Objective: To investigate the main chemical constituents of Balanophora involucrata and the mechanism of its antiinflammatory effect based on network pharmacology and molecular docking technology.
Methods: Literature reports, Materia Medica, GeneCards and other databases were searched for anti-inflammatory compounds and their targets. String database and Cytoscape 3.7.2 software were used to obtain the protein-protein interaction (PPI) network and the drug-active ingredienttargets network and for GO and KEGG enrichment analyses. Molecular docking was performed using Auto Dock Tools 1.5.6. In an inflammatory RAW264.7 cell model induced by lipopolysaccharide (LPS), the effect of 25, 50, 100, 200 μg/mL Balanophora involucrata extract was tested on the production of inflammatory cytokines and phosphorylation level of PI3K and Akt using ELISA and Western blotting.
Results: A total of 318 common targets of drugs and diseases were identified, and the core targets were Src, HSP90AA1 and PIK3CA, involving cancer, PI3K/Akt, MAPK and other signaling pathways as shown by KEGG analysis. Molecular docking showed that both the main active constituents of Balanophora involucrata could spontaneously bind to the core targets. In the inflammatory cell model, treatment with Balanophora involucrata extract significantly inhibited the production of IL-1β at the concentrations of 100 and 200 μg/mL, reduced IL-6 and TNF-α expressions at the concentrations of 50, 100, and 200 μg/mL, and lowered phosphorylation levels of PI3K and Akt proteins at the concentrations of 25, 50, 100, and 200 μg/mL (all P < 0.05).
Conclusions: The anti-inflammatory mechanism of Balanophora involucrata involves multiple targets and multiple pathways, and its effect is mediated possibly by reducing IL-1β, IL-6 and TNF-α production and inhibiting phosphorylation levels of PI3K and Akt proteins to suppress the activation of the PI3K/Akt signaling pathway.
目的: 基于网络药理学、分子对接技术及体外实验探讨文王一支笔抗炎作用的主要化学成分及潜在靶点的作用机制。
方法: 通过文献报道，本草组鉴、GeneCards等数据库获得化合物与抗炎的靶点；利用String数据库、Cytoscape 3.7.2软件得到PPI网络“、药物-活性成分-作用靶点”网络以及GO、KEGG富集分析。利用Auto Dock Tools1.5.6等软件进行分子对接。体外实验采用脂多糖（LPS）诱导的RAW264.7炎症细胞模型，分为对照组、模型组（LPS，5 μg/mL），不同浓度文王一支笔干预组（LPS 5 μg/mL+文王一支笔提取液25、50、100、200 μg/mL），ELISA法和Western blot法检测文王一支笔提取液对相关细胞因子含量和蛋白表达的影响。
结果: 药物与疾病的共有靶点318个，核心靶点为Src、HSP90AA1和PIK3CA。KEGG分析显示有癌症、PI3K/Akt、MAPK等信号通路。分子对接显示主要活性成分与核心靶点均能自发的结合。体外实验结果显示，文王一支笔提取液在100、200 μg/mL浓度下明显抑制IL-1β（P < 0.05），在50、100、200 μg/mL浓度下明显抑制IL-6和TNF-α，在25、50、100、200 μg/mL浓度下降低PI3K和Akt蛋白的磷酸化水平（P < 0.05）。
结论: 文王一支笔可以作用于多个靶点、多条通路发挥抗炎作用，其作用机制可能与降低IL-1β、IL-6和TNF-α含量，下调p-PI3K、p-Akt蛋白的表达，进而影响PI3K/Akt信号通路减轻炎症反应有关。
Keywords: Balanophora involucrata; anti-inflammation; anti-inflammatory mechanism; molecular docking; network pharmacology.