猫急性应激模型的建立及诊断标志物的筛选

doi:10.11843/j.issn.0366-6964.2023.03.036

摘要/Abstract

摘要： 本研究旨在建立猫急性应激模型，筛选诊断标志物，完善猫急性应激定义标准。选取18只猫随机分为对照组、应激组A和应激组B，每组6只。测量各组猫的生理指标，评估猫应激评分(CSS)，采血进行血常规和血生化分析；ELISA法检测血清炎性因子、激素、五羟色胺(5-HT)和热休克蛋白-70(HSP-70)浓度；微量法检测血清氧化应激指标。结果表明，与对照组相比，应激组A的体温、呼吸频率、CSS、尿素氮(BUN)含量、肌酸激酶(CK)活性、IL-1β、TNF-α、促肾上腺皮质激素释放激素(CRH)、促肾上腺皮质激素(ACTH)、皮质醇(COR)、肾上腺素(EPI)、5-HT浓度显著升高(P<0.05)，红细胞(RBC)数目、血红蛋白(HGB)、IL-10浓度、天门冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)活性、总胆固醇(TC)含量显著降低(P<0.05)，心率、白细胞(WBC)、淋巴细胞(LYM)、中性粒细胞(NEUT)数目、葡萄糖(GLU)、总蛋白(TP)、白蛋白(ALB)、肌酐(CRE)、三酰甘油(TG)、丙二醛(MDA)含量、淀粉酶(AMY)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和HSP-70浓度无显著变化(P>0.05)；应激组B的体温、呼吸频率、CSS、WBC、NEUT、RBC数目、GLU、BUN、TG含量、CK、AST、SOD、CAT活性、HGB、IL-1β、TNF-α、CRH、ACTH、EPI、HSP-70、5-HT浓度显著升高(P<0.05)，IL-10和COR浓度显著降低(P<0.05)，心率、LYM数目、ALT、AMY活性、TP、ALB、CRE、TC、MDA含量无显著变化(P>0.05)。结果提示，本研究设计的复合应激和束缚应激均能成功建立猫急性应激模型，并筛选出CRH、ACTH和CSS可用于完善猫急性应激的定义标准。

关键词: 猫, 应激模型, 诊断标志物, 定义标准, 猫应激评分

Abstract: The purpose of this study was to establish a cat acute stress model, screen diagnostic markers, and improve the definition standard of cat acute stress. Eighteen cats were randomly divided into control group, stress group A and stress group B, with six cats in each group. Physiological indexes of each group were measured, cat stress score (CSS) was assessed, blood was collected for routine blood and blood biochemical analysis; concentration of serum inflammatory factors, hormones, 5-hydroxytryptamine (5-HT) and heat shock protein-70 (HSP-70) were detected by ELISA; serum oxidative stress indexes were detected by micro-assay method. The results showed that compared with the control group, the body temperature, respiration rate, CSS, the blood urea nitrogen (BUN) content, the creatine kinase (CK) activity, the concentration of IL-1β, TNF-α, corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), cortisol (COR), epinephrine (EPI) and 5-HT were significantly increased in stress group A (P<0.05), the red blood cell (RBC) number, the concentration of hemoglobin (HGB) and IL-10, the activity of aspartic transaminase (AST) and alanine transaminase (ALT),and the total cholesterol (TC) content were significantly decreased in stress group A (P<0.05), the heart rate, the number of white blood cells (WBC), lymphocytes (LYM) and neutrophil (NEUT), the content of glucose (GLU), total protein (TP), albumin (ALB), creatinine (CRE), triglyceride (TG) and malondialdehyde (MDA), the activity of amylase (AMY), superoxide dismutase (SOD) and catalase (CAT), and the HSP-70 concentration had no significant changes in stress group A (P>0.05). Compared with the control group, the body temperature, respiration rate, CSS, the number of WBC, NEUT and RBC, the content of GLU, BUN and TG, the activity of CK, AST, SOD and CAT, the concentration of HGB, IL-1β, TNF-α, CRH, ACTH, EPI, HSP-70 and 5-HT were significantly increased in stress group B (P<0.05), the concentration of IL-10 and COR were significantly decreased in stress group B (P<0.05), the heart rate, the LYM number, the activity of ALT and AMY, the content of TP, ALB, CRE, TC and MDA had no significant changes in stress group B (P>0.05). The results suggest that the compound stress and restraint stress designed in this study can successfully establish a cat acute stress model, and screened out CRH, ACTH and CSS which can improve the definition standard of cat acute stress.

Key words: cat, stress model, diagnostic markers, definition standard, cat stress score

中图分类号:

S852.33

张海洋, 朱秋祥, 高志诚, 贾坤, 王京煜, 冀锦朝, 梁雾滢, 莫睿文, 李守军. 猫急性应激模型的建立及诊断标志物的筛选[J]. 畜牧兽医学报, 2023, 54(3): 1249-1260.

ZHANG Haiyang, ZHU Qiuxiang, GAO Zhicheng, JIA Kun, WANG Jingyu, JI Jinzhao, LIANG Wuying, MO Ruiwen, LI Shoujun. Establishment of Acute Stress Model and Screening of Diagnostic Markers in Cats[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1249-1260.

参考文献 50

[1]	DORLAND W A N. Dorland's illustrated medical dictionary[M]. 32nd ed. Philadelphia:Saunders, 2012.
[2]	DHABHAR F S, MCEWEN B S. Acute stress enhances while chronic stress suppresses cell-mediated immunity in vivo:a potential role for leukocyte trafficking[J]. Brain, Behav, Immun, 1997, 11(4):286-306.
[3]	VAN HAAFTEN K A, FORSYTHE L R E, STELOW E A, et al. Effects of a single preappointment dose of gabapentin on signs of stress in cats during transportation and veterinary examination[J]. J Am Vet Med Assoc, 2017, 251(10):1175-1181.
[4]	AMAT M, CAMPS T, MANTECA X. Stress in owned cats:behavioural changes and welfare implications[J]. J Feline Med Surg, 2016, 18(8):577-586.
[5]	RODAN I. Understanding feline behavior and application for appropriate handling and management[J]. Top Companion Anim Med, 2010, 25(4):178-188.
[6]	ELMAN I, BORSOOK D. Threat response system:parallel brain processes in pain vis-à-vis fear and anxiety[J]. Front Psychiatry, 2018, 9:29.
[7]	DHABHAR F S. The short-term stress response-Mother nature's mechanism for enhancing protection and performance under conditions of threat, challenge, and opportunity[J]. Front Neuroendocrinol, 2018, 49:175-192.
[8]	SRITHUNYARAT T, HAGMAN R, HÖGLUND O V, et al. Catestatin, vasostatin, cortisol, and visual analog scale scoring for stress assessment in healthy dogs[J]. Res Vet Sci, 2018, 117:74-80.
[9]	LIU B, XU P, BROWN P B, et al. The effect of hyperthermia on liver histology, oxidative stress and disease resistance of the Wuchang bream, Megalobrama amblycephala[J]. Fish Shellfish Immunol, 2016, 52:317-324.
[10]	ELLIS J J, STRYHN H, COCKRAM M S. Effects of the provision of a hiding box or shelf on the behaviour and faecal glucocorticoid metabolites of bold and shy cats housed in single cages[J]. Appl Anim Behav Sci, 2021, 236:105221.
[11]	KESSLER M R, TURNER D C. Stress and adaptation of cats (Felis Silvestris Catus) housed singly, in pairs and in groups in boarding catteries[J]. Anim Welfare, 1997, 6(3):243-254.
[12]	LOBERG J M, LUNDMARK F. The effect of space on behaviour in large groups of domestic cats kept indoors[J]. Appl Anim Behav Sci, 2016, 182:23-29.
[13]	王朝好,孙皓然,刘晓雅,等.不同猫粮对猫应激刺激后生理指标和血液指标的影响[J].动物营养学报, 2020, 32(11):5458-5471.WANG C H, SUN H R, LIU X Y, et al. Effects of different cat food on physiological indexes and blood indexes of cats after stress stimulation[J]. Chinese Journal of Animal Nutrition, 2020, 32(11):5458-5471.(in Chinese)
[14]	KESSLER M R, TURNER D C. Effects of density and cage size on stress in domestic cats (Felis Silvestris Catus) housed in animal shelters and boarding catteries[J]. Anim Welfare, 1999, 8(3):259-267.
[15]	GOODING M A, DUNCAN I J H, ATKINSON J L, et al. Development and validation of a behavioral acclimation protocol for cats to respiration chambers used for indirect calorimetry studies[J]. J Appl Anim Welfare Sci, 2012, 15(2):144-162.
[16]	REHNBERG L K, ROBERT K A, WATSON S J, et al. The effects of social interaction and environmental enrichment on the space use, behaviour and stress of owned housecats facing a novel environment[J]. Appl Anim Behav Sci, 2015, 169:51-61.
[17]	JOHNSTONE C P, REINA R D, LILL A. Interpreting indices of physiological stress in free-living vertebrates[J]. J Comp Physiol B, 2012, 182(7):861-879.
[18]	WESTROPP J L, KASS P H, UFFINGTON C B. In vivo evaluation of α2-adrenoceptors in cats with idiopathic cystitis[J]. Am J Vet Res, 2007, 68(2):203-207.
[19]	MCCARTY R. Learning about stress:neural, endocrine and behavioral adaptations[J]. Stress, 2016, 19(5):449-475.
[20]	FOSTER S, IJICHI C. The association between infrared thermal imagery of core eye temperature, personality, age and housing in cats[J]. Appl Anim Behav Sci, 2017, 189:79-84.
[21]	MCEWEN B S. Protective and damaging effects of stress mediators[J]. N Engl J Med, 1998, 338(3):171-179.
[22]	BURKE H M, DAVIS M C, OTTE C, et al. Depression and cortisol responses to psychological stress:a meta-analysis[J]. Psychoneuroendocrinology, 2005, 30(9):846-856.
[23]	STELLA J, CRONEY C, BUFFINGTON T. Effects of stressors on the behavior and physiology of domestic cats[J]. Appl Anim Behav Sci, 2013, 143(2-4):157-163.
[24]	DHABHAR F S, MALARKEY W B, NERI E, et al. Stress-induced redistribution of immune cells-From barracks to boulevards to battlefields:a tale of three hormones-Curt Richter Award Winner[J]. Psychoneuroendocrinology, 2012, 37(9):1345-1368.
[25]	ALUKO O M, UMUKORO S. Role of purinergic signaling pathways in the adaptogenic-like activity of methyl jasmonate in rats exposed to unpredictable chronic mild stress[J]. Drug Metab Pers Ther, 2020, 35(3):20200117.
[26]	FAZIO E, FERLAZZO A, CRAVANA C, et al. Comparison of acute versus chronic stress responses to different housing's systems of cats[J]. Acta Scientiae Vet, 2017, 45(1):9.
[27]	FAN F C, AI Y W, SUN T, et al. The role of inflammatory cytokines in anemia and gastrointestinal mucosal injury induced by foot electric stimulation[J]. Sci Rep, 2021, 11(1):3101.
[28]	ROMERO L M, BEATTIE U K. Common myths of glucocorticoid function in ecology and conservation[J]. J Exp Zool Part A:Ecol Integr Physiol, 2021, 337(1):7-14.
[29]	BRANCACCIO P, LIPPI G, MAFFULLI N. Biochemical markers of muscular damage[J]. Clin Chem Lab Med, 2010, 48(6):757-767.
[30]	LUTNICKI K, MIĘTKIEWSKA K, KUREK Ƚ. Effect of heat stress on the values of biochemical blood parameters of dairy cattle[J]. Med Weter, 2021, 77(9):456-460.
[31]	OBAYASHI K. Salivary mental stress proteins[J]. Clin Chim Acta, 2013, 425:196-201.
[32]	MARKETON J I W, GLASER R. Stress hormones and immune function[J]. Cell Immunol, 2008, 252(1-2):16-26.
[33]	ELENKOV I J, CHROUSOS G P, WILDER R L. Neuroendocrine regulation of IL-12 and TNF-alpha/IL-10 balance. Clinical implications[J]. Ann N Y Acad, 2010, 917:94-105.
[34]	KALINICHENKO L S, PERTSOV S S, KOPLIK E V. Melatonin effects on serum cytokine profiles of rats with different behavioral parameters in acute emotional stress[J]. Bull Exp Biol Med, 2014, 156(5):627-630.
[35]	GODOY L D, ROSSIGNOLI M T, DELFINO-PEREIRA P, et al. A comprehensive overview on stress neurobiology:basic concepts and clinical implications[J]. Front Behav Neurosci, 2018, 12:127.
[36]	TILBROOK A J, RIVALLAND E A T, TURNER A I, et al. Responses of the hypothalamopituitary adrenal axis and the sympathoadrenal system to isolation/restraint stress in sheep of different adiposity[J]. Neuroendocrinology, 2008, 87(4):193-205.
[37]	DICKERSON S S, KEMENY M E. Acute stressors and cortisol responses:a theoretical integration and synthesis of laboratory research[J]. Psychol Bull, 2004, 130(3):355-391.
[38]	ROSMOND R, BJÖRNTORP P. Low cortisol production in chronic stress. The connection stress-somatic disease is a challenge for future research[J]. Lkartidningen, 2000, 97(38):4120-4124.
[39]	WESTROPP J L, WELK K A, BUFFINGTON C A T. Small adrenal glands in cats with feline interstitial cystitis[J]. J Urol, 2003, 170(6):2494-2497.
[40]	WINGFIELD J C, KITAYSKY A S. Endocrine responses to unpredictable environmental events:stress or anti-stress hormones?[J]. Integr Comp Biol, 2002, 42(3):600-609.
[41]	LIGUORI I, RUSSO G, CURCIO F, et al. Oxidative stress, aging, and diseases[J]. Clin Interv Aging, 2018, 13:757-772.
[42]	RAHAL A, KUMAR A, SINGH V, et al. Oxidative stress, prooxidants, and antioxidants:the interplay[J]. Biomed Res Int, 2014, 2014:761264.
[43]	QIAO Y J, ZHAO J B, LI C, et al. Effect of combined chronic predictable and unpredictable stress on depression-like symptoms in mice[J]. Ann Transl Med, 2020, 8(15):942.
[44]	COSTANTINI D, WACHTER B, MELZHEIMER J, et al. Socioecological and environmental predictors of physiological stress markers in a threatened feline species[J]. Conserv Physiol, 2017, 5(1):cox069.
[45]	HASLBECK M, VIERLING E. A first line of stress defense:small heat shock proteins and their function in protein homeostasis[J]. J Mol Biol, 2015, 427(7):1537-1548.
[46]	ARCHANA P R, SEJIAN V, RUBAN W, et al. Comparative assessment of heat stress induced changes in carcass traits, plasma leptin profile and skeletal muscle myostatin and HSP70 gene expression patterns between indigenous Osmanabadi and Salem Black goat breeds[J]. Meat Sci, 2018, 141:66-80.
[47]	ŚLIFIRSKI G, KRÓL M, TURȽO J. 5-HT receptors and the development of new antidepressants[J]. Int J Mol Sci, 2021, 22(16):9015.
[48]	HÖGLUND E,ØVERLIØ, WINBERG S. Tryptophan metabolic pathways and brain serotonergic activity:a comparative review[J]. Front Endocrinol, 2019, 10:158.
[49]	DUNN A J. Footshock-induced changes in brain catecholamines and indoleamines are not mediated by CRF or ACTH[J]. Neurochem Int, 2000, 37(1):61-69.
[50]	SHIMIZU N, OOMURA Y, KAI Y. Stress-induced anorexia in rats mediated by serotonergic mechanisms in the hypothalamus[J]. Physiol Behav, 1989, 46(5):835-841.