猫急性应激模型的建立及诊断标志物的筛选

Establishment of Acute Stress Model and Screening of Diagnostic Markers in Cats

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。

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.