miR-92b-3p 通过靶向 HAND2 抑制血管紧张素 II 诱导的心肌细胞肥大。

MicroRNA-92b-3p suppresses angiotensin II-induced cardiomyocyte hypertrophy via targeting HAND2.

机构信息

School of Medicine, South China University of Technology, Guangzhou 510632, China; Guangdong Provincial Key Laboratory of Clinical Pharmacology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China.

Guangdong Provincial Key Laboratory of Clinical Pharmacology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China.

出版信息

Life Sci. 2019 Sep 1;232:116635. doi: 10.1016/j.lfs.2019.116635. Epub 2019 Jul 5.

DOI:10.1016/j.lfs.2019.116635

PMID:31283925

Abstract

AIMS

The pathological cardiac hypertrophy will develop into heart failure, which has no effective treatment currently. Previous studies have proved that microRNAs (miRNAs) participate in the development of cardiac hypertrophy and regulate the pathological progress. In this study, we want to investigate the role of microRNA-92b-3p (miR-92b-3p) in cardiomyocyte hypertrophy and the mechanisms involved.

MATERIALS AND METHODS

Neonatal mouse ventricular cells (NMVCs) were isolated from the hearts of 1-3-d-old newborn C57BL6 mice. The isolated NMVCs were induced hypertrophic phenotype by Angiotensin-II (Ang-II) and the cell size was examined by FITC-phalloidin staining assay. The expression of miR-92b-3p was determined by quantitative real-time PCR (qRT-qPCR). MRNA and protein level of β-MHC, ACTA1 and HAND2 in NMVCs transfected with miR-92b-3p mimic and inhibition were assessed by RT-qPCR assay and western blot assay, respectively. Dual luciferase assay was used to verify the interaction between miR-92b-3p and the 3'-untranslated region (UTR) of HAND2 gene.

KEY FINDINGS

MiR-92b-3p and HAND2 were significantly increased in Ang-II-induced NMVCs. Overexpression of miR-92b-3p can ameliorate Ang-II-induced cardiomyocyte hypertrophy. MiR-92b-3p negatively regulated HAND2 expression at the transcriptional level. Both miR-92b-3p mimic and HAND2 siRNA could efficiently inhibit Ang-II-induced hypertrophy in mouse cardiomyocytes.

SIGNIFICANCE

MiR-92b-3p inhibits Ang-II-induced cardiomyocyte hypertrophy via targeting HAND2.

摘要

目的

病理性心肌肥厚会发展为心力衰竭，目前尚无有效治疗方法。先前的研究已经证明 microRNAs（miRNAs）参与心肌肥厚的发生，并调节其病理进程。本研究旨在探讨 microRNA-92b-3p（miR-92b-3p）在心肌肥厚中的作用及其相关机制。

材料与方法

从 1-3 日龄新生 C57BL6 小鼠心脏中分离出新生鼠心室细胞（NMVCs）。用血管紧张素-II（Ang-II）诱导 NMVCs 产生肥大表型，并用 FITC-鬼笔环肽染色实验检测细胞大小。用实时定量 PCR（qRT-qPCR）检测 miR-92b-3p 的表达。用 miR-92b-3p 模拟物和抑制剂转染 NMVCs 后，通过 RT-qPCR 检测和 Western blot 检测 NMVCs 中β-MHC、ACTA1 和 HAND2 的 mRNA 和蛋白水平。用双荧光素酶报告基因实验验证 miR-92b-3p 与 HAND2 基因 3'-非翻译区（UTR）的相互作用。

主要发现

在 Ang-II 诱导的 NMVCs 中，miR-92b-3p 和 HAND2 的表达显著增加。过表达 miR-92b-3p 可改善 Ang-II 诱导的心肌细胞肥大。miR-92b-3p 可在转录水平负调控 HAND2 的表达。miR-92b-3p 模拟物和 HAND2 siRNA 均可有效抑制 Ang-II 诱导的小鼠心肌细胞肥大。