基于生物信息学探讨五味子乙素抑制铁死亡改善蛋氨酸胆碱缺乏诱导脂肪肝模型小鼠的作用机制  

Bioinformatics Reveals Mechanism of Schisandrin B in Inhibiting Ferroptosis to Ameliorate Methionine and Choline Deficiency-induced Fatty Liver Disease in Mice

作　　者：朱智峰 李文婷 曹拥军[3] 凌园园[3] 刘益飞[4] ZHU Zhifeng;LI Wenting;CAO Yongjun;LIN Yuanyuan;LIU Yifei(Medical School of Nantong University,Nantong 226000,China;Yunnan University of Chinese Medicine,Kunming 650500,China;Nantong Hospital of Traditional Chinese Medicine,Nantong 226000,China;Affiliated Hospital of Nantong University,Nantong 226000,China)

机构地区：[1]南通大学医学院,江苏南通226000 [2]云南中医药大学,昆明650500 [3]南京中医药大学南通附属医院,江苏南通226000 [4]南通大学附属医院,江苏南通226000

出　　处：《中国实验方剂学杂志》2025年第2期74-83,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省中医药科技发展计划项目(YB2020065);国家自然科学基金青年科学基金项目(82104802)。

摘　　要：目的:非酒精性脂肪性肝病(NAFLD)是一种代谢应激性肝损伤,铁死亡参与了NAFLD的发生发展,通过探究五味子乙素治疗NAFLD的疗效及作用机制,可为NAFLD的防治寻求有效方案。方法:通过检索有机小分子生物活性数据(PubChem)平台获取五味子乙素的分子结构,经小分子药物靶点预测在线平台(SwissTargetPrediction)预测相关靶点,并通过中药系统药理学数据库与分析平台(TCMSP)、高通量中药实验参考数据库(HERB)数据库进行补充,对活性成分的靶点进行预测;采用基因数据库(GeneCards)、铁死亡数据库(FerrDb)分别获取NAFLD疾病、铁死亡靶点;然后经交集映射筛选出核心靶点,并构建靶点间蛋白质-蛋白质相互作用(PPI)分析图,采用生物学信息注释数据库(DAVID)对核心靶点进行基因本体(GO)功能富集和京都基因与基因组百科全书(KEGG)通路富集分析;最后将五味子乙素-核心靶点分子对接并计算结合能。使用蛋氨酸胆碱缺乏(MCD)饲养C57BL/6小鼠建立NAFLD模型,随机分为正常组、模型组、阳性药(易善复)组、五味子乙素低剂量组、五味子乙素高剂量组,检测给药后小鼠的体质量、肝脏指数;生化试剂盒检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)及肝组织匀浆中总胆固醇(TC)、甘油三酯(TG)、丙二醛(MDA)、谷胱甘肽(GSH)、亚铁离子(Fe2+)水平,苏木素-伊红(HE)及红油O染色观察肝组织病理改变,酶联免疫吸附测定法(ELISA)检测血清细胞因子肿瘤坏死因子(TNF)-α,白细胞介素(IL)-1β、IL-6及4-羟基壬烯醛(4-HNE)水平,蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Realtime PCR)检测肝组织中铁死亡溶质载体家族7成员11(SLC7A11)、溶质载体家族3成员2(SLC3A2)、谷胱甘肽过氧化物酶4(GPX4)、转铁蛋白(Transferrin)、重链铁蛋白(FTH)相关蛋白及mRNA表达。结果:中药单体五味子乙素,共筛选得到五味子乙素作用Objective:Nonalcoholic fatty liver disease(NAFLD)is a metabolic stress liver injury.Ferroptosis is involved in the occurrence and development of NAFLD.Exploring the efficacy and mechanism of schisandrin B in treating NAFLD facilitates the development of strategies for the prevention and treatment of NAFLD.Methods:The molecular structure of schisandrin B was obtained by searching against PubChem,and the related targets were predicted by SwissTargetPrediction.The active ingredients and their targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the high-throughput experiment-and reference-guide database of traditional Chinese medicine(HERB).GeneCards and FerrDb were searched for the targets of NAFLD and ferroptosis.The common targets were taken as the core targets,and the protein-protein interaction network of the core targets was established.DAVID was used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.Finally,molecular docking was performed between schisandrin B and core targets,and the binding energy was calculated.C57BL/6 mice were fed with a methionine and cholinedeficiency(MCD)diet for the modeling of NAFLD.Mice were randomized into normal,model,positive drug(essentiale),and low-and high-dose schisandrin B groups.The body mass and liver index of mice were measured after drug administration.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the serum and those of total cholesterol(TC),triglyceride(TG),malondialdehyde(MDA),glutathione(GSH),and Fe2+in the liver homogenate were measured by biochemical assay kits.The pathological changes of the liver tissue were observed by hematoxylin-eosin(HE)and red oil O staining.Enzymelinked immunosorbent assay was employed to determine the levels of interleukin(IL)-6,IL-1β,tumor necrosis factor(TNF)-α,and 4-hydroxynonenal(4-HNE)in the serum.Western blotting and real-time PCR were employed to determine the protein and mRNA levels,respectively,of

关 键 词：非酒精性脂肪性肝病 五味子乙素 铁死亡 氧化应激 炎症 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R287