特异性阻断成纤维细胞生长因子受体信号抑制肝星状细胞激活改善小鼠肝纤维化  

Blocking of fibroblast growth factor receptor improves liver fibrosis in mice via inhibiting hepatic stellate cell activation

作　　者：周冬艳 陈双娅 何萍 李校堃 王聪 ZHOU Dongyan;CHEN Shuangya;HE Ping;LI Xiaokun;WANG Cong(School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学药学院,浙江温州325035

出　　处：《温州医科大学学报》2023年第8期603-612,共10页Journal of Wenzhou Medical University

基　　金：国家自然科学基金面上项目(82173013)。

摘　　要：目的:研究成纤维细胞生长因子受体(FGFR)信号阻断对四氯化碳(CCl_(4))诱导小鼠肝纤维化的作用及其机制。方法:腹腔注射CCl_(4)诱导小鼠肝纤维化模型,给予FGFR抑制剂AZD4547处理,比较给药前后小鼠肝脏质量、血清谷丙转氨酶(ALT)、血清谷草转氨酶(AST)变化;组织学染色分析肝脏胶原沉积、纤维化水平、肝脏炎症及肝星状细胞(HSCs)活化程度等变化;同时采用荧光定量PCR(qPCR)检测肝组织纤维化相关基因α-SMA、Col1a1的mRNA表达水平。选取人HSCs细胞株LX-2给予肿瘤坏死因子α(TNFα)诱导炎症模型,免疫荧光染色比较AZD4547处理前后Vimentin表达情况,透射电镜观察不同处理组细胞超微结构变化,JC-1染色检测线粒体膜电位,分析细胞中脂联素(ADN)mRNA和蛋白表达,蛋白质印迹(Western blot)法检测JNK和AMPK的磷酸化水平。结果:FGFR抑制剂处理不影响模型小鼠肝功能,但是可显著改善CCl_(4)诱导的肝纤维化和组织炎症反应(P<0.05),减少HSCs活化(P<0.05),损伤部位的ADN可参与补偿性保护作用(P<0.05)。同时FGFR抑制剂明显逆转TNFα诱导的HSCs形态及超微结构变化,缓解LX-2线粒体功能障碍,负反馈刺激细胞中ADN表达增加(P<0.05)及AMPK活化(P<0.05)。结论:FGFR信号阻断能够减轻肝纤维化,其机制可能与抑制HSCs炎症反应及活化有关。Objective:To study the effect of fibroblast growth factor receptor(FGFR)inhibition on carbon tetrachloride(CCl_(4))-induced liver fibrosis in mice and explore its mechanism.Methods:CCl_(4)-induced liver fibrosis mouse model was treated with FGFR inhibitor AZD4547 and the changes in liver weight,serum alanine transaminase(ALT)and serum aspertate aminotransferase(AST)concentrations were compared between groups.Histological staining was used to analyze changes in liver collagen deposition,fibrosis,liver inflammation and activation of hepatic stellate cells(HSCs).Quantitative PCR(qPCR)was used to detect the expression of fibrosis related genes(α-SMA,Col1a1 mRNA)in liver.Tumor necrosis factorα(TNFα)-induced human hepatic stellate cell line LX-2 was used as the inflammation model to investigate the effect of AZD4547.The expression of Vimentin was observed by immunofluorescence staining and the ultrastructure were checked by transmission electron microscope.JC-1 staining was applied to detect the mitochondrial membrane potential.The expression of adiponectin mRNA and protein in cells was analyzed,and the phosphorylation levels of JNK and AMPK were detected by Western blot.Results:FGFR inhibitor significantly reduced liver fibrosis,inflammation response and hepatic stellate cell activation in CCl_(4)-induced mouse model without affecting ALT and AST(P<0.05).Interestingly,inhibition of FGFR could increase adiponectin expression at the damaged site,which plays the role of compensatory protection(P<0.05).In addition,FGFR inhibitors not only markedly reversed morphology and ultrastructure of TNFα-induced hepatic stellate cells,but also downregulated JNK activation(P<0.05)and alleviated mitochondrial dysfunction in HSCs(P<0.05),where adiponectin and p-AMPK were increased,suggesting a negative feedback control by FGF signaling inhibition(P<0.05).Conclusion:Inhibition of FGFR signaling indicates therapeutic effect on liver fibrosis via inhibiting the inflammation and activation of HSCs.

关 键 词：成纤维生长因子受体 脂联素 肝星状细胞 肝纤维化 磷酸化腺苷酸活化蛋白激酶 

分 类 号：R96[医药卫生—药理学]