Xie Xueqian, Chen Hongxu, Cao Shuiling, Xu Rui, Cai Yanping, Xu Bo, Chen Yunliang, Chen Kehan, Wen Wentao, Zhao Meng, Ke Xuezhou, Yi Qi, Li Chunjing, Wang Qing, Zhou Lian, Luo Xia
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
J Ethnopharmacol. 2025 Jun 20;352:120164. doi: 10.1016/j.jep.2025.120164.
Ganoderma lucidum spore oil (GLSO) is a well-known health product that is beneficial for immuno-enhancement, which stems from a medicine fungus: Ganoderma lucidum (Curtis) P. Karst. Ganoderma lucidum has been used as tonic in China for more than 2000 years. Modern pharmacological studies have shown that it has effects with immunomodulatory, hypoglycemic, hypolipidemic, anti-oxidation, anti-aging and anti-tumor.
The immuno-enhancement ability of GLSO in hepatoma H22-bearing mice was investigated in this study, and our work aimed to reveal the potential mechanisms of the antitumor efficacy of GLSO.
The GLSO components were identified via UHPLC-Q-Orbitrap HRMS. A H22 cell subcutaneously transplanted tumor mouse model was constructed, and GLSO was preadministered. The antitumor efficacy of GLSO in hepatoma H22-bearing mice was evaluated according to tumor size, tumor growth curves, tumor inhibition rates and Ki67 level. T and B lymphocyte proliferation, delayed hypersensitivity, NK cell killing activity, macrophage phagocytotic activity and macrophage polarization, cytokine levels and CD69 molecule expression were detected to estimate immune function. Network pharmacology analysis, flow cytometry and Pro-DIA quantitative proteomics analysis were performed to investigate the potential mechanism of GLSO in tumor inhibition, which was verified by WB and RT-PCR.
Thirty-eight compounds including triterpenoids, fatty acids and esters, were identified from GLSO. Mice treated with GLSO showed the smaller initial and final tumor volumes and lower Ki67 expression, GLSO treatment could prevented tumor occurrence and inhibited tumor growth. Treatment with GLSO promoted a strong immune response including macroregulation in immune organs, enhancement of macrophage phagocytosis, NK cell cytotoxicity,T cells and B cells proliferation activity, delayed-type hypersensitivity reaction, reducing the production of M2 macrophages and regulation of cytokine secretion in hepatoma H22-bearing mice. Network pharmacology analysis and flow cytometry results showed that treatment with GLSO might have beneficial effects on improving the tumor immune microenvironment. Proteomics analysis showed that GLSO inhibited eicosanoid metabolism pathway, WB and RT-PCR re-check these results.
These findings support that GLSO enhances immunity in hepatoma H22-bearing mice and we first report that GLSO restricts tumor growth by inhibiting eicosanoid metabolism pathway.
灵芝孢子油（GLSO）是一种著名的保健品，对免疫增强有益，它源自一种药用真菌：灵芝（Curtis）P. Karst。灵芝在中国作为滋补品已使用了2000多年。现代药理学研究表明，它具有免疫调节、降血糖、降血脂、抗氧化、抗衰老和抗肿瘤等作用。
本研究考察了GLSO对荷肝癌H22小鼠的免疫增强能力，旨在揭示GLSO抗肿瘤疗效的潜在机制。
通过超高效液相色谱-四极杆-轨道阱高分辨质谱（UHPLC-Q-Orbitrap HRMS）鉴定GLSO的成分。构建H22细胞皮下移植瘤小鼠模型，并预先给予GLSO。根据肿瘤大小、肿瘤生长曲线、肿瘤抑制率和Ki67水平评估GLSO对荷肝癌H22小鼠的抗肿瘤疗效。检测T和B淋巴细胞增殖、迟发型超敏反应、NK细胞杀伤活性、巨噬细胞吞噬活性和巨噬细胞极化、细胞因子水平及CD69分子表达以评估免疫功能。进行网络药理学分析、流式细胞术和蛋白质组学分析以研究GLSO抑制肿瘤的潜在机制，并通过蛋白质免疫印迹（WB）和逆转录-聚合酶链反应（RT-PCR）进行验证。
从GLSO中鉴定出38种化合物，包括三萜类、脂肪酸和酯类。用GLSO处理的小鼠初始和最终肿瘤体积较小，Ki67表达较低，GLSO处理可预防肿瘤发生并抑制肿瘤生长。GLSO处理促进了强烈的免疫反应，包括免疫器官的宏观调节、巨噬细胞吞噬作用增强、NK细胞细胞毒性、T细胞和B细胞增殖活性、迟发型超敏反应，减少荷肝癌H22小鼠中M2巨噬细胞的产生并调节细胞因子分泌。网络药理学分析和流式细胞术结果表明，GLSO处理可能对改善肿瘤免疫微环境有有益作用。蛋白质组学分析表明GLSO抑制类花生酸代谢途径，WB和RT-PCR再次验证了这些结果。
这些发现支持GLSO增强荷肝癌H22小鼠的免疫力，并且我们首次报道GLSO通过抑制类花生酸代谢途径限制肿瘤生长。
