植物来源的miR-2096-3p靶向调控TAF15/FASN显著改善非酒精性脂肪肝病细胞模型脂肪变性  

Plant-derived miR-2096-3p targets TAF15/FASN and significantly improves steatosis in a cell model of non-alcoholic fatty liver disease

作　　者：陈辉 王佳华 CHEN Hui;WANG Jiahua(College of Marine Science,Shanghai Ocean University,Shanghai,China,201306)

机构地区：[1]上海海洋大学海洋科学学院,上海201306

出　　处：《分子诊断与治疗杂志》2025年第2期251-254,共4页Journal of Molecular Diagnostics and Therapy

基　　金：上海市科技兴农项目(2022-02-08-00-12-F01088)。

摘　　要：目的探讨植物来源的miR-2096-3p通过调控TATA-box结合蛋白相关因子15(TAF15)/脂肪酸合成酶(FASN)对非酒精性脂肪肝病(NAFLD)细胞模型脂肪变性的影响。方法利用293T细胞荧光素酶报告基因系统验证miR-2096-3p对TAF15的靶向作用。采用油酸-棕榈酸(2∶1)诱导建立HepG2细胞脂肪变性模型。实验分组包括:(1)组(即转染miR-NC的HepG2细胞)、(2)组(即转染miR-2096-3p模拟物的HepG2细胞)、阴性对照组(vector组)(即转染空载质粒的HepG2细胞)和(3)组(即共转染miR-2096-3p与TAF15模拟物的HepG2细胞)。通过油红O染色观察并比较各组细胞脂肪滴的变化,并采用实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白质印迹法(Western Blot)分别检测并比较各组细胞中TAF15和FASN的mRNA及蛋白表达水平。结果荧光素酶报告基因实验结果显示,miR-2096-3p与TAF15存在潜在靶向关系,转染miR-2096-3p的HepG2细胞WT-pmirGLO-TAF15的荧光活性明显低于转染miR-NC的HepG2细胞,差异具有统计学意义(P<0.05),而Mut-pmirGLO-TAF15的荧光活性两者比较差异无统计学意义(P>0.05)。与(1)组相比,(2)组的miR-2096-3p表达量显著升高,TAF15、FASN的mRNA和蛋白表达水平显著降低,差异有统计学意义(P<0.05)。结论过表达miR-2096-3p能通过下调TAF15/FASN,缓解NAFLD细胞模型中的脂肪滴积累。Objective To investigate the effects of plant-derived miR-2096-3p on steatosis in cell models of non-alcoholic fatty liver disease(NAFLD)through down-regulation of TATA-box binding protein-associated factor 15(TAF15)/fatty acid synthetase(FASN).Methods The 293T cell luciferase reporter system was used to confirm the targeting effect of miR-2096-3p on TAF15.The HepG2 steatosis cell model was established using a 2∶1 mixture of oleic and palmitic acids as free fatty acids(FFA).Experimental groups included:①group A(HepG2 cells transfected with miR-2096-3p),②group B(HepG2 cells transfected with Mir-2096-3p mimics),negative control group(vector group)(HepG2 cells transfected with empty plasmid),and③group C(HepG2 cell co-transfected with mi R-2096-3p and TAF15 mimics).Oil red O staining was then performed to visualize and compare the changes in fat droplet accumulation,and the mRNA and protein expression levels of TAF15 and FASN in each group were detected and compared using real-time quantitative fluorescent polymerase chain reaction(qRT-PCR)and Western Blot.Results The luciferase reporter assay showed a potential targeting relationship between miR-2096-3p and TAF15.The fluorescence activity of WT-pmirGLO-TAF15 in HepG2 cells transfected with miR-2096-3p was significantly lower than that in cells transfected with miR-NC,with a statistically significant difference(P<0.05).There was no significant difference in the fluorescence activity of Mut-pmirGLO-TAF15(P>0.05).In comparison to group①,the mRNA expression level of miR-2096-3p was significantly increased in group②.Additionally,the mRNA and protein expression levels of TAF15 and FASN were significantly decreased in group②compared to group①,with statistical significance(P<0.05).Conclusion Overexpression of miR-2096-3p can alleviate the accumulation of fat droplets in NAFLD cell models by down-regulating TAF15/FASN.

关 键 词：植物微小RNA TATA-box结合蛋白相关因子15 脂肪酸合成酶 非酒精性脂肪肝病 

分 类 号：R73[医药卫生—肿瘤]