Objective: To explore the mechanism underlying the protective effect of Lonicerae japonicae flos (LJF) extract against doxorubicin (DOX) -induced liver injury in mice.
Methods: Network pharmacology methods were used to obtain the intersection genes between LJF targets and disease targets, based on which the protein-protein interaction (PPI) network was constructed using STRING database for screening the core targets using Cytoscape software. DAVID database was used for bioinformatics analysis, and the core components and core targets were verified using molecular docking study. In a mouse model of DOX-induced liver injury, the effect of LJF extract on liver pathologies, serum levels of ALT and AST, and hepatic expressions of HYP, ROS, TNF-α, IL-6, COL-Ⅳ and P53 proteins were evaluated using HE and Masson staining, ELISA, and Western blotting.
Results: We identified 12 core targets from 43 intersection genes involving cancer pathway, IL-17 signaling pathway, and TNF signaling pathways. Molecular docking study suggested that 10 core components of LJF could bind to different core targets. The mice with DOX-induced liver injury showed elevated serum AST and ALT levels with obvious liver injury and fibrosis, increased ROS content, and enhanced expressions of TNF-α, IL-6, HYP, COL-Ⅳ and P53 proteins in the liver tissue. All these changes in the mouse models were significantly alleviated by treatment with LJF extract, suggesting obviously lowered levels of oxidative stress, inflammation and fibrosis in the liver tissues.
Conclusion: LJF extract is capable of alleviating DOX-induced liver injury in mice by downregulating Trp53, TNF and IL-6 to reduce liver oxidative stress, inflammation and fibrosis.
目的: 通过网络药理学与分子对接技术探讨金银花对阿霉素（DOX）肝脏损伤的保护作用和机制，并运用DOX诱导小鼠肝脏损伤实验进行验证。
方法: 通过网络药理学方法获得金银花靶点与疾病靶点之间的交集基因。利用STRING数据库构建交集基因PPI网络，利用Cytoscape软件进行分析，筛选核心靶点。采用DAVID数据库进行生物信息学分析，分子对接技术对核心成分和核心靶点进行验证。运用DOX诱导小鼠肝脏损伤验证网络药理学的预测结果。检测小鼠血清ALT、AST水平和肝脏组织HYP、ROS水平，HE染色和Masson染色观察肝脏组织病理变化，ELISA检测肝脏组织TNF-α、IL-6、COL-IV水平，Western blotting检测肝脏组织P53蛋白表达水平。
结果: 从交集的43个基因中筛选出12个核心靶点，涉及癌症通路、IL-17信号通路、TNF等信号通路。分子对接结果显示，10个核心成分可以与不同的核心靶点结合。小鼠实验显示，与Sham组相比，DOX组血清AST和ALT水平升高（P<0.001）；HE和Masson染色显示肝脏损伤和肝脏纤维化，ROS、TNF-α、IL-6、HYP、COL-IV和P53蛋白水平升高（P<0.001）。与DOX组相比，金银花处理组血清AST和ALT水平降低（P<0.001），肝脏损伤和肝脏纤维化改善，肝脏组织ROS、TNF-α、IL-6、HYP和COL-IV水平和P53蛋白表达降低（P<0.001），肝脏组织氧化应激、炎症和纤维化均减轻。
结论: 金银花可通过作用于Trp53、TNF、IL-6靶点减轻肝脏氧化应激、炎症和纤维化程度，减轻DOX诱导的肝脏损伤。
Keywords: Lonicerae japonicae flos; doxorubicin; inflammation; liver damage; liver fibrosis; network pharmacology; oxidative stress.