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微小RNA-214保护心肌细胞免受过氧化氢诱导的损伤。

MicroRNA-214 protects cardiac myocytes against H2O2-induced injury.

作者信息

Lv Guangwei, Shao Suxia, Dong Hua, Bian Xiaohua, Yang Xingwei, Dong Shimin

机构信息

Department of ICU, The Third Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, P.R. China.

出版信息

J Cell Biochem. 2014 Jan;115(1):93-101. doi: 10.1002/jcb.24636.

Abstract

Reactive oxygen species (ROS)-induced cardiac myocyte injury resulting from changes in the expression levels of multiple genes plays a critical role in the pathogenesis of numerous heart diseases. The purpose of this study was to determine the potential roles of microRNA-214 (miR-214) in hydrogen peroxide (H2O2)-mediated gene regulation in cardiac myocytes. In this study, we used quantitative real-time RT-PCR (qRT-PCR) to demonstrate that miR-214 was upregulated in cardiac myocytes after treatment with H2O2. We transfected cells with pre-miR-214 to upregulate miR-214 expression and transfected cells with a miR-214 inhibitor (anti-miR-214) to downregulate miR-214 expression. H2O2-induced cardiac cell apoptosis was detected by flow cytometry. The level of apoptosis was increased by the miR-214 inhibitor and decreased by pre-miR-214. Therefore, we believe that miR-214 plays a positive role in H2O2-induced cardiac cell apoptosis. Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is constitutively active and is considered to be the primary downregulator of the pro-oncogenic PI3K/Akt pathway. Western blot analysis revealed that the expression of the PTEN protein in cardiac myocytes decreased after H2O2 induction. Anti-miR-214 increased PTEN protein expression level, in contrast, pre-miR-214 decreased the PTEN protein expression level in cultured cardiac myocytes. These results indicate that PTEN is regulated by miR-214 and serves as an important target of miR-214 in cardiac myocytes. In conclusion, miR-214 is sensitive to H2O2 stimulation, and miR-214 protects cardiac myocytes against H2O2-induced injury via one of its targets, PTEN.

摘要

活性氧(ROS)诱导的心肌细胞损伤是由多个基因表达水平的变化引起的,在多种心脏病的发病机制中起关键作用。本研究的目的是确定微小RNA-214(miR-214)在过氧化氢(H2O2)介导的心肌细胞基因调控中的潜在作用。在本研究中,我们使用定量实时RT-PCR(qRT-PCR)证明,用H2O2处理后心肌细胞中miR-214上调。我们用pre-miR-214转染细胞以上调miR-214表达,并用miR-214抑制剂(抗miR-214)转染细胞以下调miR-214表达。通过流式细胞术检测H2O2诱导的心肌细胞凋亡。miR-214抑制剂增加了凋亡水平,而pre-miR-214降低了凋亡水平。因此,我们认为miR-214在H2O2诱导的心肌细胞凋亡中起积极作用。10号染色体上缺失的磷酸酶和张力蛋白同源物(PTEN)具有组成性活性,被认为是促癌PI3K/Akt途径的主要下调因子。蛋白质印迹分析显示,H2O2诱导后心肌细胞中PTEN蛋白的表达降低。相反,抗miR-214增加了PTEN蛋白表达水平,而pre-miR-214降低了培养心肌细胞中PTEN蛋白的表达水平。这些结果表明,PTEN受miR-214调控,并作为miR-214在心肌细胞中的重要靶点。总之,miR-214对H2O2刺激敏感,并且miR-214通过其靶点之一PTEN保护心肌细胞免受H2O2诱导的损伤。

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