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

doi:10.16352/j.issn.1001-6325.2024.01.0023

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (1): 23-30.doi: 10.16352/j.issn.1001-6325.2024.01.0023

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

苏拾香^1,2, 王语阳², 覃宗帅¹, 黄桂香², 徐键², 岑兰英², 覃月秋^1*

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

收稿日期:2023-09-11 修回日期:2023-11-22 出版日期:2024-01-05 发布日期:2023-12-25
通讯作者: ^*:gxbsqyq@126.com
基金资助:
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)

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

SU Shixiang^1,2, WANG Yuyang², QIN Zongshuai¹, HUANG Guixiang², XU Jian², CEN Lanying², QIN Yueqiu^1*

1. The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000;
2. Graduate School,Youjiang Medical University for Nationalities, Baise 533000,China

Received:2023-09-11 Revised:2023-11-22 Online:2024-01-05 Published:2023-12-25
Contact: ^*:gxbsqyq@126.com

摘要/Abstract

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。

关键词: miR-142-3p, 凋亡, 急性胰腺炎, 高迁移率族蛋白B1

Abstract: 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)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.

Key words: miR-142-3p, apoptosis, acute pancreatitis, high mobility group protein B1

中图分类号:

R576

苏拾香, 王语阳, 覃宗帅, 黄桂香, 徐键, 岑兰英, 覃月秋. 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[J]. 基础医学与临床, 2024, 44(1): 23-30.

SU Shixiang, WANG Yuyang, QIN Zongshuai, HUANG Guixiang, XU Jian, CEN Lanying, QIN Yueqiu. miR-142-3p inhibits cerulein-induced apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1[J]. Basic & Clinical Medicine, 2024, 44(1): 23-30.

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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

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