Cd对凡纳滨对虾血细胞活性氧含量(ROS)、细胞凋亡率及应激反应基因表达的影响  

Effects of Cd on reactive oxygen species production, apoptosis rate, and expression of stress genes in hemocytes of Litopenaeus vannamei

作　　者：李玲 吴方舟 梁金荣 陈丽芬 郭慧[1] 申玉春[1] LI Ling;WU Fangzhou;LIANG Jinrong;CHEN Lifen;GUO Hui;SHEN Yuchun(College of Fisheries,Guangdong Ocean University,Zhanjiang 524000,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524000

出　　处：《中国水产科学》2021年第2期167-175,共9页Journal of Fishery Sciences of China

基　　金：国家自然科学基金项目(31600321).

摘　　要：为探讨镉(Cd)对虾类血细胞的免疫毒性,以凡纳滨对虾(Litopenaeus vannamei)为研究对象,设置0 mg/L对照组和10.5 mg/L Cd处理组,利用流式细胞仪检测血细胞中的活性氧含量(ROS)和细胞凋亡率,并利用实时荧光定量PCR技术检测血细胞中抗氧化酶(Cu-Zn SOD、CAT和GPx)、金属硫蛋白(MT)、热休克蛋白(HSP60、HSP70和HSP90)以及caspase-3基因的表达量变化。结果显示,与对照组相比,Cd胁迫导致血细胞ROS含量在胁迫后3~48 h极显著升高(P<0.01),在24 h达到最大值;细胞凋亡率在3 h达到最大值87.99%(P<0.01),在12~48 h显著升高(P<0.05);Cu-Zn SOD和HSP60基因表达水平分别在3~48 h和12~48 h显著升高(P<0.05);GPx基因表达量在12 h和48 h显著上调(P<0.05);CAT、MT、HSP60和HSP90的基因表达水平在胁迫3 h显著下降(P<0.05),在胁迫后期显著升高(P<0.05);HSP70表达量在3~24 h无显著变化(P>0.05),在48 h显著上升(P<0.05);caspase-3基因表达水平在12~24 h显著上升(P<0.05)。综上所述,Cd诱导血细胞产生过量ROS,影响应激反应基因的表达水平,最终促进细胞凋亡。In this study,we aimed to explore the effects of Cd on the reactive oxygen species(ROS)production,apoptosis rate,and gene expression of stress responses in the hemocytes of Litopenaeus vannamei.L.vannamei,with mean weights of(10.22±0.47)g,were cultured at a salinity of 24,pH of 7.9-8.0,and temperature of(26±2)℃for 2 weeks with aeration.Shrimp of similar size and body color were then transferred into culture buckets for stress testing.Based on the results of a preliminary investigation and previous studies,two concentrations of Cd,0 and 10.5 mg/L,were tested.Twenty shrimps were placed in each bucket and each treatment was conducted in three replicates.At 0 h,3 h,12 h,24 h,and 48 h of exposure,three shrimp in each bucket were sampled.The ROS production and cell apoptosis rate in the hemocytes of the shrimp were evaluated using flow cytometry,and the expression profiles of metallothionein(MT);antioxidant genes including copper zinc superoxide dismutase(Cu-Zn SOD),glutathione peroxidase(GPx),and catalase(CAT);caspase-3;and heat shock protein genes(HSPs)including HSP60,HSP70,and HSP90 were detected by quantitative reverse-transcription PCR.Compared to the control group,ROS production significantly increased at 3-48 h(P<0.01)and peaked at 24 h.The apoptosis rate peaked to 87.99%at 3 h(P<0.01)and significantly increased at 12-48 h(P<0.05).The expression levels of Cu-Zn SOD and HSP60 significantly increased at 3-48 h and 12-48 h,respectively(P<0.05).The expression of GPx was significantly upregulated at 12 h and 48 h(P<0.05).The expression profiles of CAT,MT,HSP60,and HSP90 significantly decreased at 3 h(P<0.05),while CAT increased significantly at 12 h(P<0.05),and MT,HSP60 and HSP90 increased at 24-48 h(P<0.05).The expression of HSP70 showed no significant differences at 3-24 h(P>0.05),whereas it increased significantly at 48 h(P<0.05).Caspase-3 expression increased significantly at 12-24 h(P<0.05).This research found that Cd promoted excessive ROS production,significantly affected the expression of stress response

关 键 词：CD 凡纳滨对虾 ROS 凋亡率 应激 基因表达 

分 类 号：S966[农业科学—水产养殖]