短途运输应激对实验大鼠和小鼠的影响及干预研究
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摘要
短途运输是实验动物最常见的运输形式,但是目前针对短途运输应激对实验动物的影响及其干预的研究较少见。本项研究通过探讨短途运输应激对实验大鼠和小鼠影响的初步作用机制,探索建立短途运输应激评价的技术指标体系,确定实验大鼠和小鼠经历短途运输应激后的健康适应期,研发减轻短途运输应激不良反应的干预措施。
第一部分短途运输应激对实验大鼠的影响研究
[目的]探讨短途运输应激对啮齿类实验动物影响的可能机制,建立评价短途运输应激对啮齿类实验动物影响的技术指标体系。[方法]使Wistar大鼠分别经历不同运输时间(0.5hr、1hr和1.5hr),以及经历1.5hr运输时间到达实验室后分别恢复不同时间(24hr、48hr、72hr和96hr),测定其生长代谢、神经内分泌和免疫功能主要指标,与未经历运输的对照组比较,观察分析短途运输应激对实验大鼠的影响。[结果]短途运输导致Wistar大鼠的体重迅速下降,体重丢失率随着运输时间的延长呈增大趋势;运输结束后大鼠的体重迅速恢复,高水平的体重增长率一直持续到运输后72hr,且以24hr内的体重增长率为最高。大鼠血糖(GLU)在运输0.5hr和1hr时均升高,而在运输1.5hr时降低,运输结束后24hr时GLU再次升高,此后与对照组无差异;血清皮质酮(CORT)亦在运输0.5hr和1hr时升高而运输1.5hr时降低,但运输结束24hr起即恢复至对照组水平;血清β-内啡肽(β-EP)在经历不同运输时间以及运输结束24hr时均降低,至48hr恢复正常。不同的运输时间均使大鼠外周白细胞总数(WBC)急剧减少,下降幅度超过40%,在结束运输后的48hr内虽有回升但仍低于对照组,直至72hr才恢复;免疫球蛋白G(IgG)、白介素2(IL-2)、γ干扰素(IFN-γ)及白介素1(IL-1)的血清含量在不同时间运输后即刻均没有显著变化,但在结束运输后的48hr内均不同程度降低,直到72hr才恢复至正常水平。肝脏hsp72mRNA的表达随着运输时间的延长而极显著上调,运输结束后逐渐恢复,直到72hr才恢复至正常水平。[结论]短途运输应激对实验大鼠的生长代谢、神经内分泌和免疫功能均有不利影响,经历常规短途运输(1.5hr以内)的实验大鼠,到达目的地后的健康适应期至少应为72hr;外周血中白细胞总数(WBC)、血糖(GLU)、皮质酮(CORT)、β-内啡肽(β-EP)、免疫球蛋白G(IgG)、白介素2(IL-2)、γ干扰素(IFN-γ)及白介素1(IL-1)的含量可作为评价短途运输应激的指标,hsp72可能是短途运输应激分子机理研究的切入点。
第二部分短途运输应激对实验小鼠影响的评价研究
[目的]评价短途运输应激对近交系和封闭群实验小鼠的影响。[方法]以近交系BALB/c小鼠和封闭群ICR小鼠为研究对象,测定其经历运输1.5hr及到达实验室后分别恢复不同时间(24hr、48hr和72hr)时的代谢、神经内分泌和免疫功能主要指标,与未经历运输的对照组比较,观察分析短途运输应激对不同遗传背景实验小鼠的影响。[结果]运输结束时,BALB/c小鼠和ICR小鼠的GLU和血清中CORT、β-EP含量均降低,CORT在到达目的地后24hr已恢复正常,β-EP和GLU至抵达后48hr恢复正常。BALB/c小鼠和ICR小鼠外周血中WBC总数在运输结束时均下降,在BALB/c小鼠,WBC总数降幅超过74%,直到72hr才恢复正常,在ICR小鼠,WBC总数降幅超过55%,于48hr恢复正常。BALB/c小鼠和ICR小鼠血清IL-2的含量在运输结束时也降低,降幅分别为43%和45%,BALB/c小鼠在运输结束48hr时IL-2水平恢复正常,ICR小鼠则到72hr才恢复正常。[结论]短途运输应激对不同遗传背景实验小鼠的代谢、神经内分泌和免疫功能均会产生不利影响,其中免疫功能所受的影响较大。经历常规短途运输(1.5hr以内)的实验小鼠,到达目的地后的健康适应期至少应为72hr。外周血中WBC总数和GLU、CORT、β-EP、IL-2的含量可作为评价短途运输应激对实验小鼠影响的主要指标。
第三部分短途运输应激对实验小鼠影响的营养干预研究
[目的]研制凝胶状营养补充剂,并对其抗短途运输应激的作用效果进行评价。
[方法]研究采用BALB/c小鼠,测定运输营养干预组和运输对照组在经历运输1.5hr及到达实验室后分别恢复24hr、48hr和72hr时代谢、神经内分泌和免疫功能主要指标的变化,与未经历运输的空白对照组比较,分析凝胶状营养补充剂的干预效果。[结果]营养干预组GLU、血清β-EP、IL-2的含量都在运输结束到达目的地后的24hr内恢复正常,外周WBC总数在48hr内恢复正常,而运输对照组GLU、血清β-EP、IL-2的含量则都在结束运输到达目的地后的48hr内恢复正常,外周WBC总数则在结束运输到达目的地后的72hr内恢复正常。两组的血清CORT含量则在结束运输到达目的地后的24hr内均恢复正常。[结论]凝胶状营养补充剂能够减轻短途运输应激对动物代谢和免疫功能的不利影响,促使其较快提前恢复至正常水平,具有一定的抗短途运输应激作用。
Short-term transportation is the most commonly used means in moving and relocating laboratory animals.However nowadays few study has been conducted on the influence of stress on laboratory animal due to short-term transportation and few intervention methods have been introduced to reduce the stress.This study is aimed to explore the establishment of a technical evaluation index system on the stress due to short-term transportation;to identify the lengthen of acclimatization laboratory animals needed following transportation;to study intervention measures to relief stress;and to explore a preliminary system on the influence of stress to laboratory rats and mice due to short-term transportation.
PART ONE Study on the influence of stress on laboratory rats due to short-term transportation
[Objective]To study the possible influence of short-term transportation on laboratory rodents,to establish a technical index system to evaluate the influence of the stress on laboratory rodents.[Methods]To assign Wistar rats into 8 groups,including 3 groups experiencing different transportation time(0.5hr、1hr and 1.5hr),4 groups all experiencing 1.5hr of transportation time but were given different period of acclimatization(24hr、48hr、72h and 96hr) after end of transportation,and 1 group as control group(normal laboratory raising,not experiencing transportation).Main indexes of metabolism,neuroendocrine and immunity were tested.All the indexes of rats experiencing transportation were compared with those of the control group to analyze the impact of short-term transportation on laboratory rats.[Results]Wistar rats experienced short-term transportation saw sharp decrease in weight,the longer the transportation time,the higher the rate of weight loss.Weight increase rate rose sharply right after stop of transportation and this high weight increase rate lasted until 72hr after end of transportation.Of the 72hr,the first 24hr saw most sharp increase rate.GLU of rats increased both at the moment of 0.5hr and the moment of 1hr during transportation,while decreased at the time of 1.5hr during transportation. GLU increased again at the moment of 24hr after stop of transportation.After that, GLU had no significant difference as compared with those of control group.Serum CORT also increased both at the moment of 0.5hr and the moment of 1hr during transportation,while decreased at the moment of 1.5hr during transportation,but resumed to normal 24hr after stop of transportation,after 24hr no significant different in serum CORT was found as compared with that of the control group.Serumβ-EP saw decrease at all three different time(0.5hr、1hr and 1.5hr) of transportation as well as 24hr after transportation,until at the time of 48hr index of Serumβ-EP started to resume to normal.Total WBC in peripheral blood at all three different time(0.5hr、1hr and 1.5hr) of transportation decreased sharply,decreased by 40%,though increased within 48hr after end of transportation total WBC in peripheral blood still lower than normal,until 72hr after transportation the parameter started to resume to normal. IgG,IL-2,IFN-γ,IL-1 in serum had no significant change at time of transportation, but decreased sharply between the moment of end transportation and 48hr after transportation,until at the moment of 72hr after transportation these parameters started to resume to normal.The expression of hsp72mRNA in liver increased significantly at all three different time(0.5hr、1hr and 1.5hr) of transportation,the longer the transportation time,the higher the expression of hsp72mRNA in liver,and resumed quickly within 72hr after end of transportation.[Conclusion]Short-term transportation imposed negative impact on metabolism,neuroendocrine and immunity of laboratory rats.Laboratory rats experienced short-term transportation(1.5hr) needed at least 72hr of acclimatization after end of transportation.Parameters of total WBC in peripheral blood,GLU,CORT,β-EP,IgG,IL-2,IFN-γ,IL-1 in serum can be used as indexes to evaluate stress due to short-term transportation.The expression of hsp72mRNA is beneficial to discover the mechanism of molecular biology in the stress due to short-term transportation.
PART TWO Evaluation study on the stress to laboratory mice due to short-term transportation
[Objective]To evaluate the stress to inbred strain mice and outbred stock mice due to short-term transportation.[Methods]BALB/c mice and ICR mice were transported for 1.5hr and then taken back to laboratory.Parameters of metabolism, neuroendocrine and immunity at the moment of arrival and at different acclimatization moments(24hr,48hr,72hr respectively after end of transportation) were tested.All the parameters were compared with those of control group(normal laboratory raising,not experiencing transportation) to study and analyze the impact of stress to laboratory mice with different genetic background.[Results]Parameters of GLU and CORT andβ-EP in serum of BALB/c mice and ICR mice all decreased in rate at the moment of stop transportation.Serum CORT resumed to normal within 24hr after arrival,whileβ-EP in serum and GLU resumed to normal within 48hr after arriving at destination.Total WBC in peripheral blood of both BALB/c mice and ICR mice saw decrease at the moment of stop transportation.To BALB/c mice,total WBC in peripheral blood decreased by 74%,until 72hr started to resume to normal, while to ICR mice,total WBC in peripheral blood decreased by 55%,and resumed to normal 48hr after end of transportation.Serum IL-2 of both BALB/c mice and ICR mice also decreased at the end of transportation,decrease rate was 43%and 45% respectively.IL-2 in serum of BALB/c mice resumed to normal at the moment of 48hr after end of transportation,while IL-2 in serum of ICR mice did not recover to normal until the moment of 72hr after end of transportation.[Conclusion] Short-term transportation had negative impact on metabolism,neuroendocrine and immunity of laboratory mice with different genetic background.Among which, impact on immunity was quite significant.Laboratory mice experienced short-term transportation(within 1.5hr) needed at least 72hr of acclimatization after arriving at destination.Total WBC in peripheral blood,GLU,CORT、β-EP、IL-2 in serum could be used as key indexes to evaluate stress to laboratory mice due to short-term transportation.
PART THREE Study on nutrition intervention to the stress of laboratory mice due to short-term transportation
[Objective]To research and prescribe jelly-shaped nutrition supplements,and to evaluate their affect on the reduction of stress caused by short-term transportation.
[Methods]To divide BALB/c mice into nutrition intervention group and non nutrition intervention group and to test changes on major parameters of metabolism, neuroendocrine and immunity at the moment of arriving at laboratory after 1.5hr of transportation as well as those at the moment of 24h,48hr and 72hr after end of transport respectively.All these parameters were used to compare with those of control group(normal laboratory raising,not experiencing transportation).Analysis was made to the result of intervention by jelly-shaped nutrition supplements.
[Results]β-EP、IL-2 in serum and GLU of nutrition intervention group resumed to normal within 24hr after end of transportation.Total WBC in peripheral blood resumed to normal within 48hr after end of transportation.While to the non nutrition intervention group,β-EP、IL-2 in serum and GLU resumed to normal within 48hr after arriving at destination,and their total WBC in peripheral blood resumed to normal within 72hr after end of transportation.Serum CORT of both groups resumed to normal within 24hr after end of transportation.[Conclusion] Jelly-shaped nutrition supplements were able to reduce negative impact on metabolism and immunity of the animals experienced short-term transportation,and were able to help shorten acclimatization period.Therefore these nutrition supplement were effective in reduce stress caused by short-term transportation.
第一部分短途运输应激对实验大鼠的影响研究
[目的]探讨短途运输应激对啮齿类实验动物影响的可能机制,建立评价短途运输应激对啮齿类实验动物影响的技术指标体系。[方法]使Wistar大鼠分别经历不同运输时间(0.5hr、1hr和1.5hr),以及经历1.5hr运输时间到达实验室后分别恢复不同时间(24hr、48hr、72hr和96hr),测定其生长代谢、神经内分泌和免疫功能主要指标,与未经历运输的对照组比较,观察分析短途运输应激对实验大鼠的影响。[结果]短途运输导致Wistar大鼠的体重迅速下降,体重丢失率随着运输时间的延长呈增大趋势;运输结束后大鼠的体重迅速恢复,高水平的体重增长率一直持续到运输后72hr,且以24hr内的体重增长率为最高。大鼠血糖(GLU)在运输0.5hr和1hr时均升高,而在运输1.5hr时降低,运输结束后24hr时GLU再次升高,此后与对照组无差异;血清皮质酮(CORT)亦在运输0.5hr和1hr时升高而运输1.5hr时降低,但运输结束24hr起即恢复至对照组水平;血清β-内啡肽(β-EP)在经历不同运输时间以及运输结束24hr时均降低,至48hr恢复正常。不同的运输时间均使大鼠外周白细胞总数(WBC)急剧减少,下降幅度超过40%,在结束运输后的48hr内虽有回升但仍低于对照组,直至72hr才恢复;免疫球蛋白G(IgG)、白介素2(IL-2)、γ干扰素(IFN-γ)及白介素1(IL-1)的血清含量在不同时间运输后即刻均没有显著变化,但在结束运输后的48hr内均不同程度降低,直到72hr才恢复至正常水平。肝脏hsp72mRNA的表达随着运输时间的延长而极显著上调,运输结束后逐渐恢复,直到72hr才恢复至正常水平。[结论]短途运输应激对实验大鼠的生长代谢、神经内分泌和免疫功能均有不利影响,经历常规短途运输(1.5hr以内)的实验大鼠,到达目的地后的健康适应期至少应为72hr;外周血中白细胞总数(WBC)、血糖(GLU)、皮质酮(CORT)、β-内啡肽(β-EP)、免疫球蛋白G(IgG)、白介素2(IL-2)、γ干扰素(IFN-γ)及白介素1(IL-1)的含量可作为评价短途运输应激的指标,hsp72可能是短途运输应激分子机理研究的切入点。
第二部分短途运输应激对实验小鼠影响的评价研究
[目的]评价短途运输应激对近交系和封闭群实验小鼠的影响。[方法]以近交系BALB/c小鼠和封闭群ICR小鼠为研究对象,测定其经历运输1.5hr及到达实验室后分别恢复不同时间(24hr、48hr和72hr)时的代谢、神经内分泌和免疫功能主要指标,与未经历运输的对照组比较,观察分析短途运输应激对不同遗传背景实验小鼠的影响。[结果]运输结束时,BALB/c小鼠和ICR小鼠的GLU和血清中CORT、β-EP含量均降低,CORT在到达目的地后24hr已恢复正常,β-EP和GLU至抵达后48hr恢复正常。BALB/c小鼠和ICR小鼠外周血中WBC总数在运输结束时均下降,在BALB/c小鼠,WBC总数降幅超过74%,直到72hr才恢复正常,在ICR小鼠,WBC总数降幅超过55%,于48hr恢复正常。BALB/c小鼠和ICR小鼠血清IL-2的含量在运输结束时也降低,降幅分别为43%和45%,BALB/c小鼠在运输结束48hr时IL-2水平恢复正常,ICR小鼠则到72hr才恢复正常。[结论]短途运输应激对不同遗传背景实验小鼠的代谢、神经内分泌和免疫功能均会产生不利影响,其中免疫功能所受的影响较大。经历常规短途运输(1.5hr以内)的实验小鼠,到达目的地后的健康适应期至少应为72hr。外周血中WBC总数和GLU、CORT、β-EP、IL-2的含量可作为评价短途运输应激对实验小鼠影响的主要指标。
第三部分短途运输应激对实验小鼠影响的营养干预研究
[目的]研制凝胶状营养补充剂,并对其抗短途运输应激的作用效果进行评价。
[方法]研究采用BALB/c小鼠,测定运输营养干预组和运输对照组在经历运输1.5hr及到达实验室后分别恢复24hr、48hr和72hr时代谢、神经内分泌和免疫功能主要指标的变化,与未经历运输的空白对照组比较,分析凝胶状营养补充剂的干预效果。[结果]营养干预组GLU、血清β-EP、IL-2的含量都在运输结束到达目的地后的24hr内恢复正常,外周WBC总数在48hr内恢复正常,而运输对照组GLU、血清β-EP、IL-2的含量则都在结束运输到达目的地后的48hr内恢复正常,外周WBC总数则在结束运输到达目的地后的72hr内恢复正常。两组的血清CORT含量则在结束运输到达目的地后的24hr内均恢复正常。[结论]凝胶状营养补充剂能够减轻短途运输应激对动物代谢和免疫功能的不利影响,促使其较快提前恢复至正常水平,具有一定的抗短途运输应激作用。
Short-term transportation is the most commonly used means in moving and relocating laboratory animals.However nowadays few study has been conducted on the influence of stress on laboratory animal due to short-term transportation and few intervention methods have been introduced to reduce the stress.This study is aimed to explore the establishment of a technical evaluation index system on the stress due to short-term transportation;to identify the lengthen of acclimatization laboratory animals needed following transportation;to study intervention measures to relief stress;and to explore a preliminary system on the influence of stress to laboratory rats and mice due to short-term transportation.
PART ONE Study on the influence of stress on laboratory rats due to short-term transportation
[Objective]To study the possible influence of short-term transportation on laboratory rodents,to establish a technical index system to evaluate the influence of the stress on laboratory rodents.[Methods]To assign Wistar rats into 8 groups,including 3 groups experiencing different transportation time(0.5hr、1hr and 1.5hr),4 groups all experiencing 1.5hr of transportation time but were given different period of acclimatization(24hr、48hr、72h and 96hr) after end of transportation,and 1 group as control group(normal laboratory raising,not experiencing transportation).Main indexes of metabolism,neuroendocrine and immunity were tested.All the indexes of rats experiencing transportation were compared with those of the control group to analyze the impact of short-term transportation on laboratory rats.[Results]Wistar rats experienced short-term transportation saw sharp decrease in weight,the longer the transportation time,the higher the rate of weight loss.Weight increase rate rose sharply right after stop of transportation and this high weight increase rate lasted until 72hr after end of transportation.Of the 72hr,the first 24hr saw most sharp increase rate.GLU of rats increased both at the moment of 0.5hr and the moment of 1hr during transportation,while decreased at the time of 1.5hr during transportation. GLU increased again at the moment of 24hr after stop of transportation.After that, GLU had no significant difference as compared with those of control group.Serum CORT also increased both at the moment of 0.5hr and the moment of 1hr during transportation,while decreased at the moment of 1.5hr during transportation,but resumed to normal 24hr after stop of transportation,after 24hr no significant different in serum CORT was found as compared with that of the control group.Serumβ-EP saw decrease at all three different time(0.5hr、1hr and 1.5hr) of transportation as well as 24hr after transportation,until at the time of 48hr index of Serumβ-EP started to resume to normal.Total WBC in peripheral blood at all three different time(0.5hr、1hr and 1.5hr) of transportation decreased sharply,decreased by 40%,though increased within 48hr after end of transportation total WBC in peripheral blood still lower than normal,until 72hr after transportation the parameter started to resume to normal. IgG,IL-2,IFN-γ,IL-1 in serum had no significant change at time of transportation, but decreased sharply between the moment of end transportation and 48hr after transportation,until at the moment of 72hr after transportation these parameters started to resume to normal.The expression of hsp72mRNA in liver increased significantly at all three different time(0.5hr、1hr and 1.5hr) of transportation,the longer the transportation time,the higher the expression of hsp72mRNA in liver,and resumed quickly within 72hr after end of transportation.[Conclusion]Short-term transportation imposed negative impact on metabolism,neuroendocrine and immunity of laboratory rats.Laboratory rats experienced short-term transportation(1.5hr) needed at least 72hr of acclimatization after end of transportation.Parameters of total WBC in peripheral blood,GLU,CORT,β-EP,IgG,IL-2,IFN-γ,IL-1 in serum can be used as indexes to evaluate stress due to short-term transportation.The expression of hsp72mRNA is beneficial to discover the mechanism of molecular biology in the stress due to short-term transportation.
PART TWO Evaluation study on the stress to laboratory mice due to short-term transportation
[Objective]To evaluate the stress to inbred strain mice and outbred stock mice due to short-term transportation.[Methods]BALB/c mice and ICR mice were transported for 1.5hr and then taken back to laboratory.Parameters of metabolism, neuroendocrine and immunity at the moment of arrival and at different acclimatization moments(24hr,48hr,72hr respectively after end of transportation) were tested.All the parameters were compared with those of control group(normal laboratory raising,not experiencing transportation) to study and analyze the impact of stress to laboratory mice with different genetic background.[Results]Parameters of GLU and CORT andβ-EP in serum of BALB/c mice and ICR mice all decreased in rate at the moment of stop transportation.Serum CORT resumed to normal within 24hr after arrival,whileβ-EP in serum and GLU resumed to normal within 48hr after arriving at destination.Total WBC in peripheral blood of both BALB/c mice and ICR mice saw decrease at the moment of stop transportation.To BALB/c mice,total WBC in peripheral blood decreased by 74%,until 72hr started to resume to normal, while to ICR mice,total WBC in peripheral blood decreased by 55%,and resumed to normal 48hr after end of transportation.Serum IL-2 of both BALB/c mice and ICR mice also decreased at the end of transportation,decrease rate was 43%and 45% respectively.IL-2 in serum of BALB/c mice resumed to normal at the moment of 48hr after end of transportation,while IL-2 in serum of ICR mice did not recover to normal until the moment of 72hr after end of transportation.[Conclusion] Short-term transportation had negative impact on metabolism,neuroendocrine and immunity of laboratory mice with different genetic background.Among which, impact on immunity was quite significant.Laboratory mice experienced short-term transportation(within 1.5hr) needed at least 72hr of acclimatization after arriving at destination.Total WBC in peripheral blood,GLU,CORT、β-EP、IL-2 in serum could be used as key indexes to evaluate stress to laboratory mice due to short-term transportation.
PART THREE Study on nutrition intervention to the stress of laboratory mice due to short-term transportation
[Objective]To research and prescribe jelly-shaped nutrition supplements,and to evaluate their affect on the reduction of stress caused by short-term transportation.
[Methods]To divide BALB/c mice into nutrition intervention group and non nutrition intervention group and to test changes on major parameters of metabolism, neuroendocrine and immunity at the moment of arriving at laboratory after 1.5hr of transportation as well as those at the moment of 24h,48hr and 72hr after end of transport respectively.All these parameters were used to compare with those of control group(normal laboratory raising,not experiencing transportation).Analysis was made to the result of intervention by jelly-shaped nutrition supplements.
[Results]β-EP、IL-2 in serum and GLU of nutrition intervention group resumed to normal within 24hr after end of transportation.Total WBC in peripheral blood resumed to normal within 48hr after end of transportation.While to the non nutrition intervention group,β-EP、IL-2 in serum and GLU resumed to normal within 48hr after arriving at destination,and their total WBC in peripheral blood resumed to normal within 72hr after end of transportation.Serum CORT of both groups resumed to normal within 24hr after end of transportation.[Conclusion] Jelly-shaped nutrition supplements were able to reduce negative impact on metabolism and immunity of the animals experienced short-term transportation,and were able to help shorten acclimatization period.Therefore these nutrition supplement were effective in reduce stress caused by short-term transportation.
引文
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