摘要
目的:采用网络药理学和分子对接的方法,探讨大柴胡汤治疗非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)的作用机制,并进行动物实验验证。方法:利用TCMSP和SwissTargetPrediction数据库收集并筛选大柴胡汤的活性成分及作用靶点,通过GeneCards、DrugBank、DisGeNET、OMIM及TTD数据库获取NAFLD的相关疾病靶点,并确定药物-疾病交集靶点。使用Cytoscape 3.9.1软件构建“药物-活性成分-靶点”网络和蛋白相互作用网络(PPI),进一步筛选出核心活性成分及核心靶点。利用Metascape数据库对交集靶点进行基因本体(GO)功能和京都基因与基因组百科全书(KEGG)通路富集分析,并对大柴胡汤治疗NAFLD的核心活性成分与核心靶点进行分子对接验证。构建高脂饮食诱导的NAFLD小鼠模型,通过动物实验验证网络药理学预测结果。结果:网络药理学预测得到90个大柴胡汤治疗NAFLD的潜在靶基因,核心靶点包括TNF、IL6、IL1B、TP53和PPARG等。核心活性成分包括原卟啉、11-二十碳烯酸、10,13-二十碳二烯酸、表小檗碱、滇刺枣碱D、山柰酚(kaempferol)等。GO功能富集分析结果表明,对细菌来源分子的反应、小分子代谢过程的正向调节、分泌调节等在生物过程中富集度较高;膜筏、受体复合物、胞质核周区等在细胞组分中富集度较高;核受体活性、单羧酸结合、蛋白质结构域特异性结合等在分子功能中富集度较高。KEGG通路富集结果表明,大柴胡汤可能通过调节过氧化物酶体增殖物激活受体(PPAR)等信号通路发挥抗NAFLD的作用。分子对接结果显示,以上核心活性成分与核心靶点均具有良好的结合活性。动物实验结果表明,大柴胡汤可有效减少NAFLD模型小鼠的脂质、氧化应激和炎症因子的水平。大柴胡汤降低了血清TG、TC、LDL-C含量(P<0.05或P<0.01)、血清AST和ALT活力(P<0.05或P<0.01)、肝脏MDA、TNF-α、IL-6、IL-1β含量(P<0.05或P<0.01),升高了血清HDL-C含量(P<0.05)和肝脏SOD活力(P<0.01);HE染色及油红O染色结果显示,大柴胡汤有效减轻了NAFLD模型小鼠的肝脏脂肪变性、空泡样病变和红色脂滴;大柴胡汤明显下调了肝脏Tnfa、Il6、Il1b的mRNA表达(P<0.05或P<0.01),上调Pparγ、Akt1、Ampk mRNA表达(P<0.05)。结论:大柴胡汤可有效减轻NAFLD小鼠的脂质沉积、氧化应激及炎症反应,改善肝脏功能及减轻病理损伤,其作用机制可能与抑制TNF-α、IL-6、IL-1β的分泌及调控PPARγ信号通路有关。
Objective:To explore the mechanism of action of DaChaiHu(大柴胡)Decoction in the treatment of non-alcoholic fatty liver disease(NAFLD)by using network pharmacology and molecular docking methods,and to validate these findings through animal experiments.Methods:The active ingredients and targets of DaChaiHu Decoction were collected and screened by using the TCMSP and SwissTargetPrediction databases.Disease targets related to NAFLD were obtained from GeneCards,DrugBank,DisGeNET,OMIM,and TTD databases,and drug-disease intersecting targets were identified.The“drug-active ingredient-target”network and protein-protein interaction(PPI)network were constructed using Cytoscape 3.9.1 software to further screen core active ingredients and targets.Gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analyses of the intersecting targets were performed by using the Metascape database.Molecular docking was conducted to verify the core active ingredients and targets of DaChaiHu Decoction in the treatment of NAFLD.A high-fat diet-induced NAFLD mouse model was established,and the network pharmacology prediction results were validated through animal experiments.Results:Network pharmacology predicted 90 potential target genes for DaChaiHu Decoction in the treatment of NAFLD,including TNF,IL6,IL1B,TP53,and PPARG.The core active ingredients identified were protoporphyrin,(E)-11-gondoic acid,10,13-eicosadienoic acid,epiberberine,mauritine D,and kaempferol.GO function enrichment analysis showed significant enrichment in biological processes such as response to bacterial molecules,positive regulation of small molecule metabolic processes,and regulation of secretion,in cellular components such as membrane rafts,receptor complexes,and cytoplasmic perinuclear regions,and in molecular functions such as nuclear receptor activity,monocarboxylic acid binding,and protein domain-specific binding.KEGG pathway enrichment analysis indicated that DaChaiHu Decoction exerted anti-NAFLD effects by modulating peroxisome proliferator-activated receptor(PPAR)and other signalling pathways.Molecular docking results demonstrated that the core active ingredients had strong binding activity with the core targets.Animal experiments showed that DaChaiHu Decoction effectively reduced lipid levels,oxidative stress,and inflammatory factors in NAFLD mouse models.It decreased serum contents of TG,TC,and LDL-C(P<0.05 or P<0.01),serum AST and ALT activities(P<0.05 or P<0.01),liver MDA level(P<0.01),and contents of TNF-α,IL-6,and IL-1β(P<0.05 or P<0.01)while increasing serum HDL-C levels(P<0.05)and liver SOD activity(P<0.01).HE and Oil Red O staining results indicated that DaChaiHu Decoction effectively alleviated hepatic fatty degeneration,vacuolar lesions,and red lipid droplets in NAFLD mouse models.RT-qPCR results showed that DaChaiHu Decoction significantly down-regulated the mRNA expression levels of Tnfα,Il6,and Il1βin liver(P<0.05 or P<0.01)and up-regulated the mRNA expression levels of Pparγ,Akt1,and Ampk(P<0.05).Conclusion:DaChaiHu Decoction effectively reduces lipid deposition,oxidative stress,and inflammatory responses in NAFLD mice,improves liver function,and alleviates pathological damage.Its mechanism of action may be related to down-regulating the levels of TNF-α,IL-6,and IL-1βand regulating the PPARγsignalling pathway.
作者
钟光成
刘启华
麦灏铭
潘亚茹
李舒婷
杨虹颖
谢怀德
杨从
王奇
ZHONG Guangcheng;LIU Qihua;MAI Haoming;PAN Yaru;LI Shuting;YANG Hongying;XIE Huaide;YANG Cong;WANG Qi(Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510405;Nanfang Hospital,Southern Medical University,Guangzhou 510515;The First Affiliated Hospital of Guangxi Medical University,Nanning 530021)
出处
《中药药理与临床》
北大核心
2025年第3期2-9,共8页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金面上项目(编号:82274616)
广东省基础与应用基础研究基金企业联合基金(编号:2022A1515220121)。
作者简介
通信作者:王奇,教授,博士生导师,研究方向:老年脑病的临床与实验研究,E-mail:wangqi@gzucm.edu.cn;钟光成,在读博士,研究方向:老年脑病的临床与实验研究,E-mail:1494470737@qq.com。
