氧化苦参碱在db/db小鼠非酒精性脂肪性肝病中的抗炎机制  

The anti-inflammatory mechanism of oxymatrine in NAFLD of db/db mice

作　　者：肖雅文 彭灿 肖瑛[1] 梁丹 张帆 郭兵[1] XIAO Yawen;PENG Can;XIAO Ying;LIANG Dan;ZHANG Fan;GUO Bing(Department of Pathophysiology&Guizhou Provincial Key Laboratory of Drug Research and Pathogenesis of Common Chronic Diseases,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学病理生理学教研室贵州省常见慢性疾病发病机制及药物研究重点实验室,贵阳550025

出　　处：《免疫学杂志》2020年第6期497-503,共7页Immunological Journal

基　　金：国家自然科学基金(81860656);贵州省科技厅国际科技合作计划(黔科合平台人才[2019]5801号);贵阳市科技局基金(筑科合同[2017]15-29号)。

摘　　要：目的通过观察db/db小鼠非酒精性脂肪性肝病(NAFLD)中骨成形蛋白-激活素膜结合阻断因子(BAMBI)、NLRP3炎症小体的表达情况及氧化苦参碱(OMT)治疗后的变化,初步探讨OMT改善db/db小鼠NAFLD炎症反应的可能机制。方法db/db小鼠随机分成模型组和OMT给药组,OMT组给予60 mg/(kg·d)OMT腹腔注射,同期wt/wt小鼠为正常对照组,每组6只。油红O染色、HE染色和组织上清液ALT、AST检测评估NAFLD模型一般情况;免疫组化观察肝组织中BAMBI、NLRP3蛋白表达,qPCR观察肝组织中BAMBI、NLRP3 mRNA表达水平;Western blot观察肝组织中BAMBI、NLRP3炎症小体相关指标、Ⅳ型胶原(COLⅣ)、纤维连接蛋白(FN)蛋白水平变化;spearman秩相关分析对BAMBI和NLRP3进行相关性分析。结果NAFLD模型组HE染色、油红O染色及ALT、AST检测提示存在肝细胞损伤且肝脏有脂质积累。NAFLD模型组相比正常对照组,BAMBI表达量降低,NLRP3表达量增多,COLⅣ、FN表达量增多,OMT治疗后降低了肝脏脂质积累,小鼠肝组织中BAMBI表达量有增加,炎症及纤维化指标均有改善。统计学分析结果提示BAMBI和NLRP3呈显著负相关(P<0.05)。结论OMT可促进NAFLD肝组织中BAMBI表达水平增多,抑制炎症及纤维化反应,提示OMT可能通过影响BAMBI抑制db/db NAFLD小鼠的炎症及纤维化进程。To investigate the possible mechanism of oxymatrine(OMT)in reducing nonalcoholic fatty liver disease(NAFLD)inflammation in db/db mice,we detected the expression of BMP and the activin membrane-bound inhibitor(BAMBI)and NLRP3 inflammasome in NAFLD of db/db mice before and after OMT treatment.db/db Mice were randomly divided into NAFLD group and OMT group,while wild-type mice was set as control group.Mice in OMT group was given intraperitoneal injection of 60 mg/(kg·d)OMT.NAFLD disease model was evaluated by oil red O staining,HE staining and the levels of ALT and AST in tissue supernatant;immunohistochemistry was used to observe the protein expression of BAMBI and NLRP3 in liver tissues,and PCR was used to observe the mRNA expression of BAMBI and NLRP3 in liver tissues.Western blot was applied to detect the levels of BAMBI,NLRP3 inflammasome,collagenⅣ(COLⅣ),and fibronectin(FN)in the liver tissue.Spearman rank correlation analysis was carried out to analyze the correlation between BAMBI and NLRP3.Data showed that there were hepatic cell injury and lipid accumulation in the liver of NAFLD model group.Compared with normal group,BMABI expression in NAFLD model group decreased,but NLRP3 inflammasome,COLⅣ,FN expression increased.After OMT treatment,liver lipid accumulation decreased,and BAMBI expression in liver tissue increased,meanwhile inflammation and fibrosis indexes decreased.Statistical analysis suggested that BAMBI and NLRP3 were negatively correlated.Taken together,OMT can promote the increase of BAMBI expression level in NAFLD liver,and suppress inflammation and fibrosis,suggesting that OMT affects the inflammation and fibrosis process of NAFLD in db/db mice by regulating BAMBI.

关 键 词：非酒精性脂肪性肝病 氧化苦参碱 NOD样受体蛋白3 骨成形蛋白-激活素膜结合阻断因子 

分 类 号：R575.5[医药卫生—消化系统] R363[医药卫生—内科学]