基于代谢组学和网络药理学探讨羟基红花黄色素A治疗脓毒症急性肝损伤作用机制  

Exploration on the Mechanism of Hydroxyl Safflower Flavin A in the Treatment of Sepsis-induced Liver Injury Based on Metabolomics and Network Pharmacology

作　　者：颜诗帆 潘冰冰[2] 余婷 侯常苗 蒋宇[2] 陈芳[2] 王菁菁 刘艳娟 祝益民[2] YAN Shifan;PAN Bingbing;YU Ting;HOU Changmiao;JIANG Yu;CHEN Fang;WANG Jingjing;LIU Yanjuan;ZHU Yimin(School of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese,Changsha 410208,China;Institute of Emergency Medicine,Hunan Provincial People's Hospital,Hunan Key Laboratory of Critical Metabolomics,Changsha 410000,China;Jiangxi University of Chinese Medicine,Nanchang 330006,China)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南省人民医院急救医学研究所,急危重症代谢组学湖南省重点实验室,湖南长沙410000 [3]江西中医药大学,江西南昌330006

出　　处：《中国中医药信息杂志》2024年第2期130-137,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82102278);湖南省自然科学基金(2020JJ8010、2021JJ40292);湖南创新型省份建设专项(2019RS1067)。

摘　　要：目的采用代谢组学与网络药理学方法探讨羟基红花黄色素A(HSYA)治疗脓毒症急性肝损伤作用机制。方法将50只C57BL/6小鼠随机分为假手术组(10只)、脓毒症组(20只)和HSYA组(20只),采用盲肠结扎穿刺法构建脓毒症急性肝损伤小鼠模型,HSYA组造模后2 h皮下注射HSYA(2.25 mg/kg)。检测小鼠血清炎症因子含量及肝功能,HE染色观察肝组织病理变化,超高效液相色谱-四级杆-飞行时间质谱(UPLC-Q-TOF-MS)对肝组织进行代谢组学分析,多元数据统计方法筛选差异代谢物,采用网络药理学预测HSYA治疗脓毒症急性肝损伤潜在靶点,并对潜在靶点进行GO和KEGG通路富集分析,用Metabo Analyst 5.0数据库对模型组和HSYA组差异代谢物与潜在靶点进行匹配,构建靶点-代谢物-代谢通路网络,使用AutoDock Vina软件对HSYA与核心靶点进行分子对接,最后采用RT-qPCR验证核心基因表达。结果HSYA能降低模型小鼠血清白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α含量,恢复肝功能,减轻肝组织病理损伤。代谢组学筛选出26个向假手术组回调的差异代谢物,包括氟芬那酸、白叶藤碱、邻苯二甲酸、甲氰菊酯等,主要涉及不饱和脂肪酸的生物合成、α-亚麻酸代谢等5条代谢通路。网络药理学分析得到HSYA治疗脓毒症急性肝损伤潜在靶点81个,涉及2735个GO条目、124条信号通路;联合分析共匹配到5个差异代谢物,对应IL1B、STAT3、PTGS2、TP53等14个靶点参与HSYA对脓毒症急性肝损伤代谢紊乱的调控。分子对接结果显示,HSYA与IL1B、STAT3、PTGS2、TP53有良好的结合活性。RT-qPCR结果显示,HSYA能抑制肝组织IL1B、STAT3、PTGS2基因表达。结论HSYA可能通过调节IL1B、STAT3、PTGS2基因表达抑制炎症因子释放,维持机体代谢稳态,从而减轻脓毒症急性肝损伤。Objective To explore the mechanism of hydroxyl safflower flavin A(HSYA)in the treatment of sepsis-induced liver injury by using metabolomics and network pharmacology.Methods A total of 50 male C57BL/6 mice were randomly divided into sham-operation group(10 mice),sepsis group(20 mice)and HSYA group(20 mice).Cecal ligation and puncture was conducted to establish the sepsis-induced liver injury mouse model.The mice in HSYA group were subcutaneously injected with HSYA after 2 hours of modeling.The content of serum inflammatory factors and liver function were detected,and the pathological changes of liver tissue were observed with HE staining,UPLC-Q-TOF-MS metabolomics was used to analyze liver tissue,screening for differential metabolites using multivariate statistical methods,network pharmacology was used to predict potential targets for HSYA treatment of sepsis-induced liver injury,and conduct GO and KEGG pathway enrichment analysis on potential targets,Metabo Analyst 5.0 database was used to match differential metabolites and potential targets between the model group and HSYA group,a targets metabolite-metabolism pathway network was constructed.AutoDock Vina software was used to perform molecular docking between HSYA and core genes,and finally RT-qPCR was used to verify the expression of core genes.Results HSYA can reduce the contents of IL-6,IL-1βand TNFαin serum,restore liver function,and alleviate the morphological alternation in liver induced by sepsis.A total of 26 differential metabolites identified by metabolomics were screened out,including flufenamic acid,cryptolepine,opthalmic acid,fenpropathrin etc.,which were mainly involved in 5 metabolic pathways such as biosynthesis of unsaturated fatty acids and alpha-linolenic acid metabolism.Network pharmacology identified 81 potential targets,2735 items enriched in GO and 124 signaling pathways enriched in KEGG;a total of 5 differential metabolites were matched for joint analysis,corresponding to 14 targets including IL1B,STAT3,PTGS2,TP53,etc.,involved in the

关 键 词：脓毒症 急性肝损伤 羟基红花黄色素A 网络药理学 代谢组学 

分 类 号：R285.5[医药卫生—中药学]