Qishen Yiqi dropping pills improve cardiomyocyte hypertrophy via the lncRNA TINCR/miR-193b-3p/RORA axis

Lei Chen; Lihua Pan; Yuansheng Zhang; Yuewu Chen; Yangshen Su; Ying Luo; Zengfan Wu; Wanling Zheng; Shikang Cai; Xianxia Liu; Xiaoyan Wu

doi:10.21037/jtd-22-1322

Qishen Yiqi dropping pills improve cardiomyocyte hypertrophy via the lncRNA TINCR/miR-193b-3p/RORA axis

J Thorac Dis. 2022 Nov;14(11):4372-4383. doi: 10.21037/jtd-22-1322.

Authors

Lei Chen^#¹, Lihua Pan^#¹, Yuansheng Zhang¹, Yuewu Chen¹, Yangshen Su¹, Ying Luo¹, Zengfan Wu¹, Wanling Zheng¹, Shikang Cai¹, Xianxia Liu¹, Xiaoyan Wu¹

Affiliation

¹ Department of Cardiology, the Second Affiliated Hospital of Hainan Medical College, Haikou, China.

^# Contributed equally.

Abstract

Background: This study was designed to explore the therapeutic effect and mechanism of action of Qishen Yiqi dropping pills (QYDP) in chronic heart failure (CHF) via a long non-coding RNA (lncRNA)-microRNA (miRNA)-messenger RNA (mRNA) axis. Here, the mechanism of action of the lncRNA terminal differentiation-induced non-coding RNA (TINCR), miR-193b-3p, and RAR-related orphan receptor A (RORA) mRNA was analyzed in an angiotensin (Ang) II-induced H9C2 cardiomyocyte hypertrophy model treated with QYDP.

Methods: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to analyze the gene expression changes of lncRNA, miRNA, and mRNA in H9C2 induced by QYDP on Ang II. The Gene Expression Omnibus (GEO) was used to analyze differentially expressed genes (DEGs) potentially affecting CHF progression. Cell Counting Kit-8 (CCK-8) was used to analyze the effect of QYDP on the proliferation of H9C2, RNA pull-down was used to analyze the binding of lncRNA and miRNA, and dual luciferase was used to analyze the targeting of miRNA and lncRNA or mRNA.

Results: Ang II induced TINCR and RORA downregulation, miR-193b-3p upregulation, and hypertrophy in the H9C2 cardiomyocytes, which were alleviated by QYDP. In contrast, TINCR inhibition reversed the effects of QYDP by increasing miR-193b-3p expression and downregulating RORA expression. According to subsequent double luciferase and RNA pull-down experiments, TINCR adsorbed miR-193b-3p by acting as a competitive endogenous RNA sponge and miR-193b-3p directly targeted RORA. Lastly, we showed that the Ang-II-induced inhibition of TINCR and RORA expression and promotion of cardiac hypertrophy were both reversed by a TINCR overexpression plasmid (ov-TINCR), whereas the effects of ov-TINCR were suppressed by a miR-193b-3p mimic.

Conclusions: Administration of QYDP improves Ang II-induced H9C2 cardiomyocyte hypertrophy and increase cell proliferation rate through the TINCR/miR-193b-3p/RORA axis.

Keywords: Qishen Yiqi dropping pills; RAR-related orphan receptor A (RORA); chronic heart failure (CHF); long non-coding RNA terminal differentiation-induced non-coding RNA (lncRNA TINCR); miR-193b-3p.