中草药添加剂对热应激虹鳟部分生化指标及激素水平的影响
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摘要
将平均体质量(30±5)g的虹鳟Oncorhynchus mykiss饲养在循环水族箱(0.90m×0.50m×0.45m)中,投喂添加柴胡(18g或12g/50kg体质量)、藿香(15g或9g/50kg体质量)和茵陈(20g或30g/50kg体质量)水煎剂制成的药饵15d后,在水温15℃、缓慢升温至20℃和25℃下尾静脉采血,测定血浆中皮质醇(COR)和肾上腺素(E)的含量及肝脏中酸性磷酸酶(ACP)和碱性磷酸酶(ALP)的活性,观察肝脏组织病理切片,探究中草药对虹鳟抗热应激能力的影响。结果显示,在热应激期间,低浓度藿香组鱼血浆中皮质醇含量高于对照组(P<0.05);20℃和25℃持续升温后,高浓度藿香组血浆中肾上腺素含量高于对照组,藿香组肝脏中酸性磷酸酶活性高于对照组(P<0.05)。在热应激期间,柴胡组和藿香组肝脏中碱性磷酸酶活性变化不明显(P<0.05)。20℃时虹鳟肝脏开始出现病变,随着水温升高,肝脏组织结构破坏严重,中草药组缓解了虹鳟肝脏组织受热应激的影响。25℃持续升温后,藿香组鱼肝脏组织的破坏程度最小,表明藿香能有效缓解热应激对肝脏的损伤,提高虹鳟抗热应激能力。
Rainbow trout Oncorhynchus mykiss with body weight of(30 ± 5)g were reared in circulating aquarium tanks and fed diets containing Chinese herbal medicines Bupleurum chinensis(18 g or 12 g/50 kg body weight), Agastache rugosa(15 g or 9 g/50 kg body weight)and Artemisiae capillaris(20 g or 30 g/50 kg body weight)for 15 d. Then the blood was sampled from caudal veins of the rainbow trout exposed to water temperature of 15 ℃, slowly rising to 20 ℃ and 25 ℃ to examine contents of cortisol(COR)and adrenaline(E)in plasma and the activities of acid phosphatase(ACP)and alkaline phosphatase(ALP)in liver and to observe the liver histopathological sections to evaluate the effects of Chinese herb medicines on liver biochemical indices and hormone level of rainbow trout exposed to heat-stress. There was higher level of cortisol in plasma in the low concentration group of Agastache rugosa than that in the control group, the higher level of plasma adrenaline in the high concentration group of Agastache rugosa than in the control group(P < 0.05), continuous warming at 20 ℃ and 25 ℃. At the same time, the activities of liver acid phosphatase were found to be higher in the group of Agastache rugosa than in the control group(P < 0.05), without significant difference in the activities of liver ALP in the group of Bupleurum chinensis and the group of Agastache rugosa in the whole heat-stress period. Pathological changes were first observed in the liver of rainbow trout at 20 ℃. The damaged liver tissue of rainbow trout was seriously as the water temperature became higher and alleviated in the groups of Chinese herbal medicines under heat-stress, with the minimal liver damage in the group of Agastache rugosa at continuous rising temperature of 25 ℃. In summary, Agastache rugosa effectively alleviates the damage of liver in rainbow trout than the other Chinese herbal medicines do and is used as candidates for anti-thermal stress Chinese herbal additives.
Rainbow trout Oncorhynchus mykiss with body weight of(30 ± 5)g were reared in circulating aquarium tanks and fed diets containing Chinese herbal medicines Bupleurum chinensis(18 g or 12 g/50 kg body weight), Agastache rugosa(15 g or 9 g/50 kg body weight)and Artemisiae capillaris(20 g or 30 g/50 kg body weight)for 15 d. Then the blood was sampled from caudal veins of the rainbow trout exposed to water temperature of 15 ℃, slowly rising to 20 ℃ and 25 ℃ to examine contents of cortisol(COR)and adrenaline(E)in plasma and the activities of acid phosphatase(ACP)and alkaline phosphatase(ALP)in liver and to observe the liver histopathological sections to evaluate the effects of Chinese herb medicines on liver biochemical indices and hormone level of rainbow trout exposed to heat-stress. There was higher level of cortisol in plasma in the low concentration group of Agastache rugosa than that in the control group, the higher level of plasma adrenaline in the high concentration group of Agastache rugosa than in the control group(P < 0.05), continuous warming at 20 ℃ and 25 ℃. At the same time, the activities of liver acid phosphatase were found to be higher in the group of Agastache rugosa than in the control group(P < 0.05), without significant difference in the activities of liver ALP in the group of Bupleurum chinensis and the group of Agastache rugosa in the whole heat-stress period. Pathological changes were first observed in the liver of rainbow trout at 20 ℃. The damaged liver tissue of rainbow trout was seriously as the water temperature became higher and alleviated in the groups of Chinese herbal medicines under heat-stress, with the minimal liver damage in the group of Agastache rugosa at continuous rising temperature of 25 ℃. In summary, Agastache rugosa effectively alleviates the damage of liver in rainbow trout than the other Chinese herbal medicines do and is used as candidates for anti-thermal stress Chinese herbal additives.
引文
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[9]夏斌鹏,刘哲,周彦静,等.慢性热应激对虹鳟部分血清非特异性免疫指标的影响[J].农业生物技术学报,2017,25(7):1078-1085.
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[12]赵倩,陈玉春,高绪娜,等.枯草芽孢杆菌发酵中药制剂对鲤鱼生长性能、生化指标、抗氧化指标及抗感染能力的影响[J].中国畜牧兽医,2017,44(3):724-731.
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[14]杜强,杨兴,吴林菁,等.中草药添加剂在水产养殖中的作用机制及应用前景[J].饲料研究,2014(11):71-73.
[15]蔡东森,蒋广震,王丽娜,等.甘草次酸对团头鲂生长、脂肪沉积与抗氧化功能的影响[J].水产学报,2014,38(9):1514-1521.
[16]黄聪亮,郑佳俐,李凤林,等.茯苓多糖对糖尿病小鼠淋巴细胞DNA损伤及肾组织主要抗氧化酶活性的影响[J].黑龙江畜牧兽医,2015(7):35-37.
[17]王荻,刘红柏.施氏鲟抗热应激中草药免疫制剂的筛选[J].大连海洋大学学报,2016,31(6):635-639.
[18]管标,温海深,刘群,等.急性温度胁迫对虹鳟肝脏代谢酶活性及生长相关基因表达的影响[J].大连海洋大学学报,2014,29(6):566-571.
[19]何福林,向建国,李常健,等.水温对虹鳟血液学指标影响的初步研究[J].水生生物学报,2007,31(3):363-369.
[20]孙裔雷,王荻,刘红柏.复方中草药对虹鳟抗氧化能力的影响[J].大连海洋大学学报,2015,30(2):170-174.
[21]马爱敏,闫茂仓,常维山,等.柴胡对美国红鱼免疫机能的影响[J].海洋通报,2009,28(4):37-43.
[22] Jones S R. The occurrence and mechanisms of innate immunity against parasites in fish[J]. Developmental&Comparative Immunology, 2001, 25(8-9):841-852.
[23]宁军号,秦宇博,胡伦超,等.水温骤降和缓降胁迫对褐篮子鱼血液生理生化指标的影响[J].大连海洋大学学报,2017,32(3):294-301.
[24]韩娜娜,史成银.血液指标在鱼类学研究中的应用[J].安徽农业科学,2010,38(33):18877-18878.
[25]南京中医药大学.中药大辞典.第2版[M].上海:上海科学技术出版社,2006.
[26] Webb MA H, Allert J A, Kappenman K M, et al. Identification of plasma glucocorticoids in pallid sturgeon in response to stress[J]. General and Comparative Endocrinology, 2007, 154(1-3):98-104.
[27] Gamperl A K, Vijayan M M, Boutilier R G. Experimental control of stress hormone levels in fishes:techniques and applications[J]. Reviews in Fish Biology&Fisheries, 1994,4(2):215-255.
[28] Fevolden S E, Reed K H, Fjalestad K. A combined salt and confinement stress enhances mortality in rainbow trout(Oncorhynchus mykiss)selected for high stress responsiveness[J]. Aquaculture, 2003, 216(1):67-76.
[29]田照辉,徐绍刚,王巍,等.急性热应激对西伯利亚鲟HSP70 m RNA表达、血清皮质醇和非特异性免疫的影响[J].水生生物学报,2013,37(2):344-350.
[30]何杰,强俊,徐跑,等.低温驯化下4种不同品系罗非鱼血清皮质醇与免疫相关指标的变化[J].中国水产科学,2014,21(2):266-274.
[31]程甦,左福元.热应激对动物体内蛋白质周转代谢的内分泌途径的影响[J].中国饲料,2008(3):36-38.
[32]卢玉标,游翠红,王树启,等.浅水应激后黄斑蓝子鱼生理指标变化及牛磺酸的抗应激作用[J].水生生物学报,2014,38(1):68-74.
[33]臧元奇.刺参(Apostichopus japonicus Selenka)对典型环境胁迫的生理生态学响应及其机制研究[D].青岛:中国海洋大学,2012.
[34]康绍乐,杜启艳,王萍,等.饥饿应激对泥鳅3种组织糖原、ACP和ALP的影响[J].河南师范大学学报:自然版,2008,36(6):158-162.
[35]黄布敏,袁莉刚,曲亚玲,等.虹鳟胆管系统组织学观察[J].甘肃农业大学学报,2013,48(1):31-36.
[36]向建国.虹鳟对水温应激反应的初步研究[D].长沙:湖南农业大学,2008.
[37]崔彦婷.几种重要外源因子对草鱼肝细胞活力、抗氧化、凋亡和热应激蛋白表达的影响[D].南京:南京农业大学,2014.
[38]刘洪柏,卢彤岩.渔用中草药使用技术[M].哈尔滨:黑龙江科学技术出版社,2011.
[2]管标.急性温度胁迫对虹鳟血液成分和关键代谢的影响[D].青岛:中国海洋大学,2014.
[3] Verleih M, Borchel A, Krasnov A, et al. Impact of thermal stress on kidney-specific gene expression in farmed regional and imported rainbow trout[J]. Marine Biotechnology, 2015, 17(5):576-592.
[4] Callaghan N I, Tunnah L, Currie S, et al. Metabolic adjustments to short-term diurnal temperature fluctuation in the rainbow trout(Oncorhynchus mykiss)[J]. Physiological&Biochemical Zoology, 2016, 89(6):498-510.
[5] Faught E, Henrickson L, Vijayan M M. Plasma exosomes are enriched in Hsp70 and modulated by stress and cortisol in rainbow trout[J]. Journal of Endocrinology, 2017, 232(2):237-246.
[6]史鲲鹏,董双林,周演根,等.不同倍性虹鳟幼鱼对急性温度胁迫的抗氧化响应[J].应用生态学报,2018,29(9):3102-3110.
[7] Li Y, Huang J, Liu Z, et al. Transcriptome analysis provides insights into hepatic responses to moderate heat stress in the rainbow trout(Oncorhynchus mykiss)[J]. Gene, 2017,619:1-9.
[8]夏斌鹏.慢性热应激对虹鳟生化指标及热休克蛋白基因GRP78 m RNA表达的影响[D].兰州:甘肃农业大学,2017.
[9]夏斌鹏,刘哲,周彦静,等.慢性热应激对虹鳟部分血清非特异性免疫指标的影响[J].农业生物技术学报,2017,25(7):1078-1085.
[10]周彦静.持续热应激下虹鳟热应激蛋白基因HSP90α和HSP10 m RNA表达、组织损伤及血细胞凋亡变化[D].兰州:甘肃农业大学,2017.
[11]周彦静,刘哲,夏斌鹏,等.持续热应激对虹鳟肝脏组织形态结构的影响[J].甘肃农业大学学报,2017(6):1-5.
[12]赵倩,陈玉春,高绪娜,等.枯草芽孢杆菌发酵中药制剂对鲤鱼生长性能、生化指标、抗氧化指标及抗感染能力的影响[J].中国畜牧兽医,2017,44(3):724-731.
[13]王文博.环境胁迫、中草药及基因转植对鱼体非特异性免疫功能的影响[D].武汉:中国科学院研究生院(水生生物研究所),2005.
[14]杜强,杨兴,吴林菁,等.中草药添加剂在水产养殖中的作用机制及应用前景[J].饲料研究,2014(11):71-73.
[15]蔡东森,蒋广震,王丽娜,等.甘草次酸对团头鲂生长、脂肪沉积与抗氧化功能的影响[J].水产学报,2014,38(9):1514-1521.
[16]黄聪亮,郑佳俐,李凤林,等.茯苓多糖对糖尿病小鼠淋巴细胞DNA损伤及肾组织主要抗氧化酶活性的影响[J].黑龙江畜牧兽医,2015(7):35-37.
[17]王荻,刘红柏.施氏鲟抗热应激中草药免疫制剂的筛选[J].大连海洋大学学报,2016,31(6):635-639.
[18]管标,温海深,刘群,等.急性温度胁迫对虹鳟肝脏代谢酶活性及生长相关基因表达的影响[J].大连海洋大学学报,2014,29(6):566-571.
[19]何福林,向建国,李常健,等.水温对虹鳟血液学指标影响的初步研究[J].水生生物学报,2007,31(3):363-369.
[20]孙裔雷,王荻,刘红柏.复方中草药对虹鳟抗氧化能力的影响[J].大连海洋大学学报,2015,30(2):170-174.
[21]马爱敏,闫茂仓,常维山,等.柴胡对美国红鱼免疫机能的影响[J].海洋通报,2009,28(4):37-43.
[22] Jones S R. The occurrence and mechanisms of innate immunity against parasites in fish[J]. Developmental&Comparative Immunology, 2001, 25(8-9):841-852.
[23]宁军号,秦宇博,胡伦超,等.水温骤降和缓降胁迫对褐篮子鱼血液生理生化指标的影响[J].大连海洋大学学报,2017,32(3):294-301.
[24]韩娜娜,史成银.血液指标在鱼类学研究中的应用[J].安徽农业科学,2010,38(33):18877-18878.
[25]南京中医药大学.中药大辞典.第2版[M].上海:上海科学技术出版社,2006.
[26] Webb MA H, Allert J A, Kappenman K M, et al. Identification of plasma glucocorticoids in pallid sturgeon in response to stress[J]. General and Comparative Endocrinology, 2007, 154(1-3):98-104.
[27] Gamperl A K, Vijayan M M, Boutilier R G. Experimental control of stress hormone levels in fishes:techniques and applications[J]. Reviews in Fish Biology&Fisheries, 1994,4(2):215-255.
[28] Fevolden S E, Reed K H, Fjalestad K. A combined salt and confinement stress enhances mortality in rainbow trout(Oncorhynchus mykiss)selected for high stress responsiveness[J]. Aquaculture, 2003, 216(1):67-76.
[29]田照辉,徐绍刚,王巍,等.急性热应激对西伯利亚鲟HSP70 m RNA表达、血清皮质醇和非特异性免疫的影响[J].水生生物学报,2013,37(2):344-350.
[30]何杰,强俊,徐跑,等.低温驯化下4种不同品系罗非鱼血清皮质醇与免疫相关指标的变化[J].中国水产科学,2014,21(2):266-274.
[31]程甦,左福元.热应激对动物体内蛋白质周转代谢的内分泌途径的影响[J].中国饲料,2008(3):36-38.
[32]卢玉标,游翠红,王树启,等.浅水应激后黄斑蓝子鱼生理指标变化及牛磺酸的抗应激作用[J].水生生物学报,2014,38(1):68-74.
[33]臧元奇.刺参(Apostichopus japonicus Selenka)对典型环境胁迫的生理生态学响应及其机制研究[D].青岛:中国海洋大学,2012.
[34]康绍乐,杜启艳,王萍,等.饥饿应激对泥鳅3种组织糖原、ACP和ALP的影响[J].河南师范大学学报:自然版,2008,36(6):158-162.
[35]黄布敏,袁莉刚,曲亚玲,等.虹鳟胆管系统组织学观察[J].甘肃农业大学学报,2013,48(1):31-36.
[36]向建国.虹鳟对水温应激反应的初步研究[D].长沙:湖南农业大学,2008.
[37]崔彦婷.几种重要外源因子对草鱼肝细胞活力、抗氧化、凋亡和热应激蛋白表达的影响[D].南京:南京农业大学,2014.
[38]刘洪柏,卢彤岩.渔用中草药使用技术[M].哈尔滨:黑龙江科学技术出版社,2011.