Wu Lizheng, Lin Shuhong, Hu Yongjun, Jing Shangwen, Sun Bowen, Chen Xiaoxin, Jia Jinjin, Zeng Cheng, Pei Fangli
Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Department of Gynecology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Front Pharmacol. 2024 Jul 29;15:1395160. doi: 10.3389/fphar.2024.1395160. eCollection 2024.
Endometriosis (EMs) is characterized by ectopic growth of active endometrial tissue outside the uterus. The Luoshi Neiyi prescription (LSNYP) has been extensively used for treating EMs in China. However, data on the active chemical components of LSNYP are insufficient, and its pharmacological mechanism in EMs treatment remains unclear. This study aimed to explore the potential mechanism of LSNYP for EMs through network pharmacology based on the components absorbed into the blood.
Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used to analyze blood components, and a series of network pharmacology strategies were utilized to predict targets of these components and EMs. Protein-protein interaction (PPI) network analysis, component-target-disease network construction, gene ontology (GO) functional enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Additionally, molecular docking, molecular dynamics simulations, and and experiments were conducted to validate the HIF1A/EZH2/ANTXR2 pathway associated with hypoxic pathology in EMs.
Thirty-four absorbed components suitable for network pharmacology analysis were identified, and core targets, such as interleukin 6, EGFR, HIF1A, and EZH2, were founded. Enrichment results indicated that treatment of EMs with LSNYP may involve the regulation of hypoxia and inflammatory-related signaling pathways and response to oxidative stress and transcription factor activity. Experimental results demonstrated that LSNYP could decrease the expression of HIF1A, ANTXR2, YAP1, CD44, and β-catenin, and increased EZH2 expression in ectopic endometrial stromal cells and endometriotic tissues. Molecular docking and molecular dynamics simulations manifested that there was stable combinatorial activity between core components and key targets of the HIF1A/EZH2/ANTXR2 pathway.
LSNYP may exert pharmacological effects on EMs via the HIF1A/EZH2/ANTXR2 pathway; hence, it is a natural herb-related therapy for EMs.
子宫内膜异位症（EMs）的特征是子宫外出现活跃的子宫内膜组织异位生长。在中国，洛氏内异方（LSNYP）已被广泛用于治疗EMs。然而，关于LSNYP活性化学成分的数据不足，其治疗EMs的药理机制仍不清楚。本研究旨在通过基于血液中吸收成分的网络药理学探索LSNYP治疗EMs的潜在机制。
采用超高效液相色谱-四极杆飞行时间质谱分析血液成分，并利用一系列网络药理学策略预测这些成分和EMs的靶点。进行蛋白质-蛋白质相互作用（PPI）网络分析、成分-靶点-疾病网络构建、基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析。此外，进行分子对接、分子动力学模拟以及实验以验证与EMs低氧病理相关的HIF1A/EZH2/ANTXR2通路。
鉴定出34种适合网络药理学分析的吸收成分，并确定了白细胞介素6、表皮生长因子受体（EGFR）、缺氧诱导因子1α（HIF1A）和EZH2等核心靶点。富集结果表明，LSNYP治疗EMs可能涉及低氧和炎症相关信号通路的调节以及对氧化应激和转录因子活性的反应。实验结果表明，LSNYP可降低异位子宫内膜基质细胞和子宫内膜异位组织中HIF1A、血管生成素样蛋白2（ANTXR2）、Yes相关蛋白1（YAP1）、CD44和β-连环蛋白的表达，并增加EZH2的表达。分子对接和分子动力学模拟表明，核心成分与HIF1A/EZH2/ANTXR2通路的关键靶点之间存在稳定的组合活性。
LSNYP可能通过HIF1A/EZH2/ANTXR2通路对EMs发挥药理作用；因此，它是一种治疗EMs的天然草药相关疗法。
