基于网络药理学探讨广地龙蛋白对肌肉减少症的作用机制

Exploring the Mechanism of Action of Pheretima aspergillum Protein on Sarcopenia Based on Network Pharmacology

摘要: 目的：针对人口老龄化背景下肌肉减少症患病率持续攀升、现代医学缺乏特效靶向治疗药物的临床难题，本研究依托网络药理学与分子对接技术，系统探究广地龙蛋白干预肌肉减少症的潜在作用靶点与分子机制，为传统中药的现代化应用及肌少症防治提供理论依据。方法：本研究通过PubChem、Swiss Target Prediction、Uniprot等数据库，筛选广地龙蛋白中34种核心活性成分(包含有机酸类、氨基酸类、碱基核苷类、酶类)，并规范化收集成分对应作用靶点；利用GeneCards数据库检索肌肉减少症相关疾病靶点，采用Venny 2.1软件获取药物靶点与疾病靶点的交集。借助STRING数据库构建蛋白互作(PPI)网络，导入Cytoscape 3.10.3软件，结合CytoHubba、MCODE插件进行拓扑学分析，筛选核心作用靶点。运用AutoDock 4.2.6、PyMOL等软件开展分子对接验证，检测关键活性成分与核心靶点的结合活性。结果：本研究共筛选出广地龙蛋白干预肌肉减少症的交集靶点179个，经拓扑分析确定ESR1、IL6、EGFR、GAPDH、BCL2为五大核心靶点。分子对接结果显示，以结合能 < 0 kcal·mol⁻¹为稳定结合判定标准，广地龙蛋白内亮氨酸、赖氨酸、谷氨酸、次黄嘌呤、蚓激酶等关键成分，与核心靶点均能形成稳定结合，其中赖氨酸与EGFR结合能最低，为−9.1 kcal·mol⁻¹，结合活性最优。机制分析证实，广地龙蛋白可通过调控炎症反应、肌肉蛋白合成与分解、细胞凋亡、能量代谢、肌纤维再生等通路，发挥多成分、多靶点、多途径协同的肌肉保护作用，且具备药食同源、安全性高的应用优势。结论：本研究揭示了广地龙蛋白通过多成分、多靶点、多通路协同干预肌肉减少症的作用机制，验证了关键成分与核心靶点的强结合活性。广地龙蛋白药食同源、安全性佳，在肌肉减少症营养干预、功能食品研发领域极具应用潜力，也为中药防治老年性肌肉退行性疾病提供了科学依据与研究方向。

Abstract: Objective: In response to the clinical challenges posed by the rising prevalence of sarcopenia in the context of an aging population and the lack of specific targeted therapeutic drugs in modern medicine, this study relies on network pharmacology and molecular docking technology to systematically explore the potential targets and molecular mechanisms of Pheretima aspergillum protein intervention in sarcopenia, providing a theoretical basis for the modern application of traditional Chinese medicine and the prevention and treatment of sarcopenia. Methods: This study screened 34 core active components (including organic acids, amino acids, nucleobases, nucleosides, and enzymes) from Pheretima aspergillum protein through databases such as PubChem, Swiss Target Prediction, and Uniprot, and standardized the collection of corresponding targets for these components. The GeneCards database was used to retrieve disease-related targets associated with sarcopenia, and Venny 2.1 software was employed to obtain the intersection of drug targets and disease targets. The protein-protein interaction (PPI) network was constructed using the STRING database, imported into Cytoscape 3.10.3 software, and combined with CytoHubba and MCODE plugins for topological analysis to identify core targets. Molecular docking verification was conducted using AutoDock 4.2.6, PyMOL, and other software to detect the binding activity of key active components with core targets. Results: This study identified a total of 179 intersecting targets for Pheretima aspergillum protein intervention in sarcopenia. Through topological analysis, ESR1, IL6, EGFR, GAPDH, and BCL2 were determined as the five core targets. Molecular docking results showed that, using a binding energy < 0 kcal·mol⁻¹ as the criterion for stable binding, key components such as leucine, lysine, glutamic acid, hypoxanthine, and lumbrukinase in Pheretima aspergillum protein could form stable binding with core targets. Among them, lysine had the lowest binding energy with EGFR, at −9.1 kcal·mol⁻¹, indicating the best binding activity. Mechanism analysis confirmed that Pheretima aspergillum protein exerts a multi-component, multi-target, and multi-pathway synergistic muscle-protective effect by regulating pathways such as inflammation, muscle protein synthesis and decomposition, apoptosis, energy metabolism, and muscle fiber regeneration. It also possesses the application advantages of being both food and medicine, with high safety. Conclusion: This study reveals the mechanism of Pheretima aspergillum protein intervention in sarcopenia through multi-component, multi-target, and multi-pathway synergism, and verifies the strong binding activity of key components with core targets. Pheretima aspergillum protein, which is both food and medicine, boasts excellent safety. It holds great potential for application in nutritional intervention for sarcopenia and functional food research and development. Furthermore, it provides scientific evidence and research directions for traditional Chinese medicine in preventing and treating senile muscle degenerative diseases.