灵芝β-葡聚糖的碱提取及其抗肿瘤免疫调节研究

Alkali extraction of Ganoderma lucidum β-glucan and its anti-tumor immune regulation

采用Box-Benhnken中心组合试验设计原理和响应面分析法,得到碱法提取灵芝颗粒性β-葡聚糖(Ganoderma lucidum particulateβ-glucan,GLG)最佳条件:提取温度55℃、时间8 h和料液比1∶20,提取得率为8.57%。本研究中动物实验获得江苏省中医药研究院伦理委员会的批准。体外免疫活性研究表明,GLG能促进巨噬细胞的激活并分泌促炎细胞因子,不仅能提高巨噬细胞的吞噬能力,还能促进巨噬细胞吞噬GLG自身,持续发挥免疫刺激作用;GLG还能促进小鼠脾淋巴细胞的增殖;此外,口服GLG能够显著抑制CT26荷瘤小鼠肿瘤的增殖,提高其脾脏中产生IFN-γ的CD4^+T(Th1)细胞与CD8^+T(CTL)细胞的比例,表明GLG能够促进具有杀伤肿瘤细胞作用的Th1和CTL的增殖与分化,从而增强机体抗肿瘤免疫。

In this study,an immunostimulating particulateβ-glucan was isolated from a hot alkaline extract of the fruiting bodies of Ganoderma lucidum.The optimum conditions of 8 hours treatment time,1∶20 solid-liquid ratio and 55℃for the alkaline extract process were obtained after investigating by single-factor experiments and Box-Benhnken design in terms of the Ganoderma lucidum particulateβ-glucan(GLG)increment,and these conditions resulted in a GLG yield of 8.57%.The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine.The result showed that resident macrophages were effectively activated by GLG,such as with the up-regulation of co-stimulatory molecules,the secretion of cytokines and phagocytic uptake.GLG could also promote the proliferation of spleen lymphocytes in mice.In addition,IFN-γproduction of spleen CD4^+T cells and cytotoxic T lymphocyte(CTL)responses were significantly enhanced on GLG orally treatment,which ultimately resulted in significantly decreased tumor burden.Taken together,these data suggest that GLG might act as an immune stimulator to exert antitumor effects.