急性氨氮胁迫对红螯螯虾血细胞凋亡的影响  

Effect of acute ammonia-N stress on haemocyte apoptosis of the red claw crayfish Cherax quadricarinatus

作　　者：潘训彬 张秀霞[2] 冼健安[2] 

机构地区：[1]华南师范大学生命科学学院广东省水产健康安全养殖重点实验室,广东广州510631 [2]中国热带农业科学院热带生物技术研究所,海南海口571101

出　　处：《河北渔业》2017年第11期4-7,28,共5页

基　　金：海南省重点研发计划项目(ZDYF2016084);中国热带农业科学院基本科研业务费专项资金(1630052016011和1630052016021)

摘　　要：氨氮是水产养殖过程中的常见毒性污染物之一,为探讨氨氮对红螯螯虾(Cherax quadricarinatus)血细胞的细胞毒性机制,研究了氨氮胁迫作用下红螯螯虾血细胞凋亡状态的变化。以17.6mg/L氨氮对红螯螯虾幼虾进行急性胁迫,于胁迫后的0、48、72和96h取血淋巴,应用流式细胞术测定血细胞总数(THC)、胞内游离Ca^(2+)含量、线粒体膜电位(MMP)和血细胞凋亡率。结果显示,红螯螯虾的THC在胁迫的72h开始有显著的下降,在96h时达到最低值;血细胞胞内游离Ca^(2+)含量在48h开始有显著的升高,在96h达到最高值;血细胞MMP在72h开始有显著的下降,在96h达到最低值;血细胞凋亡率在72h开始有显著的升高,在96h时达到最高值,为9.39%。这些结果表明,胞内游离Ca^(2+)含量是对氨氮胁迫最为敏感的指标;氨氮急性胁迫诱导红螯螯虾血细胞发生了Ca^(2+)参与调节、线粒体通路相关的细胞凋亡,从而导致了THC的下降。The juvenile crayfish Cherax quadricarinatus were exposed to 17.6 mg/L ammonia-N,and then total haemocyte count(THC),cytoplasmic free-Ca^(2+)concentration,mitochondrial membrane potential(MMP)and apoptotic ratio of haemocytes were determined using flow cytometry after 0,48,72,96 and 120 hexposure.Results showed that THC reduction could be observed after 72~120 hexposure,and the THC reached the lowest value after 96 h.Cytoplasmic free-Ca^(2+)concentration of haemocytes significantly increased after 48~120 hexposure,and reached the highest level after 96 h.MMP of haemocytes decreased after 72~120 hexposure,and reached the lowest level after 96 h.Apoptotic haemocyte ratio increased after 72~120 hexposure,and reached to 9.39% which was the highest level after 96 h.These results indicated that cytoplasmic free-Ca^(2+)concentration was the most sensitive indicator against ammonia-N stress.Acute ammonia-N stress induced Ca^(2+)-mediated and mitochondrial pathway-related haemocyte apoptosis,and then caused THC reduction.

关 键 词：氨氮 红螯螯虾(Cherax quadricarinatus) 血细胞 凋亡 流式细胞术 

分 类 号：S917.4[农业科学—水产科学] X503.22[环境科学与工程—环境工程]