干扰CDH1基因对雨蛙素诱导的急性胰腺炎细胞模型凋亡和自噬的影响  

Effect of Interference with CDH1 Gene on Apoptosis and Autophagy in Cell Models of Acute Pancreatitis Induced by Caer-ulein

作　　者：史洋洋 吴雪生[1] SHI Yang-yang;WU Xue-sheng(Department of Emergency Surgery,The First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)

机构地区：[1]安徽医科大学第一附属医院急诊外科,合肥230022

出　　处：《肝胆外科杂志》2021年第5期385-389,共5页Journal of Hepatobiliary Surgery

摘　　要：目的探讨干扰上皮钙粘素(CDH1)基因对雨蛙素(Caerulein)诱导的大鼠急性胰腺腺泡细胞AR42J凋亡和自噬的影响及可能的机制.方法体外培养大鼠急性胰腺腺泡细胞AR42J,将CDH1特异性siRNA转染至AR42J细胞,成功干扰CDH1表达后以雨蛙素干预构建急性胰腺炎细胞模型,实验分为空白对照(Mock)组、雨蛙素(Cae)组、雨蛙素+转染对照(Cae+si-NC)组和雨蛙素+转染(Cae+si-CDH1)组,实时荧光定量PCR(qPCR)和蛋白免疫印迹(Western blot)检测CDH1的表达变化,噻唑蓝(MTT)分析细胞增殖能力,酶联免疫吸附法(ELISA)检测炎性介质肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)的表达,流式细胞术检测细胞凋亡率,Western blot分析凋亡相关蛋白Bcl-2相关X蛋白(Bax)和B细胞淋巴瘤/白血病-2(Bcl-2),自噬标记物Beclin-1、LC3Ⅰ和LC3Ⅱ以及β-catenin信号通路相关蛋白β-catenin和其下游靶分子c-Myc和cyc-lin D1的表达.结果与Mock组相比,Cae组细胞增殖能力降低,细胞凋亡率升高,细胞中CDH1、Bax、Beclin-1、LC3-Ⅱ、TNF-α和IL-1β、β-catenin、c-Myc和cyclin D1的表达上调,LC3-Ⅰ和Bcl-2的表达下调,差异有统计学意义(P<0.05);与Cae组相比,Cae+si-CDH1组干扰CDH1能够逆转以上各指标,差异有统计学意义(P<0.05).结论干扰CDH1基因可减轻雨蛙素诱导的AR42J细胞凋亡和自噬,其作用机制可能与阻断β-catenin信号通路的激活有关.Objective To investigate the effect of interfering epithelial cadherin(CDH1)gene on the apoptosis and autophagy of rat acute pancreatic acinar cell AR42J induced by caerulcin and its possible mechanism.Methods Rat acute pancreatic acinar cells AR42J were cultured in vitro,and CDH1 specific siRNA was transfected into AR42J cells.After successfully interfering with the expression of CDH1,the acute pancreatitis cell model was constructed with cerulein intervention.The experiment was divided into blank control(Mock)group,Cerulein(Cae)group,cerulein+transfection control(Cae+si-NC)group and cerulein+transfection(Cae+si-CDH1)group,real-time fluorescent quantitative PCR(qPCR)and Western blot Detection of CDH1 expression changes,thiazolyl blue(MTT)analysis of cell proliferation,enzyme-linked immunosorbent assay(ELISA)detection of inflammatory mediator tumor necrosis factor α(TNF-α)and interleukin-1β(IL-1β)Expression,flow cytometry to detect cell apoptosis rate,Western blot analysis of apoptosis-related protein Bcl-2 related X protein(Bax)and B-cell Iymphoma/leukemia-2(Bcl-2),autophagy marker Bec-lin-1,LC3Ⅰ and LC3Ⅱ,β-catenin signaling pathway related protein β-catenin and its downstream target molecules c-Myc and cyclin D1 expression.Results Compared with the Mock group,the cell proliferation ability of the Cae group was reduced,and the apoptosis rate was increased.CDH1,Bax,Beclin-1,LC3-Ⅱ,TNF-α and IL-1β,p-catenin,and c-Myc in the cells The expression of cyclin D1 and cyclin D1 was up-rcgulated,and the expression of LC3-Ⅰ and Bcl-2 was down-regulated,and the difference was statistically significant(P<0.05);compared with the Cae group,the interference of CDH1 in the Cae+si-CDH1 group could reverse the above indicators,and the difference was statistically significant(P<0.05).Conclusion Interference with CDH1 gene can reduce the apoptosis and autophagy of AR42J cells induced by caerulein,and its mechanism may be related to blocking the activation of β-catenin signaling pathway.

关 键 词：CDH1基因 雨蛙素 急性胰腺炎 凋亡 自噬 

分 类 号：R659[医药卫生—外科学]