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[CircRNA_005647 inhibits expressions of fibrosis-related genes in mouse cardiac fibroblasts via sponging miR-27b-3p].
- Source :
-
Nan fang yi ke da xue xue bao = Journal of Southern Medical University [Nan Fang Yi Ke Da Xue Xue Bao] 2019 Nov 30; Vol. 39 (11), pp. 1312-1319. - Publication Year :
- 2019
-
Abstract
- Objective: To investigate the effect of circRNA&#95;005647 on fibrotic phenotype of mouse cardiac fibroblasts (CFs) and explore its mechanism.<br />Methods: We used an angiotensin Ⅱ (Ang-Ⅱ) capsule pump (daily dose of 1.46 mg/kg for 2 weeks) to establish a mouse model of myocardial fibrosis in C57BL/6 mice. Masson staining was used to detect myocardial fibrosis in the myocardium. The expression of circRNA&#95;005647 in the myocardium of Ang-Ⅱ-infused mice and in Ang-Ⅱ-treated CFs were detected with real-time PCR. Actinomycin D and RNase R exonuclease digestion were used to test the stability of circRNA&#95;005647 in mouse CFs. Over- expression of circRNA&#95;005647 was achieved in the CFs by infecting the cells with a recombinant circRNA&#95;005647 adenovirus (rAd-circRNA&#95;005647), and the expressions of Col1a1, Col3a1 and Acta2 were detected in the cells with real-time PCR and Western blotting. Dual luciferase reporter assay, RNA antisense purification and RNA Pull down assay were performed to identify the interaction between circRNA&#95;005647 and miR-27b-3p.<br />Results: CircRNA&#95;005647 was up- regulated in the myocardium of Ang-Ⅱ- infused mice and in Ang-Ⅱ-treated mouse CFs ( P < 0.01). CircRNA&#95;005647 was more stable than its host gene Myosin IXA ( Myo9a ) in response to actinomycin D ( P < 0.01) and RNase R exonuclease treatment. The expressions of fibrosis-associated genes was down-regulated in the CFs over-expressing circRNA&#95; 005647 ( P < 0.05). Dual luciferase reporter assay, RNA antisense purification and RNA Pull down assay revealed the interaction between miR-27b-3p and circRNA&#95;005647. MiR-27b-3p obviously enhanced the fibrotic phenotype but reversed the inhibitory effect of circRNA&#95;005647 on the expression of fibrosis associated genes in the CFs ( P < 0.05).<br />Conclusions: CircRNA&#95;005647 is upregulated in cardiac fibrosis and inhibits the expression of fibrosis-related genes through sponging miR-27b-3p in mouse CFs.
- Subjects :
- Animals
Fibrosis
Mice
Mice, Inbred C57BL
MicroRNAs
RNA, Circular
Fibroblasts
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1673-4254
- Volume :
- 39
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Nan fang yi ke da xue xue bao = Journal of Southern Medical University
- Publication Type :
- Academic Journal
- Accession number :
- 31852652
- Full Text :
- https://doi.org/10.12122/j.issn.1673-4254.2019.11.08