皂术茵陈方对非酒精性脂肪性肝炎大鼠USP18/TAK1/NF-κB信号通路的影响  

Effects of Zaozhu Yinchen Formula on rats with non-alcoholic steatohepatitis based on USP18/TAK1/NF-κB signaling pathway

作　　者：梁惠卿[1,2] 林立 刘垚昱 吴昦辰 黄稚真 郑燕茹 吴晓纹 胡敏[3] 卢大为[1] 庄鸿莉[4] 陈少东[1] LIANG Huiqing;LIN Li;LIU Yaoyu;WU Haochen;HUANG Zhizhen;ZHENG Yanru;WU Xiaowen;HU Min;LU Dawei;ZHUANG Hongli;CHEN Shaodong(Department of Traditional Chinese Medicine,School of Medicine,Xiamen University,Xiamen 361102,China;Center for Liver Disease,Xiamen Hospital of Traditional Chinese Medicine,Xiamen 361009,China;The First School of Clinical Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 351012,China;Department of Traditional Chinese Medicine,The First Affiliated Hospital of Xiamen University,Xiamen 361005,China)

机构地区：[1]厦门大学医学院中医系,厦门361102 [2]厦门市中医院肝病中心,厦门361009 [3]福建中医药大学第一临床医学院,福州351012 [4]厦门大学附属第一医院中医科,厦门361005

出　　处：《中华中医药杂志》2023年第10期4887-4890,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81873242,No.82174141);国家中医药管理局青年岐黄学者支持项目(No.国中医药办人教函[2021]200号,国中医药办人教函[2022]256号);福建省自然科学基金项目(No.2023J011636);厦门市科技计划项目(No.3502Z20224018);厦门市医疗卫生指导性项目(No.3502Z20214ZD1314,No.3502Z20224ZD1166);厦门市扶持中医药发展专项(No.XWZY20230101,No.XWZY20230202)。

摘　　要：目的:观察皂术茵陈方对非酒精性脂肪性肝炎(NASH)大鼠的作用特点及其对肝脏蛋白质泛素化USP18-TAK1-NF-κB信号通路的影响,探讨其治疗NASH的作用机制。方法:32只SPF级SD大鼠随机分为正常组、模型组、皂术茵陈方组和吡格列酮组,每组8只。除正常组外,其余组采用高脂饮食16周建立大鼠NASH模型,在造模第9周开始给药,共治疗8周。观察大鼠一般状态;生化法检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性和肝组织甘油三酯(TG)含量;HE染色观察肝组织病理学变化;实时荧光定量聚合酶链式反应(RT-PCR)法和ELISA分别检测肝组织USP18、TAK1、NF-κB等的基因与蛋白表达。结果:与正常组比较,模型组大鼠显示出典型的NASH组织学特征,皂术茵陈方组肝细胞脂肪变性、炎症浸润较模型组减轻。与正常组比较,模型组大鼠肝湿重,血清ALT、AST活性,肝脏TG含量均显著升高(P<0.01),同时,肝组织USP18、TAK1蛋白和mRNA表达显著降低(P<0.01),NF-κB蛋白和mRNA表达显著升高(P<0.01)。与模型组比较,皂术茵陈方组肝组织USP18、TAK1蛋白和mRNA表达显著升高(P<0.05)、NF-κB蛋白和mRNA表达显著降低(P<0.05)。结论:皂术茵陈方具有改善NASH大鼠肝组织脂肪变性、气球样变性、炎症的药理效应,同时可显著升高肝组织USP18、TAK1表达,降低NF-κB表达,提示皂术茵陈方治疗NASH的作用机制与调控蛋白质泛素化USP18-TAK1-NF-κB信号通路有关。Objective:To observe the effect of Zaozhu Yinchen Formula(ZYF)on nonalcoholic steatohepatitis(NASH)and the mechanism on USP18-TAK1-NF-κB signaling pathway.Methods:Thirty-two SPF grade SD rats were randomly divided into normal group,model group,ZYF group and Pioglitazone group,with 8 rats in each group.Except for the normal group,the rat NASH model groups were established by high-fat diet for 16 weeks.The rat NASH model was established by highfat diet for 16 weeks.The drug was administered at the 9th week,and the treatment lasted for 8 weeks.The activities of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and the contents of triglyceride(TG)in liver were detected by biochemical method;HE staining was used to observe the pathological changes of liver tissue;Real time PCR and ELISA were used to detect Gene and protein expression of USP18,TAK1,NF-κB in the liver tissue.Results:Compared with the normal group,the rats in the model group showed typical histological characteristics of NASH.The steatosis and inflammatory infiltration of hepatocytes in ZYF group were reduced compared with the model group.Compared with the normal group,the liver weight,the activities of serum ALT and AST,the contents of liver TG in the model group were significantly increased(P<0.01),the protein and mRNA expressions of USP18,TAK1 were significantly decreased and the protein and mRNA expressions of NF-κB were significantly increased(P<0.01).Compared with the model group,the protein and mRNA expressions of USP18,TAK1 were significantly increased and NF-κB were significantly decreased(P<0.05).Conclusion:ZYF has good effect on the treatment for NASH,and significantly increase the expression of USP18 and TAK1,reduce the expression of NF-κB,which suggests the mechanism of ZYF in the treatment of NASH be related with the regulation of endoplasmic reticulum stress based on USP18-TAK1-NF-κB signaling pathway.

关 键 词：皂术茵陈方 非酒精性脂肪性肝炎 蛋白质泛素化 信号通路 

分 类 号：R285.5[医药卫生—中药学]