香菇多糖对鲤鱼(Cyprinus carpio)离体培养免疫细胞的活性调节作用  

Effect of Lentinan on the Activation of Immunological Cell Culture In vitro in Cyprinus carpio

作　　者：曹丽萍[1] 丁炜东[1] 张柳[2] Galina Jeney 徐跑[1] 殷国俊[1] 

机构地区：[1]中国水产科学研究院淡水渔业研究中心农业部水生动物遗传育种和养殖生物学重点开放实验室,江苏无锡214081 [2]华中农业大学,湖北武汉430070 [3]Research Institute for Fisheries,Aquaculture and Irrigation,Anna light8,Szarvas H-4440,Hungary

出　　处：《农业生物技术学报》2008年第4期616-621,共6页Journal of Agricultural Biotechnology

基　　金：中匈(匈牙利)国际合作项目(第3期)(No．CHN-27/2006);国际科学基金(No．AA13611-R);中央级公益性科研院所基本科研业务费专项资金(No．6-115009．6-115038)资助。

摘　　要：采用Percoll密度离心等技术,对鲤鱼(Cyprinus carpip)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香菇多糖后,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,香菇多糖1、10和100μg/mL作用头肾巨噬细胞24 h后能显著地诱导巨噬细胞的氧爆发活性;作用头肾巨噬细胞96 h后,其低浓度对巨噬细胞生成NO无显著影响,当作用浓度为1 000μg/mL表现出显著的抑制作用;浓度为100和1000μg/mL的香菇多糖作用外周血白细胞24 h后显著地促进鲤鱼外周血白细胞的增殖,且在作用巨噬细胞24 h后100μg/mL能显著增强IL-1β的体外诱导表达。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,是一种有潜力的鱼用免疫增强剂。The head kidney （HK） macrophages and the peripheral blood leukocytes were isolated from the head kidney and blood of Cyprinus carpio respectively by percoll gradient density centrifugation, cultured in vitro and exposed to different concentrations of lentiuan. To evaluate the immunostimulating effects of lentinan, proliferation of the peripheral blood leukocytes, respiratory burst of macrophage and IL-1β mRNA expression of macrophages were investigated by MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide）, NBT （nitroblue tetrazolium） reduction, Griess reaction and real-time PCR, respectively. Results showed that proliferation of peripheral blood leukocytes after 24 h incubation by the induction of lentinan at 100 and 1000μg/mL was markedly stimulated. Lentiuan at 1100 and 1000μg/mL could induce superoxide anion in macrophages markedly. Reduction of nitric oxide of HK macrophages after 96 h incubation lentinan showed it had no conspicuous effect on nitrogen burst activity of macrophages; moreover it inhibited the nitrogen burst activity at higher doses. The expression of IL-1β in the HK macrophages after 24 h stimulation by the polysaccharide showed lentinan stimulated IL-1β expression in carp head kidney. It suggested that lentinan can modulate the immune response of C.carpio.

关 键 词：香菇多糖 鲤鱼 巨噬细胞 IL-1Β 

分 类 号：S185[农业科学—农业基础科学] S188