m6A methyltransferase METTL3 participated in sympathetic neural remodeling post-MI via the TRAF6/NF-κB pathway and ROS production

作者：Lei Qi, Ye Wang, Hui Hu, Pingjiang Li, Hesheng Hu, Yan Li, Kang Wang, Yuepeng Zhao, Meng Feng, Hangji Lyu, Jie Yin, Yugen Shi, Yu Wang, Xiaolu Li, Suhua Yan · 发表于：Journal of Molecular and Cellular Cardiology · 年份：2022 · DOI：10.1016/j.yjmcc.2022.06.004 · 被引用次数：34 · 研究领域：RNA modifications and cancer、Cancer-related gene regulation、Ubiquitin and proteasome pathways

Objective Sudden cardiac death caused by ventricular arrhythmias (VAs) is the main cause of high mortality in patients with myocardial infarction (MI). Sympathetic neural remodeling caused by inflammation after MI is closely associated with the occurrence of VAs. METTL3, the earliest identified m 6 A methyltransferase, is critical in mediating inflammatory responses. Our aim was to investigate whether the m 6 A methyltransferase METTL3 was involved in sympathetic remodeling post-MI and its specific mechanism. Methods and results A rat MI model was established via left coronary artery ligation. The expression of METTL3, TRAF6, NOX2, and NF-κB increased at 3 days and remained elevated at 7 days after MI, as determined via Western blotting. METTL3 was primarily present in macrophages, as determined via immunofluorescence. Intramyocardial injection of lentivirus carrying METTL3-shRNA inhibited METTL3 expression in vivo. Methylated immunoprecipitation-qPCR determined the METTL3 knockdown inhibited the m 6 A level of TRAF6 mRNA 3′-UTR. The co-immunoprecipitation experiment proved that METTL3 combines with TRAF6. Western blotting showed that silencing METTL3 inhibited TRAF6 level, NF-κB activation, and ROS production; decreased cytokine release (TNF-α and IL-1β); and downregulated nerve growth factor expression. Finally, METTL3 knockdown reduced sympathetic remodeling after MI, as determined via immunofluorescence assays of tyrosine hydroxylase and growth-associated protein 43. Prog...