皮质醇对黄颡鱼头肾巨噬细胞呼吸爆发活动的影响  

Effects of Cortisol on Respiratory Burst of Head Kidney Macrophages in Pelteobagrus fulvidraco

作　　者：刘小玲[1] 葛海燕[1] 田珍[1] 顾泽茂[1] 雷曼红[1] 

机构地区：[1]华中农业大学水产学院,武汉430070

出　　处：《华中农业大学学报》2008年第6期749-754,共6页Journal of Huazhong Agricultural University

基　　金：湖北省科技攻关项目(2007AA203A01);中国水产科学研究院淡水生态与健康养殖重点开放实验室课题(2007FEA0201);华中农业大学人才科研启动项目(52204-07027)资助

摘　　要：用Percoll连续密度梯度离心法分离黄颡鱼头肾巨噬细胞,经含0.1%牛血清的L-15培养液中培养,在光镜和电镜下观察其形态、超微结构和吞噬作用;采用硝基四氮唑蓝（NBT）与多孔酶标仪检测细胞在佛波豆蔻酸乙酯（PMA）刺激下产生的呼吸爆发,并观察外源皮质醇对黄颡鱼体内外头肾巨噬细胞呼吸爆发活动的影响。结果表明：每尾黄颡鱼可分离出1.2-1.6 mL浓度为1×10^7cells/mL的高纯度头肾巨噬细胞;培养3-18 h后,贴壁纯化的头肾巨噬细胞达到95%-98%;分离培养的头肾巨噬细胞具有巨噬细胞的形态结构特征,对福尔马林灭活的金黄色葡萄球菌（F-SA）的吞噬率达95%以上。PMA刺激分离的头肾巨噬细胞产生显著的呼吸爆发活动。在较高浓度和较长作用时间情况下,外源皮质醇对黄颡鱼体内外头肾巨噬细胞的呼吸爆发均产生抑制作用。The head kidney macrophages of yellow catfish （Pelteobagrus fulvidraco） were separated by Percoll continuous density gradient centrifugation, and cultivated in L-15 medium with 0. 1% fetal bovine serum. The morphological and ultrastructural features,as well as phagocytosis were observed under light and electron microscope. Nitroblue tetrazolium （NBT） and multiscan spectrophotometer were used for the respiratory burst assay of the macrophages after stimulation by phorbol-12-myristate-13-acetate （PMA）,and the effects of exogenous cortisol on the respiratory burst activity both in vitro and in vivo were investigated. The results indicated that a large number of （1.2-1.6 mL per fish） high purity head kidney macrophages at a concentration of 1 × 10^7 cells/mL could be easily obtained. About 95 % 98% cells adhered and spread out on cover slips after 3-18 h in culture were macrophages, The obtained head kidney macrophages showed the morphological characteristics of macrophages and above 95% of the cells had the ability to phagocytose the formalin killed Staphylococcus aureus （F-SA）. PMA induced respiratory burst in the obtained macrophages,which could be inhibited both in vitro and in vivo after a relatively long exposure of the head kidney macrophages to a relatively high dose of exogenous cortisol.

关 键 词：黄颡鱼 巨噬细胞 分离培养 吞噬作用 外源皮质醇 呼吸爆发 

分 类 号：Q959.46[生物学—动物学]