miR-142-3p通过调控Hmgb1抑制雨蛙素诱导的大鼠胰腺外分泌细胞系AR42J凋亡  

miR-142-3p inhibits cerulein-induced apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1

作　　者：苏拾香 王语阳 覃宗帅 黄桂香 徐键 岑兰英 覃月秋[1] SU Shixiang;WANG Yuyang;QIN Zongshuai;HUANG Guixiang;XU Jian;CEN Lanying;QIN Yueqiu(The Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000;Graduate School,Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院附属医院,广西百色533000 [2]右江民族学院研究生院,广西百色533000

出　　处：《基础医学与临床》2024年第1期23-30,共8页Basic and Clinical Medicine

基　　金：国家自然科学基金(82260134);广西自然科学基金(2023GXNSFAA026118);右江民族医学院附属医院高层次人才科研项目(R202011702);广西研究生教育创新计划(YCSW2023506)。

摘　　要：目的探讨miR-142-3p调控Hmgb1对大鼠胰腺外分泌细胞系AR42J凋亡的影响。方法将AR42J细胞分为空白组(blank)、急性胰腺炎模型组(AP,100 nmol/L雨蛙素作用24 h),再分别用miR-142-3p mimics、mimics NC、miR-142-3p inhibitor和inhibitor NC转染模型组细胞,记为miR-142-3p mimics组、mimics NC组、miR-142-3p inhibitor组和inhibitor NC组。用RT-qPCR检测细胞中miR-142-3p表达;Western blot检测HMGB1、caspase-3、Bax、Bcl-2蛋白表达;Hoechst染色测定细胞凋亡;流式细胞测量术检测细胞凋亡率;双荧光素酶报告基因实验明确miR-142-3p和Hmgb1的靶向关系。结果与空白组相比,AP组中miR-142-3p表达水平显著下调(P<0.01),HMGB1、caspase-3蛋白表达量上调(P<0.05),Bax蛋白表达量显著上调(P<0.01),Bcl-2蛋白表达量显著降低(P<0.01),细胞凋亡率显著升高(P<0.01);与mimics NC组相比,miR-142-3p mimics组miR-142-3p水平显著上调(P<0.01),HMGB1、caspase-3、Bax蛋白表达量显著下调(P<0.01),Bcl-2蛋白表达量上调(P<0.05),细胞凋亡率显著降低(P<0.01);与inhibitor NC组相比,miR-142-3p inhibitor组miR-142-3p表达水平下调(P<0.05),HMGB1、caspase-3、Bax蛋白表达量显著上调(P<0.01),Bcl-2蛋白表达量降低(P<0.05),细胞凋亡率显著升高(P<0.01),差异均有统计学意义。双荧光素酶报告基因实验显示Hmgb1为miR-142-3p的靶基因。结论1)miR-142-3p在模型组细胞中低表达。2)miR-142-3p可靶向抑制Hmgb1表达进而抑制AR42J细胞凋亡。Objective To investigate the effect of miR-142-3p on the apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1.Methods AR42J cells were divided into blank group(blank),acute pancreatitis model group(AP,100 nmol/L cerulein for 24 h),and then transfected with miR-142-3p mimics,mimics NC,miR-142-3p inhibitor and inhibitor NC,respectively.The cells in the model group were recorded as miR-142-3p mimics group,mimics NC group,miR-142-3p inhibitor group and inhibitor NC.The expression of miR-142-3p in cells was detected by RT-qPCR.The protein expressions of HMGB1,caspase-3,Bax and Bcl-2 were detected by Western blot.Hoechst staining was used to determine cell apoptosis.The apoptosis rate of cells was detected by flow cytometry.The targeting relationship between miR-142-3p and Hmgb1 was determined by dual luciferase reporter gene assay.Results Compared with blank control group,the expression level of miR-142-3p in the AP group was significantly down-regulated(P<0.01),the expression level of HMGB1 and caspase-3 proteins was up-regulated(P<0.05),the expression level of Bax protein was significantly up-regulated(P<0.01),the expression level of Bcl-2 protein was significantly decreased(P<0.01)and the apoptosis rate increased significantly(P<0.01).Compared with the mimics NC group,the level of miR-142-3p in the miR-142-3p mimics group was significantly up-regulated(P<0.01),the expression of HMGB,caspase-3 and Bax proteins was significantly down-regulated(P<0.01),the expression of Bcl-2 protein was up-regulated(P<0.05),and the apoptosis rate decreased significantly(P<0.01).Compared with inhibitor NC group,the expression level of miR-142-3p in miR-142-3p inhibitor group was down-regulated(P<0.05),the expression levels of HMGB1,caspase-3 and Bax proteins were significantly up-regulated(P<0.01),the expression level of Bcl-2 protein was decreased(P<0.05)and the apoptosis rate increased significantly(P<0.01).The dual luciferase reporter gene assay showed that Hmgb1 was the target gene of miR-142-3p.Conclusions 1)T

关 键 词：miR-142-3p 凋亡 急性胰腺炎 高迁移率族蛋白B1 

分 类 号：R576[医药卫生—消化系统]