目的:探讨小续命汤治疗中风的药效物质及作用机制.方法:基于UHPLC-Q Exactive Plus HRMS鉴定小续命汤中的成分,网络药理学预测小续命汤治疗中风的活性成分及潜在机制,鹌鹑胚绒毛尿囊膜(qCAM)实验可视化小续命汤的促血管生成作用.结果:从小续命汤中鉴别出麻黄碱、升麻素苷和黄芩苷等68个成分,包括氨基酸类、生物碱类、黄酮类、香豆素类、萜类、丁基苯肽类等化合物;成分与中风的交集靶点建立了蛋白质-蛋白质相互作用(PPI)网络,共筛选出IL-6、TNF、EGFR、PDGFRB和PIK3CA等33个核心靶点;麻黄碱、黄芩苷和人参皂苷Rg1等57种活性成分;核心靶点的富集分析表明小续命汤治疗中风可能主要通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K-AKT)和血管生成因子(VEGF)信号通路等与血管生成相关的通路.qCAM实验中阳性对照组(Gas,0.02 mg)和小续命汤高剂量组(XXMD-H,2 mg,折合生药量16.69 mg)的血管相对面积比空白组显著增加(P＜0.05),较好地可视化了小续命汤促进血管生成.结论:本研究通过结合UHPLC-Q Exactive Plus HRMS和网络药理学的方法阐明了小续命汤治疗中风的药效物质,得出潜在作用机制可能与作用于PI3K-AKT、VEGF等信号通路促进血管生成有关,并通过qCAM实验评价其促进血管生成活性,RT-qPCR进一步验证相关靶点,为小续命汤的质量研究与进一步开发提供参考.
Pharmacodynamic Substances and Mechanism of Xiaoxuming Decoction in the Treatment of Stroke Based on Network Pharmacology and Experimental Verification
Objective:To explore the effective substances and mechanism of Xiaoxuming Decoction(XXMD)in the treatment of stroke.Methods:UHPLC-Q Exactive Plus HRMS was used to identify the components of XXMD.Network pharmacology was em-ployed to predict the active components and potential mechanism of XXMD in treating stroke.The quail embryo chorioallantoic membrane(qCAM)experiment was used to visualize the angiogenic effect of XXMD.Results:Sixty-eight components were identi-fied from XXMD,including amino acids,alkaloids,flavonoids,coumarins,terpenes,and butylphenyl peptides,such as ephedrine,cimicifugoside,and baicalin.A protein-protein interaction(PPI)network was constructed based on intersection targets of compo-nents and stroke,with 33 core targets identified,including IL-6,TNF,EGFR,PDGFRB,and PIK3CA.Fifty-seven active components were identified,including ephedrine,baicalin,and ginsenoside Rg1.Enrichment analysis of the core targets suggested that the treat-ment of stroke by XXMD may primarily involve pathways related to angiogenesis,such as the phosphatidylinositol 3-kinase/protein kinase B(PI3K-AKT)and vascular endothelial growth factor(VEGF)signaling pathways.In the qCAM experiment,the positive control group(Gas,0.02 mg)and the high-dose XXMD group(XXMD-H,2 mg,equivalent to 16.69 mg of crude drug)showed a significant increase in the relative vascular area compared to the blank group(P＜0.05),effectively visualizing the angiogenic effect of XXMD.Conclusion:This study elucidated the effective substances of XXMD in treating stroke by combining UHPLC-Q Exactive Plus HRMS and network pharmacology methods.The potential mechanism may be related to promoting angiogenesis through pathways involving PI3K-AKT and VEGF.The qCAM experiment further evaluated its angiogenic activity,and RT-qPCR confirmed related targets,providing a reference for the quality research and further development of XXMD.
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