基于网络药理学及实验探讨两性霉素B导致肝、肾损伤及过敏机制研究

Study on the Mechanism of Liver and Kidney Injury and Allergies Induced by Amphotericin B Based on Network Pharmacology and Experimen

关键词:  两性霉素B  肝、肾损伤  过敏  网络药理学  实验验证

Key words:  Amphotericin B  Liver and kidney injury  Allergies  Network pharmacology  Experimental verification

摘要:

目的 通过网络药理学、细胞实验和动物实验初步探讨两性霉素B(AmB)导致肝、肾损伤及过敏的机制。方法 对我院使用AmB的患者进行生化指标分析,利用ProTox-II进行AmB毒性预测,通过Pharm Mapper、CTD和STP数据库预测其潜在作用靶点。利用DAVID数据库,通过GO、KEGG通路富集分析AmB可能的干预通路机制。利用DisGeNET及OMIM数据库进行相关靶点检索,将潜在靶点和疾病靶点进行交集分析。使用STRING 11.0分析交集靶点的PPI网络,再通过Cytoscape 3.8.0软件进行网络拓扑分析。经不同浓度AmB处理的LAD2细胞进行β-氨基已糖苷酶、组胺、TNF-α、MCP-1、IL-4的测定。建立AmB肝、肾损伤小鼠模型,记录小鼠体重变化,取小鼠肝脏、肾脏计算脏器指数并进行病理学切片观察。结果 临床数据分析显示,使用AmB后多个生化指标发生显著变化,表明了其对肝肾功能有影响。此外,部分患者首次使用时出现寒战、发热等过敏样反应,表明其可能机制与I型过敏反应不同。通过网络药理学分析,推测肾损伤和过敏反应可能与细胞炎症及肥大细胞刺激相关。体内外实验结果显示,AmB能刺激LAD2细胞脱颗粒并释放细胞因子,同时对小鼠肝肾组织造成损伤,支持其可能引发肝肾毒性及过敏反应的假设。结论 该研究为AmB的安全性和不良反应机制提供了新的视角,强调了未来研究的必要性。

Abstract:

OBJECTIVE To investigate the mechanism of liver and kidney injury and allergies induced by Amphotericin B (AmB) through network pharmacology and animal experiments. METHODS Biochemical indicator analysis of patients using AmB in our hospital. Toxicity predictions for AmB were made using the ProTox II platform, while potential targets were predicted through the PharmMapper, CTD, and STP databases. Using the DAVID database, potential intervention mechanisms of AmB were analyzed through GO and KEGG pathway enrichment. Target retrieval was performed using the DisGeNET and OMIM databases. Intersection analysis was conducted to identify overlapping targets between potential AmB targets and known disease-related targets. The STRING 11.0 tool was used to analyze the PPI (Protein-Protein Interaction) network of the intersecting targets. Network topology analysis was subsequently performed using Cytoscape 3.8.0 software. LAD2 cells treated with varying concentrations of AmB were analyzed for the levels of β-aminoglucosidase, histamine, TNF-α, MCP-1, and IL-4. A mouse model of AmB-induced liver and kidney injury was established. Changes in mouse body weight were recorded, organ indices were calculated from liver and kidney samples, and pathological sections were observed. RESULTS Clinical data analysis revealed significant alterations in several biochemical markers, such as alanine aminotransferase and blood creatinine levels, before and after AmB administration, indicating its impact on liver and kidney function. Additionally, allergic reactions, including chills and fever, were observed in some patients during initial treatment, suggesting that the underlying mechanism may differ from typical type I allergic reactions. Network pharmacological analysis further hypothesizes that renal injury and allergic reactions may be associated with cellular inflammation and mast cell activation. In vivo and in vitro experiments demonstrated that AmB stimulates LAD2 cells to degranulate and release cytokines, while also causing liver and kidney damage in mice. These findings support the hypothesis that AmB induces both liver and kidney toxicity as well as allergic reactions. CONCLUSION This study provides new insights into the safety profile and underlying mechanisms of adverse reactions associated with AmB, emphasizing the need for further investigation.