老年糖尿病肾病患者转化生长因子β1基因表达调控区甲基化改变及机制  

Changes of methylation and mechanism of expression of transforming growth factorβ1 gene in elderly patients with diabetic nephropathy

作　　者：金琳[1] 王海萍[1] JIN Lin WANG Hai-Ping.(Department of Internal Medicine, Provincial Hospital Affiliated to Shandong University, Jinan 250000, Shandong, Chin)

机构地区：[1]山东大学附属省立医院内科及心血管中心,山东济南250000

出　　处：《中国老年学杂志》2016年第24期6133-6135,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(81200530)

摘　　要：目的观察老年糖尿病肾病患者转化生长因子(TGF)β1基因表达调控区甲基化改变情况,探讨甲基化在糖尿病肾病发病机制中的作用。方法老年2型糖尿病患者182例,分为单纯糖尿病组84例、糖尿病肾病组98例,选取同期体检的老年健康志愿者60名为健康对照组。提取3组外周血基因组DNA,进行亚硫酸氢盐修饰处理。采用甲基化特异性PCR技术初筛3组TGF-β1基因表达调控区DNA甲基化人群,用亚硫酸氢盐修饰后测序技术检测3组TGF-β1基因表达调控区DNA甲基化水平。ELISA法检测3组血清TGF-β1蛋白表达水平;分析糖尿病肾病组TGF-β1蛋白表达相关影响因素。结果单纯糖尿病组和糖尿病肾病组TGF-β1基因第一外显子区甲基化比例均明显低于健康对照组(P<0.05),糖尿病肾病组与单纯糖尿病组相比进一步降低。单纯糖尿病组甲基化水平明显低于健康对照组(P<0.05);糖尿病肾病甲基化水平明显低于健康对照组和单纯糖尿病组(P<0.05)。单纯糖尿病组和糖尿病肾病组TGF-β1蛋白水平均明显高于健康对照组(P<0.05);糖尿病肾病组与单纯糖尿病组相比进一步升高。血清TGF-β1蛋白表达水平与UACR、BUN、Scr、FBS、PBS呈明显正相关,与e GFR、基因甲基化呈明显负相关。进一步多元逐步回归分析显示,e GFR和基因甲基化与血清TGF-β1蛋白表达水平存在明显相关性,是影响TGF-β1表达的因素。血清TGF-β1蛋白表达水平与病理分级呈明显正相关;糖尿病肾病组12个Cp G位点甲基化含量与病理分级相关。结论 TGF-β1基因表达调控区甲基化是高糖诱导系膜细胞TGF-β1基因表达激活的重要机制,参与老年糖尿病肾病的发生与发展。Objective To observe the changes of methylation status in the regulation of transforming growth factor β1 gene expres- sion in elderly patients with diabetic nephmpathy （DN）, and explore the role of methylation in the pathogenesis of DN. Methods 182 cases of elderly patients with type 2 diabetes were selected and divided into diabetes （DM） group with 84 cases and DN group with 98 cases. 60 healthy elderly healthy volunteers were selected into control group. Genomic DNA from peripheral blood of the study object was extracted and modified by hydrogen sulfate. Methylation specific PCR technique was used to detect the expression of TGFq31 gene, and the DNA methylation level was detected by the sequencing technology after the modification of the TGF-β1 gene. ELISA method was used to detect the expression level of TGF-β1 protein. The correlation between the expression of TGF-β1 protein and the pathological grade was analyzed. Resuits TGFβ gene and significant promoter region methylation ratio in DM group and DN group were significantly lower than those of control group （P〈0.05） ; Compared with DM group, DN group had further reduce. Methylation level of DM group was significantly lower than that in control group （ P〈0.05 ） ; Methylation level of DN group was significantly lower than that in control group and DM group （ P〈0.05 ）. The protein levels of TGF-β1 in DM group and DN group were significantly higher than those in control group （P〈0.05）. protein levels of TGF- β1 of DN group was higher than that of DM group （P〈0.05）. The level of serum TGF-β1 protein was significantly positively correlated with UACR, BUN, Scr, FBS, PBS, and eGFR, gene methylation was significantly negatively correlated. Multiple stepwise regression analysis showed that there was a significant correlation between eGFR and gene methylation and the expression level of TGF-1 protein in serum,which was the influence factor of TGF-β1 expression. The expression level of serum TGF-β1 protein was significant

关 键 词：糖尿病肾病 DNA甲基化 转化生长因子Β1 

分 类 号：R587[医药卫生—内分泌]