Abstract:
Polysaccharide is one of the principal bioactive components found in medicinal mushrooms and has been proven to enhance host immunity. However, the possible mechanism of immunomodulatory activity of Cordyceps militaris polysaccharide is not fully understood. Hot water extraction and alcohol precipitation, DEAE-Sephadex A-25 chromatography, and Sephadex G-100 chromatography were used to isolate polysaccharide from C. militaris. A high-molecular-weight polysaccharide isolated from C. militaris was designated as HCMP, which had an Mw of 6.18 × 105 Da and was composed of arabinose, galactose, glucose, mannose, and xylose in a mole ratio of 2.00:8.01:72.54:15.98:1.02. The polysaccharide content of HCMP was 91.2% ± 0.16. The test in vitro showed that HCMP activated mouse macrophage RAW 264.7 cells by enhancing phagocytosis and NO production, and by regulating mRNA expressions of inflammation-related molecules in RAW 264.7 cells. Western blotting revealed that HCMP induced the phosphorylation of mitogen-activated protein kinases (MAPKs). Moreover, using inhibitors of MAPKs decreased the mRNA levels of inflammation-related molecules induced by HCMP. These data evidenced that the immunomodulatory effect of HCMP on RAW 264.7 macrophages was mediated via the MAPK signaling pathway. These findings suggested that HCMP could be developed as a potent immunomodulatory agent for use in functional foods and dietary supplements.
摘要:
多糖是药用蘑菇中发现的主要生物活性成分之一,已被证明可以增强宿主免疫力。然而,虫草多糖免疫调节活性的可能机制尚不完全清楚。热水提取和酒精沉淀,DEAE-SephadexA-25色谱法,采用SephadexG-100色谱法分离蛹多糖。分离自C.milaris的高分子量多糖被指定为HCMP,Mw为6.18×105Da,由阿拉伯糖组成,半乳糖,葡萄糖,甘露糖,和木糖的摩尔比为2.00:8.01:72.54:15.98:1.02。HCMP的多糖含量为91.2%±0.16。体外试验表明,HCMP通过增强吞噬作用和NO的产生来激活小鼠巨噬细胞RAW264.7细胞,并通过调节RAW264.7细胞中炎症相关分子的mRNA表达。Western印迹显示HCMP诱导丝裂原活化蛋白激酶(MAPK)的磷酸化。此外,使用MAPKs抑制剂可降低HCMP诱导的炎症相关分子的mRNA水平。这些数据证明HCMP对RAW264.7巨噬细胞的免疫调节作用是通过MAPK信号通路介导的。这些发现表明HCMP可以被开发为用于功能性食品和膳食补充剂的有效免疫调节剂。