凯里酸汤对大鼠非酒精性脂肪肝病作用的分子机制

Molecular mechanism of the effect of Kaili sour soup on non-alcoholic fatty liver disease in rats

目的探讨凯里酸汤对大鼠非酒精性脂肪肝病(NAFLD)作用的分子机制。方法结合文献数据库及TCMSP、PubChem、SEA、SwissTargetPrediction、SuperPred等数据库获取凯里酸汤的化合物成分及其潜在靶点,用Genecards及DisGeNet建立疾病靶点数据集,数据通过Venn制作工具、Cytoscape软件、蛋白质相互作用网络数据库(STRING)及注释、可视化和综合发现(DAVID)等数据库进行分析及网络可视化;18只大鼠均分为对照(ND)组(予正常饲料)、模型(HF)组(予高脂饲料)及凯里酸汤干预(HS)组(予高脂饲料加1 mL/100 g酸汤灌胃),连续12周,结束时麻醉处死各组大鼠,取心脏血及肝脏,采用生化分析仪检测各组大鼠血清天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、甘油三酯(TC)及总胆固醇(TG)水平,采用苏木精-伊红(HE)染色和油红O染色观察各组大鼠肝组织的形态学特征,采用实时荧光定量(RT-qPCR)、Western blot及免疫组织化学(IHC)染色检测各组大鼠肝组织中过氧化物酶增殖物激活受体α(PPARα)、雌激素磺基转移酶1E1(SULT1E1)及雌激素受体α(ERα)的表达。结果网络药理学方法获得凯里酸汤11个主要成分及PPARα、ERα等12个关键靶点,生物信息学分析显示凯里酸汤参与多个生物过程,并可能通过雌激素信号通路在NAFLD防治中起作用;RT-qPCR、Western blot及IHC结果显示,与ND组相比,HF组大鼠肝组织中SULT1E1表达增加(P<0.05),PPARα、ERα相对表达降低(P<0.05),HS组则相反(P<0.05)。结论凯里酸汤对大鼠NAFLD作用的分子机制可能与PPARα/SULT1E1/ERα/雌激素信号转导恢复有关。

Objective To explore molecular mechanisms of the effect of Kaili sour soup on nonalcoholic fatty liver disease(NAFLD)in rats.Methods Combined with the literatures and databases such as TCMSP,PubChem,SEA,SwissTargetPrediction,SuperPred,and etc.,compound components of Kaili sour soup and its potential targets were obtained.Genecards and DisGeNet were used to establish a disease target dataset.The data was analyzed and visualized using Venn,Cytoscape,protein-protein interaction network database(STRING),Database for Annotation,Visualization and Integrated Discovery(DAVID),and other databases.Eighteen rats were randomly divided into control(ND)group(fed with normal diet),model(HF)group(fed with high-fat diet)and Kaili sour soup intervention(HS)group(fed high-fat diet and 1 mL/100 g Kaili sour soup by gavage).After 12 week treatment,all rats were anesthetized and sacrificed.Blood taken from rat hearts and livers were collected.Biochemical analyzer was used to analyze the levels of serum aspartate aminotransferase(AST),alanine aminotransferase(ALT),triglyceride(TC),and total cholesterol(TG).The morphological characteristics of rat liver tissues in each group were observed by hematoxylin eosin(HE)staining and oil red O staining.Expression levels of peroxidase proliferator activated receptorα(PPARα),estrogen sulfotransferase(SULT1 E1),and estrogen receptorα(ERα/ESR1)in rat liver tissues in each group were detected by real-time fluorescent quantification(RT-qPCR),Western blot,and immunohistochemistry(IHC).Results Eleven major components of Kaili sour soup and 12 hub targets such as PPARαand ERαwere obtained by network pharmacological approach.Bioinformatics analysis revealed that Kaili sour soup participated in multiple biological processes and might play a role in the prevention and treatment of NAFLD through estrogen signaling pathway.The results from RT-qPCR,Western blot,and IHC showed that when compared with ND group,SULT1 E1 expression was increased in rat liver tissues of HF group,while the expression levels of PPARαand ERαwere decreased(P<0.05),opposite to HS group(P<0.05).Conclusion The mechanism of Kaili sour soup in NAFLD rats may be related to the recovery of PPARα/SULT1 E1/ERα/signal transduction.