人参多糖的分级及其免疫活性初探

Fractionation of Ginseng Polysaccharides and Primary Study of Immune Activity

目的:采用柱分离对人参多糖进行分级,并考察各级分对脾淋巴细胞增殖的刺激作用,找出活性部位。方法:用快流速Q-琼脂糖凝胶(Q-Sepharose FF)色谱将人参多糖进行分级,先用水和0.4 mol.L-1NaCl溶液洗脱,将人参多糖分成中性多糖和酸性多糖,再将酸性糖依次用0～0.4 mol.L-1NaCl溶液洗脱,得到不同的级分。取小鼠脾脏制备脾淋巴细胞,调整小鼠脾淋巴细胞密度至3.0×106/mL,加样品混合均匀,孵育48 h,考察每个级分不同浓度(25,50,100,200,400 mg.L-1)对脾淋巴细胞增殖的刺激作用。结果:从GP中分级得到GPN和GPA,GPA分级后得到GPA-1,GPA-2,GPA-3和GPA-4。GP,GPA,GPN,GPA-2,GPA-3和GPA-4对脾淋巴细胞增殖都具有促进作用,其最大刺激指数分别为1.073,1.082,1.119,1.101,1.102和1.296。GPA,GPA-2,GPA-3和GPA-4的刺激作用均随着浓度的增加而增强;GPN随着浓度的增加促进作用减弱,在大于100 mg.L-1时呈现抑制作用,且浓度越大抑制作用越强。GPA-1对脾淋巴细胞的增殖具有抑制作用,且浓度越大抑制作用越强。结论:Q-Sepharose FF色谱可将酸性不同的人参多糖逐级分离。人参多糖各级分对脾淋巴细胞增殖有不同程度的促进作用,以GPA-4的刺激作用最强。

Objective：Fractionation of ginseng polysaccharides were done by column chromatographic,and study the stimulation of collected fractions on proliferation of spleen lymphocyte to find out the active fractions.Method： Fractionation of ginseng polysaccharides was done by Q-Sepharose FF,separated GP into neutral polysaccharide and acidic polysaccharide with water and 0.4 mol·L-1 NaCl as eluents,then separated GPA into several fractions with 0-0.4 mol·L-1 NaCl as eluents.Spleen lymphocytes of mice was isolated and adjusted the spleen lymphocytes concentration to 3.0×106/mL,added sample and incubated for 48 h,the stimulation of each fractions with different concentration（25,50,100,200,400 mg·L-1）on proliferation of spleen lymphocyte was observed.Result： Obtained GPN and GPA from GP,separate GPA into GPA-1,GPA-2,GPA-3 and GPA-4.GP,GPA,GPN,GPA-2,GPA-3 and GPA-4 had the promoting function on proliferation of spleen lymphocyte,their biggest stimulating indexes were 1.073,1.082,1.119,1.101,1.102 and 1.296.The effects of GPA,GPA-2,GPA-3 and GPA-4 enhanced along with the increasing of concentration;with the increasing of concentration,the promoting action of GPN was reduced,when the concentration was over 100 mg·L-1,inhibition was found,and with the increasing of concentration,the inhibition became stronger.GPA-1 had suppression inhibitory effects on proliferation of spleen lymphocyte,and with the increasing of concentration,the inhibition became stronger.Conclusion： Different polysaccharide acid were separated from GP with Q-Sepharose FF.Fractionations of ginseng polysaccharides have different promoting function on proliferation of spleen lymphocyte,and GPA-4 had the strongest effect.