目的 结合血清药物化学和网络药理学，预测四妙勇安汤的潜在质量标志物（quality marker，Q-Marker），并在此结果基础上开展动物实验加以验证。方法 采用HPLC-Q-TOF/MS技术分析四妙勇安汤的入血成分，基于入血成分结合网络药理学预测主要作用靶点、通路，使用Cytoscape软件，构建“入血成分-复方功效-靶点通路”网络，依据拓扑学分析结果与四妙勇安汤主治热毒炽盛之脱疽的相关性，综合确定四妙勇安汤的潜在Q-Marker。在此基础上建立热毒炽盛之脱疽模型大鼠，造模成功后连续ig四妙勇安汤（57g/kg）2周，实验结束后计算各组大鼠创面愈合率，检测各组血清中丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）、γ干扰素（interferon-γ，IFN-γ）和血管内皮生长因子（vascular endothelialgrowth factor，VEGF）水平，并采用免疫组化法检测创面组织中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和白细胞介素-1β（interleukin-1β，IL-1β）蛋白表达。结果 鉴定四妙勇安汤20个入血成分，结合网络药理学筛选出四妙勇安汤治疗热毒炽盛之脱疽的42个关键功效靶点、62条相关通路，根据化合物、靶点以及信号通路之间的度值，初步预测木犀草素、异鼠李素、甘草素等9个功效相关的潜在入血成分并结合绿原酸等主要有效成分作为四妙勇安汤潜在的Q-Marker。动物实验结果表明，给予四妙勇安汤后可明显提高热毒炽盛之脱疽大鼠的创面愈合率（P＜0.05），显著回调血清中关键功效靶点（VEGF、MAPK、IFN-γ）水平（P＜0.05），明显降低创面组织中TNF信号通路相关蛋白TNF-α、IL-1β的表达（P＜0.05）。结论 通过血清药物化学结合网络药理学挖掘预测四妙勇安汤治疗热毒炽盛之脱疽的潜在Q-Marker，为整体控制和评价四妙勇安汤质量提供科学依据。动物实验初步证实了四妙勇安汤可通过调控VEGF、MAPK和IFN-γ等关键功效靶点蛋白以及TNF信号通路相关蛋白，进而发挥治疗热毒炽盛之脱疽的作用。

Objective To predict the potential quality marker (Q-Marker) of Simiao Yong’an Decoction (四妙勇安汤) by combining serum medicinal chemistry and network pharmacology, and based on these results, animal experiments were carried out to verify it. Methods HPLC-Q-TOF/MS was used to analyze the blood components of Simiao Yong’an Decoction. Based on the combination of components absorbed into blood and network pharmacology, main targets and pathways were predicted. Cytoscape software was used to construct “component absorbed into blood-efficacy of compound prescription-target pathway” network. Based on the correlation between the topological analysis results and traditional efficacy of Simiao Yong’an Decoction, potential Q-Markers of Simiao Yong’an Decoction were comprehensively determined. On this basis, rats model of gangrene due to excessive noxious heat was established. After successful modeling, rats were continuously ig Simiao Yong’an Decoction (57 g/kg) for 2 weeks. After the experiment, wound healing rate of rats in each group was calculated, levels of mitogen-activated protein kinase (MAPK), interferon-γ (IFN-γ) and vascular endothelial growth factor (VEGF) in serum of each group were detected, protein expressions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in wound tissue were detected by immunohistochemistry. Results Twenty components absorbed into blood of Simiao Yong’an Decoction were identified. Combined with network pharmacology, 42 key efficacy targets and 62 related pathways of Simiao Yong’an Decoction in treatment of gangrene due to excessive noxious heat were screened out. According to degree value between compounds, targets and signaling pathways, nine potential blood components related to efficacy such as luteolin, isorhamnetin, and liquiritigenin were preliminarily predicted, and combined with the main active components such as chlorogenic acid as Simiao Yong’an Decoction potential Q-Markers. The results of animal experiments showed that Simiao Yong’an Decoction significantly increased the wound healing rate of gangrene due to excessive noxious heat rats (P<0.05), significantly modulated the levels of key efficacy targets (VEGF, MAPK, IFN-γ) in serum (P<0.05), and significantly reduced TNF signaling pathway related protein (TNF-α, IL-1β) expressions in wounded tissues (P<0.05). Conclusion The potential Q-Marker of Simiao Yong’an Decoction in treating gangrene due to excessive noxious heat was predicted by combining serum pharmacochemistry with network pharmacology, which provided scientific basis for overall control and evaluation of Simiao Yong’an Decoction quality. Animal experiments preliminarily confirmed that Simiao Yong’an Decoction can regulate key functional target proteins such as VEGF, MAPK and IFN-γ, as well as proteins related to TNF signaling pathway, thereby play a role in treating gangrene due to excessive noxious heat.

R285.5

国家自然科学基金青年基金资助项目(81803742);四川省科技计划重点研发项目(2020YFS0567);四川省科技厅四川省科技计划项目(2021YJ0251);四川省中医药管理局中医药科研专项(2021MS109);四川省教育厅自然科学重点项目(18ZA0187);成都中医药大学杏林学者青年学者专项(QNXZ2019031);四川新绿色药业博士后科研专项(301022019)