西洋参多糖联合人参稀有皂苷发挥协同抗炎作用

Synergistic Anti-Inflammatory Action of Panax quinquefolius Polysaccharide and Rare Ginsenosides

摘要:

为探讨西洋参多糖（AGP）与人参稀有皂苷（HTS）联合对脂多糖（Lipopolysaccharide，LPS）诱导的RAW264.7细胞炎症的抗炎作用及其分子机制，研究采用CCK-8法评估不同质量浓度的AGP、HTS及其联用对细胞活力的影响，确定三者安全给药质量浓度范围及LPS最佳造模质量浓度。实验分为空白对照组、模型组、HTS组、AGP组及HTS+AGP组，处理24 h后通过倒置显微镜观察细胞形态，采用Griess法测定NO含量，ELISA检测TNF-α与IL-6水平，免疫荧光与RT-qPCR分析TNF-α、CD68和CD163的表达。结果显示，HTS+AGP组细胞存活率显著高于HTS组（IC??分别为280.50 μg·mL?1和149.40 μg·mL?1），且AGP具有促细胞增殖作用。在LPS诱导的炎症模型中，HTS+AGP干预显著抑制NO释放、TNF-α（模型组473.12 pg·mL?1；HTS+AGP组366.58 pg·mL?1）和IL-6（模型组61.92 pg·mL?1;HTS+AGP组33.07 pg·mL?1）。RT-qPCR与免疫荧光结果表明，HTS+AGP可下调M1型标志物CD68与TNF-α mRNA表达，上调M2型标志物CD163 mRNA表达，有效逆转LPS诱导的巨噬细胞M1型极化。综上所述，AGP与HTS联用可通过抑制炎症因子释放并促进巨噬细胞向M2型极化，抗炎效应增强，其机制与调控巨噬细胞极化状态相关。

Abstract:

The anti-inflammatory effects and underlying molecular mechanisms of Panax quinquefolius polysaccharide (AGP) combined with rare ginsenosides (HTS) on lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells were investigated. The cell viability was evaluated using the CCK-8 assay after treatment with various concentrations of AGP, HTS, and their combination. the safe concentration ranges of the agents and the optimal modeling concentration of LPS were determined. The experiment was divided into the following groups: the blank control group, the model group, the HTS group, the AGP group, and the HTS+AGP group. After 24 hours of treatment, cell morphology was observed under an inverted microscope. NO production was measured using the Griess method. The levels of TNF-α and IL-6 were determined by ELISA. Furthermore, the expression of TNF-α, CD68, and CD163 was analyzed by immunofluorescence and RT-qPCR. The cell survival rate in the HTS+AGP group was found to be significantly higher than that in the HTS group alone, with IC?? values determined to be 280.50 μg·mL?1 and 149.40μg·mL?1, respectively. Furthermore, AGP alone was observed to promote cell proliferation. In the LPS-induced inflammatory model, the release of NO was significantly inhibited by HTS+AGP intervention. Similarly, the levels of TNF-α (Model:473.12 pg·mL?1; HTS+AGP:366.58 pg·mL?1) and IL-6 (Model:61.92 pg·mL?1; HTS+AGP:33.07 pg·mL?1) were also markedly suppressed. As indicated by RT-qPCR and immunofluorescence results, the mRNA expression of M1 markers (CD68 and TNF-α) was significantly downregulated, while the mRNA expression of the M2 marker (CD163) was upregulated by HTS+AGP treatment, effectively reversing the LPS-induced shift toward M1 macrophage polarization. In summary, the combined application of AGP and HTS enhances the anti-inflammatory effect by inhibiting the release of inflammatory factors and promoting the polarization of macrophages toward the M2 phenotype. This mechanism is associated with the regulation of macrophage polarization status.