基于网络药理学与代谢组学和分子对接的整合药理学策略研究温胆汤治疗慢性阻塞性肺疾病的作用机制

Research on the Mechanism of Wendan Decoction in Treating Chronic Obstructive Pulmonary Disease through an Integrated Pharmacological Strategy Based on Network Pharmacology,Metabolomics,and Molecular Docking

目的探讨温胆汤治疗慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)的作用机制。方法首先,基于网络药理学方法,利用中药系统药理学数据库与分析平台(TCMSP)、Swiss Target Prediction和Metascape等数据库,揭示温胆汤治疗COPD的潜在机制;其次,采用代谢组学方法,研究温胆汤对COPD诱发内源性代谢物紊乱的干预作用;最后,借助MetScape算法,整合网络药理学与代谢组学研究结果,并采用分子模拟对接和分子生物学方法,深入研究温胆汤治疗COPD的作用机制。结果网络药理学结果揭示温胆汤的102个潜在活性成分及其治疗COPD的98个潜在靶点。进一步分析显示温胆汤的11个潜在药效物质,包括3-叔丁基己二酸、6-甲基姜辣二醇双乙酸酯和甘草异黄烷酮等可能通过调节脂质与动脉粥样硬化、环磷腺苷(cyclic adenosine monophosphate,cAMP)信号通路、磷脂酰肌醇3激酶-蛋白激酶B(phosphoinositide 3-kinase-protein kinase B,PI3K-Akt)信号通路和白介素17(interleukin-17,IL-17)信号通路等发挥疗效。另一方面代谢组学研究显示,丙二醇、N-乙酰神经氨酸、苏氨酸、原胆酸、2-氧异戊酸酯、乙酰磷酸、柠檬酸盐和异丁酸等9种内源性物质,可能是温胆汤治疗COPD的关键代谢物。最后整合分析显示,脂质代谢与氨基酸代谢可能是温胆汤治疗COPD的关键过程,黄嘌呤脱氢酶(xanthine dehydrogenase,XDH)、激酶插入区受体(kinase insert domain receptor,KDR)、磷酸二酯酶5A(phosphodiesterase 5A,PDE5A)、细胞色素P450家族2亚家族C成员9(cytochrome P450 family 2 subfamily C member 9,CYP2C9)和成员19(cytochrome P450 family 2 subfamily C member 19,CYP2C19)等同氧化和炎症相关的基因可能是温胆汤发挥治疗作用的关键靶点;并且结果显示温胆汤可有效改善COPD大鼠肺部充血、水肿和炎性细胞浸润等多种病理状态,显著降低COPD大鼠肺组织中白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)含量和丙二醛(malonaldehyde,MDA)活性,提高超氧化物歧化酶(superoxide dismutase,SOD)活性;而且分子对接结果也揭示了潜在药效物质与核心靶点基因之间的关联性。结论基于代谢组学、网络药理学和分子对接的整合药理学研究表明温胆汤可能通过调控炎症反应、氧化应激、脂质代谢和氨基酸代谢等途径实现治疗COPD的作用。

OBJECTIVE To explore the mechanism of Wendan Decoction in the treatment of chronic obstructive pulmonary disease(COPD).METHODS Firstly,based on the network pharmacology method,the potential mechanism of Wendan Decoction in the treatment of COPD was revealed by using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Swiss Target Prediction and Metascape databases.Secondly,the intervention effect of Wendan Decoction on endogenous metabolite disorder induced by COPD was studied by metabolomics method.Finally,the results of network pharmacology and metabolomics were integrated through MetScape algorithm,and molecular simulation docking and molecular biology methods were used to study the mechanism of Wendan Decoction in the treatment of COPD.RESULTS The results of network pharmacology revealed 102 active components of Wendan Decoction and 98 potential targets for the treatment of COPD.Further analysis showed that 11 potential pharmacodynamic substances of Wendan Decoction,including 3-tert-butyladipic acid,6-methylgingediacetate and licoisoflavanone may play a role in regulating lipid and atherosclerosis,CAMP signaling pathway,PI3K-Akt signaling pathway and IL-17 signaling pathway.On the other hand,metabolomics studies have shown that 9 endogenous substances,such as 2-propanol,N-acetylneuraminic acid,threonine,protocholic acid,2-oxoisovalerate,acetylphosphate,citrate and isobutyric acid,may be the key metabolites of Wendan Decoction in the treatment of COPD.Finally,the integrated analysis showed that lipid metabolism and amino acid metabolism might be the key processes of Wendan Decoction in the treatment of COPD.XDH,KDR,PDE5 A,CYP2C9 and CYP2C19,which were related to oxidation and inflammation,were the potential key targets of Wendan Decoction in the treatment of COPD.The results showed that Wendan Decoction could effectively improve the pathological state of pulmonary congestion,edema and inflammatory cell infiltration in COPD rats,significantly reduce the content of IL-6,TNF-αand MDA activity in lung tissue of COPD rats,and increase SOD activity.Moreover,the molecular docking results also revealed the correlation between potential pharmacodynamic substances and core target genes.CONCLUSION The integrated pharmacological studies based on metabolomics,network pharmacology and molecular docking showed that Wendan Decoction might achieve the effect of treating COPD by regulating inflammatory response,oxidative stress,lipid metabolism and amino acid metabolism.