蛴螬多糖的提取分离和体外对小鼠免疫细胞增殖的影响

Isolation and proliferation effect of polysaccharide extracted from grub on the immunocytes of mice in vitro

以中国药用昆虫白星花金龟(Protaetia brevitarsis)幼虫(蛴螬)为研究对象,通过匀浆、乙醇沉淀、Sevag法去除蛋白,得到蛴螬总多糖,其得率为57%。用苯酚硫酸法进行多糖浓度测定。经MTT法检测表明,终质量浓度为5和20μg.mL-1的总多糖对小鼠脾细胞的生长增殖率分别为48.8%和62.2%,对巨噬细胞生长增殖率分别为240%和516%。用DEAE-Sephadex A-25、LPLC-Sephadex G-75层析法分离纯化总多糖,得到5种蛴螬多糖,其中相对分子质量分别为42 000和26 000的2种蛴螬多糖对脾细胞生长增殖率较高,终质量浓度为5μg.mL-1的该2种多糖对脾细胞生长增殖率分别为25.2%和41.4%。

The polysaecharides were extracted from Protaetia brevitarsis larva （grub） with homogenization, ethanol and Sevag methods. The concentration of total polysaccharides of grub was measured with phenol-sulfuric acid method. MTT assay showed that the rate of proliferation of total polysaccharides with concentration of 5 μg · mL^-1 and 20 μg · mL^-1 on mouse spleen cell was 48.8% and 62. 2% , respectively, and the rate of proliferation of total polysaccharides with concentration of 5 μg · mL^-1 and 20 μg· mL^-1 on maerophages was 240% and 516% , respectively. Five polysaccharides were isolated and purified from total polysac- charides using DEAE-Sephadex A-25 and LPLC-Sphadex G-75, and the two kinds of polysaccharides were associated with the activity to improve mouse splenocytes proliferation in Htro, whose molecular weight were 42 000 and 26 000, respectively. The rate of prolif- eration of the two kinds of polysaccharides with concentration of 5 μg · mL^-1 on spleen cell was 25.2% and 41.4% , respectively, which indicated that the two kinds of polysaccharide could significantly improve the proliferation of the immunocytes of mice in vitro.