ABSTRACT Currently there is tremendous interest in the discovery of natural polysaccharide immunomodulators for treating immune-related diseases. Herein, the immunomodulatory activities of a polysaccharide fraction isolated from blackberry seeds (BSP-2) were examined in RAW264.7 cells. The results revealed that BSP-2 at 37.5–300 µg/mL could significantly enhance the phagocytic activity of RAW264.7 cells and their ability to release NO, TNF-α and IL-6 cytokines. BSP-2 could significantly up-regulate the mRNA expression of iNOS, TNF-α and IL-6, as well as the phosphorylation of p65, p38, ERK and JNK proteins. The mechanisms underlying the immunomodulatory actions of BSP-2 likely involve the activation of the NF-κB and MAPK signaling pathways and up-regulation of the phosphorylation of key proteins involved in these pathways. These findings suggest that BSP-2 could be applied as a polysaccharide immunotherapeutic adjuvant with the effective dose as low as 70 µg/mL (BSP-2 may be cytotoxic at levels > 600 μg/mL). Highlights Immunostimulatory potential of a blackberry seed polysaccharide (BSP-2) was examined. BSP-2 at 37.5–300 µg/mL could enhance the phagocytic activity of RAW264.7 cells. BSP-2 could significantly up-regulate the mRNA expression of iNOS, TNF-α and IL-6. BSP-2 up-regulated the phosphorylation of p65, p38, ERK and JNK proteins. Immunostimulatory BSP-2 acted via activating the NF-κB and MAPK signalling pathways.

中文翻译：

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来自黑莓种子的免疫调节多糖及其通过激活 NF-κB/MAPK 通路对 RAW 264.7 细胞的作用

摘要 目前，人们对发现用于治疗免疫相关疾病的天然多糖免疫调节剂有着极大的兴趣。在此，在 RAW264.7 细胞中检查了从黑莓种子 (BSP-2) 中分离的多糖部分的免疫调节活性。结果表明，37.5–300 µg/mL 的 BSP-2 可显着增强 RAW264.7 细胞的吞噬活性及其释放 NO、TNF-α 和 IL-6 细胞因子的能力。BSP-2 可显着上调 iNOS、TNF-α 和 IL-6 的 mRNA 表达，以及 p65、p38、ERK 和 JNK 蛋白的磷酸化。BSP-2 免疫调节作用的潜在机制可能涉及激活 NF-κB 和 MAPK 信号通路以及上调参与这些通路的关键蛋白质的磷酸化。这些发现表明 BSP-2 可用作多糖免疫治疗佐剂，有效剂量低至 70 µg/mL（BSP-2 在水平 > 600 µg/mL 时可能具有细胞毒性）。亮点检测了黑莓种子多糖 (BSP-2) 的免疫刺激潜力。37.5–300 µg/mL 的 BSP-2 可以增强 RAW264.7 细胞的吞噬活性。BSP-2可显着上调iNOS、TNF-α和IL-6的mRNA表达。BSP-2 上调 p65、p38、ERK 和 JNK 蛋白的磷酸化。免疫刺激 BSP-2 通过激活 NF-κB 和 MAPK 信号通路起作用。亮点检测了黑莓种子多糖 (BSP-2) 的免疫刺激潜力。37.5–300 µg/mL 的 BSP-2 可以增强 RAW264.7 细胞的吞噬活性。BSP-2可显着上调iNOS、TNF-α和IL-6的mRNA表达。BSP-2 上调 p65、p38、ERK 和 JNK 蛋白的磷酸化。免疫刺激 BSP-2 通过激活 NF-κB 和 MAPK 信号通路起作用。亮点检测了黑莓种子多糖 (BSP-2) 的免疫刺激潜力。37.5–300 µg/mL 的 BSP-2 可以增强 RAW264.7 细胞的吞噬活性。BSP-2可显着上调iNOS、TNF-α和IL-6的mRNA表达。BSP-2 上调 p65、p38、ERK 和 JNK 蛋白的磷酸化。免疫刺激 BSP-2 通过激活 NF-κB 和 MAPK 信号通路起作用。