Wu Jingyu, Ou Yi, Yao Min, Liu Jiaquan, Ran Hengxing, Wu Zhengrong, Wu Rihui, Gan Lishe, Li Dongli, Jin Jingwei
School of Pharmacy and Food Engineering, Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, Wuyi University, Jiangmen, China.
International Healthcare Innovation Institute, Jiangmen, China.
Front Pharmacol. 2025 Apr 25;16:1514284. doi: 10.3389/fphar.2025.1514284. eCollection 2025.
The flavonoids found in exhibit a wide range of pharmacological activities, with their immunostimulatory effects emerging as a significant area of research in recent years. However, the underlying mechanism of their immunostimulatory activity remains unclear.
To investigate the immunostimulatory effects and elucidate the specific mechanisms of flavonoids both and .
The immunostimulatory effects and underlying mechanisms of flavonoids from were evaluated using a variety of techniques, including cell viability assays, flow cytometry, real-time reverse transcription-quantitative polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), molecular docking, plasmid recombination and transformation, recombinant protein expression, surface plasmon resonance (SPR), and NF-κB/SEAP assays. To investigate the immune response in animal experiments, flavonoids were compared with traditional adjuvants, utilizing biochemical analysis and flow cytometry.
flavonoids, primarily composed of icaritin, icariin I and icariin II, were observed to significantly enhance the expression of surface co-stimulatory molecules (CD40, CD80, CD86) and major histocompatibility complex (MHC-I, MHC-II) in bone marrow-derived dendritic cells (BMDCs) and RAW 264.7 cells. Additionally, the production of chemokines and pro-inflammatory cytokines was significantly increased in RAW 264.7 cells. , the findings demonstrated that the vaccine adjuvant containing flavonoids significantly increased the serum concentration of total OVA-specific IgG compared to the control group. SPR analysis revealed that icariin II exhibited the highest binding response to TLR7, while icariin I and icariin II showed the strongest interactions with TLR8 protein, even surpassing the positive control drug, Resiquimod. The NF-κB/SEAP assay further confirmed that icaritin, icariin I, and icariin II enhanced NF-κB activity and stimulated SEAP secretion through TLR7/8 activation.
[植物名称]中发现的黄酮类化合物具有广泛的药理活性，近年来其免疫刺激作用成为一个重要的研究领域。然而，其免疫刺激活性的潜在机制仍不清楚。
研究[植物名称]黄酮类化合物的免疫刺激作用，并阐明其体内外的具体机制。
采用多种技术评估[植物名称]黄酮类化合物的免疫刺激作用及其潜在机制，包括细胞活力测定、流式细胞术、实时逆转录定量聚合酶链反应（qRT-PCR）、酶联免疫吸附测定（ELISA）、分子对接、质粒重组与转化、重组蛋白表达、表面等离子体共振（SPR）和NF-κB/SEAP测定。为了在动物实验中研究免疫反应，将[植物名称]黄酮类化合物与传统佐剂进行比较，采用生化分析和流式细胞术。
观察到主要由淫羊藿素、淫羊藿苷I和淫羊藿苷II组成的[植物名称]黄酮类化合物能显著增强骨髓来源的树突状细胞（BMDCs）和RAW 264.7细胞表面共刺激分子（CD40、CD80、CD86）和主要组织相容性复合体（MHC-I、MHC-II）的表达。此外，RAW 264.7细胞中趋化因子和促炎细胞因子的产生显著增加。此外，研究结果表明，与对照组相比，含有[植物名称]黄酮类化合物的疫苗佐剂显著提高了总OVA特异性IgG的血清浓度。SPR分析显示，淫羊藿苷II对TLR7的结合反应最高，而淫羊藿苷I和淫羊藿苷II与TLR8蛋白的相互作用最强，甚至超过阳性对照药物瑞喹莫德。NF-κB/SEAP测定进一步证实，淫羊藿素、淫羊藿苷I和淫羊藿苷II通过激活TLR7/8增强NF-κB活性并刺激SEAP分泌。
