Next Generation Sequencing Combined Analysis of DNA Methylome and Transcriptome Reveal Novel Candidate Genes of Neuroprotective with Response to Ischemic Preconditioning

Ischemic preconditioning (IPC) is a phenomenon in which a short-term sublethal ischemic exposure induces tolerance to a subsequent lethal ischemic insult; however, the detailed mechanism underlying IPC-induced neuroprotection remains unclear. Here, we applied middle cerebral artery occlusion (MCAO) as a preconditioning ischemic insult mouse model to investigate the molecular mechanism underlying cerebral IPC. RNA-seq and whole-genome bisulfite sequencing (WGB-Seq) were performed to explore the gene expression profile and DNA methylation changes after cerebral IPC treatment. In this study, we identified 636 differentially expressed genes enriched for several pathways that were partially overlapping or interconnected. The involvement of several genes involved in IPC induced neuroprotection were first reported. Genes induced by IPC including Arid5a, Nptx2 and Stc2 were validated with a neuroprotective effect against oxygen-glucose deprivation (OGD)-induced neurotoxicity in vitro. Moreover, Arid5a and Nptx2 were two DMR related genes and showed hypo-methylation in their regulative regions. Thus, our findings provide new insights into IPC signaling pathways and offer a new therapeutic strategy for the treatment of stroke. RNA-seq and whole-genome bisulfite sequencing (WGBS) were performed to explore the gene expression profile and DNA methylation changes after cerebral IPC treatment, in triplicate, using Illumina HiSeq 2000 platform.