微RNA-372-3p负调控FBXO11参与心肌成纤维细胞活化的机制
miR-372-3p Anti-fibroblast Activation by Targeting FBXO11 in Transforming Growth Factor-β1-Induced Human Cardiac Fibroblasts
柯晓 1易明 2陈俊羽 1郑欣馨 3徐延路3
作者信息
- 1. 中国医学科学院阜外医院深圳医院心血管内科,广东深圳 518057
- 2. 湖南中医药大学附属第二中西医结合医院心血管内科,长沙 410300
- 3. 中国医学科学院 北京协和医学院 国家心血管病中心 阜外医院心血管内科,北京 100037
- 折叠
摘要
目的 探讨在心肌成纤维细胞(CFs)活化过程中,微RNA(miRNA)对FBXO11表达的调控及其分子机制.方法 正常心肌成纤维细胞(hCFs)经miR-372-3p mimics转染和转化生长因子β1(TGF-β1)诱导后,采用qRT-PCR检测FBXO11及hCFs活化标志物的表达;通过双荧光素酶报告基因实验检测FBXO11与miR-372-3p的结合情况.结果 hCFs活化后,FBXO11表达呈剂量依赖性上调,且与hCFs活化标志物表达水平呈正相关.敲除FBXO11后,hCFs活化标志物表达下调.在hCFs体外纤维化诱导模型中,miR-372-3p表达下调,且与FBXO11呈显著负相关.miR-372-3p通过与FBXO11 3'非翻译区(3'UTR)的特定位点结合,对FBXO11进行转录后调控.过表达miR-372-3p可以抑制TGF-β1诱导的FBXO11高表达和hCFs活化标志物表达水平升高.结论 miR-372-3p通过负调控FBXO11表达参与hCFs活化.
Abstract
Objective To investigate the mechanism of FBXO11 expression regulated by microRNA(miRNA)and its molecular mechanism in the activating process of cardiac fibroblasts.Methods Human normal cardiac fibroblasts(hCFs)were transfected with miR-372-3p mimics and induced by transforming growth factor-β1(TGF-β1),and qRT-PCR was used to detect FBXO11 as well as the expression of hCFs activation markers;the binding of FBXO11 to MiR-372-3p was detected by dual luciferase reporter gene assay.Results Upon hCFs activation,the expression of FBXO11 was upregulated in dose-dependent manner and positively correlated with the activation markers of hCFs.After the knockdown of FBXO11,the expression of activation markers of hCFs was downregulated.In vitro,in a TGF-β1-induced fibrosis model of hCFs,MiR-372-3p expression was down-regulated and significantly negatively correlated with FBXO11.MiR-372-3p was up-regulated in an in TGF-β1-induced fibrosis model of hCFs by binding to the FBXO11 3'untranslated region(3'UTR).Overexpression of miR-372-3p inhibited TGF-β1-induced high FBXO11 expression and elevated levels of hCFs activation markers.Conclusion miR-372-3p is involved in the anti-hCFs activation by negatively regulating FBXO11 expression.
关键词
FBXO11/微RNA-372-3p/心肌成纤维细胞/转化生长因子β1Key words
FBXO11/miR-372-3p/Cardiac fibroblasts/Transforming growth factor-β1引用本文复制引用
基金项目
广东省基础与应用基础研究基金面上项目(2021A1515010178)
深圳市科技计划项目(JCYJ20210324124605015)
湖南省自然科学基金(2024JJ9572)
出版年
2024
NETL
NSTL
万方数据