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低温胁迫对日本对虾抗氧化系统和细胞凋亡的影响
于振兴,任宪云,邵慧鑫,刘萍,李健
1.水产科学国家级实验教学示范中心 上海海洋大学 上海 201306;2.青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 山东 青岛 266071;3.中国水产科学研究院黄海水产研究所 农业农村部海洋渔业可持续发展重点实验室 山东 青岛 266071
摘要:
日本对虾(Marsupenaeus japonicus)适宜生长温度为23℃~32℃,由于北方地区冬季温度较低,严重影响日本对虾养殖周期,因此,培育耐低温的新品种非常重要。将(14.24±1.12) g的日本对虾暴露于10℃、16℃、22℃水体中72 h,以28℃水体中的对虾为对照组。结果显示,对虾鳃和肝胰腺的总抗氧化能力、超氧化物歧化酶(SOD)活性、还原型谷胱甘肽含量总体分别呈先下降后升高再降低的变化趋势,而丙二醛含量、天冬氨酸特异性半胱氨酸蛋白酶3 (Caspase-3)活性一直呈上升趋势;SOD和Caspase-3基因的相对表达量分别呈先下降后升高再降低和持续升高的趋势,与各自对应的酶活变化趋势相似。TUNEL检测发现,随胁迫时间的延长,机体细胞凋亡率显著增加,其中,对照组、3 h 10℃组和72 h 10℃组鳃组织细胞凋亡率分别为2.03%、6.20%和26.27%,肝胰腺细胞凋亡率分别为1.06%、25.65%和42.33%。综上所述,低温胁迫降低了日本对虾的抗氧化能力,破坏了内环境原有的平衡,进而导致机体出现氧化损伤。本研究可为耐低温新品种的培育提供参考。
关键词:  日本对虾  低温胁迫  总抗氧化能力  细胞凋亡  TUNEL检测
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Effect of Low Temperature Stress on Antioxidant System and Apoptosis of Marsupenaeus japonicus
YU Zhenxing1,2, REN Xianyun2,3,4, SHAO Huixin1,2, LIU Ping2,3,4, LI Jian2,3,4
1.National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;2.Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao, Shandong 266071, China;3.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs;4.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, Shandong 266071, China
Abstract:
Marsupenaeus japonicus is an important cultured species of marine shrimp in China. Its characteristics include a varied diet, fast growth, and tolerating periods out of water, as well as being suitable for live shrimp marketing and long-distance transportation. The suitable growth temperature of M. japonicus is 23℃~32℃. The culture cycle of M. japonicus is seriously affected by low winter temperatures in northern China, which increases the costs of parent shrimp overwintering. Therefore, it is necessary to breed new varieties with low temperature tolerances. M. japonicus was exposed to water at 10℃, 16℃, and 22℃ for 72 h, with M. japonicus exposed to water at 28℃ used as the control group. The results showed that the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) enzyme activity, and GSH content in the gill and hepatopancreas decreased first, then increased and then decreased, while the MDA content and caspase-3 enzyme activity increased over time. The relative expression of the SOD and Caspase-3 genes decreased first, then increased, then decreased, and continued to increase, which was similar to their corresponding changes in enzyme activity. In order to prove that low temperature stress caused damage to the body of M. japonicus, a TUNEL test was performed. It was found that the apoptosis rate increased significantly with the extension of the stress time. The apoptotic rates of the gill control group, 3 h at 10℃, and 72 h at 10℃ were 2.03%, 6.20%, and 26.27%, respectively. The apoptotic rates of the hepatopancreas cells were 1.06%, 25.65%, and 42.33% for these exposure times, respectively. In conclusion, low temperature stress reduced the antioxidant capacity of M. japonicus, destroyed the original balance of the internal environment, and then led to oxidative damage. In this study, the changes in cell apoptosis and the antioxidant status of M. japonicus under low temperature conditions provided data that support the need to cultivate new varieties with low temperature tolerances.
Key words:  Marsupenaeus japonicus  Low temperature stress  T-AOC  Apoptotic  TUNEL