Department of Cardiovascular Medicine, Chengdu Fifth People's Hospital.
Department of Day Surgery Ward, Chengdu Fifth People's Hospital.
Int Heart J. 2021;62(4):891-899. doi: 10.1536/ihj.20-298.
Long-chain noncoding RNA (lncRNA) is a new class of molecular regulators in heart development and disease. However, the role of specific lncRNA in cardiac fibrosis remains to be fully explored. This study aimed to investigate the role and potential mechanism of lncRNA MHRT in myocardial fibrosis after myocardial infarction (MI).Cardiac fibroblasts (CFs) were isolated from a mouse model of MI. The expression levels of MHRT and miR-3185 in the hearts of MI and CFs mice treated with transforming growth factor beta 1 (TGF-β1) were analyzed by qRT-PCR. The collagen expression was assessed using qRT-PCR and Western blot. Cell proliferation was assessed by performing MTT and EdU assays. The direct interaction between lncRNA and miRNA was analyzed by luciferase assay, RNA-binding protein immunoprecipitation (RIP) assay, and RNA pull-down assay.The expression levels of MHRT were raised in MI and CFs mice treated with TGF-β1. Overexpression of MHRT promoted collagen production and CF proliferation, while silencing of MHRT showed the opposite effect. MiR-3185 was a target gene of MHRT. In addition, overexpression of MHRT reduced the expression levels of miR-3185, and siMHRT reversed the inhibitory effect of TGF-β1 on the expression of miR-3185. Overexpression of miR-3185 inhibited the upregulation of Col I and Col III induced by TGF-β1.MHRT promoted cardiac fibrosis after MI through miR-3185 and increased myocardial collagen deposition and promoted myocardial fibrosis.
长链非编码 RNA(lncRNA)是心脏发育和疾病中一种新的分子调控因子。然而,特定 lncRNA 在心肌纤维化中的作用仍有待充分探索。本研究旨在探讨 lncRNA MHRT 在心肌梗死后心肌纤维化中的作用及其潜在机制。
从心肌梗死后小鼠模型中分离心肌成纤维细胞(CFs)。通过 qRT-PCR 分析 MHRT 和 miR-3185 在 MI 小鼠和 TGF-β1 处理的 CFs 小鼠心脏中的表达水平。通过 qRT-PCR 和 Western blot 评估胶原表达。通过 MTT 和 EdU 测定评估细胞增殖。通过荧光素酶测定、RNA 结合蛋白免疫沉淀(RIP)测定和 RNA 下拉测定分析 lncRNA 和 miRNA 之间的直接相互作用。
MHRT 的表达水平在 MI 小鼠和 TGF-β1 处理的 CFs 小鼠中升高。MHRT 的过表达促进了胶原产生和 CF 增殖,而 MHRT 的沉默则表现出相反的效果。miR-3185 是 MHRT 的靶基因。此外,MHRT 的过表达降低了 miR-3185 的表达水平,而 siMHRT 逆转了 TGF-β1 对 miR-3185 表达的抑制作用。miR-3185 的过表达抑制了 TGF-β1 诱导的 Col I 和 Col III 的上调。
MHRT 通过 miR-3185 促进 MI 后心肌纤维化,增加心肌胶原沉积并促进心肌纤维化。