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OACSTPCD
miR-142-3p inhibits cerulein-induced apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1
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 signifi-cantly(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 signifi-cantly 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)The expression of miR-142-3p was low in the model group.2)miR-142-3p can inhibit the apoptosis of AR42J cells by inhibiting the expression of Hmgb1.
苏拾香;王语阳;覃宗帅;黄桂香;徐键;岑兰英;覃月秋
右江民族医学院附属医院, 广西 百色 533000||右江民族学院 研究生院, 广西 百色 533000右江民族学院 研究生院, 广西 百色 533000右江民族医学院附属医院, 广西 百色 533000
临床医学
miR-142-3p凋亡急性胰腺炎高迁移率族蛋白B1
miR-142-3papoptosisacute pancreatitishigh mobility group protein B1
《基础医学与临床》 2024 (001)
23-30 / 8
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)
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