成纤维细胞生长因子21对非酒精性脂肪性肝炎的作用及机制的研究

Effect of Fibroblast Growth Factor 21 on Non-alcoholic Steatohepatitis and Its Mechanism

目的:探索成纤维细胞生长因子21(FGF-21)对非酒精性脂肪性肝炎(NASH)的作用及其机制。方法:用浓度为500μmol/L的油酸和棕榈酸混合物(摩尔比=2:1)诱导HepG2细胞建立NASH细胞模型,实验分为5组:正常对照组(Control)、模型组(Model)、低剂量FGF-21组(LFGF-21,0.5μmol/L)、中剂量FGF-21组(MFGF-21,1.0μmol/L)和高剂量FGF-21组(HFGF-21,2.0μmol/L),油红O染色法观察细胞内脂滴,全自动生化分析仪检测细胞内ALT、AST、TC、TG的水平,Real-time PCR和Western blot分别检测细胞内核因子E2相关因子-2(Nrf-2)、核苷酸结合寡聚化结构域样受体3(NLRP3)的m RNA和蛋白水平,ELISA检测IL-1β、TNF-α水平。结果:NASH细胞造模成功,油红O染色结果显示对照组细胞无明显脂滴蓄积,模型组细胞内可见大量橘红色脂滴,并出现融合现象。不同浓度FGF-21治疗组的细胞内红色脂滴明显减少,并呈剂量依赖性。模型组ALT、AST、TC、TG、IL-1β、TNF-α和NLRP3的水平均高于对照组(P<0.05),FGF-21治疗组其水平低于模型组(P<0.05)。模型组Nrf2的水平低于对照组(P<0.05),FGF-21治疗组Nrf2的水平高于模型组(P<0.05)。结论:FGF-21通过促进Nrf-2、抑制NLRP3减少非酒精性脂肪性肝炎细胞的脂质沉积,减轻炎症反应,对NASH有保护作用。

Objective:To investigate the effect of fibroblast growth factor 21(FGF-21)on non-alcoholic steatohepatitis(NASH)and the mechanism of its action.Methods:Culturing HepG2 cells with DMEM medium supplemented with 10%fetal bovine serum.HepG2 cells were randomly divided into five groups:control group,model group(500μmol/L FFA),low dose FGF-21(LFGF-21,0.5μmol/L),middle dose FGF-21(MFGF-21,1.0μmol/L)and high dose FGF-21(HFGF-21,2.0μmol/L).Observing intracellular lipid droplets by oil red O staining,the levels of AST,ALT,TC and TG were measured by automatic chemistry analyzer.The m RNA contents and protein levels of Nrf-2 and NLRP3 were determined by Real-time PCR and Western blot.The protein levels of IL-1βand TNF-αwere measured by ELISA kit.Results:The NASH cell model was established.The oil red O staining showed that no lipid droplets were found in the control group,and there were mass lipid droplets in the model group,and some were fused,and those in the treatment groups decreased significantly compared with the model group on dose-dependent.The levels of AST,ALT,TC,TG,IL-1β,TNF-αand NLRP3 in the model group were significantly higher than those in the control group(P<0.05),and those in the treatment group decreased significantly compared with the model group(P<0.05).The levels of Nrf-2 in the model group were significantly lower than those in the control group(P<0.05),and that in the treatment group increased significantly compared with the model group(P<0.05).Conclusion:FGF-21 can reduce lipid deposition and inhibit inflammation through activating Nrf-2 and inhibiting NLRP3 to attenuate steatohepatitis.