| 王艳鹏,黄世恩,陈志明.miR-100过表达通过增强AMPK-mTOR-p70S6K信号通路介导的自噬而发挥抗脓毒症作用的研究[J].浙江中西医结合杂志,2022,32(2): |
| miR-100过表达通过增强AMPK-mTOR-p70S6K信号通路介导的自噬而发挥抗脓毒症作用的研究 |
| miR-100 overexpression inhibits sepsis through enhance of AMPK-mTOR-p70S6K signaling-mediated autophagy |
| 投稿时间：2021-05-05 修订日期：2022-01-21 |
| DOI： |
| 中文关键词: 脓毒症 miR-100 自噬 AMPK-mTOR-p70S6K通路 |
| 英文关键词:sepsis miR-100 cell autophag AMPK-mTOR-p70S6K pathway |
| 基金项目:浙江省医药卫生科技计划项目(No.2020KY734) |
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| 中文摘要: |
| 目的：探讨miR-100过表达通过增强AMPK-mTOR-p70S6K信号通路介导的自噬,进而发挥抗脓毒症的作用机制研究。方法：利用脂多糖(LPS)刺激大鼠心肌细胞H9C2构建脓毒症体外细胞模型；细胞通过转染miR-100的激动剂或抑制剂使miR-100在细胞内过表达或敲低,并用qRT-PCR检测细胞内miR-100的表达水平的改变；ELISA检测不同细胞处理组上清中炎症因子IL-1β、IL-6和TNF-α的释放；细胞免疫荧光检测LC3蛋白在细胞中的表达；Western blot检测自噬相关蛋白(p62、LC3和beclin-1)和AMPK-mTOR-p70S6K通路相关蛋白(AMPK、p-AMPK、mTOR、p-mTOR、p70S6K和p-p70S6K)的表达。结果：在LPS处理心肌细胞H9C2的脓毒症细胞模型中,细胞上清炎症因子分泌显著增加,细胞自噬增强,炎症增加；而上调miR-100表达能够诱导增强细胞自噬,并且减弱炎症反应；相反,下调miR-100表达减弱了自噬,并且增强了炎症反应；进一步研究发现miR-100能够引起自噬相关信号通路AMPK-mTOR-p70S6K中相关蛋白表达变化。结论：本研究发现miR-100过表达可以通过上调AMPK-mTOR-p70S6K信号通路介导的自噬水平,发挥抗炎作用,进而在脓毒症中发挥保护作用。 |
| 英文摘要: |
| Objective: To explore the mechanism of miR-100 inhibiting sepsis through enhance of AMPK-mTOR-p70S6K signaling-mediated autophagy. Methods: Rat cardiomyocytes H9C2 were stimulated with lipopolysaccharide (LPS) to establish sepsis cell model in vitro; intracellular overexpression or knockdown of miR-100 was constructed by transfection of miR-100 agonists or inhibitors; the expression of IL-1β, IL-6 and TNF-α was detected by ELISA; the expression of miR-100 was measured by qRT-PCR; the LC3 protein was detected by immunofluorescence; autophagy related proteins (p62, LC3-I, LC3-II and beclin-1) and AMPK-mTOR-p70S6K pathway related proteins (AMPK, p-AMPK, mTOR, p-mTOR, p70S6K and p-p70S6K) were detected by Western blot. Results: LPS treatment of H9C2 significantly increased the secretion of pro-inflammatory factors, and strengthened cell autophagy. At the same time, up-regulation of miR-100 expression can furtherly enhance cell autophagy, and reduce inflammatory response; on the contrary, down-regulation can weaken autophagy, and enhance inflammatory response; it was found that miR-100 can cause AMPK-mTOR-p70S6K pathway related protein expression changes which was related to cell autophagy in the further study. Conclusions: This study revealed that miR-100 can play an anti-inflammatory role by inducing cell autophagy mediated by AMPK-mTOR-p70S6K pathway, and then play a protective role in sepsis. |
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