通经活络精油治疗类风湿性关节炎的网络药理机制及工艺研究  

Mechanism of Tongjing Huoluo Essential Oil in Treating Rheumatoid Arthritis Based on Network Pharmacology and Extraction Optimization

作　　者：张琪 李铁[1] 赵晋莹[1] 彭锐 刘武[2,3] 张楠 王富春[1] ZHANG Qi;LI Tie;ZHAO Jinying;PENG Rui;LIU Wu;ZHANG Nan;WANG Fuchun(Changchun University of Chinese Medicine,Changchun 130117,China;Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430065,China)

机构地区：[1]长春中医药大学,长春130117 [2]湖北中医药大学,武汉430065 [3]湖北时珍实验室,武汉430065

出　　处：《世界中医药》2024年第22期3420-3430,共11页World Chinese Medicine

基　　金：国家自然科学基金项目(82374600);湖北省中医药管理局项目(ZY2023M067);湖北省自然科学基金项目(2024AFD292);第七批吉林省青年科技人才托举工程项目(QT202319);吉林省发改委产业技术研究与开发项目(2022C42-3)。

摘　　要：目的:运用网络药理学和分子对接技术探讨通经活络方治疗类风湿性关节炎(RA)的作用机制,优化其挥发油组分的提取工艺,并验证药理作用。方法:利用中药系统药理学分析平台(TCMSP)数据库收集通经活络方成分,构建活性成分-靶点网络;筛选RA疾病靶点,架构蛋白互作网络;确定潜在治疗靶点,并进行基因本体(GO)及京都基因和基因组百科全书(KEGG)富集分析;构建“药物-活性成分-靶点-通路”网络,解析通经活络方治疗RA的核心成分和靶点并进行分子对接验证。采用单因素实验确定考察因素及范围,运用Box-Behnken响应面法以通经活络精油得率为响应指标,优化提取工艺。通过1,1-二苯基-2-三硝基苯肼(DPPH)、2,2-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS+)自由基清除法测定通经活络精油抗氧化能力。结果:从通经活络方中筛选出182个活性成分,作用于215个靶点,其中通经活络方改善RA的核心活性成分为槲皮素、芹菜素、木犀草素、山柰酚、杨梅素,核心靶点为前列腺素内过氧化物合酶2(PTGS2)、核转录因子-κB(RELA)、AKT丝氨酸/苏氨酸激酶(AKT1)、热休克蛋白90AB1(HSP90AB1)、胱天蛋白酶-3(CASP3);涉及氧化应激、炎症、糖脂代谢等多条信号通路;分子对接显示其核心活性成分与核心靶点具有高亲和力。通经活络精油的最佳提取工艺为粉碎粒度60目,液料比10 mL/g,提取时间3.5 h,提取温度110℃,得率可达0.483%。此条件下浓度为200μg/mL的通经活络精油具有较高氧化清除活性。结论:通经活络方可通过多成分、多通路、多靶点协同作用,发挥抗炎、抗氧化、抗细胞凋亡等药理机制,以改善RA症状。最优工艺条件下所得的通经活络精油经验证具有较高的体外抗氧化活性,值得进一步研究和利用。Objective:To explore the mechanism of action of Tongjing Huoluo Formula in treating rheumatoid arthritis(RA)using network pharmacology and molecular docking techniques,optimize the extraction process of its essential oil components,and verify its pharmacological effects.Methods:Active components of Tongjing Huoluo Formula were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and an active component-target network was constructed.RA disease targets were screened,and a protein-protein interaction(PPI)network was built.Potential therapeutic targets were identified,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.A“drug-active component-target-pathway”network was constructed to analyze the core components and targets of Tongjing Huoluo Formula in treating RA and verify them using molecular docking.Single-factor experiment was used to determine the factors and ranges for the extraction process.The Box-Behnken response surface method was employed to optimize the extraction process with essential oil yield as the response indicator.The antioxidant capacity of Tongjing Huoluo Essential Oil was measured using the 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)(ABTS+)radical scavenging methods.Results:A total of 182 active components were screened from Tongjing Huoluo Formula,interacting with 215 targets.The core active components that improved RA were quercetin,apigenin,luteolin,kaempferol,and myricetin.Core targets included prostaglandin-endoperoxide synthase 2(PTGS2),nuclear factor kappa B(RELA),AKT serine/threonine kinase 1(AKT1),heat shock protein 90AB1(HSP90AB1),and caspase-3(CASP3).The formula involved multiple signaling pathways,such as oxidative stress,inflammation,and glucose-lipid metabolism.Molecular docking revealed that the core active components had high affinity with the core targets.The optimal extraction process for Tongjing Huoluo Essential

关 键 词：通经活络精油 类风湿性关节炎 网络药理学 分子对接 响应面法 工艺优化 抗氧化 氧化应激 

分 类 号：R285[医药卫生—中药学]