【目的】 基于网络药理学和转录组学探讨内异膏治疗子宫内膜异位症的作用机制。【方法】 采用自体移植法复制子宫内 膜异位症SD大鼠模型。将造模成功的大鼠随机分为模型组，内异膏低、中、高剂量组，地诺孕素组，每组10只。另设假手 术组（10只大鼠）。连续 4周干预后，测量大鼠异位病灶体积大小，采用苏木素-伊红（HE）染色法观察大鼠病灶体积病理变 化。提取大鼠病灶RNA进行转录组学测序，并对组间差异基因进行基因本体论（GO）富集分析及京都基因与基因组百科全书 （KEGG）功能富集分析。在TCMSP等数据库收集并筛选内异膏的主要活性成分及其靶点，通过OMIM等数据库收集子宫内膜 异位症疾病靶点，利用Venn图获得药物成分靶点与疾病之间的交集靶点，利用基因富集分析在线工具（Metascape）对药物成 分与疾病的交集靶点进行基因的GO富集分析和KEGG通路富集分析。将网络药理学富集通路与转录组学富集通路进行交集， 采用实时定量聚合酶链反应（qPCR）法和Western Blot法分别对应检测交集通路上关键靶点的mRNA、蛋白表达。【结果】 与假 手术组比较，模型组病灶体积显著增大（P < 0.01）；与模型组比较，给药组大鼠病灶体积显著缩小（P < 0.05）。共得到内异膏 活性成分 341种，子宫内膜异位症疾病相关靶点 2 178个，药物靶点与疾病靶点交集为 278个；假手术组与模型组共筛选出 189个差异基因；模型组与内异膏高剂量组共筛选出255个差异基因；假手术组与内异膏高剂量组共筛选出740个差异基因， 其中上调基因390个，下调基因350个。网络药理学与转录组学KEGG富集通路取交集，主要包括P53信号通路、FOXO信号 通路、细胞周期通路等。qPCR及Western Blot验证结果显示内异膏可以抑制子宫内膜异位症大鼠子宫内膜间质细胞增殖，上 调子宫内膜异位症大鼠 BAX、Caspase3 的 mRNA、蛋白表达水平（P < 0.05 或 P < 0.01），下调 BCL-2 mRNA、蛋白表达水平 （P < 0.05或P < 0.01）。【结论】 内异膏可能通过调控P53信号通路发挥治疗子宫内膜异位症的作用。

Objective To explore the mechanism of action of Neiyi Soft Extract in the treatment of endometriosis （EMS）based on network pharmacology and transcriptomics. Methods A model of SD rat with EMS was replicated by autotransplantation method. After successful modeling，the rats were randomly divided into model group，Neiyi Soft Extract low -，medium - and high - dose groups and dienogest group，with 10 rats in each group. Another sham-operated group（10 rats）was set up. After four consecutive weeks of intervention，the volume size of the endometriotic lesions was measured， and its pathological changes were detected by hematoxylin-eosin （HE） staining. RNA was extracted from the rat lesions for transcriptomic sequencing， and Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG）functional enrichment analysis were performed on the differential genes among various groups. The main active ingredients and their targets of Neiyi Soft Extract were collected and screened in databases such as TCMSP， the disease targets of EMS were collected through databases such as OMIM， the intersection targets between the drug ingredient targets and the diseases were obtained by using Venn diagrams，GO enrichment analysis and KEGG pathway enrichment analysis of genes in the intersection targets of the drug ingredients and the diseases were performed by using gene enrichment analysis online tool（Metascape）. The network pharmacology enrichment pathway and transcriptomics enrichment pathway were taken for intersection，the mRNA and protein expression levels of the key targets on the intersection pathway were correspondingly detected by real-time quantitative polymerase chain reaction（qPCR）and Western Blot，respectively. Results The volume of lesions in the model group was significantly increased compared with that of the sham-operated group（P < 0.01）；the volume of lesions in rats in the drug-administered groups was significantly reduced compared with that of the model group（P < 0.05）. A total of 341 active ingredients were obtained from Neiyi Soft Extract，and there were 2 178 disease-related targets of EMS，and 278 intersections between drug targets and disease targets；189 differential genes were screened in the sham-operated group and the model group；255 differential genes were screened in the model group and Neiyi Soft Extract high-dose group； and 740 differential genes were screened in the sham-operated group and the Neiyi Soft Extract high-dose group， including 390 up-regulated genes and 350 down-regulated genes. The result of intersection of KEGG enrichment pathways between the network pharmacology and transcriptomics showed that the distriution mainly included P53 signaling pathway， FOXO signaling pathway， cell cycle， etc. The qPCR and Western Blot validation results showed that Neiyi Soft Extract could inhibit the proliferation of endometrial stromal cells and up-regulated the mRNA and protein expression levels of BAX，Caspase3 in EMS rats（P < 0.05 or P < 0.01），and down-regulated BCL-2 mRNA and protein expression levels（P < 0.05 or P < 0.01）. Conclusion Neiyi Soft Extract may play a therapeutic role in the treatment of EMS by regulating the P53 signaling pathway.

R285.5

广东省基础与应用基础研究基金项目（编号：2022A1515220178）；佛山市科技创新项目（编号：2016AG100531）；佛山市科技 创新项目（编号：2017AG100211）；邬素珍广东省名中医传承工作室