冬虫夏草多糖单糖组成及免疫活性研究

The Monosaccharides Composition and Immune Activity of Polysaccharides from Cordyceps sinensis

以鲜冬虫夏草为原料,采用水提醇沉、Sevag法脱蛋白制备鲜冬虫夏草粗多糖(FCSP),通过苯酚硫酸法、考马斯亮蓝法、高效凝胶渗透色谱联用蒸发光散射检测器(HPGPC-ELSD)、傅立叶红外变换光谱(FT-IR)及离子色谱-脉冲安培检测器联用法(HPAEC-PAD)对FCSP的纯度、分子量分布、单糖组成进行分析,并考察FCSP刺激巨噬细胞增殖和释放NO、TNF-α的免疫活性。结果表明,粗多糖FCSP由葡萄糖(59.13%)、甘露糖(21.73%)和半乳糖(19.14%)三种单糖组成,且含有1.186×10^4 kDa(3.510%)、1.070×10^3 kDa(24.232%)和2.115×10 kDa(72.258%)三个多糖组分;体外细胞实验表明,FCSP可显著增强巨噬细胞增殖活性,有效刺激细胞释放NO和TNF-α(P<0.05)。FCSP良好的免疫调节作用可为后期药品的开发以及冬虫夏草新资源食品的申报提供参考。

FCSP was extracted with hot water from fresh Cordyceps sinensis,and precipitated using ethanol.Then the proteins were removed via Sevag method.Afterwards,the phenol sulfuric acid method,Coomassie brilliant blue method and high performance gel permination chromatography coupled evaporative light-scattering detector(HPGPC-ELSD),Fourier transform infrared spectroscopy(FT-IR)and high performance anion exchange chromatography with pulsed amperometric detection(HPAEC-PAD)were applied to analyze the purity,molecular weight distribution and monosaccharides composition of FCSP.The immunological activity of FCSP was estimated by proliferation,NO and TNF-α releasing of macrophages.Results showed that FCSP was made up of glucose(59.13%),mannose(21.73%)and galactose(19.14%),which included 1.186×10^4 kDa(3.510%),1.070×10^3 kDa(24.232%)and 2.115×10 kDa(72.258%)polysaccharides.In vitro cell experiments showed that FCSP could significantly enhance the activity of macrophages on proliferation and release NO or TNF-α(P<0.05).Because of its excellent immunoregulatory activity,FCSP could provide the reference for the certain drug development and the application of new resource food.