EFFECTS OF HEAT STRESS ON SOME NON-SPECIFIC IMMUNE FACTORS AND BLOOD CELL APOPTOSIS IN SCHIZOTHORAX PRENATI
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摘要: 为研究热应激及恢复对齐口裂腹鱼（Schizothorax prenanti）机体损伤、非特异性免疫功能和细胞凋亡的影响，测定了热应激前（19℃，对照组）、热应激（27℃，0、2h、4h、8h、12h，实验组）及恢复适温6h后（19℃，恢复组）的非特异性免疫相关生理生化指标及细胞凋亡率。结果表明，超氧化物歧化酶（Superoxide dismutase，SOD）在应激0、4h、8h、12h后活性上升，应激0和12h与应激前差异极显著（P<0.01）；髓过氧化物酶（Myeloper oxidase，MPO）在热应激4h达到最大值；谷草转氨酶（Glutamic-oxalacetic transaminase，GOT）的活性实验组与对照组差异不显著，恢复6h后GOT活性极显著高于对照组（P<0.01）；谷丙转氨酶（Glutamic-pyruvic transaminase，GPT）在应激0、4h、12h和恢复到适温6h后的活性极显著高于对照组（P<0.01）；丙二醛（Malondialdehyde，MDA）含量在热应激2h、4h和12h时极显著高于对照组（P<0.01），8h时显著高于应激前（P<0.05），应激12h MDA含量达到最大值，恢复6h后MDA含量恢复至应激前水平（P<0.05）；补体3（Complement 3，C3）含量升高，在热应激12h时显著高于对照组（P<0.05），恢复6h C3含量下降，与对照组差异不显著（P>0.05）。持续27℃热应激不同时间后，各实验组的细胞凋亡率都显著高于对照组（P<0.05），且在处理4h时细胞凋亡率达到最高值（52.42%），恢复19℃ 6h后细胞凋亡率显著降低（P<0.05）。研究表明，持续27℃热应激改变了机体非特异性免疫力，导致鱼体出现炎症，损伤细胞，恢复到适温后，鱼体各项机能逐渐恢复。研究结果可为深入研究齐口裂腹鱼高温适应调控机理提供基础数据，也为齐口裂腹鱼养殖、运输等过程中的温度控制提供科学依据。Abstract: Schizothorax prenati, an important cold-water fish in China, is a precious commercial fish in the upper Yangtze River. The current study investigated the effects of heat stress and recovery on the damage of organism, the functions of non-specific immunity and apoptosis in Schizothorax prenati under 3 conditions:before heat stress (19℃, control group), after heat stress (27℃ for 0, 2, 4, 8 and 12h, respectively; experimental groups) and 6h after heat stress recovery (19℃ recovery group). The results indicated that the activity of superoxide dismutase (SOD) increased at 0, 4, 8 and 12h after stress, and it increased significantly at 0, 12h compared with the pre-stress (P<0.01). The activity of myeloperoxidase (MPO) reached the maximum level at 4h. The activity of glutamic-oxalacetic transaminase (GOT) in all experimental groups had no significant difference compared with the control group, and it was significantly higher than that in the control group at 6h after recovery (P<0.01). The activity of glutamic-pyruvic transaminase (GPT) at 0, 4, 12h after stress and 6h after recovery was significantly higher than that in the control group (P<0.01). The content of malonaldehyde (MDA) at 2, 4, and 12h after heat stress was significantly higher than that in the control group (P<0.01), and it was significantly higher than that in the pre-stress at 8h (P<0.05), which reached the maximum level at 12h after stress, and the content of MDA at 6h after recovery returned to pre-stress levels (P<0.05). C3 complement (C3) level kept rising and was significantly higher than that of the control group at 12h under heat stress (P<0.05), but it decreased at 6h after recovery having significant difference from the control group (P>0.05). Compared with the pre-stress group, apoptosis rate in all experimental groups was significantly higher after heat stress (27℃) (P<0.05) and reached the maximum level (52.42%) at 4h after treatment, while declined markedly at 6h after recovery. These results indicated that heat stress changed the non-specific immunity of the organism and induced inflammation and induced cells damage, but all functions can recover gradually after the temperature returning to the moderate level. The results provided basic data for further research on the mechanism of high temperature adaption in Schizothorax prenati, as well as a scientific basis for temperature control during cultivation and transit of this species.
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Keywords:
- Schizothorax prenati /
- Heat stress /
- Non-specific immunity /
- Apoptosis
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图 1 热应激对血清SOD、MPO活性的影响
Figure 1. The effects of heat stress on serum SOD and MPO activity
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图 2 热应激对血清GOT、GPT活性的影响
Figure 2. The effects of heat stress on serum GOT and GPT activity
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图 3 热应激对血清MDA、C3含量的影响
Figure 3. The effects of heat stress on the content of serum MDA and C3
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[1] 许友卿, 曹占旺, 丁兆坤, 等. 高温对鱼类的影响及其预防研究. 水产科学, 2010, 29(4):235-242 http://www.cnki.com.cn/Article/CJFDTOTAL-CHAN201004015.htm Xu Y Q, Cao Z W, Ding Z Y, et al. Effects and prevention of high temperature on fishes[J]. Fisheries Science, 2010, 29(4):235-242
许友卿, 曹占旺, 丁兆坤, 等. 高温对鱼类的影响及其预防研究. 水产科学, 2010, 29(4):235-242 http://www.cnki.com.cn/Article/CJFDTOTAL-CHAN201004015.htm[2] Scaion D, Belhomme M, Sébert P. Pressure and temperature interactions on aerobic metabolism of migrating European silver eel[J]. Respiratory Physiology &Neurobiology, 2008, 164(3):319-322 http://cn.bing.com/academic/profile?id=2106310687&encoded=0&v=paper_preview&mkt=zh-cn
[3] August S M, Hicks B J. Water temperature and upstream migration of glass eels in New Zealand:implications of climate change[J]. Environmental Biology of Fishes, 2007, 81(2):195-205 doi: 10.1007/s10641-007-9191-z
[4] Pörtner H O, Berdal B, Blust R, et al. Climate induced temperature effects on growth performance, fecundity and recruitment in marine fish:developing a hypothesis for cause and effect relationships in Atlantic cod (Gadusmorhua) and common eelpout (Zoarces viviparus)[J]. Continental Shelf Research, 2001, 21(18-19):1975-1997 doi: 10.1016/S0278-4343(01)00038-3
[5] 王文博, 李爱华. 环境胁迫对鱼类免疫系统影响的研究概况. 水产学报, 2002, 26(4):368-374 http://www.cnki.com.cn/Article/CJFDTOTAL-SCKX200204013.htm Wang W B, Li A H. The effect of environmental stress to fish immune system[J]. Journal of Fisheries of China, 2002, 26(4):368-374
王文博, 李爱华. 环境胁迫对鱼类免疫系统影响的研究概况. 水产学报, 2002, 26(4):368-374 http://www.cnki.com.cn/Article/CJFDTOTAL-SCKX200204013.htm[6] 周玉, 郭文场, 杨振国, 等. 鱼类血液学指标研究的进展. 上海水产大学学报, 2001, 10(2):163-165 http://www.cnki.com.cn/Article/CJFDTOTAL-SSDB200102012.htm Zhou Y, Guo W C, Yang Z G, et al. Advances in the study of haemotological indices of fish[J]. Journal of Shanghai Fisheries University, 2001, 10(2):163-165
周玉, 郭文场, 杨振国, 等. 鱼类血液学指标研究的进展. 上海水产大学学报, 2001, 10(2):163-165 http://www.cnki.com.cn/Article/CJFDTOTAL-SSDB200102012.htm[7] 武云飞, 吴翠珍. 青藏高原鱼类. 成都:四川科学技术出版社. 1992, 361-365 Wu Y F, Wu C Z. Fish in the Tibetan Plateau[M]. Chengdu:Sichuan Science and Technology Press. 1992, 361-365
武云飞, 吴翠珍. 青藏高原鱼类. 成都:四川科学技术出版社. 1992, 361-365[8] 陈毅峰, 曹文宣, 乐佩琦. 中国动物志·硬骨鱼纲·鲤形目(下)裂腹鱼亚科. 北京:科学出版社. 2000, 292-294 Chen Y F, Cao W X, Yue P Q. Chinese Journal of Animal Osteichthyes Common Carp (2) Schizothoracinae[M]. Beijing:Science Press. 2000, 292-294
陈毅峰, 曹文宣, 乐佩琦. 中国动物志·硬骨鱼纲·鲤形目(下)裂腹鱼亚科. 北京:科学出版社. 2000, 292-294[9] Geng Y, Wang K Y, Huang X L, et al. Streptococcus agalactiae, an emerging pathogen for cultured Ya-fish, Schizothorax prenanti, in China[J]. Transboundary and Emerging Diseases, 2012, 59(4):369-375 doi: 10.1111/tbed.2012.59.issue-4
[10] Lam P, Harvie E A, Huttenlocher A. Heat shock modulates neutrophil motility in zebrafish[J]. PLoS One, 2013, 8(12):e84436 doi: 10.1371/journal.pone.0084436
[11] Haugland G T, Jakobsen R A, Vestvik N, et al. Phagocytosis and respiratory burst activity in Lumpsucker (Cyclopterus lumpus L.) leucocytes analysed by flow cytometry[J]. PLoS One, 2012, 7(10):e47909 doi: 10.1371/journal.pone.0047909
[12] Liu X, Li K, Du J, et al. Growth rate, catalase and superoxide dismutase activities in rock carp (Procypris rabaudi Tchang) exposed to supersaturated total dissolved gas[J]. Journal of Zhejiang University Science B (Biomed &Biotechnology), 2011, 12(11):909-914 http://cn.bing.com/academic/profile?id=2000015642&encoded=0&v=paper_preview&mkt=zh-cn
[13] Klebanoff S J. Myeloperoxidase[J]. Proceedings of the Association of American Physicians, 1999, 111(5):383-389
[14] Kutter D. Prevalence of myeloperoxidase deficiency:population studies using Bayer-Technicon automated hematology[J]. Journal of Molecular Medicine, 1998, 76(10):669-675 doi: 10.1007/s001090050266
[15] Cooper L C, Dial E J, Lichtenberger L M. Effects of milk, prostaglandin, and antacid on experimentally induced duoden it is the rat Use of myeloperoxidase as an index of inflammation[J]. Digestive Diseases &Sciences, 1990, 35(10):1211-1216 http://cn.bing.com/academic/profile?id=2032758359&encoded=0&v=paper_preview&mkt=zh-cn
[16] 王艳妮, 刘哲, 康玉军, 等. 热应激对虹鳟部分非特异性免疫指标的影响. 农业生物技术学报, 2015, 23(5):634-642 http://www.cnki.com.cn/Article/CJFDTOTAL-NYSB201505009.htm Wang Y N, Liu Z, Kang Y J, et al. Effects of heat stress on some nonspecific immunity parameters in rainbow Trout (Oncorhynchus mykiss)[J]. Journal of Agricultural Biotechnology, 2015, 23(5):634-642
王艳妮, 刘哲, 康玉军, 等. 热应激对虹鳟部分非特异性免疫指标的影响. 农业生物技术学报, 2015, 23(5):634-642 http://www.cnki.com.cn/Article/CJFDTOTAL-NYSB201505009.htm[17] 孙学亮, 邢克智, 陈成勋, 等. 急性温度胁迫对半滑舌鳎血液指标的影响. 水产科学, 2010, 29(7):387-392 http://www.cnki.com.cn/Article/CJFDTOTAL-CHAN201007003.htm Sun X L, Xing K Z, Chen C X, et al. The effects of acute temperature stress on blood parameters in half-smooth tongue-sole (Cynoglossus semilaevis)[J]. Fisheries Science, 2010, 29(7):387-392
孙学亮, 邢克智, 陈成勋, 等. 急性温度胁迫对半滑舌鳎血液指标的影响. 水产科学, 2010, 29(7):387-392 http://www.cnki.com.cn/Article/CJFDTOTAL-CHAN201007003.htm[18] 周鑫, 董云伟, 王芳等. 草鱼hsp70和hsp90对温度急性变化的响应. 水产学报, 2013, 37(2):216-221 doi: 10.3724/SP.J.1231.2013.37965 Zhou X, Dong Y W, Wang F, et al. Effect of rapid temperature change on expression of hsp70 and hsp90 in grass carp (Ctenopharyngodon idella)[J]. Journal of Fisheries of China, 2013, 37(2):216-221
周鑫, 董云伟, 王芳等. 草鱼hsp70和hsp90对温度急性变化的响应. 水产学报, 2013, 37(2):216-221 doi: 10.3724/SP.J.1231.2013.37965[19] Heredia-Middleton P, Brunelli O, 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 doi: 10.1016/j.cbpb.2007.05.012
[20] Demers N E. The acute effects of stress on plasma proteins of rainbow trout (Oncorhynchus mykiss)[D]. Oregon State University, Supervisor:Bayne Christopher J. 1994
[21] Ndong D, Chen Y Y, Lin Y H, et al. The immune response of tilapia Oreochromis mossambicus and its susceptibility to Streptococcus iniae under stress in low and high temperatures[J]. Fish &Shellfish Immunology, 2007, 22(6):686-694 http://cn.bing.com/academic/profile?id=2034021671&encoded=0&v=paper_preview&mkt=zh-cn
[22] Sakai D K. Repertoire of complement in immunological defense mechanisms of fish[J]. Annual Research of Fish Disease, 1992, 2:223-247 doi: 10.1016/0959-8030(92)90065-6
[23] Jyothi B, Narayan G. Effect of phorate on certain protein profiles of serum in freshwater fish Claries batrachus[J]. Journal of Environmental Biology, 1997, 18:137-140
[24] 陈晨, 黄峰, 舒秋艳, 等. 共轭亚油酸对草鱼生长、肌肉成分、谷草转氨酶及谷丙转氨酶活性的影响. 水生生物学报, 2010, 34(3):647-651 http://ssswxb.ihb.ac.cn/EN/Y2010/V34/I3/647 Chen C, Huang F, Shu Q Y, et al. Effects of dietary conjugated linoleic acids on growth performance, muscle composition, AST and ALT activities in serum in juveniles grass carp (Ctenopharyngodon idellus)[J]. Acta Hydrobiologica Sinica, 2010, 34(3):647-651
陈晨, 黄峰, 舒秋艳, 等. 共轭亚油酸对草鱼生长、肌肉成分、谷草转氨酶及谷丙转氨酶活性的影响. 水生生物学报, 2010, 34(3):647-651 http://ssswxb.ihb.ac.cn/EN/Y2010/V34/I3/647[25] Fuchs Y, Steller H. Programmed cell death in animal development and disease[J]. Cell, 2011, 147(4):742-758 doi: 10.1016/j.cell.2011.10.033
[26] Cao J, Chen J, Wang J, et al. Effects of fluoride on liver apoptosis and Bcl-2, Bax protein expression in freshwater teleost, Cyprintis carpio[J]. Chemosphere, 2013, 91(8):1203-1212 doi: 10.1016/j.chemosphere.2013.01.037
[27] Taleb M, Brandon C S, Lee F, et al. Hsp70 inhibits aminoglycosideinduced hair cell death and is necessary for the protective effect of heat shock[J]. Journal of the Association for Research in Otolaryngology, 2008, 9:277-289 doi: 10.1007/s10162-008-0122-2
[28] Long L L, Han Y L, Sheng Z, et al. Expression analysis of HSP70 in the testis of Octopus tankahkeei under thermal stress[J]. Comparative Biochemistry and Physiology, Part A, 2015, 187:150-159 doi: 10.1016/j.cbpa.2015.05.022
[29] 王枫, 李泊, 杨宝平, 等. 预热诱导K562细胞热应激蛋白70高表达对高温引起细胞凋亡的影响. 卫生研究, 2001, 30(2):65-67 http://www.cnki.com.cn/Article/CJFDTOTAL-WSYJ200102000.htm Wang F, Li B, Yang B P, et al. Effects of heat shock protein 70 overexpression on apoptosis of K-562cell caused by hyperthermia[J]. Journal of Hygiene Research, 2001, 30(2):65-67
王枫, 李泊, 杨宝平, 等. 预热诱导K562细胞热应激蛋白70高表达对高温引起细胞凋亡的影响. 卫生研究, 2001, 30(2):65-67 http://www.cnki.com.cn/Article/CJFDTOTAL-WSYJ200102000.htm
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