Circular RNA circ_0036176 Inhibiting Myocardial Fibrosis Via Sponging miRNA-218-5p
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2022.0108
- VernacularTitle:环状RNA circ_0036176结合miR-218-5p发挥抑制心肌纤维化的作用
- Author:
Zhi-qi HUANG
1
;
Yu-min YAN
2
;
Jing GUO
3
;
Ji-shen GUO
4
;
Jie-ning ZHU
5
;
Xian-hong FANG
5
;
Jin-dong XU
5
;
Zhi-xin SHAN
1
Author Information
1. School of Biology and Biological Engineering， South China University of Technology， Guangzhou 510006， China
2. School of Pharmacy， Southern Medical University， Guangzhou 510515， China
3. School of Medicine， South China University of Technology， Guangzhou 510006， China
4. The Second School of Clinical Medicine， Southern Medical University， Guangzhou 510280， China
5. Guangdong Cardiovascular Institute， Guangdong Provincial People’s Hospital， Guangdong Academy of Medical Sciences， Guangzhou 510080， China.
- Publication Type:Journal Article
- Keywords:
circRNA;
cardiac fibrosis;
cardiac fibroblasts;
miRNA
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2022;43(1):61-69
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the biological effect of circ_0036176 on myocardial fibrosis. MethodsLevels of circ_0036176 and its host gene Myo9a were determined by real-time quantitative polymerase chain reaction （RT-qPCR） assay in human myocardial tissue， including 22 healthy organ donors and 26 patients with heart failure （HF）. A cell model of angiotensin Ⅱ （Ang-Ⅱ）-induced fibrosis in human atrial fibroblasts （HAFs） was achieved. To test the typical ring structure of circ_0036176， actinomycin D treatment and RNase R exonuclease digestion were performed. The expression of fibrosis-related gene in HAFs with overexpression of circ_0036176 was detected at mRNA and protein level. To select miRNAs that can effectively bind to circ_0036176， dual luciferase reporter gene assay and RNA antisense purification assay （RAP） were conducted， respectively. The neonatal mouse cardiac fibroblasts （mCFs） were used to study whether mir-218-5p mediates the effect of circ_0036176 on myocardial fibrosis phenotype. ResultsThe expression of circ_0036176 was up-regulated in the myocardium of HF patients （P＜0.001）， and the expression of circ_0036176 and the host gene Myo9a was down-regulated in Ang-Ⅱ-induced HAFs （P＜0.01）. In response to actinomycin D treatment and RNase R exonuclease digestion， circ_0036176 was more stable than Myo9a mRNA. The expression of COL1A1， COL3A1， TGF-β1 and ACTA2 was down-regulated in HAFs with overexpression of circ_0036176 （P＜0.05）. Results of dual luciferase reporter gene assay and RAP assay confirmed the interaction between miR-218-5p and circ_0036176. Overexpression of miR-218-5p could promote the expression of fibrosis-related genes， and attenuate the inhibitory effect of circ_0036176 on cardiac fibrosis. ConclusionsCirc_0036176 is up-regulated in the myocardium of HF patients， and circ_0036176 inhibits the expression of fibrosis-related gene through sponging miR-218-5p in CFs.
