Triterpenoids, as the main active ingredient of Ganoderma lucidum fermented extract, exert multiple pharmacological activities, including immunomodulatory properties. Our study aimed to reveal the pharmacological effects and potential mechanisms of Ganoderic acid C2 (GAC) against cyclophosphamide (CY)-associated immunosuppression. Target genes were collected from several public databases, including the DisGeNET, Comparative Toxicogenomics Database, GeneCards, and PharmMapper. STRING database was used to construct the protein-protein interaction of network. Subsequently, molecular docking was carried out to visualize the protein-GAC interactions. Experimental validations, including ELISA and qRT-PCR were performed to confirm the pharmacological activities of GAC on CY-induced immunosuppression model. A total of 56 GAC-related targets were identified to be closely associated with CY-induced immunosuppression. Enrichment analyses results revealed that these targets were mainly involved in immune and inflammatory response-related pathways. STAT3 and TNF were identified as the core targets of GAC. Molecular docking indicated that GAC combined well with STAT3 and TNF protein. In addition, animal experiments indicated that GAC improved immunity as well as STAT3 and TNF genes expression in CY-induced immunosuppression, which further verified the prediction through bioinformatics analysis and molecular docking. We successfully revealed the potential therapeutics mechanisms underlying the effect of GAC against CY-induced immunosuppression based on the combination of bioinformatics analysis, molecular docking, and animal experiments. Our findings lay a theoretical foundation for the in-depth development and utilization of Ganoderma lucidum fermentation product in the future, and also provide theoretical guidance for the development of innovative drugs that assist in improving immunity.

中文翻译：

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灵芝酸 C2 对环磷酰胺诱导的免疫抑制发挥药理作用：一项涉及分子对接和实验验证的研究。

三萜类化合物作为灵芝发酵提取物的主要活性成分，具有多种药理活性，包括免疫调节特性。我们的研究旨在揭示灵芝酸C2（GAC）对抗环磷酰胺（CY）相关免疫抑制的药理作用和潜在机制。目标基因从多个公共数据库收集，包括 DisGeNET、比较毒理学数据库、GeneCards 和 PharmMapper。使用STRING数据库构建蛋白质-蛋白质相互作用网络。随后，进行分子对接以可视化蛋白质-GAC 相互作用。进行了包括 ELISA 和 qRT-PCR 在内的实验验证，以确认 GAC 对 CY 诱导的免疫抑制模型的药理活性。总共确定了 56 个 GAC 相关靶点与 CY 诱导的免疫抑制密切相关。富集分析结果显示，这些靶点主要涉及免疫和炎症反应相关途径。STAT3和TNF被确定为GAC的核心靶点。分子对接表明GAC与STAT3和TNF蛋白结合良好。此外，动物实验表明，GAC可提高免疫力，并在CY诱导的免疫抑制中提高STAT3和TNF基因的表达，通过生物信息学分析和分子对接进一步验证了这一预测。基于生物信息学分析、分子对接和动物实验相结合，我们成功揭示了GAC对抗CY诱导的免疫抑制的潜在治疗机制。我们的研究结果为今后灵芝发酵产品的深入开发利用奠定了理论基础，也为辅助提高免疫力的创新药物的开发提供了理论指导。