李燚,丛硕,唐娟,刘邵雯,赵珮伶,刘咏梅
LI Yi,CONG Shuo,TANG Juan,LIU Shaowen,ZHAO Peiling,LIU Yongmei

• 1:贵州医科大学附属医院临床检验中心
• 2:贵州医科大学医学检验学院

摘要(Abstract)：

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 receptor α(PPARα), estrogen sulfotransferase(SULT1 E1), and estrogen receptor α(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 PPARα and ERα were 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 PPARα and ERα were 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.

关键词(KeyWords)： 雌激素受体α;酸汤;非酒精性脂肪肝病;网络药理学;过氧化物酶增殖物激活受体α;雌激素磺基转移酶1E1
estrogen receptor α(ERα/ESR1);sour soup;non-alcoholic fatty liver disease(NAFLD);network pharmacology;peroxidase proliferator activated receptor alpha(PPARα);estrogen sulfotransferase1 E1(SULT1E1)

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81960118);; 凯里市科技计划项目(凯教科通[2019]44);; 贵州医科大学博士启动基金项目(校博合J字[2020] 16)

作者(Author): 李燚,丛硕,唐娟,刘邵雯,赵珮伶,刘咏梅
LI Yi,CONG Shuo,TANG Juan,LIU Shaowen,ZHAO Peiling,LIU Yongmei

DOI: 10.19367/j.cnki.2096-8388.2023.01.001

参考文献(References)：

• [1]IOANNOU G N.Epidemiology and risk-stratification of NAFLD-associated HCC[J].Journal of Hepatology,2021,75(6):1476-1484．
• [2]FERGUSON D,FINCK B N.Emerging therapeutic approaches for the treatment of NAFLD and type 2 diabetes mellitus[J].Nature Reviews.Endocrinology,2021,17(8):484-495．
• [3]吴文燕，胡萍，李娟，等．苗族酸汤的研究进展[J]．食品与发酵科技，2021,57(6):83-88．
• [4]龙四红，陈谢花，李红洲，等．离子色谱法测定酸汤中13种有机酸的分析方法[J]．食品科技，2022,47(8):256-263．
• [5]吴映梅，徐龙泉，杨倩，等．凯里红酸汤中番茄红素异构体含量及与有机酸的相关性分析[J]．中国调味品，2022,47(6):23-26．
• [6]潘季红，秦礼康，文安燕，等．贵州红酸汤营养品质及呈味特征分析[J]．中国调味品，2020,45 (6):43-48．
• [7]周绍琴，李凤兰，吴映梅，等．红酸汤的抗氧化活性研究[J]．中国调味品，2021,46(7):17-21．
• [8]SHIN M K,YANG S M,HAN I S.Capsaicin suppresses liver fat accumulation in high-fat diet-induced NAFLDmice[J].Animal Cells and Systems,2020,24 (4):214-219．
• [9]UGBAJA R N,UGWOR E I,DOSUMU O A,et al.Lycopene abrogates obesity-provoked hyperactivity of neurosignalling enzymes,oxidative stress and hypothalamic inflammation in female Wistar rats[J].Neurochemistry International,2021,149:105125．
• [10]CONG S,LI Z,YU L,et al.Integrative proteomic and lipidomic analysis of Kaili Sour Soup-mediated attenuation of high-fat diet-induced nonalcoholic fatty liver disease in a rat model[J].Nutrition&Metabolism,2021,18(1):26．
• [11]SZKLARCZYK D,GABLE A L,NASTOU K C,et al.The STRING database in 2021:customizable protein-protein networks,and functional characterization of user-uploaded gene/measurement sets[J].Nucleic Acids Research,2021,49(1):605-612．
• [12]NOGALES C,MAMDOUH Z M,LIST M,et al.Network pharmacology:curing causal mechanisms instead of treating symptoms[J].Trends in Pharmacological Sciences,2022,43(2):136-150．
• [13]LUUKKONEN P K,QADRI S,AHLHOLM N,et al.Distinct contributions of metabolic dysfunction and genetic risk factors in the pathogenesis of non-alcoholic fatty liver disease[J].Journal of Hepatology,2022,76 (3):526-535．
• [14]FRANCQUE S,SZABO G,ABDELMALEK M F,et al.Nonalcoholic steatohepatitis:the role of peroxisome proliferator-activated receptors[J].Nature Reviews.Gastroenterology&Hepatology,2021,18(1):24-39．
• [15]FOUGERAT A,MONTAGNER A,LOISEAU N,et al.Peroxisome proliferator-activated receptors and their novel ligands as candidates for the treatment of non-alcoholic fatty liver disease[J].Cells,2020,9(7):1638．
• [16]FRANCQUE S,VERRIJKEN A,CARON S,et al.PPARα gene expression correlates with severity and histological treatment response in patients with non-alcoholic steatohepatitis[J].Journal of Hepatology,2015,63(1):164-173．
• [17]ZHU L,BROWN W C,CAI Q,et al.Estrogen treatment after ovariectomy protects against fatty liver and may improve pathway-selective insulin resistance[J].Diabetes,2013,62(2):424-434．
• [18]LEE C,KIM J,JUNG Y.Potential therapeutic application of estrogen in gender disparity of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis[J].Cells,2019,8(10):1259．
• [19]MEYER M R,CLEGG D J,PROSSNITZ E R,et al.O-besity,insulin resistance and diabetes:sex differences and role of oestrogen receptors[J].Acta Physiologica,2011,203(1):259-269．
• [20]LONARDO A,NASCIMBENI F,BALLESTRI S,et al.Sex differences in nonalcoholic fatty liver disease:state of the art and identification of research gaps[J].Hepatology,2019,70(4):1457-1469．
• [21]WORM N.Beyond body weight-loss:dietary strategies targeting intrahepatic fat in NAFLD[J].Nutrients,2020,12(5):1316．
• [22]WAN X,YANG Z,JI H,et al.Effects of lycopene on abdominal fat deposition,serum lipids levels and hepatic lipid metabolism-related enzymes in broiler chickens[J].Animal Bioscience,2021,34(3):385-392．
• [23]XU Y,YANG X,WANG Z,et al.Estrogen sulfotransferase (SULT1E1) regulates inflammatory response and lipid metabolism of human endothelial cells via PPARγ[J].Molecular and Cellular Endocrinology,2013,369(1-2):140-149．
• [24]BARBOSA A C S,FENG Y,YU C,et al.Estrogen sulfotransferase in the metabolism of estrogenic drugs and in the pathogenesis of diseases[J].Expert Opinion on Drug Metabolism&Toxicology,2019,15(4):329-339．
• [25]MATSUSHITA N,HASSANEIN M T,MARTINEZ-CLE-MENTE M,et al.Gender difference in NASH susceptibility:roles of hepatocyte Ikkβ and Sult1e1[J].PLo SOne,2017,12(8):e0181052．
• [26]IHUNNAH C A,WADA T,PHILIPS B J,et al.Estrogen sulfotransferase/SULT1E1 promotes human adipogenesis[J].Molecular and Cellular Biology,2014,34(9):1682-1694．
• [27]LI Y,XU Y,LI X,et al.Effects of PPAR-α agonist and IGF-1 on estrogen sulfotransferase in human vascular endothelial and smooth muscle cells[J].Molecular Medicine Reports,2013,8(1):133-139．
• [28]GORRES B K,BOMHOFF G L,GUPTE A A,et al.Altered estrogen receptor expression in skeletal muscle and adipose tissue of female rats fed a high-fat diet[J].Journal of Applied Physiolog,2011,110 (4):1046-1053．
• [29]YU Y M,ZHOU B H,YANG Y L,et al.Estrogen deficiency aggravates fluoride-induced liver damage and lipid metabolism disorder in rats[J].Biological Trace Element Research,2022,200(6):2767-2776.

文章评论(Comment)：