利拉鲁肽通过干扰素基因刺激因子途径改善代谢相关脂肪性肝病炎症反应及机制的研究

Liraglutide improves the inflammatory response in metabolic associated fatty liver disease through stimulator of interferon genes pathway

目的探讨利拉鲁肽通过干扰素基因刺激因子(STING)信号通路改善代谢相关脂肪性肝病(MAFLD)炎症反应及机制。方法20只雄性C57BL/6J小鼠随机分为正常对照(NC)组、利拉鲁肽干预普食组(NC+Lir组)、高脂饮食(HFD)组、利拉鲁肽干预高脂饮食组(HFD+Lir组),每组各5只。小鼠原代肝细胞(MPHs)分为正常对照(Con组)、利拉鲁肽干预对照组(Con+Lir组)、棕榈酸(PA)诱导组(PA组)、利拉鲁肽干预棕榈酸组(PA+Lir组)。检测各组小鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)及肝脏中TG,HE、油红O染色观察肝脏的病理变化并计算MAFLD活动度评分(NAS),qRT-PCR检测各组小鼠和细胞中STING、IL-1β、TNF-αmRNA表达,Western blot法检测各组小鼠和细胞中STING、磷酸化干扰素调节因子3(p-IRF3)、干扰素β(IFN-β)蛋白表达。结果HFD组体重、肝脏重量、ALT、AST、肝脏TG、脂肪变性、小叶炎症和气球样变NAS高于NC组(P<0.05或P<0.01)。HFD+Lir组体重、肝脏重量、ALT、AST、肝脏TG、脂肪变性、小叶炎症和气球样变NAS低于HFD组(P<0.05或P<0.01)。HFD组肝脏中STING、IL-1β、TNF-αmRNA表达,STING、p-IRF3、IFN-β蛋白表达高于NC组(P<0.05)。HFD+Lir组STING、IL-1β、TNF-αmRNA表达,STING、p-IRF3、IFN-β蛋白表达低于HFD组(P<0.05)。PA组STING、IL-1β、TNF-αmRNA表达,STING、p-IRF3、IFN-β蛋白表达高于Con组(P<0.01)。PA+Lir组STING、IL-1β、TNF-αmRNA表达,STING、p-IRF3、IFN-β蛋白表达低于PA组(P<0.05或P<0.01)。结论利拉鲁肽通过影响STING信号通路改善MAFLD中高脂诱导的炎症反应。

Objective To investigate the mechanism by which Liraglutide improves the inflammatory response in metabolic associated faty liver disease(MAFLD)by regulating the interferon gene stimulating factor(STING)signaling pathway.Methods20 male C57BL/6J mice were randomly divided into normal diet group(NC),Liraglutide intervention group(NC+Lir group),high fat diet group(HFD group)and Liraglutide intervention high fat diet group(HFD+Lir group),with 5 in each group.Mouse primary hepatocytes(MPHs)were divided into normal control(Con)group,Liraglutide intervention group(Con+Lir group),palmitic acid group(PA group)and Liraglutide intervention PA group(PA+Lir group).The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)in serum and triglyceride(TG)contents in liver were detected.HE and oil red O staining were used to observe the pathological changes in the liver and to calculate the MAFLD activity score(NAS).The mRNA expression levels of STING,IL-1βand TNF-αin tissues and cells were detected by qRT-PCR.The protein expression levels of STING,p-IRF3 and IFN-βwere detected by Western blot.Results Body weight,liver tissue weight,serum ALT,AST,liver TG,steatosis,lobular inflammation and balloon-like NAS in HFD group were higher than those in NC group(P<0.05 or P<0.01).Body weight,liver tissue weight,serum ALT,AST,liver TG,steatosis,lobular inflammation and balloon-like NAS in HFD+Lir group were lower than those in HFD group(P<O.05 or P<0.01).The mRNA expressions of STING,IL-1β,TNF-αand the protein expressions of STING,p-IRF3 and IFN-β in liver of HFD group were higher than those of NC group(P<0.05).The mRNA expressions of STING,IL-1β,TNF-αand the protein expressions of STING,p-IRF3 and IFN-β in HFD+Lir group were lower than those in HFD+Lir group(P<0.05).The mRNA expressions of STING,IL-1β,TNF-αand the protein expressions of STING,p-IRF3 and IFN-β in PA group were higher than those in Con group(P<0.O1).The mRNA expressions of STING,IL-1β,TNF-αand the protein expressions of STING,p-IRF3 and IFN-βin PA+Lir group were lower than those in PA group(P<0.05 or P<0.01).Conclusion Liraglutide ameliorates the high-fat-induced inflammation responses in MAFLD by regulating the STING signaling pathways.