MiR-144 affects proliferation and apoptosis of high glucose-induced AC16 cardiomyocytes by regulating CTRP3/JNK signaling

Yanling Song; Huade Mai; Yunyun Lin; Yachun Wang; Xiaoxi Wang; Shenhong Gu

MiR-144 affects proliferation and apoptosis of high glucose-induced AC16 cardiomyocytes by regulating CTRP3/JNK signaling

Int J Clin Exp Pathol. 2020 Feb 1;13(2):142-152. eCollection 2020.

Authors

Yanling Song¹, Huade Mai¹, Yunyun Lin¹, Yachun Wang¹, Xiaoxi Wang¹, Shenhong Gu¹

Affiliation

¹ Department of General Practice, The First Affiliated Hospital of Hainan Medical University Haikou, Hainan Province, China.

PMID: 32211094
PMCID: PMC7061800

Abstract

Background: Diabetic cardiomyopathy (DCM) is a common complication of diabetes and can lead to heart failure, arrhythmia, and sudden death. microRNAs (miRNAs) are reportedly involved in many human disease, including DCM. However, little is known about the biologic functions of miR-144 in DCM progression.

Methods: The expression levels of miR-144 and C1q/TNF-related protein-3 (CTRP3) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot was used to determine the protein levels of CTRP3, phosphorylated c-Jun amino-terminal kinase (p-JNK), JNK, Bax, Bcl-2, and cleaved-caspase-3. Cell proliferation and apoptosis were detected by Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. The potential binding sites between miR-144 and CTRP3 were predicted by microRNA.org databases and further determined using a dual-luciferase assay. AC16 cardiomyocytes were cultured in high glucose (HG, 30 mmol/L) to mimic hyperglycemia.

Results: MiR-144 expression level was enhanced, while CTRP3 expression was reduced in HG-induced AC16 cardiomyocytes. Knockdown of miR-144 or overexpression of CTRP3 dramatically promoted cell proliferation and reduced apoptosis of AC16 cardiomyocytes treated with HG. Inhibition of miR-144 evidently decreased the protein levels of Bax and p-JNK, but elevated Bcl-2 expression in HG-induced AC16 cardiomyocytes. Moreover, CTRP3 was a direct target of miR-144, and its abrogation reversed the effects of miR-144 knockdown on proliferation and apoptosis in HG-induced AC16 cardiomyocytes. SP600125 (a JNK inhibitor, 10 μmol/L) attenuated the si-CTRP3-mediated inhibition of proliferation and promotion of apoptosis in AC16 cardiomyocytes transfected with anti-miR-144 and stimulated with HG.

Conclusion: MiR-144 regulates proliferation and apoptosis of HG-induced AC16 cardiomyocytes through targeting the CTRP3/JNK signaling pathway, providing a novel avenue for treatment of DCM.

Keywords: CTRP3; Diabetic cardiomyopathy; JNK signaling pathway; miR-144.