浒苔多糖的分离纯化及其对小鼠淋巴细胞活性的影响

Isolation and purification of polysaccharides in Enteromorpha and its effect on lymphocyte activity of mice

目的比较浒苔(Enteromorpha)多糖4种组分EP,EP1、EP2和EP3对体外培养的小鼠T、B淋巴细胞增殖的影响,并观察作用效果最佳组分EP2在T、B淋巴细胞中的分布及其对细胞RNA合成的影响。方法采用Sevage法,Sephacryl-300分子筛层析以及乙醇分级沉淀法分离纯化获得4个浒苔多糖组分EP,EP1、EP2和EP3,通过体外细胞增殖实验评价不同组分的免疫学活性,另外,利用PAS染色法和甲基绿-派洛宁染色法显示细胞内多糖分布及RNA合成情况。结果 EP、EP1、EP2和EP3中单糖、糖醛酸、硫酸根含量存在差异,均能促进B淋巴细胞以及ConA或LPS诱导的T、B淋巴细胞增殖,且EP2的作用效果最好;经EP2处理的T、B淋巴细胞或同时用EP2和丝裂原(ConA或LPS)处理的T、B淋巴细胞体积明显增大,RNA合成增加,B淋巴细胞有母细胞化趋势。结论浒苔多糖能调节小鼠淋巴细胞活性,促进丝裂原诱导的淋巴细胞分化和免疫应答,有望成为1种新的动物免疫增强剂。

Objective To investigate the effects of four polysaccharide fractions of Enteromorpha on mouse T,B lymphocyte proliferation in vitro,and to observe the influence of EP2 on polysaccharide distribution and RNA synthesis of T,B lymphocytes.Methods Sevage method,gel filtration chromatography（Sephacryl-300）,followed by ethanol precipitation were used to isolate and purify polysaccharide fromEnteromorpha,and four polysaccharide fractions EP,EP1,EP2 and EP3were obtained.Lymphocyte proliferation assay was employed to test their immune-modulating activities.Besides polysaccharide distribution and RNA synthesis of T,B lymphocyte were also observed with PAS staining and Methyl green-Pyranine staining.Results The contents of monosaccharide,uronic acid and sulfate radical were different among the four fractions,the cell proliferation of B lymphocytes and T,B lymphocyte stimulated by mitogen ConA or LPS were significantly promoted by the four polysaccharide fractions above,and EP2 had the best effect than that of EP,EP1 and EP3.Furthermore,the T,B lymphocytes treated with EP2 alone or co-treated with EP2 and mitogen（ConA or LPS）showed the increased cell volume and RNA synthesis,and the B lymphocytes tend to be blastogenic cells.Conclusion The polysaccharides of Enteromorphacould be used to promote the lymphocyte proliferation and differentiation,was expected to be a new immunostimulants of animal.