糖尿病肾病患者NTN4、MTHFR、FZD2、AKR1B1基因DNA甲基化水平与危险因素分析

Correlation of DNA methylation of NTN4, MTHFR, FZD2, AKR1B1 genes with risk of diabetic nephropathy

目的探讨NTN4、MTHFR、FZD2、AKR1B1基因DNA甲基化水平与糖尿病肾病(DKD)的关系。方法选择2016年10月至2017年10月杭州市红十字会医院收治的单纯糖尿病(DM)患者20例为DM组;糖尿病肾病患者20例为DKD组。另择同期体检健康者20名作为对照组。检测3组对象的血糖、肌酐、糖化血红蛋白(HbA1C)、BMI、尿微量白蛋白与肌酐比值(UACR)等指标,采用甲基化特异性PCR(MS-PCR)及非甲基化特异性PCR(UMS-PCR)方法检测3组对象NTN4、MTHFR、FZD2、AKR1B1基因DNA甲基化水平,采用Image J软件分析MS-PCR条带灰度值(M)和UMS-PCR条带灰度值(U),计算DNA甲基化率[M/(M+U)]并进行比较;对DKD组AKR1B1基因DNA甲基化的危险因素进行相关性分析及多元线性回归分析。结果对照组、DM组、DKD组FZD2基因DNA甲基化率分别为(48.82±11.25)%、(55.25±15.35)%、(77.38±25.26)%,AKR1B1基因DNA甲基化率分别为(33.88±7.77)%、(50.02±9.07)%、(62.14±7.10)%,均DKD组>DM组>对照组,差异均有统计学意义(均P<0.05);DKD患者的HbA1C、UACR是AKR1B1基因DNA甲基化的影响因素(β=-0.517、-0.471,均P<0.05)。3组NTN4、MTHFR基因DNA甲基化率差异均无统计学意义(均P>0.05)。结论FZD2、AKR1B1基因在血液中的DNA甲基化可能参与了DKD的发病过程。

Objective To explore the relationship between DNA methylation levels of NTN4,MTHFR,FZD2 and AKR1B1 genes and diabetic nephropathy.Methods From December 2016 to December 2017,20 healthy subjects(control group),20 simple diabetic patients(DM group)and 20 diabetic kidney disease patients(DKD group)in Hangzhou Red Cross Hospital were recruited in the study.The blood glucose,creatinine,HbA1C,BMI,urinary albumin-creatinine ratio(UACR)and other indicators were detected in three groups.Methylation-specific PCR(MS-PCR)and non-methylation-specific PCR(UMS-PCR)were used to detect the DNA methylation levels of NTN4,MTHFR,FZD2 and AKR1B1 genes in all subjects.Image J software was used to analyze the gray value(M)of MS-PCR bands and UMS-PCR bands.Correlation analysis and regression analysis were performed for the related factors of AKR1B1 gene methylation in DKD group.Results The DNA methylation rates of FZD2 gene in control,DM and DKD groups were(48.82±11.25)%,(55.25±15.35)%and(77.38±25.26)%,respectively,and those of AKR1B1 gene were(33.88±7.77)%,(50.02±9.07)%and(62.14±7.10)%,respectively.DKD group>DM group>contrd group(all P<0.05).HbA1C and UACR in DKD group were negatively correlated with DNA methylation of AKR1B1 gene(β=-0.517、-0.471,all P<0.05).There was no significant difference in DNA methylation rate of NTN4 and MTHFR among the three groups(all P>0.05).Conclusion The DNA methylation levels of FZD2 and AKR1B1 genes are increased in patients with diabetic nephropathy,indicating that DNA methylation of FZD2 and AKR1B1 may be involved in the pathogenesis of DKD.