草鱼血红蛋白通过诱导头肾巨噬细胞表达的炎症因子激活免疫反应  

Hemoglobin stimulate inflammatory cytokine expression in grass carp(Ctenopharyngodon idella)head-kidney macrophages

作　　者：秦真东 詹凡玢 李亚男 施斐 杨敏璇 卢志杰 李军 林蠡 QIN Zhendong;ZHAN Fanbin;LI Yanan;SHI Fei;YANG Minxuan;LU Zhijie;LI Jun;LIN Li(Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center,Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding,College of Animal Sciences and Technology,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China)

机构地区：[1]仲恺农业工程学院动物科技学院,广东省水环境与水产品安全工程技术研究中心,广州市水产病害与水禽养殖重点实验室,广东广州510225

出　　处：《水产学报》2023年第3期165-174,共10页Journal of Fisheries of China

基　　金：国家自然科学基金(31902409,31872606);广东省海洋经济发展专项(粤农2019A1)。

摘　　要：为了探究鱼类体内过量游离血红蛋白对鱼体的影响,本实验以草鱼为研究对象,通过体内注射血红蛋白模拟体内出血,探究血红蛋白对组织和组织中巨噬细胞的免疫调控作用。体内注射血红蛋白的实验结果显示,血红蛋白的刺激导致头肾和中肾组织中出现明显的血红蛋白沉淀,同时提高了鱼体血清中的抗氧化相关酶活性,促进了头肾组织中炎症因子基因TNF-α、IL-1β和IL-10的mRNA表达水平。普鲁士蓝和间接免疫荧光实验结果显示,头肾巨噬细胞对血红蛋白表现出了吞噬活性。荧光定量PCR检测结果显示,血红蛋白的刺激显著激活了头肾巨噬细胞中CD68、CD86、CSF和MHC-Ⅱ的mRNA表达水平。进一步的细胞因子检测结果显示,血红蛋白刺激12 h后,头肾巨噬细胞中的炎症因子基因(TNF-α、IL-1β和IL-4)、趋化因子基因(CCL20和CCL4)、IFN-α和TLR4的mRNA表达水平被显著上调表达。研究表明,草鱼头肾巨噬细胞对血红蛋白具有吞噬能力,同时血红蛋白也激活了巨噬细胞中多种细胞因子的表达。本研究结果首次阐述了草鱼血红蛋白与巨噬细胞的相互作用关系,丰富了鱼类血液基础免疫学理论,同时也为鱼类健康养殖提供了新的参考。In general,disease caused by the hemorrhagic pathogen infections,results in hemolysis,in series the hemoglobin(Hb)is discharged into the tissues.Consecutively,oxidation of Hb while exposed to air provokes serious high oxidative damage levels in the body and inducing a large amount of inflammation to release additional proinflammatory mediators.Promptly,it is critical to clear the highly cytotoxic Hb.Macrophages play a key role not only under stress conditions,but also in sustaining homeostasis by promoting erythropoiesis and hemoglobin(Hb)clearance.However,the complete mechanism of the oxidized Hb in teleost fish and the effects of macrophages on it not yet clear.In this study,head-kidney macrophages and Hb of the C.idella were taken as the research model to examine the interaction between Hb and macrophages.Stimulation of the Hb during in vivo studies led to the visible deposition of Hb in the head and middle of the kidney and elevated the levels of antioxidant-related enzymes in serum,and further promoted the expression levels of inflammatory cytokine,such as TNF-α,IL-1β,and IL-10 in head kidney.Thereafter,the indirect immunofluorescence and Prussian blue staining analyses indicated that the macrophages of the head kidney exhibited intense phagocytic activity to Hb.Analysis of gene expression level by quantitative real-time PCR,macrophage markers,including CD68,CD86,CSF,and MHC-Ⅱgenes were significantly elevated through the stimulation of Hb.Likewise,the expression levels of inflammatory cytokines(TNF-α,IL-1β,and IL-4),chemokines(CCL20 and CCL4),IFN-αand TLR4 genes were significantly up-regulated in the head kidney macrophages after 12 h of Hb stimulation.In summary,these results indicate that C.idella head kidney macrophages possess the phagocytic ability towards Hb,and in turn,Hb also activates the expression levels of macrophage-related markers and associated cytokines.Taken together,our findings explicitly improve the understanding of Hb macrophage interactions of C.idella and provided a new referenc

关 键 词：草鱼 血红蛋白 巨噬细胞 细胞因子 

分 类 号：S941.4[农业科学—水产养殖]