摘要
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.
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第14期162-166,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家科技部"973"计划项目(2010CB735604)