基于FABP4/PPAR-γ/NF-κB信号通路探讨芪参汤对非酒精性脂肪性肝病小鼠肝脏炎症的影响  

Effects of Qishen Decoction on Liver Inflammation of Mice with Non-alcoholic Fatty Liver Disease Through FABP4/PPAR-γ/NF-κB Signaling Pathway

作　　者：李琨 王炳予[2] 张雅丽[2] 刘长发[2] 袁星星[2,3] LI Kun;WANG Bingyu;ZHANG Yali;LIU Changfa;YUAN Xingxing(Beijing Open University,Beijing 100081,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150006,China)

机构地区：[1]北京开放大学,北京100081 [2]黑龙江中医药大学,黑龙江哈尔滨150040 [3]黑龙江省中医药科学院,黑龙江哈尔滨150006

出　　处：《中国中医药信息杂志》2022年第10期59-66,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：黑龙江省自然科学基金联合引导项目(LH2019H095);黑龙江省卫生健康委科研项目(20212121020016);黑龙江省中医药科研项目(ZHY19-062、ZHY2020-041);黑龙江省博士后面上项目(LBH-Z21218)。

摘　　要：目的观察芪参汤对非酒精性脂肪性肝病(NAFLD)小鼠肝脏炎症的影响,从FABP4/PPAR-γ/NF-κB信号通路探讨其作用机制。方法通过GEO数据库筛选NAFLD差异表达基因。以高脂饲料喂养构建NAFLD小鼠模型,将小鼠分为空白组、模型组、芪参汤组和二甲双胍组,分别予相应药物干预,于第12、16周采用HE染色、油红O染色检测肝组织病理形态,生化分析仪检测血清天冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)水平,ELISA检测血清白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α及CXC趋化因子受体3(CXCR3)、趋化因子C-C-基元配体4(CCL4)含量,Western blot和qRT-PCR检测肝组织脂肪酸结合蛋白4(FABP4)表达。以棕榈酸诱导建立RAW264.7细胞炎症模型,将细胞分为空白组、模型组、芪参汤组和抑制剂组,加入相应药物培养后,ELISA检测细胞上清液IL-1β、TNF-α、CXCR3、CCL4含量,Western blot检测FABP4/PPAR-γ/NF-κB信号通路相关蛋白表达,流式细胞术检测巨噬细胞亚群比例。结果共筛选出1246个差异表达基因(493个上调、753个下调),其中脂肪酸结合蛋白4(FABP4)表达在NAFLD发展过程中有显著差异。动物实验结果显示,与空白组比较,模型组小鼠肝细胞肿胀,呈气球样变,肝小叶结构紊乱,有明显炎症细胞浸润和大量脂滴形成,第12、16周NAFLD活动积分和油红O染色面积显著增加(P<0.01),血清AST、ALT、IL-1β、TNF-α、CXCR3、CCL4含量显著增加(P<0.01),肝组织FABP4蛋白和mRNA表达显著升高(P<0.01);与模型组比较,芪参汤组小鼠肝脏脂肪变程度、炎症细胞浸润和脂滴明显减少,第12、16周NAFLD活动积分和油红O染色面积显著减少(P<0.01),血清AST、ALT、IL-1β、TNF-α、CXCR3、CCL4含量显著减少(P<0.05,P<0.01),肝组织FABP4蛋白和mRNA表达显著降低(P<0.01)。细胞实验结果显示,与空白组比较,模型组细胞上清液IL-1β、TNF-α、CXCR3、CCL4含量显著增加(P<0.01),细胞FABP4、p-κB抑制因�Objective To observe the effects of Qishen Decoction on liver inflammation in non-alcoholic fatty liver disease(NAFLD)mice;To discuss its mechanism through FABP4/PPAR-γ/NF-κB signaling pathway.Methods Differentially expressed genes of NAFLD were screened through GEO database,and then the NAFLD mouse model was established by feeding with high-fat diet.The mice were divided into blank group,model group,Qishen Decoction group and metformin group,and were given corresponding drug intervention respectively.At the 12th and 16th week,HE staining and oil red O staining were used to detect the pathological morphology of liver tissue;biochemical analyzer was used to detect serum AST and ALT levels;ELISA was used to detect serum contents of IL-1β,TNF-α,CXCR3 and CCL4,Western blot and qRT-PCR were used to detect FABP4 expression in liver tissue.The RAW264.7 cell inflammation model was induced by palmitic acid,and the cells were divided into blank group,model group,Qishen Decoction group and inhibitor group.After adding the corresponding drugs for culture,the contents of IL-1β,TNF-α,CXCR3 and CCL4 in cell supernatant were detected by ELISA;Western blot was used to detect the expression of related proteins in FABP4/PPAR-γ/NF-κB signaling pathway;flow cytometry was used to detect the proportion of macrophage subsets.Results A total of 1246 differentially expressed genes(493 upregulated and 753 down-regulated)were screened.Among them,FABP4 expression was significantly different during the development of NAFLD.The results of animal experiments showed that compared with the blank group,the hepatocytes of mice in the model group were swollen and balloon-like,and the structure of the hepatic lobules was disordered,with obvious inflammatory cell infiltration and the formation of a large number of lipid droplets.The NAFLD activity score and the oil red O staining area at the 12th and 16th weeks increased significantly(P<0.01),serum AST,ALT,IL-1β,TNF-α,CXCR3,CCL4 content significantly increased(P<0.01),and FABP4 protein and m

关 键 词：芪参汤 非酒精性脂肪性肝病 巨噬细胞 脂肪酸结合蛋白4 炎症 

分 类 号：R285.5[医药卫生—中药学]