摘要
为系统研究草鱼血红蛋白对草鱼肾(CIK)细胞的损伤机制,实验探究了血红蛋白和血红素刺激CIK细胞后的生长情况;还检测了血红蛋白和血红素刺激对CIK细胞内铁代谢相关基因的表达情况。为进一步检测血红蛋白对CIK细胞的毒性作用,实验还检测了血红蛋白刺激CIK细胞后,细胞内铁离子和活性氧(ROS)的变化情况;以及CIK细胞受血红蛋白和血红素刺激后,细胞凋亡相关基因的表达情况。结果显示,血红蛋白和血红素明显抑制了CIK细胞的正常生长。血红蛋白和血红素的刺激不同程度地上调了铁代谢相关基因的表达。通过检测被血红蛋白和血红素刺激后的CIK细胞炎症和抗氧化酶相关基因的表达情况,发现血红蛋白和血红素可以通过NF-κB途径激活下游炎症因子的表达,如促炎因子TNF-α、IL-1β和IL-6,抑炎因子IL-10以及趋化因子IL-4和IL-8等,同时也激活了超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等抗氧化酶基因的表达水平。血红蛋白的刺激显著增加了细胞内铁离子和ROS的含量,血红蛋白和血红素都在不同程度上显著激活了CIK细胞凋亡相关基因的表达水平。研究表明,高氧化活性的血红蛋白可以激活细胞内铁代谢和炎症因子相关基因的表达,同时增加细胞内铁离子和ROS水平导致CIK细胞凋亡基因的高表达。本研究结果丰富了鱼类血液免疫基础理论,同时也为水产健康养殖提供新的参考。
This study aims to explore the damage mechanism of hemoglobin on Ctenopharyngodon idella kidney(CIK)cells.Firstly,the growth of CIK cells stimulated by hemoglobin and heme was observed,and the results showed that hemoglobin and heme significantly inhibited the growth of CIK cells.Secondly,the expression of iron metabolism-related genes in CIK cells was detected after stimulated by hemoglobin and hem.The results showed that the stimulation of hemoglobin and heme up-regulated the expression of iron metabolism-related genes to dif-ferent degrees.The expression of inflammation and antioxidant-related enzymes in CIK cells after stimulated by hemoglobin and heme was also examined.The results suggested that hemoglobin and heme could activate the expression of inflammatory factors through NF-κB pathway,such as pro-inflammatory cytokines TNF-α,IL-1βand IL-6,the anti-inflammatory factor IL-10 and chemokines IL-4 and IL-8,and also activate the expression levels of three antioxidant enzymes,including Superoxide dismutase(SOD),Catalase(CAT)and Glutathione peroxidase(GSH-Px).In order to further detect the toxic effect of hemoglobin on CIK cells,this study also detected the intra-cellular iron and ROS in CIK cells after hemoglobin stimulation.The results showed that hemoglobin stimulation significantly increased intracellular iron and ROS.Finally,we also examined the expression of apoptosis-related genes in CIK cells after stimulated by hemoglobin and heme,and the results showed that both hemoglobin and heme significantly activated the expression levels of apoptosis-related genes in CIK cells.In conclusion,the res-ults of this study suggest that the high oxidative activity of hemoglobin can activate the expression of iron metabol-ism-and inflammation-related genes,as well as increasing intracellular iron and ROS levels,leading to up-regu-lated expression of apoptosis genes in CIK cells.
作者
秦真东
杨敏璇
卢志杰
施斐
李亚男
詹凡玢
万全元
李军
林蠡
QIN Zhendong;YANG Minxuan;LU Zhijie;SHI Fei;LI Yanan;ZHAN Fanbin;WAN Quanyuan;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)
出处
《水产学报》
CAS
CSCD
北大核心
2023年第7期136-147,共12页
Journal of Fisheries of China
基金
国家自然科学基金(31902409,31872606)
广州市科技计划(201804010096)。
关键词
草鱼
血红蛋白
氧化损伤
凋亡
Ctenopharyngodon idella
hemoglobin
oxidative damage
apoptosis
