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| 氨氮胁迫对锦绣龙虾代谢酶和抗氧化酶活力的影响 |
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周钱森1,2, 任宪云2,3, 徐垚2,4, 刘萍2,3, 李健2,3
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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.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;3.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;4.江苏海洋大学 江苏省海洋生物资源与环境重点实验室/江苏省海洋生物技术重点实验室 江苏 连云港 222005
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| 摘要: |
| 为了研究急性氨氮胁迫对锦绣龙虾(Panulirus ornatus)抗氧化系统及氨氮代谢机制的影响，通过设置对照[(0.24±0.07) mg/L]、低浓度[(1.04±0.08) mg/L]、中浓度[(9.75±0.21) mg/L]和高浓度[(19.87±0.46) mg/L]氨氮胁迫方法对锦绣龙虾进行48 h急性实验，测定鳃与肝胰腺中抗氧化及氨氮代谢酶的活性，同时测定相关基因mRNA的相对表达变化情况。结果显示，氨氮胁迫下锦绣龙虾鳃和肝胰腺组织总抗氧化能力(T-AOC)、超氧化物歧化(SOD)活力有不同程度的升高，6~12 h均显著高于对照组(P<0.05)；高浓度组胁迫48 h时T-AOC、SOD活力则均被抑制；过氧化脂质(LPO)含量随胁迫时间的延长逐渐增加，48 h均显著高于对照组(P<0.05)。氨代谢相关酶谷氨酰胺合成酶(GS)、谷氨酸脱氢酶(GDH)、谷草转氨酶(GOT)、黄嘌呤氧化还原酶(XOD)的酶活在氨氮胁迫下与对照组相比均有不同程度的上调，表明其共同参与离子氨的代谢转运。肝胰腺中GDH、GOT基因表达量在12~24 h显著高于对照组(P<0.05)，在鳃组织中变化趋势基本相同。由此可见，不同浓度的氨氮胁迫会对锦绣龙虾抗氧化酶活性造成不同程度的诱导；锦绣龙虾主要通过在肝胰腺GS、GDH等代谢相关酶的共同作用下合成无毒的谷氨酰胺以避免氨在体内过量积累。本研究结果可为锦绣龙虾氨解毒代谢机制的深入研究提供理论依据。 |
| 关键词: 锦绣龙虾 氨氮胁迫 抗氧化 氨代谢酶 |
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| Effects of Ammonia Stress on Metabolic and Antioxidant Enzyme Activities in Panulirus ornatus |
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ZHOU Qiansen1,2, REN Xianyun2,3, XU Yao2,4, LIU Ping2,3, LI Jian2,3
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1.National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;2.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao, Shandong 266071, China;3.Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine
Science and Technology (Qingdao), Qingdao, Shandong 266071, China;4.Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang, Jiangsu 222005, China
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| Abstract: |
| In this study, in order to explore the effects of acute ammonia stress on the antioxidant and ammonia-nitrogen metabolism systems of Panulirus ornatus, lobsters were exposed to different concentrations of ammonia (0.24±0.07, 1.04±0.08, 9.75±0.21, and 19.87±0.46 mg/L) for up to 48 h. The activity of antioxidant and ammonia-nitrogen metabolizing enzymes and related gene expression were assessed in the gills and hepatopancreas at 0, 6, 12, 24, and 48 h after ammonia stress. From 6 to 12 h, total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity in the gills and hepatopancreas of P. ornatus under ammonia nitrogen stress increased to varying degrees, and was significantly higher than that of the control group (P<0.05). In the high-concentration group, T-AOC and SOD activity was inhibited for 48 h. The LPO content gradually increased with prolonged stress and was significantly higher than that of the control group (P<0.05) at 48 h. The activities of the ammonia metabolism enzymes glutamine synthetase (GS), glutamate dehydrogenase (GDH), glutamic-oxalacetic transaminase (GOT), and xanthine oxidoreductase (XOD) were all up-regulated to varying degrees under ammonia stress compared with those of the control group, indicating that they are jointly involved in the metabolic transport of NH4+. Additionally, from 12 to 24 h, the expression of GDH and GOT genes in the hepatopancreas was significantly higher than that of the control group (P<0.05); the same trend was identified in gill tissue. In brief, different concentrations of ammonia stress induce different levels of antioxidant enzyme activity. Panulirus ornatus mainly synthesizes glutamine under the combined action of hepatopancreas GS, GDH, and other metabolic enzymes to avoid excessive ammonia in the body. |
| Key words: Panulirus ornatus Ammonia nitrogen stress Antioxidant Ammonia metabolic enzyme |