Based On DNA Hydroxymethylation Modification To Regulate MRNA Expression, Explore The Susceptibility Genes Related To Blood Stasis Syndrome Of Early-onset Coronary Heart Disease

Objective: According to the state of DNA hydroxymethylation and the m RNA expression of whole genome chip in peripheral blood of patients of premature coronary heart disease(PCHD)with blood stasis syndrome(BSS),the differential genes of the two were screened and the intersection was taken to explore the susceptible factors of PCHD with BSS and the mechanism of disease occurrence and development.Methods:(1)Firstly,accorded to the diagnostic criteria for PCHD and TCM syndrome,422 samples were selected,which include 20 patients of PCHD with BSS,20 patients of PCHD with non-BSS,20 patients of PCHD with non-BSS and 20 healthy people;it was divided into 4target groups,6 patients in each group were selected,and 3 m L peripheral blood was taken from each case.Base on DNA hydroxymethylation,the different genes regulating blood stasis syndrome of PCHD were screened.(2)Secondly,it was divided into 2 target groups including PCHD with BSS group and control group.6 patients in each group were selected,and 2 m L peripheral blood was taken from each case.According to whole-genome chip m RNA expression,the different genes regulating blood stasis syndrome of PCHD were screened.Finally,combined DNA hydroxymethylation and m RNA expression,the different genes regulating blood stasis syndrome of PCHD were screened.Results:(1)The results of h Me DIP-chip showed that there were 4980 differential hydroxymethylation genes,which were mainly distributed in the promoter region of high Cp G density.Compared the BSS in PCHD group with the controled group,the results showed that DNA hydroxymethylation genes expression was up-regulated by 224 and down-regulated by 649.The whole-genome chip m RNA expression was up-regulated by 112 and down-regulated by293.(2)Based on DNA hydroxymethylation and m RNA expression levels,there were 11 genes with common differences,4 were up-regulated and 7 were down-regulated.(3)Through GO and KEGG,more than 20 pathways was mainly related to Nucleotide excision repair,Endocytosis and so on.(4)The levels of DNA hydroxymethylation and m RNA expression of LRIG1 and TUBB2 A genes was lower in PCHD with BSS groups(P <0.05 or P <0.01).Conclusions:(1)Compared the BSS in PCHD group with the controled group,the h Me DIP-chip showed down-regulated of 694 genes,and whole-genome chip m RNA showed down-regulated of 293 genes.There were 11 commom differential genes in the two tests,included 7 down-regulated.So the results suggested that PCHD with BSS was related to low gene expression,and down-regulation of differential gene expression may be one of the mechanisms that leaded to the occurrence and development of PCHD with BSS.(2)Through GO and KEGG,all of genes and 20 pathways were mainly related to vascular endothelial damage,inflammation,plaque and lipid metabolism.It is suggested that the above pathway was related to the mechanism of PCHD with BSS.(3)The levels of DNA hydroxymethylation and m RNA expression of LRIG1 and TUBB2 A genes was lower in PCHD with BSS(P <0.05 or P <0.01),and the interaction with EGFR gene may be due to the inhibition of TUBB2 A and LRIG1 mutation results in low expression,which weakened the negative regulation of EGFR and caused high expression,which interfered with the normal transmission of epidermal growth factor signals,affected the signal transduction of myocardial and vascular smooth muscle cell reconstruction,and leaded to PCHD with BSS.Therefore,it may be one of the risk factors of the occurrence and development in PCHD with blood stasis syndrome that the DNA hydroxymethylation of LRIG1 and TUBB2 A gene regulated the low expression of the mRNA.