Comprehensive analysis of m6A methylation and demethylation in endothelial cell dysfunction by MeRIP sequencing
Vascular endothelial dysfunction is one of the main causes of vasculopathy development in patients with type 2 diabetes mellitus (T2DM). During the development of vascular lesions, endothelial cell dysfunction occurs when endothelial cells are stimulated by external conditions such as hyperglycaemia and inflammation, leading to the loss of the typical phenotype of endothelial cells, the acquisition of mesenchymal features, and the occurrence of endothelial-to-mesenchymal transition (EndMT). It also promotes endothelial cell proliferation and migration and induces vascular injury. n6-methyladenosine (m6A) is the most prevalent post-transcriptional modification in eukaryotic mrna and long-chain non-coding rna. In this process, the m6A methylation modifying enzyme METTL3 with with the m6A demethylase FTO is involved in the development of T2DM vascular lesions. However, the mechanism by which METTL3 with FTO is involved in T2DM vasculopathy is unclear. In this study, we used methylated RNA immunoprecipitation sequencing (MeRIP-seq) to elucidate the role of aberrant m6A modification and mRNA expression in endothelial dysfunction, and MeRIP-qPCR combined with m6A site prediction to validate the methylation levels of downstream targets.