大鼠恐惧应激模型中行为与心血管指标变化
摘要
目的:建立条件恐惧应激致大鼠心血管损伤防治药物药效评价实验动物模型,观察恐惧应激对大鼠心血管功能的损伤效应,并对心血管损伤相关备选指标进行综合的检测、分析、评价。筛选有意义的评价抗恐惧药物药效备选指标,以备今后建立评价体系使用。
方法: 64只健康雄性wistar大鼠随机分为恐惧模型组与对照组,每组32只。自由摄食饮水饲养10天后,第11天起恐惧模型组每日给予2s噪音+3s足底电流刺激,电流强度为1mA,间隔1min,共30次,第12、13天重复实验;对照组每日只给予2s噪音,无足底电流刺激,间隔1min,共30次,第12、13天重复实验。第14天摄像记录两组大鼠5min噪音之后第6—30min内行为变化,评定造模是否成功。自造模成功之后第二日算起分别于1、2、3、4周每周周中(第18、25、32、39天),对两组大鼠进行体重、自发活动、血压心率以及大鼠血清高敏C反应蛋白、肌酸激酶同工酶、心肌钙蛋白I含量测定。
结果: (1)第14天摄像观察两组大鼠5min噪音之后行为学反应,采用三人评定法,恐惧模型组冻结时间显著高于对照组,差异有统计学意义(P<0.05),证明造模成功。(2)两组大鼠体重均称自然增长态势,第1周两组大鼠体重无显著性差异,第2周(P<0.05)、3周(P<0.05)、4周(P<0.01)两组大鼠体重比较均差异显著。(3)两组大鼠自发活动比较,恐惧模型组活动次数在第1周(P<0.05)、2周(P<0.01)、3周(P<0.01)、4周(P<0.05)均显著少于对照组,差异显著。(4)两组大鼠血压值比较,恐惧模型组大鼠血压趋势呈持续升高,对照组无明显变化。第1周两组大鼠血压无显著性差异,第2周(P<0.05)、3周(P<0.01)、4周(P<0.01)两组大鼠血压比较均差异显著。(5)两组大鼠心率比较,第1周两组大鼠心率无显著性差异,第2周(P<0.05)、3周(P<0.05)、4周(P<0.01)两组大鼠心率比较均差异显著。(6)两组大鼠血清C反应蛋白含量比较,恐惧模型组在第1周(P<0.05)、2周(P<0.01)、3周(P<0.01)、4周(P<0.01)均显著高于对照组;(7)两组大鼠血清肌酸激酶同工酶含量比较,恐惧模型组大鼠血清肌酸激酶同工酶在第1、2、3周均无显著差异,第4周(P<0.01)显著高于对照组,差异有统计学意义。(8)两组大鼠血清心肌钙蛋白I含量比较,恐惧模型组大鼠心肌钙蛋白I含量在第1周(P<0.05)、2周(P<0.01)、3周(P<0.01)、4周(P<0.01)均显著高于对照组,差异有统计学意义。
结论:本研究结果表明经过改良的Takashi法可以成功的制造大鼠恐惧应激模型。恐惧应激可以提高大鼠应激水平进而影响大鼠的体重、血压、心率、行为以及心血管指标的改变。部分实验备选指标可以为抗恐惧应激药物药效评价提供相关依据。
Obstract To establish a conditioned fear stress model of rats which causing cardiovascular injury for drug efficacy evaluation , observe the damage effects of fear stress to cardiovascular function of rats. Analysis the related indicators of cardiovascular damage and select meaningful alternative indicators of anti-terror drug efficacy, in order to establish the evaluation system for future use.
Mathods The experiment was carried out with conditioned fear stress model of rats, 64 adult male wistar rats were divided into fear model group and control group. After 10 days of free feeding foods and water, at the 8th day given the fear stress model group daily 2s +3 s of foot electrical stimulation, current intensity 1mA, interval 1min, 30 times everyday; control group given daily 2s noise, no foot electrical stimulation, 1min regular intervals, 30 times everyday. At the 14th days after the 6-30min 5min noise, camera recorded the two groups of rats’behavioral changes, assess the success of modeling. At the middle of week 1,2,3,4 (18th、25 th、32 th、39 th days), test two groups of rats of body weight,locomotor activity,blood pressure and heart rate ,corticosterone, high-sensitivity C-reactive protein and creatine kinase MB, Troponin-I.
Result (1) At the 14th days after the 6-30min 5min noise, camera recorded the two groups of rats’behavioral changes, use the matherd of three assessment, freezeing time of the fear model group was significantly higher, the difference was statistically significant (P <0.05), proving the model was successful maded.(2) With the increase of stress time, the blood pressure of fear stress model group was significantly higher than the control group(week 2:p<0.05; week 3: p<0.05; week 4: p<0.01). (3) compared the spontaneous activity of rats of both groups ,the fear model group were significantly higher than the control group: week1 (P <0.05), week 2 (P <0.01), week 3 (P <0.05), week 4 (P <0.05); the number of activities in the fear model group were significantly lower than the control group week1 (P <0.05), week 2 (P <0.01), week 3 (P <0.01), week 4 (P <0.05) week, the difference were significantly different. (4) With the increase of stress time, the blood pressure of fear stress model group was significantly higher than the control group(week 2:p<0.05; week 3:p<0.01; week 4: p<0.01);(5) The heart rate of conditioned fear stress group,increased significantly than the contral group(week 2:p<0.05;week 3:p<0.05;week 4: p<0.01); (6) The serum corticosterone level of fear stress group were significantly higher than the control group:week 1 (P <0.05), week 2 (P <0.01), week 3 (P <0.05), week 4 (P <0.05) ;(7) The serum C-reactive protein level of fear stress group were significantly higher than the control group:week 1 (P <0.05), week 2 (P <0.01), week 3 (P <0.01), week 4 (P <0.01) ;(8) The serum creatine kinase MB level of fear stress group were significantly higher than the control group at week 4 (P <0.01), week 1、2、3 have no significantly difference; (9)The serum Troponin-I level of fear stress group were significantly higher than the control group:week 1 (P <0.05), week 2 (P <0.01), week 3 (P <0.01), week 4 (P <0.01) ; (10),The myocardial biopsy of HE staining showed that with the prolonged of experiment the fear stress model group have some pathological changes.
Conclusion The results show that the improved of Takashi method can successfully manufactured fear stress model in rats. The fear stress can increase the level of stress and then affect the rat's body weight, blood pressure, heart rate, changes in behavior and cardiovascular measurements. Partially of the indicators can be use for the drug efficacy evaluation of anti-terror stress model.
方法: 64只健康雄性wistar大鼠随机分为恐惧模型组与对照组,每组32只。自由摄食饮水饲养10天后,第11天起恐惧模型组每日给予2s噪音+3s足底电流刺激,电流强度为1mA,间隔1min,共30次,第12、13天重复实验;对照组每日只给予2s噪音,无足底电流刺激,间隔1min,共30次,第12、13天重复实验。第14天摄像记录两组大鼠5min噪音之后第6—30min内行为变化,评定造模是否成功。自造模成功之后第二日算起分别于1、2、3、4周每周周中(第18、25、32、39天),对两组大鼠进行体重、自发活动、血压心率以及大鼠血清高敏C反应蛋白、肌酸激酶同工酶、心肌钙蛋白I含量测定。
结果: (1)第14天摄像观察两组大鼠5min噪音之后行为学反应,采用三人评定法,恐惧模型组冻结时间显著高于对照组,差异有统计学意义(P<0.05),证明造模成功。(2)两组大鼠体重均称自然增长态势,第1周两组大鼠体重无显著性差异,第2周(P<0.05)、3周(P<0.05)、4周(P<0.01)两组大鼠体重比较均差异显著。(3)两组大鼠自发活动比较,恐惧模型组活动次数在第1周(P<0.05)、2周(P<0.01)、3周(P<0.01)、4周(P<0.05)均显著少于对照组,差异显著。(4)两组大鼠血压值比较,恐惧模型组大鼠血压趋势呈持续升高,对照组无明显变化。第1周两组大鼠血压无显著性差异,第2周(P<0.05)、3周(P<0.01)、4周(P<0.01)两组大鼠血压比较均差异显著。(5)两组大鼠心率比较,第1周两组大鼠心率无显著性差异,第2周(P<0.05)、3周(P<0.05)、4周(P<0.01)两组大鼠心率比较均差异显著。(6)两组大鼠血清C反应蛋白含量比较,恐惧模型组在第1周(P<0.05)、2周(P<0.01)、3周(P<0.01)、4周(P<0.01)均显著高于对照组;(7)两组大鼠血清肌酸激酶同工酶含量比较,恐惧模型组大鼠血清肌酸激酶同工酶在第1、2、3周均无显著差异,第4周(P<0.01)显著高于对照组,差异有统计学意义。(8)两组大鼠血清心肌钙蛋白I含量比较,恐惧模型组大鼠心肌钙蛋白I含量在第1周(P<0.05)、2周(P<0.01)、3周(P<0.01)、4周(P<0.01)均显著高于对照组,差异有统计学意义。
结论:本研究结果表明经过改良的Takashi法可以成功的制造大鼠恐惧应激模型。恐惧应激可以提高大鼠应激水平进而影响大鼠的体重、血压、心率、行为以及心血管指标的改变。部分实验备选指标可以为抗恐惧应激药物药效评价提供相关依据。
Obstract To establish a conditioned fear stress model of rats which causing cardiovascular injury for drug efficacy evaluation , observe the damage effects of fear stress to cardiovascular function of rats. Analysis the related indicators of cardiovascular damage and select meaningful alternative indicators of anti-terror drug efficacy, in order to establish the evaluation system for future use.
Mathods The experiment was carried out with conditioned fear stress model of rats, 64 adult male wistar rats were divided into fear model group and control group. After 10 days of free feeding foods and water, at the 8th day given the fear stress model group daily 2s +3 s of foot electrical stimulation, current intensity 1mA, interval 1min, 30 times everyday; control group given daily 2s noise, no foot electrical stimulation, 1min regular intervals, 30 times everyday. At the 14th days after the 6-30min 5min noise, camera recorded the two groups of rats’behavioral changes, assess the success of modeling. At the middle of week 1,2,3,4 (18th、25 th、32 th、39 th days), test two groups of rats of body weight,locomotor activity,blood pressure and heart rate ,corticosterone, high-sensitivity C-reactive protein and creatine kinase MB, Troponin-I.
Result (1) At the 14th days after the 6-30min 5min noise, camera recorded the two groups of rats’behavioral changes, use the matherd of three assessment, freezeing time of the fear model group was significantly higher, the difference was statistically significant (P <0.05), proving the model was successful maded.(2) With the increase of stress time, the blood pressure of fear stress model group was significantly higher than the control group(week 2:p<0.05; week 3: p<0.05; week 4: p<0.01). (3) compared the spontaneous activity of rats of both groups ,the fear model group were significantly higher than the control group: week1 (P <0.05), week 2 (P <0.01), week 3 (P <0.05), week 4 (P <0.05); the number of activities in the fear model group were significantly lower than the control group week1 (P <0.05), week 2 (P <0.01), week 3 (P <0.01), week 4 (P <0.05) week, the difference were significantly different. (4) With the increase of stress time, the blood pressure of fear stress model group was significantly higher than the control group(week 2:p<0.05; week 3:p<0.01; week 4: p<0.01);(5) The heart rate of conditioned fear stress group,increased significantly than the contral group(week 2:p<0.05;week 3:p<0.05;week 4: p<0.01); (6) The serum corticosterone level of fear stress group were significantly higher than the control group:week 1 (P <0.05), week 2 (P <0.01), week 3 (P <0.05), week 4 (P <0.05) ;(7) The serum C-reactive protein level of fear stress group were significantly higher than the control group:week 1 (P <0.05), week 2 (P <0.01), week 3 (P <0.01), week 4 (P <0.01) ;(8) The serum creatine kinase MB level of fear stress group were significantly higher than the control group at week 4 (P <0.01), week 1、2、3 have no significantly difference; (9)The serum Troponin-I level of fear stress group were significantly higher than the control group:week 1 (P <0.05), week 2 (P <0.01), week 3 (P <0.01), week 4 (P <0.01) ; (10),The myocardial biopsy of HE staining showed that with the prolonged of experiment the fear stress model group have some pathological changes.
Conclusion The results show that the improved of Takashi method can successfully manufactured fear stress model in rats. The fear stress can increase the level of stress and then affect the rat's body weight, blood pressure, heart rate, changes in behavior and cardiovascular measurements. Partially of the indicators can be use for the drug efficacy evaluation of anti-terror stress model.
引文
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