干露及恢复对中间球海胆抗氧化能力、细胞凋亡和免疫相关基因表达的影响  

Effects of air exposure and recovery on antioxidant capacity,apoptosis and immune related gene expression in coelomocytes of sea urchin(Strongylocentrotus intermedius)

作　　者：林威港 孙琦 海航瑜 刘亚婷 赵鹏[1] 常亚青[1] 马得友 LIN Weigang;SUN Qi;HAI Hangyu;LIU Yating;ZHAO Peng;CHANG Yaqing;MA Deyou(Key Laboratory of Mariculture&Stock Enhancement in North Chinas Sea,Ministry of Agriculture and Rural Affairs,Dalian Ocean University,Dalian 116023,China;Shanghai Fisheries Research Institute,Shanghai 200433,China;College of Marine Life Sciences,Ocean University of China,Qingdao 266000,China)

机构地区：[1]大连海洋大学农业农村部北方海水增养殖重点实验室,辽宁大连116023 [2]上海市水产研究所,上海200433 [3]中国海洋大学海洋生命学院,山东青岛266000

出　　处：《大连海洋大学学报》2024年第1期66-73,共8页Journal of Dalian Ocean University

基　　金：国家重点研发计划项目(2019YFD0900405);辽宁省教育厅高等学校基本科研项目(20230059);辽宁省“兴辽英才计划”项目(XLYC2002107)。

摘　　要：为研究干法运输对中间球海胆(Strongylocentrotus intermedius)活力的影响,采用低温(4℃)干露胁迫12、24、36 h后再浸润恢复12 h的处理方法,测定了湿质量为(3.57±0.63)g的中间球海胆体腔细胞超氧化物歧化酶(T-SOD)活力、丙二醛(MDA)含量和总抗氧化能力(T-AOC)水平,通过流式细胞仪检测了细胞凋亡情况,利用实时定量PCR分析了不同时间干露胁迫下免疫相关基因LYZ、TLR1及凋亡相关基因Caspase-3和Caspase-9在体腔细胞中的表达情况。结果表明:干露试验组海胆体腔液T-SOD活力、MDA含量和T-AOC水平随干露时间的延长呈现先升高后降低的变化趋势,干露12 h时各指标值均达到最高,随后下降,并于复水12 h时接近对照组(P>0.05);免疫相关基因LYZ和TLR1总体上随干露时间的延长呈先减弱后增强的表达趋势,分别在干露24、36 h时达到峰值,入水恢复12 h时两基因的表达量显著低于对照组(P<0.05);发生凋亡的体腔细胞比率随干露时间延长的变化趋势与T-AOC水平的变化趋势相似,干露12 h时达到最高值,而Caspase-3、Caspase-9基因的表达量具有时间累积效应,分别在干露24、36 h时显著高于对照组和其他处理组(P<0.05)。研究表明,中间球海胆具有一定的耐干露能力,通过抗氧化酶系统维持活性氧自由基的动态平衡,LYZ、TLR1等固有免疫分子和Caspase依赖性细胞凋亡因子在应对干露胁迫、机体维持内稳态的过程中发挥重要作用。To evaluate the effect of air exposure transportation on the survival of sea urchin(Strongylocentrotus in-termedius),the total superoxide dismutase(T-SOD)activity,malondialdehyde(MDA)content and total antioxi-dant capacity(T-AOC)were determined during 12,24 and 36 h air exposure stress and subsequent 12 h recovery,and apoptosis was detected by flow cytometry,and the expression levels of lysozyme gene LYZ,TLR1,Caspase-3 and Caspase-9 in coelomocytes were investigated using real-time quantitative PCR.The main results showed that the T-SOD activity,MDA content,and T-AOC level generally increased first and then decreased during exposure to air,with the maximum values observed at 12 h air exposure stress and no significant differences between control and 12 recovery(P>0.05).Air exposure had significant effects on the expression of immune related genes LYZ and TLR1,reaching the maximum values at 24 and 36 h of air exposure,which were significantly higher than those in other stress groups(P<0.05).Their expression levels after 12 h recovery following exposure times 36 h were also significantly different from those in controls(P<0.05).It was identified that the change trend of apoptosis rates was similar to that of the T-AOC levels,reaching the maximum value at 12 h air exposure.However,the expression levels of Caspase-3 and Caspase-9 genes presented significant differences between control and time intervals(P<0.05),with the maximum values observed at 24 h and 36 h of air exposure,respectively,suggesting response of the two important apoptosis-related Caspases had time cumulative effect.These findings indicated that sea urchins S.intermedius have certain ability on air exposure tolerance,using antioxidant enzyme system to conserve the dy-namic balance of reactive oxygen species(ROS).Innate immune molecules such as lysozyme and TLR1,as well as Caspase dependent apoptosis,play important roles in maintaining homeostasis in response to air exposure stress.

关 键 词：中间球海胆 干露 抗氧化 免疫 细胞凋亡 

分 类 号：S917.4[农业科学—水产科学]