Investigation Into DNA Methylation In Primary Gout

Purpose:To investigate the correlation between DNA(ABCG2,SLC2A9 and P2RX7)CpG island methylation and primary gout,and using the iTRAQ labeling technique combined with liquid chromatography-tandem mass spectrometry to screen out the differential proteins of primary gout,and further to analyze the methylation level of gene(TUBB4A)which encoded the differential protein(TBB4A)in primary gout.Then to provide a theoretical basis for the early diagnosis and early treatment of primary gout.methods:1.DNA methylation was detected by pyrosequencing method in primary gout group and healthy control group.Analysis of methylation levels were performed using t-test or nonparametric test.2.The differential proteins were screened out by using iTRAQ labeling technique combined with liquid chromatography tandem mass spectrometry,and meaningful differential proteins were found through biological processes,cell localization,molecular function,KEGG pathway and protein interaction analysis of bioinformatics analysis.3.Proteins were detected with Elisa technology,and total protein was detected with BCA method.Analysis of protein levels were performed using ttest or nonparametric test.Results:1.The methylation levels of ABCG2 CpG(pos1,2,4,5,6),SLC2A9 CpG(pos1,3,4,5,7)and P2RX7 CpG pos2 were statistically significant between primary gout group and control group(all P < 0.05).2.95 differentially expressed proteins(50 up-regulated and 45 downregulated)and 20 significant KEGG pathways were identified.Among them,glyceraldehyde-3-phosphate dehydrogenase,?-enolase,phosphoglycerate kinase,glucose-6-phosphate isomerase,and moesin are likely to play a key role in the pathogenesis of primary gout.3.The level of TBB4 A in the primary gout was significantly higher than the healthy control group(P = 0.023).4.The methylation levels of TUBB4 A CpG(pos4)was statistically significant between primary gout group and control group(P = 0.036).Conclusion:1.The methylation levels of ABCG2,SLC2A9 and P2RX7 were associated with primary gout,but the mechanism of ABCG2 gene is still unknown.2.The expression of TBB4 A was related to primary gout.3.TUBB4 A gene methylation was associated with primary gout,but the mechanism is still unknown.