Genome-Wide Analysis of Electroacupuncture in Restoring Splicing Regulation after Myocardial Ischemia-reperfusion Injury

Background: Pre-mRNA alternative splicing (AS) is an important post-transcriptional regulation mechanism. The abnormal splicing of genes can lead to cardiovascular diseases. Acupuncture has been shown to alleviate myocardial ischemia-reperfusion injury (MIRI), but most studies have focused on the regulation of gene expression by acupuncture. Previously, we reported that electroacupuncture (EA) can relieve angina by regulating the AS of GABARG2; however, the genome-wide regulation of AS by EA remains unknown. Methods: We explored the protective effects of EA on MIRI. We then studied the AS profiles retrieved from the previously submitted to Gene Expression Omnibus (GEO) database and analyzed the data using the replicate multivariate analysis of transcript splicing (rMATS) tool. Subsequently, we conducted validation experiments on splicing regulatory factors and their target genes modulated by EA, as identified through bioinformatics analysis. Results: The results showed that EA at PC6 point could effectively alleviate MIRI. More than 200 differential alternative splicing events (ASEs) changed following MIRI. The differential ASEs underwent protein-protein interaction (PPI) network analysis, gene ontology (GO) enrichment, and pathway analysis, and were shown to be involved in distinct biological functions, especially in the maintenance of synaptic structure. Enrichment analysis also identified several pathways that are potentially associated with the progression of MIRI. Importantly, we identified critical ASEs and pathways that could be completely rescued by EA treatment. In the validation experiments, we found that AS of the key gene CAMK2G, associated with synaptic function, is regulated by the expression level of the MBNL1 protein modulated by EA. Conclusion: Our results indicate that EA is not only an effective procedure to protect against MIRI by rescuing gene expression but also rescues abnormal ASEs. This study is a major contribution to a genome-wide comprehensive analysis of the regulation of AS by EA in MIRI.