单细胞测序揭示 LRRc17 介导的心脏成纤维细胞铁死亡调控在心肌梗死后心肌纤维化中的作用。

Single-cell sequencing reveals LRRc17-mediated modulation of cardiac fibroblast ferroptosis in regulating myocardial fibrosis after myocardial infarction.

机构信息

Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi Province, China; Department of Cardiology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, China; Department of Cardiovascular Medicine, Jiangxi Provincial People's Hospital, The First Affiliated Hospital of Nanchang Medical College, Nanchang, Jiangxi Province, China.

Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi Province, China; Department of Cardiology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, China.

出版信息

Cell Signal. 2024 Sep;121:111293. doi: 10.1016/j.cellsig.2024.111293. Epub 2024 Jul 10.

DOI:10.1016/j.cellsig.2024.111293

PMID:38996956

Abstract

PURPOSE

Myocardial fibrosis after myocardial infarction (MI) is one of the main causes of death in patients, but there is no effective treatment for myocardial fibrosis at present. Hence, it is important to elucidate the pathogenesis of fibrosis after MI and find therapeutic targets for regulating ventricular remodeling after MI.

METHODS

Differentially expressed gene analysis, weighted Gene co-expression network analysis (WGCNA) and differential gene analysis in cardiac fibroblasts (CFs) were performed on the MI-related data (GSE153485 and GSE210159) from the GEO database to screen the hub genes related to ferroptosis in MI. After the establishment of MI model in vivo and in vitro, the myocardial CFs were observed by Masson staining, hematoxylin-eosin staining, CCK-8, and Transwell, and the changes of LRRc17 and ferroptosis-related proteins were detected by qRT-PCR and Western blotting. The expression of ROS was detected by fluorescence dye method.

RESULTS

Three DEGs were identified in MI related to ferroptosis, among which LRRc17 was selected for subsequent study. In both in vitro and in vivo models of MI, we found a sustained downregulation of LRRc17 expression and the expression of ferroptosis-related proteins GPX-4 and xCT, but increased ROS expression and enhanced migration and viability of CFs. After oe-LRRc17 treatment, the expression levels of GPX-4 and xCT were restored, while ROS levels were inhibited, and the migration and viability of CFs were inhibited. After treatment with ferroptosis inducer Erastin, there were down-regulated expressions of GPX-4 and xCT, increased expression of ROS, and enhanced migration and viability of CFs.

CONCLUSION

LRRc17 affects ventricular remodeling by mediating ferroptosis in CFs to regulate the degree of fibrosis after MI.

摘要

目的

心肌梗死后心肌纤维化是患者死亡的主要原因之一，但目前尚无有效的心肌纤维化治疗方法。因此，阐明心肌梗死后纤维化的发病机制，寻找调节心肌梗死后心室重构的治疗靶点非常重要。

方法

从 GEO 数据库中 MI 相关数据集（GSE153485 和 GSE210159）中进行差异表达基因分析、加权基因共表达网络分析（WGCNA）和心脏成纤维细胞（CFs）差异基因分析，筛选与 MI 相关的铁死亡相关的关键基因。在体内和体外建立 MI 模型后，通过 Masson 染色、苏木精-伊红染色、CCK-8 和 Transwell 观察心肌 CFs 的变化，通过 qRT-PCR 和 Western blot 检测 LRRc17 和铁死亡相关蛋白的变化，通过荧光染料法检测 ROS 的表达。

结果

在 MI 相关的与铁死亡相关的 3 个 DEGs 中，选择 LRRc17 进行后续研究。在 MI 的体外和体内模型中，我们发现 LRRc17 表达持续下调，铁死亡相关蛋白 GPX-4 和 xCT 的表达下调，但 ROS 表达增加，CFs 的迁移和活力增强。过表达 LRRc17 后，GPX-4 和 xCT 的表达水平恢复，ROS 水平受到抑制，CFs 的迁移和活力受到抑制。用铁死亡诱导剂 Erastin 处理后，GPX-4 和 xCT 的表达下调，ROS 表达增加，CFs 的迁移和活力增强。