Overexpression of miR‐383‐3p protects cardiomyocytes against hypoxia/reoxygenation injury via regulating PTEN/PI3K/AKT signal pathway

• Published in: Journal of biochemical and molecular toxicology 2022-12, Vol.36 (12), p.e23205-n/a
• Main Authors: Zeng, Huan, Li, Ying, Liu, Xinzong, Li, Xinxin, Zhou, Tian, Cao, Shanshan, Wang, Mingjuan, Ju, Mingfei
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; 3' Untranslated regions; AKT protein; Apoptosis; Apoptosis - genetics; Binding sites; Biomarkers; Cardiomyocytes; Cardiovascular diseases; Caspase; Caspase 3 - metabolism; Cell viability; Chromosome 3; Enzyme-linked immunosorbent assay; Flow cytometry; H/R‐induced injury; H9c2; Humans; Hypoxia; Hypoxia - metabolism; Immunofluorescence; Injuries; L-Lactate dehydrogenase; Lactate dehydrogenase; Lactic acid; MicroRNAs - metabolism; miRNA; miR‐383‐3p; Myocytes, Cardiac - metabolism; Oxidative stress; Phosphatidylinositol 3-Kinases - metabolism; PI3K/AKT signal pathway; Proto-Oncogene Proteins c-akt - metabolism; PTEN; PTEN Phosphohydrolase - genetics; PTEN Phosphohydrolase - metabolism; PTEN protein; Reactive oxygen species; Reporter gene; Signal Transduction; Superoxide dismutase; Therapeutic targets
• ISSN: 1095-6670; 1099-0461
• DOI: 10.1002/jbt.23205

MicroRNAs are widely reported as biomarkers and therapeutic targets in cardiovascular diseases. This study is aimed to expound on the regulatory responsibility of miR‐383‐3p in H/R‐induced injury of H9c2 cells. In this study, H9c2 cells were administrated with H/R. MiR‐383‐3p expression was measured using qRT‐PCR. ELISA was used to determine lactate dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA) levels. Reactive oxygen species (ROS) were detected with 2,7‐Dichlorodihydrofluorescein diacetate probe. 3‐(4,5)‐dimethylthiahiazo (‐z‐y1)‐3,5‐di‐ phenytetrazoliumromide, flow cytometry, and TUNEL experiments were conducted to measure cell viability and apoptosis. Cleaved caspase‐3, caspase‐3, Bax, Bcl‐2, PTEN, PI3K, p‐PI3K, Akt, p‐AKT expression levels were examined by Western blot. Cleaved caspase‐3 expression was also measured by immunofluorescence staining. Dual‐luciferase reporter gene assay was applied to validate the binding sites in miR‐383‐3p and the 3′UTR of PTEN. We reported that, miR‐383‐3p expression in H9c2 cells treated with H/R was remarkably decreased. MiR‐383‐3p overexpression ameliorated oxidative stress and apoptosis and promoted cell viability in H9c2 cells treated with H/R, while miR‐383‐3p inhibitor showed the reverse effects. PTEN was identified as a target gene of miR‐383‐3p. Additionally, enhancement of PTEN expression abolished the influences of miR‐383‐3p on H9c2 cells. MiR‐383‐3p mimics could significantly decrease PTEN expression in H9c2 cells while increasing p‐PI3K expression and p‐AKT expression, while the miR‐383‐3p inhibitors showed the opposed effects. In conclusion, miR‐383‐3p protected H9c2 cells from H/R‐induced injury via regulating PTEN/PI3K/AKT signal pathway.