目的 通过超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF-MS）技术结合网络药理学研究消瘀散精简方新组方贴膏（简称新组方贴膏）治疗骨关节炎的药效基础与作用靶点。方法 以 UPLC-Q-TOF-MS 技术结合文献检索获取消瘀散精简方新组方贴膏的活性成分信息，使用 SwissTargetPrediction 和 Similarity ensemble approach 数据库进行作用靶点预测；利用 GeneCards 和 OMIM 数据库检索与骨关节炎相关的疾病靶点，并与活性成分作用靶点进行合并去重，通过 Cytoscape 3.10.2 软件构建“活性成分-有效靶点”网络；将交集靶点导入STRING 数据库构建蛋白互作网络；将交集靶点用 R（4.2.1）语言进行 GO 富集分析和 KEGG 通路富集分析；采用 CB-dock2 在线分析软件进行关键成分与有效靶点的分子对接研究。结果 通过应用 UPLC-Q-TOF-MS 技术，从消瘀散精简方新组方贴膏中共鉴别出 54 个活性成分，与骨关节炎交集靶点 332 个。预测消瘀散精简方新组方贴膏可能通过菊苣酸、迷迭香酸、咖啡酸、大黄酸、壬二酸、秦皮乙素、藁本内酯、新隐丹参酮和阿魏酸等主要活性成分，作用于蛋白酪氨酸磷酸酶 11（PTPN11）、磷脂酰肌醇 3-激酶调节亚基 1（PIK3R1）、磷脂酰肌醇-4，5-二磷酸 3-激酶催化亚基 α（PIK3CA）、表皮生长因子受体（EGFR）、Janus 酪氨酸激酶 2（JAK2）等关键靶点，调控肿瘤坏死因子（TNF）、磷酯酰肌醇 3 激酶（PI3K）-蛋白激酶 B（Akt）、表皮生长因子受体（EGFR）酪氨酸激酶抑制剂耐药等信号通路发挥免疫调节与抗炎镇痛作用；其中 PIK3CA 与咖啡酸（Caffeic acid）结合能最高，为-9.7 kJ•mol-1 ，表明所筛选的活性成分与核心靶点之间具有较好的结合亲和力。结论 消瘀散精简方新 组方贴膏可能通过菊苣酸、迷迭香酸、咖啡酸、大黄酸、壬二酸、秦皮乙素、藁本内酯、新隐丹参酮和阿魏酸等主要活性成分，作用于 PTPN11、PIK3R1、PIK3CA、EGFR、JAK2 等关键靶点，调控 TNF、PI3K-Akt、EGFR 等信号通路发挥免疫调节与抗炎镇痛作用。

Objective To investigate the pharmacodynamic basis and targets of the new formula patch of simplify Xiaoyusan for treating osteoarthritis by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS） coupled with network pharmacology. Methods The active ingredients of the new formula patch of simplify Xiaoyusan were obtained by UPLC-Q-TOF-MS combined with literature search， and the SwissTargetPrediction and Similarity ensemble approach databases were used for target prediction. GeneCards and OMIM databases were used to search for disease targets related to osteoarthritis， which were merged with targets of active ingredients and removed duplicates. The “active ingredients-effective targets” network was constructed by Cytoscape 3.10.2 software. The intersected targets were imported into STRING database to construct the protein interactions network， the intersected targets were further subjected to GO enrichment analysis and KEGG pathway enrichment by using the R （4.2.1） language. The molecular docking of key components and effective targets was carried out by using CB-dock2 online analysis software. Results A total of 54 active ingredients were identified from the new formula patch of simplify Xiaoyusan by applying UPLC-Q-TOF-MS technology， with 332 targets intersecting with osteoarthritis. It was predicted that the main components from the new formula patch of simplify Xiaoyusan，such as chicoric acid，rosmarinic acid，caffeic acid，rhubarb acid，azelaic acid，esculetin，ligustilide，neocryptotanshinone， and ferulic acid might act on key targets，including protein tyrosine phosphatase 11 （PTPN11），phosphoinositide 3- kinase regulatory subunit 1 （PIK3R1）， phosphatidylinositol-4，5-bisphosphate 3-kinase （PI3K） catalytic subunit alpha （PIK3CA）， epidermal growth factor receptor （EGFR） and Janus kinase 2 （JAK2）， and display immunomodulatory， anti-inflammatory and analgesic effects through regulating TNF， PI3K-Akt， EGFR tyrosine kinase inhibitor resistance and other signaling pathways. The screened potential active components showed good binding affinity with the core target. The binding energy of PIK3CA and caffeic acid （-9.7 kJ•mol-1 ） was the highest. Conclusion The study predicted that the main components （chicoric acid，rosmarinic acid，caffeic acid，rhubarb acid， azelaic acid， esculetin， ligustilide， neocryptotanshinone， and ferulic acid） from the new formula patch of simplify Xiaoyusan might act on core targets （PTPN11， PIK3R1， PIK3CA， EGFR， JAK2） via regulating TNF， PI3K-Akt，EGFR signaling pathways to play an immunomodulatory，anti-inflammatory andanalgesic effects.

R285.5

上海市科委项目（20S21900200）；上海市卫健委项目[2022QN050，ZY（2021-2023）-03-02-25，2024002]