2种多糖对鲤鱼离体培养免疫细胞活性的影响

Effect of the activation of immunological cells in the commom carp after stimulation by Lentinan and Astragalus polysaccharides

采用Percoll密度离心等技术,对鲤鱼的头肾巨噬细胞和外周血白细胞进行分离纯化,并离体培养。体外暴露不同浓度的黄芪多糖(Astragalus polysaccharides,APS)和香菇多糖(Lentinan,LNT)后,分别采用MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)法测定它们对鲤鱼外周血白细胞增殖的影响;NBT(nitroblue tetrazolium)还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸暴发的影响。结果显示,香菇多糖能显著诱导巨噬细胞的氧暴发活性,黄芪多糖则没有显著的诱导作用;香菇多糖低浓度时对细胞的氮呼吸暴发活性无显著影响,黄芪多糖能显著诱导氮呼吸暴发活性,随着两者作用浓度的增加均表现为高浓度抑制作用;香菇多糖和黄芪多糖都能显著促进鲤鱼外周血淋巴细胞的增殖。结果表明黄芪多糖和香菇多糖对离体培养鲤鱼免疫细胞有明显活性作用,对鲤鱼非特异性免疫和特异性免疫具有促进作用,是有潜力的鱼用免疫增强剂。

The head kidney macrophages and peripheral blood granulocytes isolated from the head kidney of Cyprinus carpio were purified by means of Percoll gradient density centrifugation and were exposed to Astragalus polysaccharides and Lentinan in vitro. The effects of the two polysaccharides on the peripheral blood gTanulocytes proliferation and respiratory burst of macrophage were investigated by using MTT（3-（4,5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide） , NBT（ nitroblue tetrazolium） reduction and Griess reagent coloration respectively, to evaluate the immunostimulant of the two polysaccharides and compare their immune activities with the control group. The results showed that Astragalus polysaccharides and Lentinan stimulated markedly the proliferation of the peripheral blood granulocytes. Lentinan could induce superoxide anion in macrophages markedly, while Astragalus polysaccharides had no effect. Astragalus polysaccharides had conspicuous effects on nitrogen burst activity of macrophages but Astragalus polysaccharides had no effect, moreover, both Astragalus polysaccharides and Lentinan inhibited the nitrogen burst activity at higher doses. It is suggested that Astragalus polysaccharides and Lentinan could modulate the immune response of Cyprinus carpio.