RNAm6A去甲基化酶FTO在体外培养肝细胞中的脂肪代谢作用研究  

Study on fat metabolism of RNAm6A demethylase FTO in hepatocytes cultured in vitro

作　　者：王倩 折瑞莲 沙文琼 林湖滨 WANG Qian;ZHE Ruilian;SHA Wenqiong;LIN Hubin(Department of Obstetrics,Shenzhen People’s Hospital,Guangdong,Shenzhen 518020,China)

机构地区：[1]深圳市人民医院产科,广东深圳518020

出　　处：《中国医药科学》2022年第21期15-19,共5页China Medicine And Pharmacy

基　　金：广东省深圳市科技计划项目(JCYJ20190807144403563)。

摘　　要：目的研究脂肪与肥胖相关基因(FTO)编码的RNAm6A去甲基化酶在肝细胞中的代谢作用及FTO如何调节肝细胞代谢。方法构建FTO过表达载体FTO-OE及干扰载体FTO-sh,研究CCK-8分析甲基激活剂三甲基甘氨酸(TMG)与抑制剂环亮氨酸(cycloleucine)分别对肝细胞的增殖效果;试剂盒分析甘油产生水平;qPCR实验分析TMG与cycloleucine对肝细胞脂肪代谢相关基因表达水平(脂肪分解相关基因ATGL、成脂分化基因c/EBPβ、脂肪酸从头合成相关基因FAS);甲基化检测试剂盒分析细胞总RNA甲基化水平。结果FTO过表达显著促进脂肪细胞增殖(P<0.05)、促进其甘油水平提高(P<0.05)、引起代谢基因ATGL水平降低(P<0.05)、c/EBPβ与FAS水平升高(P<0.05)、降低RNAm6A甲基化水平(P<0.05)。结论FTO具有调控肝细胞脂肪代谢,其通过m6A调控肝细胞脂肪代谢。Objective To study the metabolism of RNAm6A demethylase encoded by the fat mass and obesity-associated(FTO)gene in hepatocytes and how FTO regulates the metabolism of hepatocytes.Methods FTO overexpression vector FTO-OE and interference vector FTO-sh were constructed,and CCK-8 assay was conducted to analyze the proliferation effects of methyl activator trimethylglycine(TMG)and inhibitor cycloleucine on hepatocytes,in which the CCK-8 kit was used to analyze glycerol production level.qPCR experiment was conducted to analyze the effects of TMG and cycloleucine on the expression levels of genes related to fat metabolism of hepatocytes(lipolysis-related gene ATGL,adipogenic differentiation-related gene c/EBPβ,fatty acid de novo synthesis-related gene FAS),in which the methylation assay kit was used to analyze the methylation level of total RNA in cells.Results Over-expression of FTO significantly promoted the proliferation of adipocytes(P<0.05),increased their glycerol level(P<0.05),decreased the level of metabolic gene ATGL(P<0.05),increased the levels of metabolic genes c/EBPβand FAS(P<0.05),and decreased the methylation level of RNAm6A(P<0.05).Conclusion FTO can regulate hepatocyte fat metabolism through m6A modification.

关 键 词：去甲基化酶FTO 代谢 脂肪 肝细胞 

分 类 号：R575.5[医药卫生—消化系统]