声刺激对斜带石斑鱼血清酶活力及肝组织HSP70表达量、病理变化的影响
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摘要
为研究声刺激对斜带石斑鱼Epinephelus coioides生理生化的影响,以体质量为(42.54±6.03)g、体长为(13.26±0.78)cm的斜带石斑鱼为研究对象,将其暴露于正弦波纯音[600 Hz,130 dB(re 1μPa)]信号下,分别在声刺激0 (对照)、2、6、12、24、72、120 h时测定斜带石斑鱼血清中酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、谷草转氨酶(GOT)和谷丙转氨酶(GPT)活力及肝脏中热休克蛋白70(HSP70)表达量的变化,并对肝脏组织进行病理染色检测分析。结果表明:在试验周期内,斜带石斑鱼无死亡现象;声刺激0～120 h内,斜带石斑鱼血清中ACP酶活力呈现先降低后升高再恢复至初始(0 h)水平的变化趋势,AKP酶活力总体呈现先下降后恢复至初始水平的变化趋势,ACP酶活力在声刺激24 h时达到最高值(15.04 U/100 mL)(P<0.05),AKP酶活力在声刺激48 h时达到最低值(6.22 U/100 mL)(P<0.05),两种酶均在72 h后恢复至初始水平(P>0.05);GOT和GPT酶活力均呈现先升高后下降最后恢复至初始水平的变化趋势,GOT和GPT酶活力分别在6、12 h时达到最高水平(P<0.05),均在48 h后恢复至初始水平(P>0.05);Western Blot检测显示,声刺激后斜带石斑鱼肝脏中HSP70表达量均有所升高,声刺激12 h时HSP70表达量最高(P<0.05);肝组织切片观察结果显示,声刺激后斜带石斑鱼肝细胞胞质内脂质含量增加,细胞多呈空泡状,在试验后期,肝细胞出现肝窦间隙增大,肝板消失,细胞核偏移甚至消失等变化,12、48、72 h时有红细胞聚集现象,但肝细胞均未见明显的坏死、炎症和纤维化等病理改变。研究表明,声刺激会导致斜带石斑鱼产生应激反应,且生理生化指标的变化具有一定时间效应。
Activities of acid phosphatase(ACP), alkaline phosphatase(AKP), aspartate aminotransferase(GOT), alanine aminotransferase(GPT) in the serum and changes in expression of heat shock protein 70(HSP70) in liver were determined in grouper Epinephelus coioides with body weight of(42.54±6.03)g, and body lengthof(13.26±0.78)cm exposed to sine wave pure tone[600 Hz, 130 dB(re 1 μPa)]0, 2, 6, 12, 24, 72, and 120 h, and the liver tissue was analyzed by pathological staining to investigate the effect of acoustic stimulation on physiological and biochemical characteristics of the grouper. The results showed that the activities of serum ACP in the grouper with the acoustic exposure were decreased first and then increased to the initial level, without death during the test period. The activities of AKP enzyme were found to be decreased first and then returned to the initial level, with the maximal activities of ACP and AKP(15.04 U/100 mL) at 24 h and the minimal value(6.22 U/100 mL) at 48 h(P<0.05), finally, both being recovery to original levels at 72 h(P>0.05). The activities of GOT and GPT were shown to be increased first and then returned to the initial level. Western Blot showed that the expression of HSP70 in the grouper was increased, with the maximum at 12 h(P<0.05). The histological sections showed that there was increase in lipid content in the cytoplasm of hepatocytes, the most vacuolated cells, after sonic stimulation. In the later stage of the experiment, the hepatic sinus gaps were increased, the liver plates were disappeared, and the nucleus was even shifted. There were blood cell aggregations at 12, 48, and 72 h, without obvious pathological changes including necrosis, inflammation and fibrosis in hepatocytes. The findings indicated that acoustic stimulation led to stress response and to a certain time effect on changes in physiological and biochemical indicators in the grouper.
Activities of acid phosphatase(ACP), alkaline phosphatase(AKP), aspartate aminotransferase(GOT), alanine aminotransferase(GPT) in the serum and changes in expression of heat shock protein 70(HSP70) in liver were determined in grouper Epinephelus coioides with body weight of(42.54±6.03)g, and body lengthof(13.26±0.78)cm exposed to sine wave pure tone[600 Hz, 130 dB(re 1 μPa)]0, 2, 6, 12, 24, 72, and 120 h, and the liver tissue was analyzed by pathological staining to investigate the effect of acoustic stimulation on physiological and biochemical characteristics of the grouper. The results showed that the activities of serum ACP in the grouper with the acoustic exposure were decreased first and then increased to the initial level, without death during the test period. The activities of AKP enzyme were found to be decreased first and then returned to the initial level, with the maximal activities of ACP and AKP(15.04 U/100 mL) at 24 h and the minimal value(6.22 U/100 mL) at 48 h(P<0.05), finally, both being recovery to original levels at 72 h(P>0.05). The activities of GOT and GPT were shown to be increased first and then returned to the initial level. Western Blot showed that the expression of HSP70 in the grouper was increased, with the maximum at 12 h(P<0.05). The histological sections showed that there was increase in lipid content in the cytoplasm of hepatocytes, the most vacuolated cells, after sonic stimulation. In the later stage of the experiment, the hepatic sinus gaps were increased, the liver plates were disappeared, and the nucleus was even shifted. There were blood cell aggregations at 12, 48, and 72 h, without obvious pathological changes including necrosis, inflammation and fibrosis in hepatocytes. The findings indicated that acoustic stimulation led to stress response and to a certain time effect on changes in physiological and biochemical indicators in the grouper.
引文
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[14] 虞顺年,魏小岚,韦芳三,等.不同运动强度对黑鲷生长、血清和肝脏抗氧化指标的影响[J].水生生物学报,2018,42(4):255-263.
[15] 谭树华,李玉峰,刘文海,等.十二烷基硫酸钠(SDS)对克氏原螯虾抗氧化功能和酸性磷酸酶活性的影响[J].生态与农村环境学报,2014,30(3):392-397.
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[19] 崔前进,陈冰,邱丽华,等.低盐胁迫对钝吻黄盖鲽幼鱼鳃Na+/K+-ATP酶、肝脏抗氧化酶和非特异免疫酶的影响[J].广东海洋大学学报,2017,37(6):26-32.
[20] 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.
[21] 陈玉春,顾雪飞,刘敏.5种中草药对鲤血清谷丙转氨酶、谷草转氨酶及红细胞过氧化氢酶活性的影响[J].淡水渔业,2007,37(5):11-13.
[22] 王涛,苗亮,李明云,等.突降盐度胁迫对大黄鱼(Pseudosciaena crocea)血清生理生化及鳃丝Na+/K+-ATP酶活性的影响[J].海洋与湖沼,2013,44(2):421-426.
[23] Heredia-Middleton P,Brunelli J,Drew R E,et al.Heat shock protein (HSP70) RNA expression differs among rainbow trout (Oncorhynchus mykiss) clonal lines[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2008,149(4):552-556.
[24] O’Connor W A,Lawler N F.Salinity and temperature tolerance of embryos and juveniles of the pearl oyster,Pinctada imbricata R?ding[J].Aquaculture,2004,229(1-4):493-506.
[25] Ruscoe I M,Shelley C C,Williams G R.The combined effects of temperature and salinity on growth and survival of juvenile mud crabs (Scylla serrata Forsk?l)[J].Aquaculture,2004,238(1-4):239-247.
[26] Soria G,Merino G,von Brand E.Effect of increasing salinity on physiological response in juvenile scallops Argopecten purpuratus at two rearing temperatures[J].Aquaculture,2007,270(1-4):451-463.
[27] Castelo Branco N A,Rodriguez E,Alves-Pereira M,et al.Vibroacoustic disease:some forensic aspects[J].Aviation,Space,and Environmental Medicine,1999,70(3):A145-A151.
[28] Alves-Pereira M,Castelo Branco N A A.Vibroacoustic disease:biological effects of infrasound and low-frequency noise explained by mechanotransduction cellular signalling[J].Progress in Biophysics and Molecular Biology,2007,93(1-3):256-279.
[29] Oliveira M J R,Freitas D,Carvalho A P O,et al.Exposure to industrial wide band noise increases connective tissue in the rat liver[J].Noise & Health,2012,14(60):227-229.
[30] 区又君,刘奇奇,温久福,等.急性低温胁迫对四指马鲅幼鱼肝脏、肌肉以及鳃组织结构的影响[J].生态科学,2018,37(05):53-59.
[31] 彭士明,施兆鸿,李杰,等.运输胁迫对银鲳血清皮质醇、血糖、组织中糖元及乳酸含量的影响[J].水产学报,2011,35(6):831-837.
[2] Hawkins A,Popper A.Effects of noise on fish,fisheries,and invertebrates in the U.S.Atlantic and Arctic from energy industry sound-generating activities[R].Bedford N H:Bureau of Ocean Energy Management,2012.
[3] Ren Huiying,Halvorsen M B,Deng Z D,et al.Aquatic acoustic metrics interface utility for underwater sound monitoring and analysis[J].Sensors,2012,12(6):7438-7450.
[4] Vasconcelos R O,Amorim M C P,Ladich F.Effects of ship noise on the detectability of communication signals in the Lusitanian toadfish[J].Journal of Experimental Biology,2007,210(12):2104-2112.
[5] McDonald J I,Wilkens S L,Stanley J A,et al.Vessel generator noise as a settlement cue for marine biofouling species[J].Biofouling:The Journal of Bioadhesion and Biofilm Research,2014,30(6):741-749.
[6] Popper A N,Fay R R,Platt C,et al.Sound detection mechanisms and capabilities of teleost fishes[M]//Collin S P,Marshall N J.Sensory Processing in Aquatic Environments.New York:Springer,2003:3-38.
[7] Voellmy I K,Purser J,Flynn D,et al.Acoustic noise reduces foraging success in two sympatric fish species via different mechanisms[J].Animal Behaviour,2014,89:191-198.
[8] Voellmy I K,Purser J,Simpson S D,et al.Increased noise levels have different impacts on the anti-predator behaviour of two sympatric fish species[J].PLoS One,2014,9(7):e102946.
[9] Holles S,Simpson S D,Radford A N,et al.Boat noise disrupts orientation behaviour in a coral reef fish[J].Marine Ecology Progress Series,2013,485:295-300.
[10] Wysocki L E,Dittami J P,Ladich F.Ship noise and cortisol secretion in European freshwater fishes[J].Biological Conservation,2006,128(4):501-508.
[11] Slabbekoorn H,Bouton N,van Opzeeland I,et al.A noisy spring:the impact of globally rising underwater sound levels on fish[J].Trends in Ecology & Evolution,2010,25(7):419-427.
[12] Hoelzel A R.Marine Mammal Biology:An Evolutionary Approach[M].Oxford:Wiley-Blackwell,2002.
[13] 何海琪,孙凤.中国对虾酸性和碱性磷酸酶的特性研究[J].海洋与湖沼,1992,23(5):555-560.
[14] 虞顺年,魏小岚,韦芳三,等.不同运动强度对黑鲷生长、血清和肝脏抗氧化指标的影响[J].水生生物学报,2018,42(4):255-263.
[15] 谭树华,李玉峰,刘文海,等.十二烷基硫酸钠(SDS)对克氏原螯虾抗氧化功能和酸性磷酸酶活性的影响[J].生态与农村环境学报,2014,30(3):392-397.
[16] 尤宏争,石洪玥,贾磊,等.短途运输胁迫对珍珠龙胆石斑鱼血清酶活力及葡萄糖含量的影响[J].经济动物学报,2018,22(2):72-77,84.
[17] 韩振兴,王韫沛,张硕,等.电磁场对鲫血清4种生化指标的影响研究[J].大连海洋大学学报,2018,33(3):359-364.
[18] 郑榕辉,王重刚,李岩,等.三丁基锡暴露对褐菖鲉碱性磷酸酶活性的影响[J].厦门大学学报:自然科学版,2005,44(S1):203-206.
[19] 崔前进,陈冰,邱丽华,等.低盐胁迫对钝吻黄盖鲽幼鱼鳃Na+/K+-ATP酶、肝脏抗氧化酶和非特异免疫酶的影响[J].广东海洋大学学报,2017,37(6):26-32.
[20] 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.
[21] 陈玉春,顾雪飞,刘敏.5种中草药对鲤血清谷丙转氨酶、谷草转氨酶及红细胞过氧化氢酶活性的影响[J].淡水渔业,2007,37(5):11-13.
[22] 王涛,苗亮,李明云,等.突降盐度胁迫对大黄鱼(Pseudosciaena crocea)血清生理生化及鳃丝Na+/K+-ATP酶活性的影响[J].海洋与湖沼,2013,44(2):421-426.
[23] Heredia-Middleton P,Brunelli J,Drew R E,et al.Heat shock protein (HSP70) RNA expression differs among rainbow trout (Oncorhynchus mykiss) clonal lines[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2008,149(4):552-556.
[24] O’Connor W A,Lawler N F.Salinity and temperature tolerance of embryos and juveniles of the pearl oyster,Pinctada imbricata R?ding[J].Aquaculture,2004,229(1-4):493-506.
[25] Ruscoe I M,Shelley C C,Williams G R.The combined effects of temperature and salinity on growth and survival of juvenile mud crabs (Scylla serrata Forsk?l)[J].Aquaculture,2004,238(1-4):239-247.
[26] Soria G,Merino G,von Brand E.Effect of increasing salinity on physiological response in juvenile scallops Argopecten purpuratus at two rearing temperatures[J].Aquaculture,2007,270(1-4):451-463.
[27] Castelo Branco N A,Rodriguez E,Alves-Pereira M,et al.Vibroacoustic disease:some forensic aspects[J].Aviation,Space,and Environmental Medicine,1999,70(3):A145-A151.
[28] Alves-Pereira M,Castelo Branco N A A.Vibroacoustic disease:biological effects of infrasound and low-frequency noise explained by mechanotransduction cellular signalling[J].Progress in Biophysics and Molecular Biology,2007,93(1-3):256-279.
[29] Oliveira M J R,Freitas D,Carvalho A P O,et al.Exposure to industrial wide band noise increases connective tissue in the rat liver[J].Noise & Health,2012,14(60):227-229.
[30] 区又君,刘奇奇,温久福,等.急性低温胁迫对四指马鲅幼鱼肝脏、肌肉以及鳃组织结构的影响[J].生态科学,2018,37(05):53-59.
[31] 彭士明,施兆鸿,李杰,等.运输胁迫对银鲳血清皮质醇、血糖、组织中糖元及乳酸含量的影响[J].水产学报,2011,35(6):831-837.