MDM2通过p53/Bcl-2/Bax轴调控H2O2诱导的肺泡Ⅱ型上皮细胞损伤

扫码查看

原文链接

NETL
NSTL
万方数据

中文摘要：目的探索H2O2诱导的氧化损伤时MDM2的功能及与p53的关系.方法使用0.5 mmol/L H2O2建立MLE-12 HALI细胞模型,分为:健康对照组、H2O2损伤组、H2O2+MDM2 overexpressed组、H2O2+MDM2-shRNA组.通过腺病毒载体感染MLE-12细胞过表达、沉默MDM2;采用免疫共沉淀(Co-IP)分析MDM2与p53的相互作用;采用Western blotting检测HALI建模后MDM2、p53、Bcl-2、Bax和cleaved caspase-3蛋白表达水平;检测各组细胞凋亡率.结果通过转录组测序后发现p53信号通路与HALI密切相关.与正常组相比,H2O2损伤组MDM2表达较低(P＜0.05);与H2O2损伤组相比,MDM2过表达后MLE-12细胞凋亡率降低(P＜0.05),p53、Bax、cleaved caspase-3 蛋白表达水平下降,MDM2、Bcl-2 蛋白表达上调(P＜0.05).与H2O2损伤组相比,当MDM2沉默时,细胞凋亡率增加(P＜0.05),p53、Bax、cleaved caspase-3蛋白表达水平上调,MDM2、Bcl-2蛋白表达下降(P＜0.05).Co-IP实验显示MDM2与p53蛋白结合.结论这些结果表明,MDM2可通过p53/Bcl-2/Bax轴抑制MLE-12细胞凋亡从而发挥对HALI的保护作用.

外文标题：MDM2 regulates H2O2 induced alveolar type Ⅱ epithelial cell injury through p53/Bcl-2/Bax axis

外文摘要：Objective To explore the function of MDM2 and its relationship with p53 at the cellular level during H2O2 induced oxidative damage.Methods MLE-12 HALI cell models were established using 0.5 mmol/L H2O2,and were divided into three groups:normal control group,H2O2 injury group,H2O2+MDM2 overexpressed group,and H2O2+MDM2 shRNA group.Infection of MLE-12 cells with adenovirus vector overexpressing and silencing MDM2;Using immunoprecipitation(Co-IP)to analyze the interaction between MDM2 and p53;Western blotting was used to detect the protein expression levels of MDM2,p53,Bcl-2,Bax,and cleared caspase-3 after HALI modeling;Measure the apoptosis rate of cells in each group.Results After transcriptome sequencing,the p53 signaling pathway closely related to HALI.Compared with the normal group,the expression of MDM2 in the H2O2 injury group was lower(P＜0.05);Compared with the H2O2 injury group,overexpression of MDM2 resulted in a decrease in the apoptosis rate of MLE-12 cells(P＜0.05),a decrease in the expression levels of p53,Bax,and cleared caspase-3 proteins,and an upregulation of MDM2 and Bcl-2 protein expression(P＜0.05).Compared with the H2O2 injury group,when MDM2 was silenced,the cell apoptosis rate increased(P＜0.05),and the expression levels of p53,Bax,and cleared caspase-3 proteins were upregulated,while the expression levels of MDM2 and Bcl-2 proteins decreased(P＜0.05).Co-IP experiments showed that MDM2 binds to p53 protein.Conclusions MDM2 can exert a protective effect on HAL1 by inhibiting MLE-12 cell apoptosis through the p53/Bcl-2/Bax axis.

外文关键词：

ApoptosisMDM2Hyperoxic acute lung injuryp53

作者：

郑杰、陈博文、梅鸿、刘鑫鑫、廖贞亮、余琨、余虹、冯帮海、陈淼、傅小云、覃松

展开 >

作者单位：

遵义医科大学附属医院重症医学科,遵义 563000

贵州医科大学附属医院重症医学科,贵阳 550004

遵义市中医院重症医学科,遵义 563000

关键词：

凋亡鼠双微立体基因2 高氧急性肺损伤 p53

基金：

国家自然科学基金贵州省科技计划贵州省卫生健康委科学技术基金贵州省卫生健康委科学技术基金遵义市科技与大数据局科学技术项目遵义市科技与大数据局科学技术项目Science and Technology Fund Project of Guizhou Provincial Health CommissionScience and Technology Fund Project of Zunyi Science and Technology and Big Data Bureau:Science and Technology Fund Project of Zunyi Science and Technology and Big Data Bureau:

项目编号：

81960362ZK-2023-544gzwkj2024-310gzwjkj2019-1-0682023-2212023-199gzwkj2024-3102023-2212023-199

出版年：

2024

DOI：

10.3760/cma.j.issn.1671-0282.2024.08.007

中华急诊医学杂志

中华医学会

中华急诊医学杂志

CSTPCD北大核心

影响因子：1.556

ISSN：1671-0282

年,卷(期)：2024.33(8)