摘要
目的探讨糖尿病肾病(DN)患者基因组DNA及结缔组织生长因子(CTGF)启动子甲基化在DN发病中的作用。方法根据1999年WHO糖尿病诊断标准,选取我院2型糖尿病患者90例及同期健康体检者30例,分为糖尿病组48例、DN组42例和健康对照组30例。提取各组受检者外周血白细胞DNA,应用高效液相色谱法检测基因组DNA整体甲基化水平;甲基化特异性PCR及测序方法检测CTGF启动子甲基化状态;用酶联免疫吸附法检测各组血清CTGF蛋白水平。结果DN组基因组DNA甲基化水平为5.23%±0.09%,与糖尿病组(4.71%±0.03%)和健康对照组(4.37%±0.01%)比较,差异均无统计学意义(P〉0.05)。DN组CTGF基因启动子甲基化水平为22.02%±12.90%,明显低于糖尿病组(49.18%±8.01%,P=0.019)及健康对照组(72.18%_±19.30%,P=0.000),差异均有统计学意义。DN组血清CTGF蛋白水平为(193.44±11.9)mg/L,显著高于糖尿病组[(127.65±10.30)mg/L,P=0.0311及健康对照组[(95.84±5.1)mg/L,P=0.001]。结论DN患者CTGF启动子的甲基化程度明显低于糖尿病非肾病患者,而CTGF表达则高于非肾病患者,提示CTGF基因启动子的低甲基化可能参与DN的发生发展。
Objective To investigate the role of methylation of genomic DNA and connective tissue growth factor (CTGF) gene promoter in the pathogenesis of diabetic nephropathy (DN). Methods According to the WHO 1999 guideline for diabetes mellitus diagnosis and classification standard, 90 patients diagnosed as diabetes mellitus and 30 healthy volunteers were enrolled in this study. All the participants were divided into diabetes mellitus without DN (DM) group (n=48), DN group (n=42) and healthy control group (n=30) accordingly. DNA was extracted from the peripheral blood and high pressure liquid chromatography (HPLC) was used to measure the overall methylation level of genomic DNA. The methylation status of CTGF gene promoter was determined by PCR and sequencing analysis. Serum CTGF protein level was measured by ELISA. Results The overall methylation level of genomic DNA was 5.23%±0.09% for DN group, 4.71% ±0.03% for DM group, and 4.37%±0.01% for healthy control group, with no significant differences among three groups (all P〉0.05). The CTGF promoter methylation level in DN group (22.02%± 12.90%) was significantly decreased, compared to DM group (49.18%±8.01%, P=0.019) or healthy control group (72.18%±19.30%, P=0.000). Moreover, the serum CTGF protein level in DN group [(193.44 ± 11.90) mg/L] was significantly increased, compared to DM group [(127.65 _± 10.30) mg/L, P=0.031] and healthy control group [(95.84±5.10) nag/L, P=0.001]. Conclusion In DN patients, CTGF gene promoter methylation level is significantly decreased, but CTGF protein level is higher as compared to non-DN patients, which indicates that CTGF gene promoter demethylation may be involved in the pathogenesis of diabetic nephropathy.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2012年第3期189-193,共5页
Chinese Journal of Nephrology
基金
国家自然科学基金青年科学基金项目(81100516)
教育部高等院校博士点基金新教师项目(200805331159)
中南大学中央高校基本科研业务费专项资金资助项目(2011QNZT200)
关键词
糖尿病肾病
结缔组织生长因子
DNA
甲基化
Diabetic nephropathy
Connective tissue growth factor
DNA
Methylation