目的 基于中药复方专利数据库，整合专利挖掘、时空多组学及计算模拟技术系统筛选缺血性脑卒中的相关核心药组及候选作用靶点，阐释中药复方“通腑醒神”治则的科学内涵。方法 检索国家知识产权局专利数据库，采用聚类分析筛选出核心药组并通过网络药理学及分子对接、动力学模拟筛选出中药复方治疗缺血性脑卒中的候选作用靶点，结合单细胞RNA测序、空间转录组及虚拟敲降分析进行多维度关联分析。结果 筛选出远志、石菖蒲、大黄、郁金候选核心药组并获得1-羟基菖蒲烯酮、马兜铃酮、菖蒲二醇、β-细辛醚、α-细辛醚、山柰酚、百里香酚、丁香酚及咖啡酸9个代表性活性成分和肿瘤坏死因子（tumor necrosis factor，TNF）、白细胞介素-6（interleukin-6，IL-6）、蛋白激酶B1（protein kinase B1，AKT1）、表皮生长因子受体（epidermal growth factor receptor，EGFR）、丝裂原活化蛋白激酶1（mitogen-activated protein kinase 1，MAPK1）、前列腺素内过氧化物合酶2（prostaglandin-endoperoxide synthase 2，PTGS2）共6个候选作用靶点，分子对接及分子动力模拟提示部分代表性活性成分与候选作用蛋白之间具有结合倾向，其中山柰酚-PTGS2和山柰酚-EGFR复合物在模拟条件下表现出一定稳定性；进一步通过单细胞与空间转录组分析显示，候选靶点高表达区域与巨噬细胞与星形胶质细胞等细胞群的空间分布具有一定一致性；并且CellOracle结果提示，Jun/Fos、信号转导和转录激活因子3（signal transducer and activator of transcription 3，STAT3）等相关转录因子可能参与缺血后炎症应答、细胞状态转换及修复相关调控过程。结论 从国家专利中药复方筛选出远志、石菖蒲、大黄、郁金核心药组，并提出PTGS2、IL-6及EGFR等可能是缺血性脑卒中炎症微环境调控中的重要候选靶点，研究结果初步构建了核心药组、核心成分、候选靶点及时空验证相结合的机制证据链，并从炎症微环境调控、细胞通讯和神经血管重塑角度为通腑醒神治则治疗缺血性脑卒中的现代机制解释及后续实验验证提供参考。

Objective To systematically identify the candidate core herb combination and potential therapeutic targets for ischemic stroke by integrating patent mining, spatiotemporal multi-omics analysis, and computational simulation, and elucidate the scientific connotation of the traditional Chinese medicine therapeutic principle of “Tongfu Xingshen” based on the patent database of traditional Chinese medicine compound prescriptions. Methods The China National Intellectual Property Administration patent database was searched. Cluster analysis was used to screen the candidate core herb combination. Network pharmacology, molecular docking, and molecular dynamics simulations were then performed to identify and evaluate candidate therapeutic targets of traditional Chinese medicine compound prescriptions for ischemic stroke. Single-cell RNA sequencing, spatial transcriptomics, and CellOracle-based in silico knockdown analysis were further integrated to perform multidimensional association analysis. Results A candidate core herb combination consisting of Yuanzhi (Polygalae Radix), Shichangpu (Acori Tatarinowii Rhizoma), Dahuang (Rhei Radix et Rhizoma), and Yujin (Curcumae Radix) was identified. Nine representative active components, including 1-hydroxyacoronene, aristolone, calamendiol, β-asarone, α-asarone, kaempferol, thymol, eugenol, and caffeic acid, and six candidate therapeutic targets, including tumor necrosis factor (TNF), interleukin-6 (IL-6), protein kinase B1 (AKT1), epidermal growth factor receptor (EGFR), mitogen-activated protein kinase 1 (MAPK1), and prostaglandin-endoperoxide synthase 2 (PTGS2), were obtained. Molecular docking and molecular dynamics simulations suggested that some representative active components exhibited certain binding tendencies with the candidate target proteins. Among them, the kaempferol-PTGS2 and kaempferol-EGFR complexes showed a degree of stability under simulation conditions. Single-cell and spatial transcriptomic analyses further showed that the regions with high expression of candidate targets were spatially consistent with the distribution of macrophages, astrocytes, and other cell populations. CellOracle analysis suggested that Jun/Fos, signal transducer and activator of transcription 3 (STAT3), and other related transcription factors may be involved in post-ischemic inflammatory responses, cell-state transitions, and repair-related regulatory processes. Conclusion This study identified a candidate core herb combination consisting of Polygalae Radix, Acori Tatarinowii Rhizoma, Rhei Radix et Rhizoma, and Curcumae Radix from national patent traditional Chinese medicine compound prescriptions, and proposed that PTGS2, IL-6, EGFR, and related targets may serve as important candidate targets in the regulation of the inflammatory microenvironment in ischemic stroke. These findings preliminarily construct a mechanistic evidence chain integrating the core herb combination, core components, candidate targets, and spatiotemporal validation, and provide a reference for the modern mechanism explanation and subsequent experimental verification of the “Tongfu Xingshen” therapeutic principle in ischemic stroke from the perspectives of inflammatory microenvironment regulation, cell-cell communication, and neurovascular remodeling.

R285

北京市属医院科研培育计划（PZ2024007）