基于超高效液相色谱-串联质谱技术结合网络药理学研究四土汤的抗炎机制  

Anti-inflammatory Mechanism of Situ Decoction Based on UPLC-Q-TOF-MS Combined with Network Pharmacology

作　　者：郭奥奥 周贤[1,2] 何丽姗 田代志 李娟[1,2] 胡俊杰[1,2] 陈新 刘松林[2] GUO Aoao;ZHOU Xian;HE Lishan;TIAN Daizhi;LI Juan;HU Junjie;CHEN Xin;LIU Songlin(Hubei Key Laboratory of Resources and Chemistry of Chinese Medicine,School of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430075,China;Hubei Provincial Drug Administration Accreditation Centre,Wuhan 430071,China)

机构地区：[1]湖北中医药大学中药资源与中药化学湖北省重点实验室,武汉430065 [2]湖北时珍实验室,武汉430075 [3]湖北省药品监督管理局评审中心,武汉430071

出　　处：《世界中医药》2025年第2期198-210,共13页World Chinese Medicine

基　　金：国家自然科学基金项目(82004253);湖北省中医药重点学科建设项目(鄂中医通〔2023〕2号);湖北省自然科学基金项目(2023AFD146)。

摘　　要：目的:探讨四土汤中的抗炎活性成分及其作用机制。方法:采用超高效液相色谱-串联质谱(UPLC-Q-TOF-MS)技术检测四土汤中的化学成分。利用网络药理学筛选活性成分与核心靶点,预测四土汤抗炎作用机制。运用分子对接技术验证活性成分与核心靶点的结合活性,结合基因表达综合数据库(GEO)验证数据集绘制受试者工作特征曲线(ROC)评估核心靶点与炎症性疾病的相关性,进一步制备角叉菜胶致炎大鼠模型进行结果验证。结果:UPLC-Q-TOF-MS共鉴定出85个化学成分,主要包括黄酮类、蒽醌类和酚酸类等。网络药理学结果显示,四土汤能够通过槲皮素、木犀草素、大黄素、落新妇苷等活性成分作用于肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)等炎症靶点,调节磷脂酰肌醇3-激酶/蛋白激酶B(PI3K-AKT)、肿瘤坏死因子-α(TNF-α)、Toll样受体(Toll-like)等信号通路发挥抗炎作用。分子对接结果显示,13种活性成分与10个核心靶点的结合能均<-5 kcal/mol。GEO数据挖掘结果显示,核心靶点在3个炎症疾病数据集中均具有较高的ROC值。动物实验结果显示,与模型组比较,四土汤给药组大鼠足肿胀度显著性降低(P<0.05),足跖皮下组织病理损伤有所改善,足肿胀组织髓过氧化物酶(MPO)、一氧化氮(NO)含量显著性降低(P<0.01),大鼠血清中TNF-α、IL-1β、IL-6显著性降低(P<0.05),IL-10显著性升高(P<0.05)。结论:本研究初步验证了四土汤的抗炎药理活性,并揭示了可能的分子机制,为进一步全面阐释四土汤的药效物质基础及其药物开发提供了实验依据。Objective:To investigate the anti-inflammatory active components and mechanisms of Situ Decoction.Methods:Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was used to detect the chemical components of Situ Decoction.Network pharmacology was employed to screen active components and core targets,predicting the anti-inflammatory mechanisms of Situ Decoction.Molecular docking technology was used to verify the binding activity between active components and core targets.The Gene Expression Omnibus(GEO)database was utilized to generate receiver operating characteristic(ROC)curves to assess the correlation between core targets and inflammatory diseases.Furthermore,a carrageenan-induced rat inflammation model was established for experimental validation.Results:A total of 85 chemical components were identified via UPLC-Q-TOF-MS,mainly including flavonoids,anthraquinones,and phenolic acids.Network pharmacology analysis indicated that Situ Decoction exerted anti-inflammatory effects through active components such as quercetin,luteolin,emodin,and astilbin,targeting inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and interleukin-10(IL-10).These interactions regulated signaling pathways including phosphatidylinositol 3-kinase/protein kinase B(PI3K-AKT),TNF-α,and Toll-like receptor pathways.Molecular docking results showed that the binding energies of 13 active components with 10 core targets were all below-5 kcal/mol.GEO data mining revealed that core targets had high ROC values in three inflammatory disease datasets.In animal experiments,compared with the model group,Situ Decoction significantly reduced paw swelling in rats(P<0.05),alleviated histopathological damage in subcutaneous plantar tissue,and significantly decreased myeloperoxidase(MPO)and nitric oxide(NO)levels in swollen foot tissues(P<0.01).Additionally,serum levels of TNF-α,IL-1β,and IL-6 were significantly reduced(P<0.05),while IL-10 levels w

关 键 词：四土汤 抗炎 超高效液相色谱-串联质谱 药效物质基础 网络药理学 分子对接 基因表达综合数据库数据挖掘 大鼠足肿胀模型 

分 类 号：R285.5[医药卫生—中药学]