J胰腺腺泡细胞在LipofectamineTM 2000介导下转染PIAS1-siRNA和阴性siRNA.将细胞分为PIAS1-siRNA＋雨蛙肽组、阴性siRNA＋雨蛙肽组、脂质体＋雨蛙肽组、PBS＋雨蛙肽组和对照组.以DNA Ladder、Hoechst 33258染色检测细胞凋亡情况,流式细胞术测定细胞周期和细胞凋亡率,RT-PCR和蛋白质印迹法检测p53、Bax、Bcl-2、caspase-3 mRNA和蛋白表达.结果：与其余各组相比,PIAS1-siRNA＋雨蛙肽组DNA梯度裂解条带明显增加,荧光着色阳性细胞数增多;G1期细胞数增多,S期细胞数减少,细胞凋亡率增加;p53、Bax、caspase-3 mRNA和蛋白表达明显上调,Bcl-2 mRNA和蛋白表达下调（P均〈0.05）.结论：PIAS1基因沉默可增强雨蛙肽活化的caspase-3凋亡途径诱导的胰腺腺泡细胞凋亡,为通过调控PIAS1表达治疗急性胰腺炎提供新的理论依据.
Background： Silencing of protein inhibitor of activated STAT 1 （PIAS1） gene may enhance the inflammatory responses of eerulein stimulated pancreatic acinar cells. Aims： To investigate the effects of PIAS1 gene silencing on apoptosis of pancreatic acinar cells with cernlein stimulation. Methods： AR42J cells were transfected with PIASI-siRNA or negative siRNA by LipofectamineTM 2000. Cells were divided into PIASl-siRNA＋cerulein group, negative siRNA＋cerulein group, LipofectamineTM 2000＋cerulein group, PBS＋cerulein group and PBS treated control group. Cell apoptosis was determined by DNA Ladder and Hoechst 33258 staining. Cell cycle and cell apoptosis rate were measured by flow cytometry, mRNA and protein expressions of p53, Bax, Bcl-2, caspase-3 were analyzed by RT-PCR and Western blotting, respectively. Results： Compared with the other groups, PIASI-siRNA ＋cendein group resulted in a significant formation of DNA Ladder, more positive cells stained with Hoechst 33258; larger population of sub-G1 cells, less population of sub-S cells, and increased cell apoptosis rate; mRNA and protein expressions of p53, Bax, caspase-3 increased significantly, while those of Bcl-2 were decreased significantly （all P〈0.05）. Conclusions： PIAS1 gene silencing may enhance the activity of caspase-3 apoptotic pathway and promote apoptosis in pancreatic acinar cells with cerulein stimulation, which may provide evidences that PIAS1 could become a new therapeutic target for acute pancreatitis.
出处
《胃肠病学》
2010年第7期395-399,共5页
Chinese Journal of Gastroenterology
