Deptor protects against myocardial ischemia-reperfusion injury by regulating the mTOR signaling and autophagy.

IF 6.1 2区生物学 Q1 CELL BIOLOGY Cell Death Discovery Pub Date : 2024-12-19 DOI:10.1038/s41420-024-02263-1

Qunjun Duan, Weijun Yang, Xian Zhu, Zhanzeng Feng, Jiangwei Song, Xiaobin Xu, Minjian Kong, Jiayan Mao, Jian Shen, Yuqin Deng, Rujia Tao, Hongfei Xu, Wei Chen, Weidong Li, Aiqiang Dong, Jie Han

{"title":"Deptor protects against myocardial ischemia-reperfusion injury by regulating the mTOR signaling and autophagy.","authors":"Qunjun Duan, Weijun Yang, Xian Zhu, Zhanzeng Feng, Jiangwei Song, Xiaobin Xu, Minjian Kong, Jiayan Mao, Jian Shen, Yuqin Deng, Rujia Tao, Hongfei Xu, Wei Chen, Weidong Li, Aiqiang Dong, Jie Han","doi":"10.1038/s41420-024-02263-1","DOIUrl":null,"url":null,"abstract":"<p><p>Deptor knockout mice were constructed by crossing Deptor Floxp3 mice with myh6 Cre mice, establishing a myocardial ischemia-reperfusion (I/R) model. Deptor knockout mice exhibited significantly increased myocardial infarction size and increased myocardial apoptosis in vivo. ELISA analysis indicated that the expression of CK-MB, LDH, and CtnT/I was significantly higher in the Deptor knockout mice. Deptor siRNA significantly reduced cell activity and increased myocardial apoptosis after I/R-induced in vitro. Deptor siRNA also significantly up-regulated the expression of p-mTOR, p-4EBP1, and p62, and down-regulated the expression of LC3 after I/R induction. Immunofluorescence indicated that LC3 dual fluorescence was weakened by Deptor knockout, and was enhanced after transfection with Deptor-overexpression plasmids. Treatment with OSI027, a co-inhibitor of mTORC1 and mTORC2, in either Deptor knockout mice or Deptor knockout H9C2 cells, resulted in a significant reduction in infarction size and apoptotic cardiomyocytes. ELISA analysis also showed that the expression of CK-MB, LDH, and CtnT/I were significantly down-regulated by treatment with OSI027. CCK-8 cell viability indicated that cell viability was enhanced, and the number of apoptotic cells was decreased in vitro following treatment with OSI027. These results revealed that OSI027 exerts a protective effect on myocardial ischemia/reperfusion injury in both an in vivo and in an in vitro model of I/R. These findings demonstrate that Deptor protects against I/R-induced myocardial injury by inhibiting the mTOR pathway and by increasing autophagy.</p>","PeriodicalId":9735,"journal":{"name":"Cell Death Discovery","volume":"10 1","pages":"508"},"PeriodicalIF":6.1000,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659626/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cell Death Discovery","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1038/s41420-024-02263-1","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CELL BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Deptor knockout mice were constructed by crossing Deptor Floxp3 mice with myh6 Cre mice, establishing a myocardial ischemia-reperfusion (I/R) model. Deptor knockout mice exhibited significantly increased myocardial infarction size and increased myocardial apoptosis in vivo. ELISA analysis indicated that the expression of CK-MB, LDH, and CtnT/I was significantly higher in the Deptor knockout mice. Deptor siRNA significantly reduced cell activity and increased myocardial apoptosis after I/R-induced in vitro. Deptor siRNA also significantly up-regulated the expression of p-mTOR, p-4EBP1, and p62, and down-regulated the expression of LC3 after I/R induction. Immunofluorescence indicated that LC3 dual fluorescence was weakened by Deptor knockout, and was enhanced after transfection with Deptor-overexpression plasmids. Treatment with OSI027, a co-inhibitor of mTORC1 and mTORC2, in either Deptor knockout mice or Deptor knockout H9C2 cells, resulted in a significant reduction in infarction size and apoptotic cardiomyocytes. ELISA analysis also showed that the expression of CK-MB, LDH, and CtnT/I were significantly down-regulated by treatment with OSI027. CCK-8 cell viability indicated that cell viability was enhanced, and the number of apoptotic cells was decreased in vitro following treatment with OSI027. These results revealed that OSI027 exerts a protective effect on myocardial ischemia/reperfusion injury in both an in vivo and in an in vitro model of I/R. These findings demonstrate that Deptor protects against I/R-induced myocardial injury by inhibiting the mTOR pathway and by increasing autophagy.

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

detor通过调节mTOR信号传导和自噬来预防心肌缺血再灌注损伤。

将Deptor Floxp3小鼠与myh6 Cre小鼠杂交构建Deptor敲除小鼠，建立心肌缺血-再灌注（I/R）模型。detor基因敲除小鼠体内心肌梗死面积明显增加，心肌凋亡明显增加。ELISA分析显示，敲除detor小鼠CK-MB、LDH和CtnT/I的表达显著升高。体外I/ r诱导后，detor siRNA显著降低细胞活性，增加心肌凋亡。在I/R诱导后，detor siRNA也显著上调p-mTOR、p-4EBP1和p62的表达，下调LC3的表达。免疫荧光显示，敲除Deptor后LC3双荧光减弱，转染Deptor过表达质粒后LC3双荧光增强。在detor基因敲除小鼠或detor基因敲除H9C2细胞中，使用OSI027 （mTORC1和mTORC2的共同抑制剂）治疗可显著减少梗死大小和心肌细胞凋亡。ELISA分析还显示，OSI027处理后，CK-MB、LDH和CtnT/I的表达均显著下调。体外CCK-8细胞活力显示，OSI027处理后细胞活力增强，凋亡细胞数量减少。上述结果表明，OSI027对体内和体外I/R模型心肌缺血再灌注损伤均有保护作用。这些发现表明，detor通过抑制mTOR通路和增加自噬来保护I/ r诱导的心肌损伤。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Cell Death Discovery Biochemistry, Genetics and Molecular Biology-Cell Biology

CiteScore

8.30

自引率

1.40%

发文量

468

审稿时长

9 weeks

期刊介绍： Cell Death Discovery is a multidisciplinary, international, online-only, open access journal, dedicated to publishing research at the intersection of medicine with biochemistry, pharmacology, immunology, cell biology and cell death, provided it is scientifically sound. The unrestricted access to research findings in Cell Death Discovery will foster a dynamic and highly productive dialogue between basic scientists and clinicians, as well as researchers in industry with a focus on cancer, neurobiology and inflammation research. As an official journal of the Cell Death Differentiation Association (ADMC), Cell Death Discovery will build upon the success of Cell Death & Differentiation and Cell Death & Disease in publishing important peer-reviewed original research, timely reviews and editorial commentary. Cell Death Discovery is committed to increasing the reproducibility of research. To this end, in conjunction with its sister journals Cell Death & Differentiation and Cell Death & Disease, Cell Death Discovery provides a unique forum for scientists as well as clinicians and members of the pharmaceutical and biotechnical industry. It is committed to the rapid publication of high quality original papers that relate to these subjects, together with topical, usually solicited, reviews, editorial correspondence and occasional commentaries on controversial and scientifically informative issues.