温度对大口黑鲈幼鱼不同组织抗氧化能力及免疫相关蛋白表达的影响
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摘要
将体质量(110±5 g)的大口黑鲈幼鱼饲养在室内100 L白色PVC水桶中,每个桶6尾,在水温21, 25, 29℃和33℃的环境下驯养2周,测定肌肉、肝脏、鳃和肠道中抗氧化酶活性以及丙二醛(MDA)、谷胱甘肽(GSH)、热休克70蛋白(HSP70)和金属硫蛋白(MT)含量,以研究温度对大口黑鲈幼鱼不同组织抗氧化能力及免疫相关蛋白表达的影响。结果显示,在21—33℃条件下,实验鱼超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性均在鳃中最高且变化较大,整体随温度升高而先降低后升高,在29℃组最低。MDA含量在鳃中较高,肠中次之,均在21℃组含量最高。肝脏,鳃和肠中均含有较高GSH,整体随温度升高而呈先降低后升高的趋势。肝脏中总抗氧化能力(T-AOC)处于较高,肠中次之,肌肉中最低,整体随温度升高呈波动性变化,在29℃组最低。HSP70含量在肌肉、肝脏和肠中较高,整体随着温度升高而先升高后降低。MT含量在肌肉和肝脏中较高,不同温度组中MT含量变化不大。结果表明,长期温度胁迫对大口黑鲈幼鱼产生一定应激损伤,引起其机体免疫水平降低,并且高温对其应激损伤大于低温;大口黑鲈幼鱼肝脏、鳃和肠道组织对温度胁迫更敏感。
Juvenile Micropterus salmoides with initial body weight of(110±5 g) g was randomly allocated into 4 temperature groups(21, 25,29℃ and 33℃)in 2 weeks to investigate the effects of temperature on antioxidant capacity and immune associated protein expression in different tissues and organs(white muscle, liver, gill and intestine) of juvenile M. salmoides. Parameters analyzed included superoxide dismutase(SOD), catalase(CAT) activity, malondialdehyde(MDA), glutathione(GSH), total antioxidant capacity(T-AOC), heat shock protein70(HSP70) and metallothionein(MT) content. Results showed that as the temperature ranged from 21℃ to 33℃, the SOD and CAT activity presented earlier decreased and later increased trend, and obtained the minimum value at 29℃. The content of MDA was highest at 21℃ and accumulated in gill and intestine. GSH expressed in various tissues of juvenile M. salmoides, including liver, gill and intestine and presented upward parabola trend. T-AOC also obtained the minimum value at 29℃ and presented a decreasing tendency from liver to intestine and to muscle. HSP 70 was higher in liver, gill and intestine, and revealed in the tendency of parabola. MT was higher in muscle and liver, and showed no great change between different temperature groups. In conclusion, temperature stress does have some effects on liver,gill and intestine of juvenile M. salmoides, leading decreased immunity. High temperature causes more severe stress damage than low temperature. Liver, gill and intestine seem to be more sensitive to temperature stress than muscle.
Juvenile Micropterus salmoides with initial body weight of(110±5 g) g was randomly allocated into 4 temperature groups(21, 25,29℃ and 33℃)in 2 weeks to investigate the effects of temperature on antioxidant capacity and immune associated protein expression in different tissues and organs(white muscle, liver, gill and intestine) of juvenile M. salmoides. Parameters analyzed included superoxide dismutase(SOD), catalase(CAT) activity, malondialdehyde(MDA), glutathione(GSH), total antioxidant capacity(T-AOC), heat shock protein70(HSP70) and metallothionein(MT) content. Results showed that as the temperature ranged from 21℃ to 33℃, the SOD and CAT activity presented earlier decreased and later increased trend, and obtained the minimum value at 29℃. The content of MDA was highest at 21℃ and accumulated in gill and intestine. GSH expressed in various tissues of juvenile M. salmoides, including liver, gill and intestine and presented upward parabola trend. T-AOC also obtained the minimum value at 29℃ and presented a decreasing tendency from liver to intestine and to muscle. HSP 70 was higher in liver, gill and intestine, and revealed in the tendency of parabola. MT was higher in muscle and liver, and showed no great change between different temperature groups. In conclusion, temperature stress does have some effects on liver,gill and intestine of juvenile M. salmoides, leading decreased immunity. High temperature causes more severe stress damage than low temperature. Liver, gill and intestine seem to be more sensitive to temperature stress than muscle.
引文
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[2]农业部渔业渔政管理局.中国渔业统计年鉴[J].北京:中国农业出版社,2017
[3]曾慧慧.胶州湾近岸水域鱼类群落结构特征及多样性研究[D].青岛:中国海洋大学,2012.
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[5]ROESSIG J M,WOODLEY C M,CECH J J,et al.Effects of global climate change on marine and estuarine fishes and fisheries[J].Reviews in Fish Biology&Fisheries,2004,14(2):251-275.
[6]AN M I,CHOI C Y.Activity of antioxidant enzymes and physiological responses in ark shell,Scapharca broughtonii,exposed to thermal and osmotic stress:effects on hemolymph and biochemical parameters[J].Comparative Biochemistry&Physiology Part B Biochemistry&Molecular Biology,2010,155(1):34-42.
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[8]GARCíAG A,HERRERAA J,OCHOA J L.Effect of superoxide dismutase from bovine erythrocytes on different activity parameters in adjuvant-induced arthritis[J].Archives of Medical Research,1999,30(2):132-137.
[9]WINSTON G W,GIULIO R T D.Prooxidant and antioxidant mechanisms in aquatic organisms[J].Aquatic Toxicology,1991,19(2):137-161.
[10]LUSHCHAK V I.Environmentally induced oxidative stress in aquatic animals[J].Aquatic Toxicology,2011,101(1):13-30.
[11]MADEIRA D,NARCISO L,CABRAL H N,et al.HSP70production patterns in coastal and estuarine organisms facing increasing temperatures[J].Journal of Sea Research,2012,73(3):137-147.
[12]杨丽芳.金属硫蛋白的心肌保护机制研究[D].西安:第四军医大学,2014.
[13]SáNCHEZ A,PASCUAL C,SáNCHEZ A,et al.Hemolymph metabolic variables and immune response in Litopenaeus setiferus,adult males:the effect of acclimation[J].Aquaculture,2001,198(1):13-28.
[14]ROBERT E R.Biological indicators of stress in fish[J].Transactions of the American Fisheries Society,1992,121(2):274-276.
[15]罗胜玉,徐冬冬,楼宝,等.低温胁迫对黄姑鱼(Nibea albiflora)抗氧化酶、Na+/-K+-ATP酶及Hsp70蛋白含量的影响[J].海洋通报,2017,36(2):189-194.
[16]LUSHCHAK V I,BAGNYUKOVA T V.Temperature increase results in oxidative stress in goldfish tissues.1.Indices of oxidative stress[J].Comparative Biochemistry&Physiology Part C Toxicology&Pharmacology,2006,143(1):30-35.
[17]KIM J H,PARK H J,KIM K W,et al.Growth performance,oxidative stress,and non-specific immune responses in juvenile sablefish,Anoplopoma fimbria,by changes of water temperature and salinity[J].Fish Physiology&Biochemistry,2017,43(1):1421-1431.
[18]高权新,谢明媚,彭士明,等.急性温度胁迫对银鲳幼鱼代谢酶、离子酶活性及血清离子浓度的影响[J].南方水产科学,2016,12(2):59-66.
[19]谢妙.低温胁迫对斜带石斑鱼生理、生化、脂肪酸的影响[D].湛江:广东海洋大学,2012.
[20]俞丹,沈中源,张智,等.温度驯化对尖头鱥热耐受特征的影响[J].水生生物学报,2017,41(3):538-542.
[21]MADEIRA A,CAMPS M,ZORZANO A,et al.Biophysical assessment of human aquaporin-7 as a water and glycerol channel in 3T3-L1 adipocytes[J].Plos One,2013,8(12):e83442.
[22]HALLIWELL B,GUTTERIDGE J M C.Free radicals in biology and medicine[J].Journal of free radicals in biology&medicine,1985,1(4):331-332.
[23]沈凡,樊启学,杨凯,等.不同溶氧条件下黄颡鱼免疫机能及抗病力的研究[J].淡水渔业,2010,40(4):44-49.
[24]KOMMALY ONXAYVIENG.不同养殖密度对黄颡鱼组份和抗氧化系统产生的相关影响[D].武汉:华中农业大学,2016.
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[26]SEVCIKOVA M,MODRA H,SLANINOVA A,et al.Metals as a cause of oxidative stress in fish:a review[J].VeterinárníMedicína,2011,56(11):537-546.
[27]KIM J H,KANG J C.Oxidative stress,neurotoxicity,and non-specific immune responses in juvenile red sea bream,Pagrus major,exposed to different waterborne selenium concentrations[J].Chemosphere,2015,135:46-52.
[28]杨健,陈刚,黄建盛,等.温度和盐度对军曹鱼幼鱼生长与抗氧化酶活性的影响[J].广东海洋大学学报,2007,27(4):25-29.
[29]王海贞,王辉,李瑞伟,等.温度和盐度对吉富罗非鱼幼鱼肠道两种抗氧化酶活力的联合效应[J].广东海洋大学学报,2012,32(1):47-53.
[30]徐冬冬,楼宝,詹炜,等.高温胁迫对褐牙鲆生长及肝脏抗氧化酶活性的影响[J].水产学报,2010,34(7):1099-1105.
[31]刘峰,刘阳阳,楼宝,等.温度对小黄鱼体内抗氧化酶及消化酶活性的影响[J].海洋学报,2016,38(12):76-85.
[32]王新安,郭黎,马爱军,等.基于响应面法分析温度和盐度及其交互作用对大菱鲆幼鱼抗氧化酶活性的影响[J].海洋科学,2014,38(6):17-23.
[33]LUSHCHAK V I,BAGNYUKOVA T V.Temperature increase results in oxidative stress in goldfish tissues.2.Antioxidant and associated enzymes[J].Comparative Biochemistry&Physiology Part C,2006,143(1):36-41.
[34]FERNO D,DENISSE R A,GONZáLEZ R A,et al.Temperature preference and oxygen consumption of the largemouth bass Micropterus salmoides(Lacépède)acclimated to different temperatures[J].Aquaculture Research,2010,38(13):1387-1394.
[35]雷衍之.养殖水环境化学[M].北京:中国农业出版社,2004.
[36]PATTERSON J T,MIMS S D,WRIGHT R A.Effects of body mass and water temperature on routine metabolism of American paddlefish Polyodon spathula[J].Journal of Fish Biology,2013,82(4):1269-1280.
[37]王刚,李加儿,区又君,等.温度、盐度、p H对卵形鲳鲹幼鱼离体鳃组织耗氧量的影响[J].南方水产科学,2011,07(5):37-42.
[38]VIARENGO A,CANESI L,MARTINEZ P G,et al.Pro-oxidant processes and antioxidant defence systems in the tissues of the Antarctic scallop(Adamussium colbecki)compared with the Mediterranean scallop(Pecten jacobaeus)[J].Comparative Biochemistry&Physiology Part B Biochemistry&Molecular Biology,1995,111(1):119-126.
[39]曾令清,张耀光,付世建,等.双向急性变温对南方鲇幼鱼静止耗氧率和临界游泳速度的影响[J].水生生物学报,2011,35(2):276-282.
[40]BLACK M C,MILLSAP D S,MCCARTHY J F.Effects of acute temperature change on respiration and toxicant uptake by Rainbow Trout,Salmo gairdneri(Richardson)[J].Physiological Zoology,1991,64(1):145-168.
[41]GIESEG S P,CUDDIHY S,HILL J V,et al.A comparison of plasma vitamin C and E levels in two Antarctic and two temperate water fish species[J].Comparative Biochemistry&Physiology Part B Biochemistry&Molecular Biology,2000,125(3):371-378.
[42]罗胜玉,徐冬冬,楼宝,等.低温胁迫对黄姑鱼(Nibea albiflora)抗氧化酶、Na+/-K+-ATP酶及Hsp70蛋白含量的影响[J].海洋通报,2017,36(2):189-194.
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