过氧化物酶体增殖物活化受体–α基因表达与甲基化的关系  

The Relationship between the Methylation of the Peroxisome Proliferator-activated Receptors-alpha and the Expression of PPAR-α mRNA

作　　者：屈飞 黄浩辉 郑晓筠 吴晓晓 QU Fei;HUANG Hao-hui;ZHENG Xiao-jun;WU Xiao-xiao(The Hedong Department of Guangzhou First People’s Hospital,Guangdong Guangzhou 510180)

机构地区：[1]广州市第一人民医院鹤洞分院,广东广州510180

出　　处：《深圳中西医结合杂志》2021年第14期36-38,F0003,共4页Shenzhen Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的:探讨过氧化物酶体增殖物激活受体(PPAR)–α的表达与甲基化的关系及检测甲基化位点。方法:将人正常肝细胞L02细胞株(L02细胞)分为正常组、油酸组、正常+5–氮杂–2′–脱氧胞苷(5–Aza–CdR)组、油酸+5–Aza–CdR组。生化仪检测细胞内三酰甘油(TG)、总胆固醇(TC)及上清液TG、TC、谷丙转氨酶(ALT)、谷草转氨酶(AST)含量;油红O染色观察细胞脂肪沉积量;逆转录聚合酶链反应(RT–PCR)检测PPAR–α信使核糖核酸(mRNA)表达;焦磷酸测序检测PPAR–α启动子区甲基化水平。结果:(1)与正常组比较,油酸组细胞内TG、TC及上清液ALT、AST升高,油红染色可见明显脂滴沉积,PPAR–α mRMA表达下降,PPAR–α启动子区–273、–234位点甲基化率升高,差异具有统计学意义(P<0.05)。(2)与油酸组比较,油酸+5–Aza–CdR组细胞内TG、TC及上清液ALT、AST下降,油红染色脂滴沉积减少,PPAR–α mRMA表达增加,在PPAR–α启动子区–273、–234位点甲基化率降低,差异具有统计学意义(P<0.05)。结论:5–Aza–CdR可通过降低PPAR–α启动子区甲基化率,促进PPAR–α表达,改善非酒精性脂肪性肝病(NAFLD)的脂肪变性。Objective To investigate the relationship between the methylation of the Peroxisome proliferator-activated receptors-alpha(PPAR-α) and the expression of PPAR-αmRNA,and to detect the methylation site.Methods The normal human hepatocytes(L02 cells) were divided into 4 groups:normal group,oleic acid group,normal + 5–Aza–2′–deoxycytidine (5-Aza-CdR)group,oleic acid+5–Aza–2′–deoxycytidine group.Biochemical assays were used to assess the TG,TC contents in cells and the TG,TC,ALT,AST levels in supernatant.Oil red staining was used to observe the droplets accumulation in cells.The level of PPAR-αmRNA expression was measured by real time reverse transcription-polymerase chainreaction(RT-PCR).The DNA methylation level was analyzed by pyro-sequencing.Results (1) Compared with the normal group,there are significant increase was observed in TG,TC contents in cells and the ALT,AST levels in supernatant,and more droplets accumulation in cells in oleic acid group,and so was the down-regulation of the expression of PPAR-αmRNA and the high-regulation of the DNA methylation of CpG-273,-234(P<0.05).(2) Compared with the oleic acid group,there are significant reduction was observed in TG,TC contents in cells and the ALT,AST levels in supernatant,and less droplets accumulation in cells in oleic acid+5–Aza–2′–deoxycytidine group,and so was the up-regulation of PPAR-αmRNA and the down-regulation of the DNA methylation of CpG-273,-234 (P<0.05).Conclusion 5–Aza–2′–deoxycytidine can increases the expression of PPAR-α mRNA through down-regulates the methylation on CpG promoters in PPAR-α,thereby ameliorating hepatocytestestosis in nonacoholic fatty liver disease.

关 键 词：非酒精性脂肪性肝病 过氧化物酶体增殖物激活受体–α 脱氧核糖核酸甲基化 

分 类 号：R575.5[医药卫生—消化系统] S432.22[医药卫生—内科学]