Based On Hydroxymethylation Modification To Explore The Relationship Between Early-onset Coronary Heart Disease Blood Stasis Syndrome And ARF6, PSD2 Susceptibility Genes

Objective: Through distribution of 5-hydroxymethylcytosine(5hmC)in the global genome,to investigate the effect of DNA hydroxymethylation on susceptibility genes related to blood stasis syndrome(BSS)in premature coronary heart disease(PCHD),in order to provide a basis for epigenetic research on the pathogenesis of BSS in PCHD.Methods:1.Four target groups including PCHD with BSS group,PCHD with non-BSS group,the non-PCHD with BSS group,controled group.Using Arraystar Human 4x180 K Refseq Promoter Microarray--DNA hydroxymethylated immunoprecipitation chip(hMeDIP-Chip)to detect 5hmC distribution in CpG islands in the promoter region of global genome.Concluded and analized the gene express spectrums of differential hydroxymethylation of BSS in PCHD through comparison between 4 groups.2.To verified the differential hydroxymethylation gene of ADP-ribosylation factor 6(ARF6),pleckstrin and Sec7 domain containing 2(PSD2)by use hMeDIP-qPCR.The possible physiological and pathological relationships were also analyzed that the degree of hydroxymethylation with ARF6 and PSD2 genes of BSS in PCHD.Results: There were 128 hydroxymethylation genes and 30 pathways associated with BSS in PCHD.All of genes and pathways is mainly related to energy metabolism of myocardial cells,inflammation,plaque formation,endothelial injury,immune response and lipid metabolism.When compared controled group with BSS in PCHD group,there were significant differences in pathways related with differentially hydroxymethylated genes that included AMPK signaling pathway(P=0.031),Endocytosis(P=0.011),Nucleotide excision repair(P=0.016),Tight junction(P=0.045),Longevity regulating pathway(P=0.047).According to PeakScore,PeakDMvalue,gene function,pathway relationship,physiological and pathological characteristics of PCHD,we found that the P value of pathway related of ARF6(PeakScore=2.79,PeakDMvalue=0.364),PSD2(PeakScore=2.01,PeakDMvalue=0.132)was significant and the hydroxymethylation expression of ARF6,PSD2 genes was low.Compared with normal people,the DNA hydroxymethylation degree of ARF6 and PSD2 genes of BSS in PCHD showed degree decreased(P < 0.01 or P < 0.05).This is correspond to the results of hMeDIP-Chip,which shows that the results of the chip have high reliability.Conclusions: 1.There were 128 differential hydroxymethylation genes and 30 pathways related with BSS in PCHD.All of genes and pathways is mainly related to energy metabolism of myocardial cells,inflammation,plaque formation,endothelial injury,immune response and lipid metabolism.2.There were significant differences in the pathways associated with BSS in PCHD that included AMPK signaling pathway,Endocytosis,Nucleotide excision repair,Tight junction,Longevity regulating pathway.3.The degree of hydroxymethylation of ARF6 and PSD2 genes was lower in PCHD with BSS,which may be the lower hydroxymethylation level of PSD2 gene through PSD2-GEFs-ARF6-small GTPase ARF6 signal transduction pathway regulatesthe expression of low hydroxymethylation of ARF6 gene.Therefore,ARF6 and PSD2 genes participate in the regulation of risk factors such as lipid accumulation,vascular endothelial injury with BSS in PCHD.