The Relationship Between The Methylation Of Hcy Metabolism-related Genes And Ischemic Stroke

ObjectiveTo investigate the asscoiation of cerebral infarction(CI)with the methylation of Hcy metabolism-related genes in the promoter region such as S-adenosylhomocysteine hydrolase gene(AHCY),cystathionine ?-synthase gene(CBS),dihydrofolate reductase(DHFR),methylenetetrahydrofolate dehydrogenase gene 1(MTHFD1)and serine hydroxymethyltransferase gene 1(SHMT1).And evaluate that the diagnostic value of AHCY,CBS,DHFR,MTHFD1,SHMT1 methylation in cerebral infarction.MethodsWe recruit that 152 cerebral infarction patients and 152 gender and age matched healthy controls.Using quantitative methylation-specific polymerase chain reaction(qMSP)method to evaluate AHCY,CBS,DHFR,MTHFD1 and SHMT1 promoter methylation levels methylation and analyze its relationship with cerebral infarction.Results1.The DNA methylation level of AHCY promoter region was 0.16%(0.07%,0.32%)in the case group and 0.25%(0.02%,0.67%)in the control group,the difference was not statistically significant(P=0.115,Z=-1.577).2.The DNA methylation level of CBS promoter region was 70.20%(49.19%,94.87%)in the case group and 104.10%(74.65%,132.30%)in the control group,the difference was statistically significant(P=3.71E-10,Z=-6.266).According to the subgroup analysis by gender,the DNA methylation level of CBS promoter region in male was 64.33%(47.72%,85.66%)in the case group and 105.00%(73.23%,128.85%)in the control group,the difference was statistically significant(P=2.667E-08,Z=-5.562).And the DNA methylation level of CBS promoter region in female was 78.05%(51.81%,107.90%)in control group and 102.80%(86.11%,149.90%)in control group,the difference was statistically significant(P=0.003,Z=-2.990).In the subgroup analysis by age,the DNA methylation level of CBS promoter region in the youth group(?44 years old)was(69.97 ± 13.61)% in the case group and(114.71 ± 59.62)% in the control group,the difference was statistically significant(P=0.015,t=-2.636).The DNA methylation level of CBS promoter region in the middle-aged group(45-59 years)was 56.04%(48.07%,76.23%)in the case group and 91.71%(68.28%,118.58%)in the control group,the difference was statistically significant(P=6.744E-06,Z=-4.502).The DNA methylation level of CBS promoter region in the elderly group(?60 years old)was 81.60%(49.37%,127.45%)in the case group and 119.35%(87.71%,167.90%)in the control group,the difference was statistically significant(P=2.644E-04,Z=-3.648).Based on the DNA methylation levels of CBS promoter region in the case and control groups,we established the ROC curve.The the area under the ROC curve(AUC)of 0.713(95% CI: 0.652-0.773)and we obtained the optimal sensitivity of 67.4% and specificity of 74.0% with the cutoff of 87.1%.3.The DNA methylation level of DHFR promoter region was 28.37%(13.40%,50.82%)in the case group and 44.22%(19.55%,77.78%)in the control group,the difference was statistically significant(P=1.648E-04,Z=-3.768).According to the subgroup analysis by gender,the DNA methylation level of DHFR promoter region in male was 27.06%(9.55%,50.83%)in the case group and 44.81%(23.47%,77.74%)in the control group,the difference was statistically significant(P=9.455E-05,Z=-3.904).And the DNA methylation level of DHFR promoter region in female was 31.35%(17.64%,50.70%)in control group and 40.30%(8.98%,78.07%)in control group,the difference was not statistically significant(P=0.375,Z=-0.887).In the subgroup analysis by age,the DNA methylation level of DHFR promoter region in the youth group(?44 years old)was 14.36%(0.44%,25.11%)in the case group and 59.11%(42.47%,104.92%)in the control group,the difference was statistically significant(P=9.068E-04,Z=-3.318).The DNA methylation level of DHFR promoter region in the middle-aged group(45-59 years)was 24.77%(5.90%,45.91%)in the case group and 44.99%(14.81%,76.63%)in the control group,the difference was statistically significant(P=0.007,Z=-2.700).The DNA methylation level of DHFR promoter region in the elderly group(?60 years old)was 42.87%(19.75%,55.19%)in the case group and 39.38%(23.12%,74.29%)in the control group,the difference was not statistically significant(P=0.268,Z=-1.107).4.The DNA methylation level of MTHFD1 promoter region was 3.48%(2.47%,8.68%)in case group and 4.66%(1.88%,7.00%)in control group,the difference was not statistically significant(P=0.827,Z=-0.219).5.The DNA methylation level of SHMT1 promoter region was 39.53%(29.05%,58.61%)in the case group and 41.46%(25.87%,56.17%)in the control group,the difference was not statistically significant(P=0.683,Z=-0.409).Conclusions1.The methylation of the AHCY and MTHFD1 may be not related to cerebral infarction.2.The hypomethylation of CBS and DHFR promoter was associated with the pathogenesis of cerebral infarction,and the hypomethylation of CBS promoter may serve as a potential biomarker for the diagnosis of cerebral infarction.3.The methylation of the SHMT1 was not related to cerebral infarction.