Transcriptome-wide identification of altered RNA m6A profiles in cardiac tissue of rats with LPS-induced myocardial injury

Abstract

Purpose: Myocardial injury is a common complication in patients with endotoxaemia/sepsis, especially in children. Moreover, it develops through an unclear pathophysiological mechanism, and effective therapies are lacking. Recently, RNA modification, particularly N ⁶-methyladenosine (m⁶A) modification, has been found to be involved in various physiological processes and to play important roles in many diseases. However, the role of m⁶A modification in endotoxaemia/sepsis-induced myocardial injury is still in its infancy. Therefore, we attempted to construct the m⁶A modification map of myocardial injury in a rat model treated by lipopolysaccharide (LPS) and explore the role of m⁶A modification in LPS-induced myocardial injury.

Method: Myocardial injury adolescent rat model was constructed by intraperitoneal injection of LPS. m⁶A RNA Methylation Quantification Kit was used to detect overall level of m⁶A modification in rat cardiac tissue. m⁶A-specific methylated RNA immunoprecipitation followed by high-throughput sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) were conducted to identify the altered m⁶A-modified genes and differentially expressed genes in cardiac tissue of rats treated by LPS and control rats (6 versus. 6). Bioinformatics was used to analyze the functions of differentially m⁶A modified genes, differentially expressed genes, and genes with both differential m⁶A modification and differential expression. qPCR was used to detect expression of m⁶A modification related enzymes.

Result: We found that the overall level of m⁶A modification in cardiac tissue of the LPS group was up-regulated compared with that of the control group. MeRIP-seq and RNA-seq results showed that genes with differential m⁶A modification, genes with differential expression and genes with both differential m⁶A modification and differential expression were closely associated with inflammatory responses and apoptosis. In addition, we found that m⁶A-related enzymes (Mettl16, Rbm15, Fto, Ythdc2 and Hnrnpg) were differentially expressed in the LPS group versus. the control group.

Conclusion: m⁶A modification is involved in the pathogenesis process of LPS-induced myocardial injury, possibly through the regulation of inflammatory response and apoptosis-related pathways. These results provide valuable information regarding the potential pathogenic mechanisms underlying LPS-induced myocardial injury.

Keywords: apoptosis; cardiac; endotoxaemia; inflammation; m6A.