去甲基化酶FTO在缺氧复氧心肌细胞能量代谢紊乱中的保护机制

Abstract

New Findings What is the central question of this study? What is the effect of fat mass and obesity-associated protein (FTO) on energy metabolism in hypoxia–reoxygenation (H/R)-induced cardiomyocytes? What is the main finding and its importance? FTO modification of N 6 -methyladenosine (m 6 A) is associated with myocardial cell energy metabolism disorder. FTO reduced the m 6 A level of sarcoplasmic/endoplasmic reticulum calcium ATPase 2a (SERCA2a) mRNA through demethylation, thus promoting SERCA2a expression, maintaining calcium homeostasis, and improving energy metabolism of H/R cardiomyocytes. Energy metabolism disorder is the initial physiological link of myocardial ischaemia–reperfusion injury. Fat mass and obesity-associated protein (FTO) is an N 6 -methyladenosine (m 6 A) demethylase implicated in several cardiac defects. This study sought to investigate the effect of FTO on energy metabolism in hypoxia–reoxygenation (H/R)-induced cardiomyocytes. FTO and sarcoplasmic/endoplasmic reticulum calcium ATPase 2a (SERCA2a) expression in H/R-induced cardiomyocytes were determined. Cardiomyocyte viability, cytotoxicity and apoptosis were measured. The total RNA and polyA + RNA contents were isolated from cells. The m 6 A level of RNA and the enrichment of m 6 A of SERCA2a mRNA were calculated. Several indices such as the glycolytic potential, reactive oxygen species (ROS), mitochondrial activity and ATP content were evaluated. The concentration of calcium in cardiomyocytes was determined. FTO and SERCA2a were poorly expressed in H/R-induced cardiomyocytes. There was an elevated m 6 A level in total RNA and enrichment of m 6 A in SERCA2a mRNA. H/R treatment reduced the cell viability, mitochondrial membrane potential and ATP content in cardiomyocytes, but increased the cytotoxicity, apoptosis, ROS content and calcium concentration. Upregulation of FTO reversed the preceding findings with downregulation of the m 6 A level of SERCA2a mRNA. Downregulation of SERCA2a annul