香菇多糖和黄芪多糖对鲤免疫细胞的活性和IL-1β体外诱生表达的影响
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其它中匈(牙利)国际合作项目(第三期)(CHN-27/2006)和国际科学基金(AA13611-R)和中央级公益性科研院所基本科研业务费专项资金(中国水产科学研究院淡水渔业研究中心)资助项目(6-115009,6-115038)


Effects of the activation of immunological cells and the expression of interleukin-1βin the commom carp after stimulation by Lentinan and Astragalus polysaccharides*
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    摘要:

    为了探讨香菇和黄芪多糖对鲤免疫细胞的免疫活性作用,采用Percoll密度离心等技术,对鲤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,从细胞和分子水平上研究了香菇和黄芪多糖的免疫调节作用。巨噬细胞和外周血白细胞分别体外暴露不同浓度的黄芪多糖和香菇多糖后,采用 MTT法测定它们对鲤外周血白细胞增殖的影响;NBT还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸爆发的影响;实时定量PCR法测定头肾巨噬细胞细胞因子IL-1β诱导表达的影响。结果显示,黄芪和香菇多糖作用头肾巨噬细胞24 h后,香菇多糖浓度为1,10,100μg•mL-1时能显著诱导巨噬细胞的氧爆发活性,黄芪多糖则没有显著的诱导作用;黄芪和香菇多糖作用头肾巨噬细胞96h后,香菇多糖低浓度时对细胞的氮呼吸爆发活性无显著影响,黄芪多糖浓度10 μg•mL-1能显著诱导氮呼吸爆发活性,而在浓度为 1 000 μg•mL-1两者均表现为高浓度抑制作用;两者作用外周血白细胞24 h后,香菇多糖浓度为100,1 000 μg•mL-1和黄芪多糖浓度为10,100 μg•mL-1时都能显著促进鲤外周血淋巴细胞的增殖;两种多糖作用头肾巨噬细胞24 h后能显著增强IL-1β的体外诱导表达。结果表明黄芪多糖和香菇多糖对离体培养鲤免疫细胞有明显活性作用,对鲤非特异性免疫和特异性免疫具有促进作用。

    Abstract:

    To investigate the in vitro immunostimulating effects of Lentinan and Astragalus polysaccharides in cellular and molecular levels in the Cyprinus carpio, the macrophages and the peripheral blood leukocytes were isolated from the head kidney and blood of Cyprinus carpio respectively through Percoll gradient density centrifugation, cultured and exposed to different concentrations of Astragalus polysaccharides and Lentinan. The proliferation of the peripheral blood granulocytes, respiratory burst of macrophage and IL-1βmRNA expression of macrophages were analyzed by MTT ,NBT reduction, Griess reaction and Real time PCR respectively. Results showed that proliferation of peripheral blood leukocytes after 24 h incubation with Astragalus polysaccharides at 10,100µg·mL-1 and Lentinan at 100,1000µg·mL-1 was markedly stimulated. Lentinan at 1, 100,1000µg·mL-1 could induce superoxide anion in macrophages markedly, while Astragalus polysaccharides had no effect. Reduction of nitric oxide of head kidney macrophages (HK macrophages) after 96 h incubated by polysaccharides showed Astragalus polysaccharides at 10µg·mL-1 had conspicuous effect on nitrogen burst activity of macrophages but Astragalus polysaccharides had no effect, moreover both Astragalus polysaccharides and Lentinan at 1000µg·mL-1 inhibited the nitrogen burst activity at higher doses. The expression of interleukin-1βin the HK macrophages after 24 h stimulation by the two polysaccharides showed Astragalus polysaccharides and Lentinan stimulated IL-1βexpression in carp head kidney . It is suggested that Astragalus polysaccharides and Lentinan could modulate the immune response of Cyprinus carpio.

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曹丽萍,丁纬东,张柳,谢俊,殷国俊.香菇多糖和黄芪多糖对鲤免疫细胞的活性和IL-1β体外诱生表达的影响[J].水产学报,2008,32(4):628~635

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  • 收稿日期:2007-10-11
  • 最后修改日期:2008-02-21
  • 录用日期:2008-05-27
  • 在线发布日期: 2008-07-07
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