研究论文

高温暴露对黑线仓鼠能量代谢和组织氧化应激的影响

  • 张锐涵 ,
  • 罗丹 ,
  • 罗欣欣 ,
  • 胡陈晓 ,
  • 林珑 ,
  • 陈倩 ,
  • 曹静 ,
  • 赵志军
展开
  • 温州大学生命与环境科学学院, 浙江省水环境与海洋生物资源保护重点实验室, 温州 325035
张锐涵(2002-),女,本科生,主要从事动物生理生态学研究.

收稿日期: 2023-06-30

  修回日期: 2023-08-07

  网络出版日期: 2023-08-07

基金资助

国家自然科学基金(31670417);国家级大学生创新创业训练计划项目(202310351047);浙江省大学生科技创新活动计划暨新苗人才计划项目(2023R451028)

Effect of exposure to high temperature on energy metabolism and oxidative stress in striped hamsters

  • ZHANG Ruihan ,
  • LUO Dan ,
  • LUO Xinxin ,
  • HU Chenxiao ,
  • LIN Long ,
  • CHEN Qian ,
  • CAO Jing ,
  • ZHAO Zhijun
Expand
  • College of Life and Environmental Science, Zhejiang Provincial Key Laboratory for Water Environment and Marine Biological Resources Protection, Wenzhou University, Wenzhou 325035, China

Received date: 2023-06-30

  Revised date: 2023-08-07

  Online published: 2023-08-07

摘要

动物能量代谢的适应性调节影响动物生长、发育、繁殖、衰老等生活史特征。代谢率和组织线粒体呼吸率与自由基水平有关,是影响机体衰老的重要因素。为了探讨高温环境下能量代谢、主要代谢活性器官组织呼吸率、自由基水平和抗氧化能力的内在联系,我们将室温(21℃)和暖温(32.5℃)驯化4周的黑线仓鼠(Cricetulus barabensis)分别进行急性高温暴露(37℃)48 h,分别测定摄入能、代谢率,以及褐色脂肪组织(brown adiposetissue,BAT)、肝脏和肌肉的线粒体呼吸率,解偶联蛋白(uncoupling protein,UCP)基因(ucp)表达,蛋白羰基和丙二醛(malondialdehyde,MDA)水平,以及超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性。结果显示,暖温驯化和急性高温暴露使摄入能、消化能、基础代谢率和非颤抖性产热显著降低。暖温驯化后BAT、肝脏和肌肉的细胞色素c氧化酶(cytochrome c oxidase,COX)活性分别降低了84.2%、50.0%和56.6%,急性高温暴露后肝脏COX活性降低了23.3%,但BAT和肌肉COX活性未发生显著变化。21℃组黑线仓鼠在急性高温暴露后BAT中ucp1表达下调89.5%、肝脏ucp2下调76.2%、肌肉ucp3下调58.8%,而32.5℃组在急性高温暴露后未发生显著变化。暖温驯化使心脏、肺脏、肾脏和肌肉组织MDA水平显著降低,而未影响肝脏和脑MDA水平;使肾脏蛋白羰基水平显著升高,但对其他组织影响不显著。急性高温暴露对各组织MDA水平的影响不显著,但显著降低了肝脏和心脏蛋白羰基水平。暖温驯化使肝脏和肌肉SOD活性显著降低,但未影响心脏、肺脏、肾脏和脑组织SOD活性。暖温驯化使心脏、肺脏、肾脏GSH-Px活性显著降低,使肌肉GSH-Px活性显著升高,而对肝脏无显著影响。结果表明:(1)急性高温暴露对能量代谢和线粒体呼吸率的影响与动物生存的环境温度有关,生存的环境温度越低,受高温的影响越大;(2)在暖温驯化和急性高温暴露条件下,机体代谢率与线粒体呼吸率降低,但心脏、肝脏、肺脏、肾脏、肌肉和脑未出现显著的氧化应激和损伤,与“自由基假说”不一致。

本文引用格式

张锐涵 , 罗丹 , 罗欣欣 , 胡陈晓 , 林珑 , 陈倩 , 曹静 , 赵志军 . 高温暴露对黑线仓鼠能量代谢和组织氧化应激的影响[J]. 兽类学报, 2023 , 43(6) : 710 -722 . DOI: 10.16829/j.slxb.150822

Abstract

The adaptive regulation of animal energy metabolism affects life history characteristics such as growth, development, reproduction, and aging. Metabolic rate and tissue mitochondrial respiration rate are related to free radical levels, which are important factors influencing the aging of the organism. In this study, we investigated the intrinsic link among energy metabolism, tissue respiration rates of major metabolically active organs, free radical levels, and antioxidant capacity in response to high ambient temperature. We used as a model species striped hamsters (Cricetulus barabensis) that were previously acclimated to room temperature (21 ℃) and warm temperature (32.5 ℃) for 4 weeks and then were acutely exposed to extremely high temperature (37 ℃) for 48 h. The energy intake, metabolic rate, body temperature, mitochondrial respiration rate, and uncoupling protein (UCP) gene (ucp) expression of brown adipose tissue (BAT), liver and skeletal muscle were measured. The levels of protein carbonyl and malondialdehyde (MDA), and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also determined. The results showed that the acclimation to 32.5℃ and acute exposure to 37℃ resulted in significant reductions in gross and digestive energy intake, basal metabolic rate, and non-shivering thermogenesis. Cytochrome c oxidase (COX) activity was reduced after acclimation to 32.5℃ by 84.2%, 50.0%, and 56.6% in BAT, liver, and muscle, respectively. The liver COX activity decreased further by 23.3% after exposure to 37℃, but COX activity in BAT and muscle was not significantly changed. The acute exposure to 37℃ deceased BAT ucp1 expression by 89.5%, liver ucp2 by 76.2%, and muscle ucp3 by 58.8% in the hamster previously acclimated to 21℃, but it had no significant effect on the gene expression in the hamsters previously acclimated to 32.5℃. Acclimation to 32.5℃ significantly decreased MDA levels in heart, lung, kidney, and muscle, but not in liver and brain, and it significantly increased protein carbonyl levels in the kidney only. Acute exposure to 37℃ significantly reduced protein carbonyl levels in liver and heart, but it had no significant effect on MDA levels in all tissues. In addition, acclimation to 32.5℃ significantly reduced SOD activity in liver and muscle, but did not affect heart, lung, kidney, or brain; and it decreased GSH-Px activity in heart, lung, and kidney, but increased GSH-Px activity in muscle, and had no effect in liver. These findings suggest that (1) the effects of acute exposure to high temperatures on energy metabolism and mitochondrial respiration rate are related to habitat ambient temperature, with the animals at lower ambient temperature being more affected by high temperature; (2) after being acclimated to the warm and acutely exposed to high temperature, the metabolic rate of the organism and mitochondria respiratory decreased, while the liver, heart, lung, kidneys, skeletal muscle, and brain do not show considerable oxidative stress and damage, being inconsistent with the‘free radical hypothesis’ .

参考文献

Balaban R S, Nemoto S, Finkel T. 2005. Mitochondria, oxidants, and aging. Cell, 120 (4): 483-495.
Bao M H, Chen L B, Hambly C, Speakman J R, Zhao Z J. 2020. Exposure to hot temperatures during lactation in Swiss mice stunted offspring growth and decreased the future reproductive performance of female offspring. The Journal of Experimental Biology, 223: jeb223560.
Barja G, Herrero A. 1998. Localization at complex I and mechanism of the higher free radical production of brain nonsynaptic mitochondria in the short-lived rat than in the longevous pigeon. Journal of Bioenergetics and Biomembranes, 30 (3): 235-243.
Bi Z Q, Wen J, Shi L L, Tan S, Xu X M, Zhao Z J. 2018. Effect of temperature and high fat diet on metabolic thermogenesis and body fat content in striped hamsters. Acta Theriologica Sinica, 38(4): 384-392. (in Chinese)
Brunet-Rossinni A K, Austad S N. 2004. Ageing studies on bats: a review. Biogerontology, 5 (4): 211-222.
Chen K X, Wang C M, Wang G Y, Zhao Z J. 2014. Energy budget, oxidative stress and antioxidant in striped hamster acclimated to moderate cold and warm temperatures. Journal of Thermal Biology, 44: 35-40.
Chen K X, Wang G Y, Zhao Z J. 2015. Effects of cold temperature on energy metabolism, anitioxidants and oxidative stress in striped hamsters. Acta Theriologica Sinica, 35 (4): 412-421. (in Chinese)
Davidović M. 1999. Genetic stability: the key to longevity? Med Hypotheses, 53 (4): 329-332.
Duan L J, Liu Y S, Zhu X. 2005. DNPH colorimetric method: a simple method for determining protein carbonyl content. Journal of Toxicology, 19 (4): 320-322. (in Chinese)
Dubey A, Forster M J, Lal H, Sohal R S. 1996. Effect of age and caloric intake on protein oxidation in different brain regions and on behavioral functions of the mouse. Archives Biochemistry Biophysics, 333 (1): 189-197.
Echtay K S, Roussel D, St-Pierre J, Jekabsons M B, Cadenas S, Stuart J A, Harper J A, Roebuck S J, Morrison A, Pickering S, Clapham J C, Brand M D. 2002. Superoxide activates mitochondrial uncoupling proteins. Nature, 415 (6867): 96-99.
Fridell Y W, Sánchez-Blanco A, Silvia B A, Helfand S L. 2005. Targeted expression of the human uncoupling protein 2 (hUCP2) to adult neurons extends life span in the fly. Cell Metabolism, 1 (2):145-152.
Furness L J, Speakman J R. 2008. Energetics and longevity in birds. Age (Dordrecht, Netherlands), 30 (2-3): 75-87.
Hammond K A, Szewczak J, Król E. 2001. Effects of altitude and temperature on organ phenotypic plasticity along an altitudinal gradient. The Journal of Experimental Biology, 204 (Pt 11):1991-2000.
Hauck A K, Huang Y, Hertzel A V, Bernlohr D A. 2019. Adipose oxidative stress and protein carbonylation. The Journal of Biological Chemistry, 294 (4): 1083-1088.
Heldmaier G, Buchberger A. 1985. Sources of heat during nonshivering thermogenesis in Djungarian hamsters: a dominant role of brown adipose tissue during cold adaptation. Journal of Comparative Physiology, 156 (2): 237-245.
Holmes D J, Fluckiger R, Austad S N. 2001. Comparative biology of aging in birds: an update. Experimental Gerontology, 36 (4-6):869-883.
Hulbert A J, Else P L, Manolis S C, Brand M D. 2002. Proton leak in hepatocytes and liver mitochondria from archosaurs (Crocodiles) and allometric relationships for ectotherms. Journal of Comparative Physiology, 172 (5): 387-397.
Huo D L, Liao S S, Cao J, Zhao Z J. 2022. The energy budget of striped hamsters in response to food shortage at different temperatures. Acta Theriologica Sinica, 42 (1): 58-68. (in Chinese)
Iossa S, Mollica M P, Lionetti L, Crescenzo R, Tasso R, Liverini G. 2004. A possible link between skeletal muscle mitochondrial efficiency and age-induced insulin resistance. Diabetes, 53 (11):2861-2866.
Jacobs P J, Oosthuizen M K, Mitchell C, Blount J D, Bennett N C. 2020. Heat and dehydration induced oxidative damage and antioxidant defenses following incubator heat stress and a simulated heat wave in wild caught four-striped field mice Rhabdomys dilectus. PLoS ONE, 15 (11): e0242279.
Jacobs P J, Oosthuizen M K, Mitchell C, Blount J D, Bennett N C. 2021. Oxidative stress in response to heat stress in wild caught Namaqua rock mice, Micaelamys namaquensis. Journal of Thermal Biology, 98: 102958.
Jastroch M, Buckingham J A, Helwig M, Klingenspor M, Brand M D. 2007. Functional characterisation of UCP1 in the common carp:uncoupling activity in liver mitochondria and cold-induced expression in the brain. Journal of Comparative Physiology, 177 (7):743-752.
Kankofer M. 2001. Antioxidative defence mechanisms against reactive oxygen species in bovine retained and not-retained placenta:activity of glutathione peroxidase, glutathione transferase, catalase and superoxide dismutase. Placenta, 22 (5): 466-472.
Kauffman A S, Cabrera A, Zucker I. 2001. Energy intake and fur in summer- and winter-acclimated Siberian hamsters (Phodopus sungorus). American Journal of Physiology, 281 (2): R519-R527.
Khakisahneh S, Zhang X Y, Nouri Z, Hao S Y, Chi Q S, Wang D H. 2019. Thyroid hormones mediate metabolic rate and oxidative, anti-oxidative balance at different temperatures in Mongolian gerbils (Meriones unguiculatus). Comparative Biochemistry and Physiology, 216: 101-109.
Li X S, Wang D H. 2007. Photoperiod and temperature can regulate body mass, serum leptin concentration, and uncoupling protein 1 in Brandt's voles (Lasiopodomys brandtii) and Mongolian gerbils(Meriones unguiculatus). Physiological and Biochemical Zoology, 80 (3): 326-334.
Liu Q S, Zhang Z Q, Caviedes-Vidal E, Wang D H. 2013. Seasonal plasticity of gut morphology and small intestinal enzymes in free-living Mongolian gerbils. Journal of Comparative Physiology, 183 (4): 511-523.
Ołdakowski Ł, Taylor J R E. 2018. Oxidative damage and antioxidant defense are assay and tissue-dependent both in captive and wild-caught bank voles (Myodes glareolus) before and after reproduction. Ecology and Evolution, 8 (15): 7543-7552.
Osorio R A, Christofani J S, D’ Almeida V, Russo A K, Piçarro I C. 2003. Reactive oxygen species in pregnant rats: effects of exercise and thermal stress. Comparative Biochemistry and Physiology, 135 (1): 89-95.
Padalko V I. 2005. Uncoupler of oxidative phosphorylation prolongs the lifespan of Drosophila. Biochemistry Biokhimiia, 70 (9):986-989.
Pamplona R, Portero-Otin M, Sanz A, Ayala V, Vasileva E, Barja G. 2005. Protein and lipid oxidative damage and complex I content are lower in the brain of budgerigar and canaries than in mice. Relation to aging rate. Age (Dordrecht, Netherlands), 27 (4):267-280.
Parker N, Crichton P G, Vidal-Puig A J, Brand M D. 2009. Uncoupling protein-1 (UCP1) contributes to the basal proton conductance of brown adipose tissue mitochondria. Journal of Bioenergetics and Biomembranes, 41 (4): 335-342.
Selman C, McLaren J S, Collins A R, Duthie G G, Speakman J R. 2002. Antioxidant enzyme activities, lipid peroxidation, and DNA oxidative damage: the effects of short-term voluntary wheel running. Archives of Biochemistry and Biophysics, 401 (2):255-261.
Selman C, McLaren J S, Collins A R, Duthie G G, Speakman J R. 2013. Deleterious consequences of antioxidant supplementation on lifespan in a wild-derived mammal. Biology Letters, 9 (4):20130432.
Simons M J, Reimert I, van der Vinne V, Hambly C, Vaanholt L M, Speakman J R, Gerkema M P. 2011. Ambient temperature shapes reproductive output during pregnancy and lactation in the common vole (Microtus arvalis): a test of the heat dissipation limit theory. The Journal of Experimental Biology, 214 (Pt 1): 38-49.
Sohal R S, Agarwal S, Sohal B H. 1995. Oxidative stress and aging in the Mongolian gerbil (Meriones unguiculatus). Mechanisms of Ageing and Development, 81 (1): 15-25.
Sohal R S. 2002. Oxidative stress hypothesis of aging. Free Radical Biology and Medicine, 33 (5): 573-574.
Song Z G, Wang D H. 2006. Basal metabolic rate and organ size in Brandt's voles (Lasiopodomys brandtii): effects of photoperiod, temperature and diet quality. Physiology & Behavior, 89 (5):704-710.
Speakman J R, Selman C. 2011. The free-radical damage theory: Accumulating evidence against a simple link of oxidative stress to ageing and lifespan. BioEssays, 33 (4): 255-259.
Speakman J R, Talbot D A, Selman C, Snart S, McLaren J S, Redman P, Krol E, Jackson D M, Johnson M S, Brand M D. 2004. Uncoupled and surviving: individual mice with high metabolism have greater mitochondrial uncoupling and live longer. Aging Cell, 3 (3): 87-95.
Trayhurn P, Ashwell M, Jennings G, Richard D, Stirling D M. 1987. Effect of warm or cold exposure on GDP binding and uncoupling protein in rat brown fat. The American Journal of Physiology, 252 (2 Pt 1): E236-E243.
Venditti P, De Rosa R, Portero-Otin M, Pamplona R, Di Meo S. 2004. Cold-induced hyperthyroidism produces oxidative damage in rat tissues and increases susceptibility to oxidants. The International Journal of Biochemistry & Cell Biology, 36 (7): 1319-1331.
Wang D H. 2011. Some progress in mammalian physiological ecology in China. Acta Theriologica Sinica, 31 (1): 15-19. (in Chinese)
Wang J M, Wang D H. 2006. Comparison of nonshivering thermogenesis induced by dosages of norepinephrine from 3 allometric equations in Brandt's voles (Lasiopodomys brandtii). Acta Theriologica Sinica, 26 (1): 84-88. (in Chinese)
Ward S, Moller U, Rayner J M V, Jackson D M, Nachtigall W, Speakman J R. 2004. Metabolic power of European starlings Sturnus vulgaris during flight in a wind tunnel, estimated from heat transfer modelling, doubly labelled water and mask respirometry. The Journal of Experimental Biology, 207 (Pt 24): 4291-4298.
Xing X, Tang G B, Sun M Y, Yu C, Song S Y, Liu X Y, Yang M, Wang D H. 2016. Leptin regulates energy intake but fails to facilitate hibernation in fattening Daurian ground squirrels (Spermophilus dauricus). Journal of Thermal Biology, 57: 35-43.
Xing X, Yang M, Wang D H. 2015. The expression of leptin, hypothalamic neuropeptides and UCP1 before, during and after fattening in the Daurian ground squirrel (Spermophilus dauricus). Comparative Biochemical Physiology, 184: 105-112.
Xu D L, Xu M M, Wang D H. 2019a. Effect of temperature on antioxidant defense and innate immunity in Brandt's voles. Zoological Research, 40 (4): 305-316.
Xu D L, Xu M M, Wang D H. 2019b. Effects of air temperatures on antioxidant defense and immunity in Mongolian gerbils. Journal of Thermal Biology, 84: 111-120.
Xu Y C, Yang D B, Speakman J R, Wang D H. 2014. Oxidative stress in response to natural and experimentally elevated reproductive effort is tissue dependent. Functional Ecology, 28 (2): 402-410.
Yang D B, Li L, Wang L P, Chi Q S, Hambly C, Wang D H, Speakman J R. 2013. Limits to sustained energy intake. XIX. A test of the heat dissipation limitation hypothesis in Mongolian gerbils (Meriones unguiculatus). The Journal of Experimental Biology, 216(Pt 17): 3358-3368.
Zhang L, Liu P F, Zhu W L, Cai J H, Wang Z K. 2012. Variations in thermal physiology and energetics of the tree shrew (Tupaia belangeri) in response to cold acclimation. Journal of Comparative Physiology, 182 (1): 167-176.
Zhang X Y, Sukhchuluun G, Bo T B, Chi Q S, Yang J J, Chen B, Zhang L, Wang D H. 2018. Correction to: Huddling remodels gut microbiota to reduce energy requirements in a small mammal species during cold exposure. Microbiome, 6 (1): 126.
Zhang Z Q, Wang D H. 2007. Seasonal changes in thermogenesis and body mass in wild Mongolian gerbils (Meriones unguiculatus). Comparative Biochemistry and Physiology, 148 (2): 346-353.
Zhao X Y, Zhang J Y, Cao J, Zhao Z J. 2015. Oxidative damage does not occur in striped hamsters raising natural and experimentally increased litter size. PLoS ONE, 10 (10): e0141604.
Zhao Z J, Wang D H. 2005. Short photoperiod enhances thermogenic capacity in Brandt’s voles. Physiology & Behavior, 85 (2):143-149.
Zhao Z J, Chi Q S, Cao J, Wang D H. 2014. Seasonal changes of body mass and energy budget in striped hamster: the role of leptin. Physiological and Biochemical Zoology, 87 (2): 245-256.
Zhao Z J, Cao J, Meng X L, Li Y B. 2010. Seasonal variations in metabolism and thermoregulation in the striped hamster (Cricetulus barabensis). Journal of Thermal Biology, 35: 52-57.
Zhao Z J, Hambly C, Shi L L, Bi Z Q, Cao J, Speakman J R. 2020. Late lactation in small mammals is a critically sensitive window of vulnerability to elevated ambient temperature. Proceedings of the National Academy of Sciences of the United States of America, 117 (39): 24352-24358.
Zhao Z J, Yang R, Li M, Bao M H, Huo D L, Cao J, Speakman J R. 2022a. Effects of ambient temperatures between 5 and 35℃ on energy balance, body mass and body composition in mice. Molecular Metabolism, 64: 101551.
Zhao Z J, Cao J, Niu C Q, Bao M H, Xu J Q, Huo D L, Liao S S, Liu W, Speakman J R. 2022b. Body temperature is a more important modulator of lifespan than metabolic rate in two small mammals. Nature Metabolism, 4 (3): 320-326.
王建梅, 王德华. 2006. 不同去甲肾上腺素剂量下布氏田鼠非颤抖性产热比较. 兽类学报, 26 (1): 84-88.
王德华. 2011. 我国哺乳动物生理生态学的一些进展和未来发展的建议. 兽类学报, 31 (1): 15-19.
毕中强, 闻靖, 施璐璐, 谭松, 徐小明, 赵志军. 2018. 温度和高脂食物对黑线仓鼠代谢产热和体脂累积的影响. 兽类学报, 38 (4):384-392.
陈可新, 王桂英, 赵志军. 2015. 低温对黑线仓鼠能量代谢、抗氧化能力和氧化应激的影响. 兽类学报, 35 (4): 412-421.
段丽菊, 刘英帅, 朱燕, 杨旭. 2005. DNPH 比色法: 一种简单的蛋白质羰基含量测定方法. 毒理学杂志, 19 (4): 320-322.
霍达亮, 廖莎莎, 曹静, 赵志军. 2022. 不同温度下黑线仓鼠应对食物短缺的能量学对策. 兽类学报, 42 (1): 58-68.
文章导航

/

〈 〉