糖尿病肾病患者TGFB1基因表达调控区甲基化水平变化  

Transformation of DNA demethylation in the regulatory region of TGFBI gene in patients with diabetic nephropathy

作　　者：张倩[1] 杨丽铃[1] 吴琼[1] 曾薇[1] 牟娇[1] 章波[1] 陈雪丹[1] 刘东擘[1] 冯兵[1] 

机构地区：[1]解放军第三军医大学新桥医院肾内科,重庆400037

出　　处：《中华肾脏病杂志》2016年第2期99-105,共7页Chinese Journal of Nephrology

基　　金：国家自然科学基金（81370820）

摘　　要：目的探讨糖尿病肾病（DN）患者转化生长因子β1（TGF-β1）基因TGFB1表达调控区甲基化改变及其甲基化在DN发病中的作用。方法根据1999年世界卫生组织（WHO）糖尿病诊断标准选取2013年6月至2015年5月期间入住本院的2型糖尿病患者91例，其中单纯糖尿病组（DM组）42例，DN组49例。并选人同期体检健康者30例作为健康对照组（Con组）。提取所有研究对象外周血基因组DNA并进行亚硫酸氢钠修饰处理。采用甲基化特异性PCR法初筛各组TGFB1基因表达调控区DNA甲基化人群，亚硫酸氢盐修饰后测序法检测各组TGFB1基因表达调控区DNA甲基化水平。ELISA检测各组血清TGF-β1蛋白水平。全自动生化分析仪检测血尿素氮、肌酐、空腹血糖、餐后血糖、糖化血红蛋白及尿微量白蛋白／肌酐水平。采用Pearson相关分析和多元逐步回归分析对DN组TGF-β1相关影响因素进行分析，并分析血清TGF-β1水平与病理分级的相关性。结果DN组TGFB1基因表达调控区DNA甲基化人群为12．2％，低于DM组的42．8％及Con组的73．3％（均P〈0．05）。DN组TGFB1基因表达调控区DNA甲基化水平为12．5％±8．1％，明显低于DM组的35．6％±6．0％及Con组的66．7％±9．1％（均P〈0．05）。DN组患者血清TGF-β1水平为（2885．73±411．36）ng／L，显著高于DM组的（1367．22±126．13）ng／L和Con组的（296．38±74．37）ng／L（均P〈0．01）。DN组患者eGFR（β=-0．690，P〈0．01）和甲基化水平（β=-0．302，P〈0．01）是血清TGF-β1水平影响因素，血清TGF-β1水平与甲基化水平呈负相关（β=-0．925，P〈0．01），与肾脏病理改变的严重程度呈正相关（rs=0．847，P〈0．01）。同时DN组甲基化水平和病理分级相关（X^2=23．667，P=0．04）。结论TGFB1基因表达调控区去甲基化可能是高糖诱导系膜细胞TGFB1基因表达激活的重要机制，进而参与DN的发生和发展。Objective To investigate the role of DNA Inethylation changes in the regulatory region of TGFB1 gene in patients with diabetic nephropathy （DN）. Methods According to the WHO 1999 guideline for diabetes mellitus diagnosis and classification standard, 91 patients who were hospitalized in June 2013 to May 2015 and diagnosed as diabetes mellitus were selected, including 42 patients with diabetes mellitus （DM group） and 49 with diabetic nephropathy （DN group）. Thirty cases with health examination were selected as healthy control group （Con group）. DNA was extracted fromall the subjects＇ peripheral blood and modified by sodium bisulfite. DNA methylation status of TGFB1 gene regulatory region was screened by methylation specific PCR and the DNA methylation level was detected by bisulfite sequencing PCR. ELISA was used to test serum TGF-β1. Blood urea nitrogen, creatinine, fasting blood sugar, postprandial blood sugar, glycosylated hemoglobin and urinary albumin to creatinine ratio （UACR） were detected by automatic biochemistry analyzer. Pearson correlation analysis and muhiple stepwise regression were used to analysis TGF-β1-related factors, the correlation between the level of serum TGF-131 and the pathological grade was analyzed in DN patients. Results There were 12.2% patients in DN group with DNA methylation of TGFB1 gene regulation region, lower than those in DM group （42.8%） with Con group （73.3%） （all P 〈 0.05）. The methylation level of TGFB1 regulatory region was 12.5%＋8.1% in DN group, significantly lower than those in DM group （55.6%±6.0%） and Con group （66.7%±9.1%） （all P 〈 0.05）. Moreover, compared with that in DM group （1367.22± 126.15 ng/L） and Con group [（296.58±74.57） ng/L], TGF-β1 expression was increased significantly in DN group [（2885.73±411.56 ng/L] （all P 〈 0.01）. In DN patients serum TGF-β1 was correlated with eGFR （β=-0.690, P 〈 0.01） and the methylation （β=-0.302, P 〈 0.01）, and the serum TGF-β1 was negat

关 键 词：糖尿病肾病 转化生长因子131 DNA甲基化 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]