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1、农业生物技术学报JournalofAgriculturalBiotechnology2008,16(4):616~621·研究论文·香菇多糖对鲤鱼（）离体培养免疫细胞的活性调节作用*1, 丁炜东1, 张柳2,GalinaJeney 3, 徐跑1, 殷国俊1**曹丽萍（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，无锡214081；2.华中农业大学，武汉430070；3.ResearchInstituteforFisheries,AquacultureandIrrigation, Annalight8,SzarvasH4440,H
2、ungary）摘要：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香菇多糖后，分别采用MTT(3(4,5dimethylthiazol2yl)2,5diphenyltetrazoliumbromide)法测定它们对鲤鱼外周血白细胞增殖的影响；NBT(nitrobluetetrazolium)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IL1茁诱导表达的影响。结果显示，香菇多糖1、10和100滋g/mL作用头肾巨噬细胞24h后能显著地诱导巨噬细
3、胞的氧爆发活性；作用头肾巨噬细胞96h后，其低浓度对巨噬细胞生成NO无显著影响，当作用浓度为1000滋g/mL表现出显著的抑制作用；浓度为100和1000滋g/mL的香菇多糖作用外周血白细胞24h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24h后100滋g/mL能显著增强IL1茁的体外诱导表达。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。关键词：香菇多糖；鲤鱼；巨噬细胞；中图分类号:S185,S188文献标识码:A 文章编号:10061304(2008)04061606EffectofL
4、entinanontheActivationofImmunological CellsCulturedinCAOLiping 1,DINGWeidong 1,ZHANGLiu 2,GalinaJeney 3,XuPao 1,YINGuojun 1** Theheadkidney(HK)macrophagesandtheperipheralbloodleukocyteswereisolatedfromtheheadkidneyandbloodofrespectivelybypercollgradientdensitycentrifugation,culturedandexp
5、osedtodifferentconcentrationsoflentinan.Toevaluatetheimmunostimulatingeffectsoflentinan,proliferationoftheperipheralbloodleukocytes,respiratoryburstofmacrophageandmRNAexpressionofmacrophageswereinvestigatedbyMTT(3(4,5dimethylthiazol2yl)2,5diphenyl tetrazoliumbromide),NBT(nitrobluetetrazol
6、ium)reduction,GriessreactionandrealtimePCR,respectively.Resultsshowedthat proliferationofperipheralbloodleukocytesafter24hincubationbytheinductionoflentinanat100and1000滋g/mLwasmarkedly stimulated.Lentinanat1,100and1000滋g/mLcouldinducesuperoxideanioninmacrophagesmarkedly.Reductionofnitrico
7、xideofHKmacrophagesafter96hincubationlentinanshowedithadnoconspicuouseffectonnitrogenburstactivityofmacrophages; moreoveritinhibitedthenitrogenburstactivityathigherdoses.TheexpressionofintheHKmacrophagesafter24hstimulationbythepolysaccharideshowedlentinanstimulatede
