四君子汤对训练小鼠血清和肝脏自由基变化影响的实验研究
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摘要
研究目的
本实验在中医基础理论的指导下,对处于运动训练状态下的小鼠灌服传统中药制剂四君子汤,观察四君子汤抗疲劳和提高运动能力的作用,并通过对不同功能状态下小鼠血清和肝脏自由基代谢、抗氧化酶活力等指标的测定,探讨四君子汤和运动训练对机体自由基代谢的影响与提高运动能力的机制,并为开发一种能延缓运动性疲劳,提高运动能力,安全有效且有别于兴奋剂的复方中药制剂奠定基础,以期为指导人们合理利用中药抗疲劳和指导运动训练提供科学的理论和实验依据。
研究方法
选取健康、雄性、昆明小鼠128只,4周龄,18-20g左右,适应性饲养一周后,随机分为四组,分别为安静对照组(A组)、单纯运动组(B组)、安静服药组(C组)、运动服药组(D组)。C、D组每天灌服一定剂量四君子汤中药制剂,A、B组灌服等量蒸馏水;B、D组进行6周游泳训练,A、C组每天保持正常生理活动,干预6周,第7周,各组小鼠再随机分成安静状态组、一定时间(游泳90min)组、力竭即刻组、力竭恢复24小时组4个亚组,并于各组相应时刻摘眼球取血后非抗凝分离制备血清,断脊椎处死小鼠,取肝脏组织,置于冰生理盐水中洗净血渍,用滤纸吸干。血清和肝脏置于-30℃冰柜保存待测SOD、GSH-PX活性和MDA含量。用秒表记录力竭时间。结果以平均值士标准差(x士s)表示。
实验结果:
(1)安静状态下,小鼠血清和肝脏中MDA含量B组和D组与A组相比,有下降的趋势,但差异不显著(p﹥0.05),GSH-PX活性和SOD的活性B、D组均显著高于A组﹙p﹤0.05﹚。
(2)一定时间运动后,血清和肝脏MDA含量B、C、D组均显著低于A组﹙p﹤0.05﹚; SOD活性B、C、D组都显著高于A组﹙p﹤0.05﹚;血清GSH-PX活性D组显著高于A组﹙p﹤0.05﹚,肝脏GSH-PX活性B、C、D组都显著高于A组﹙p﹤0.05﹚,D组显著高于B组﹙p﹤0.05﹚。
(3)力竭运动即刻,血清和肝脏中MDA含量C、D组显著低于A组,D组又显著低于B组﹙p﹤0.05﹚,SOD活性C、D组显著高于A组,且D组显著高于B组﹙p﹤0.05﹚;GSH-PX活性D组显著高于A组﹙p﹤0.05﹚,且肝脏中C组也显著高于A组,D组显著高于B组﹙p﹤0.05﹚。
(4)力竭恢复24小时后,血清和肝脏MDA含量B、C、D组显著低于A组,D组显著低于B组﹙p﹤0.05﹚,SOD活性,C、D组显著高于A组﹙p﹤0.05﹚;GSH-PX活性的变化,B、C组和D组均显著高于A组﹙p﹤0.05﹚,且肝脏中,D组还显著高于B组﹙p﹤0.05﹚。
(5)D组小鼠力竭时间相比另外三组非常显著性延长﹙p﹤0.01﹚; B和C两组与A组对比也显著性延长﹙p﹤0.05﹚。
结论:
1.四君子汤能够辅助运动训练显著延长小鼠游泳的力竭时间,具有明显的提高机体运动能力和抗疲劳能力的作用。
2.四君子汤能够有效的降低小鼠血清及肝脏安静时、一定时间、力竭运动即刻及恢复24小时后MDA含量,提高SOD、GSH-PX浓度,增强机体上述状态下的抗氧化能力。
3.运动训练能降低小鼠血清和肝脏内MDA含量,抑制脂质过氧化,提高小鼠血清和肝脏内SOD和GSH-PX活性,增强机体的抗氧化能力。
Research Aims
Under the academic instruction of herbalist doctor basic,by filling rats which were in training states with sijunzi formula of traditional preparation, to observe the effects of sijunzi formula on resisting fatigue and enhancing exercise ability,By mensurating SOD vigor and GSH-PX vigor about enzyme of antioxidation,and the content of MDA in the offspring of free radical metabolism in both the serum and the liver in rats after different exercising states ,to discuss the effects of sijunzi formula and exercise training on rats economy and on the mechanism of enhancing exercising ability,and to establish a base to develop a compound preparation of chinese traditional medicine,which would delay exercise-induced fatigue,improve exercise ability,differ from incitant but safety and efficiency,expecting to provide scientific theory and experimental grounds for people to use chinese traditional medicine reasonably to resist fatigue,and to instruct trainning.
Research methods
select128 health and male Kunming rats,4 weeks old,18-20g,after a week’s adaptabe breeding,the rats were divided into 4 groups randomly.they were contrast group(A)、exercise group(B)、sijunzi formula group(C)、exercise+ sijunzi formula group(D),Rats in group A keep quiet and be filled with distilled water、Rats in group B take a swimming exercise and be filled with distilled water、Rats in group C keep quiet and be filled with sijunzi formula、Rats in group D take a swimming exercise and be filled with sijunzi formula,The experiment lasts 6 weeks,and when finishing the experimentation,rats in each group will be divided into 4 different exercising states, there were states of quiet、fixed time(90min’s swimming)、exhaustion instantly,and exhaustion recovery 24 hours later,at the end of each exercising states, taking some coagulant blood from eyeball to separate serum, and then disconnecting spondyle of the rats to take liver tissue,purging off blood with ice PBS,bloting up blood with filter paper. serum and livers were stord in -30℃to detect SOD、GSH-PX activities,and MDA content,expressing the results with average±stdev(x士s).
Research results
(1)state of quiet,compared with group A,there were descending trends of MDA contents in rats’serum and liver of groups B and D ,but not markedly(p﹥0.05);there were remarkably increase of GSH-PX,SOD activities in groups B and D(p﹤0.05).
(2)at the end of fixed time(90min’s swimming), compared with group A, there were remarkably descending of MDA contents in rats’serum and liver of groups B、C and D (p﹤0.05); there were remarkably increase of SOD activities in rats’serum and liver of groups B、C and D (p﹤0.05); there was remarkably increase of GSH-PX activity in rats’serum of group D (p﹤0.05);there were remarkably increase of GSH-PX activity in rats’liver of groups B、C and D (p﹤0.05).
(3)exhaustion instantly, compared with group A, there were remarkably descending of MDA contents and remarkably increase of SOD activities in rats’serum and liver of groups C and D (p﹤0.05),compared with group B,there were remarkably descending of MDA contents and remarkably increase of GSH-PX and SOD activities in group D(p﹤0.05); compared with group A,there was remarkably increase of GSH-PX activity in rats’serum and liver of group D(p﹤0.05),there was remarkably increase of GSH-PX activity in rats’liver of group C(p﹤0.05).
(4)exhaustion recovery 24 hours later, compared with group A, there were remarkably descending of MDA contents in rats’serum and liver of groups B、C and D(p﹤0.05),of which group D is much lower than that of group B(p﹤0.05);compared with group A, there were remarkably increase of SOD activities in rats’serum and liver of groups C and D(P﹤0.05),there were remarkably increase of GSH-PX activities in rats’liver of groups B、C and D(p﹤0.05);compared with group B there was also remarkably increase of GSH-PX activity in rats’liver of group D(p﹤0.05).
(5)compared with group D, the exhaustion time was remarkably prolonged in groups A、Band C(p﹤0.01); compared with group A,the exhaustion time was also remarkably prolonged in groups Band C(p﹤0.05).
Conclusions
(1)Sijunzi formula can cooperate with exercise training to prolong swimming time exhaustion of rats,there is a obvious effect of sijunzi formula on increasing exercise ability and resisting fatigue.
(2)Sijunzi formula can reduce MDA content in rats’serum and liver,restrain lipid oxygenation,and reduce free radical content efficiently.Sijunzi formula can enhance enzyme activities of SOD、GSH-PX in rats’serum and liver,reminding that,sijunzi formula can enhance antioxidation abilities in different training states.
(3)Exercise training can reduce MDA content in rats’serum and liver、restrain lipid peroxide、enhance enzyme activities of SOD、GSH-PX ,and boost up antioxidation abilities.
本实验在中医基础理论的指导下,对处于运动训练状态下的小鼠灌服传统中药制剂四君子汤,观察四君子汤抗疲劳和提高运动能力的作用,并通过对不同功能状态下小鼠血清和肝脏自由基代谢、抗氧化酶活力等指标的测定,探讨四君子汤和运动训练对机体自由基代谢的影响与提高运动能力的机制,并为开发一种能延缓运动性疲劳,提高运动能力,安全有效且有别于兴奋剂的复方中药制剂奠定基础,以期为指导人们合理利用中药抗疲劳和指导运动训练提供科学的理论和实验依据。
研究方法
选取健康、雄性、昆明小鼠128只,4周龄,18-20g左右,适应性饲养一周后,随机分为四组,分别为安静对照组(A组)、单纯运动组(B组)、安静服药组(C组)、运动服药组(D组)。C、D组每天灌服一定剂量四君子汤中药制剂,A、B组灌服等量蒸馏水;B、D组进行6周游泳训练,A、C组每天保持正常生理活动,干预6周,第7周,各组小鼠再随机分成安静状态组、一定时间(游泳90min)组、力竭即刻组、力竭恢复24小时组4个亚组,并于各组相应时刻摘眼球取血后非抗凝分离制备血清,断脊椎处死小鼠,取肝脏组织,置于冰生理盐水中洗净血渍,用滤纸吸干。血清和肝脏置于-30℃冰柜保存待测SOD、GSH-PX活性和MDA含量。用秒表记录力竭时间。结果以平均值士标准差(x士s)表示。
实验结果:
(1)安静状态下,小鼠血清和肝脏中MDA含量B组和D组与A组相比,有下降的趋势,但差异不显著(p﹥0.05),GSH-PX活性和SOD的活性B、D组均显著高于A组﹙p﹤0.05﹚。
(2)一定时间运动后,血清和肝脏MDA含量B、C、D组均显著低于A组﹙p﹤0.05﹚; SOD活性B、C、D组都显著高于A组﹙p﹤0.05﹚;血清GSH-PX活性D组显著高于A组﹙p﹤0.05﹚,肝脏GSH-PX活性B、C、D组都显著高于A组﹙p﹤0.05﹚,D组显著高于B组﹙p﹤0.05﹚。
(3)力竭运动即刻,血清和肝脏中MDA含量C、D组显著低于A组,D组又显著低于B组﹙p﹤0.05﹚,SOD活性C、D组显著高于A组,且D组显著高于B组﹙p﹤0.05﹚;GSH-PX活性D组显著高于A组﹙p﹤0.05﹚,且肝脏中C组也显著高于A组,D组显著高于B组﹙p﹤0.05﹚。
(4)力竭恢复24小时后,血清和肝脏MDA含量B、C、D组显著低于A组,D组显著低于B组﹙p﹤0.05﹚,SOD活性,C、D组显著高于A组﹙p﹤0.05﹚;GSH-PX活性的变化,B、C组和D组均显著高于A组﹙p﹤0.05﹚,且肝脏中,D组还显著高于B组﹙p﹤0.05﹚。
(5)D组小鼠力竭时间相比另外三组非常显著性延长﹙p﹤0.01﹚; B和C两组与A组对比也显著性延长﹙p﹤0.05﹚。
结论:
1.四君子汤能够辅助运动训练显著延长小鼠游泳的力竭时间,具有明显的提高机体运动能力和抗疲劳能力的作用。
2.四君子汤能够有效的降低小鼠血清及肝脏安静时、一定时间、力竭运动即刻及恢复24小时后MDA含量,提高SOD、GSH-PX浓度,增强机体上述状态下的抗氧化能力。
3.运动训练能降低小鼠血清和肝脏内MDA含量,抑制脂质过氧化,提高小鼠血清和肝脏内SOD和GSH-PX活性,增强机体的抗氧化能力。
Research Aims
Under the academic instruction of herbalist doctor basic,by filling rats which were in training states with sijunzi formula of traditional preparation, to observe the effects of sijunzi formula on resisting fatigue and enhancing exercise ability,By mensurating SOD vigor and GSH-PX vigor about enzyme of antioxidation,and the content of MDA in the offspring of free radical metabolism in both the serum and the liver in rats after different exercising states ,to discuss the effects of sijunzi formula and exercise training on rats economy and on the mechanism of enhancing exercising ability,and to establish a base to develop a compound preparation of chinese traditional medicine,which would delay exercise-induced fatigue,improve exercise ability,differ from incitant but safety and efficiency,expecting to provide scientific theory and experimental grounds for people to use chinese traditional medicine reasonably to resist fatigue,and to instruct trainning.
Research methods
select128 health and male Kunming rats,4 weeks old,18-20g,after a week’s adaptabe breeding,the rats were divided into 4 groups randomly.they were contrast group(A)、exercise group(B)、sijunzi formula group(C)、exercise+ sijunzi formula group(D),Rats in group A keep quiet and be filled with distilled water、Rats in group B take a swimming exercise and be filled with distilled water、Rats in group C keep quiet and be filled with sijunzi formula、Rats in group D take a swimming exercise and be filled with sijunzi formula,The experiment lasts 6 weeks,and when finishing the experimentation,rats in each group will be divided into 4 different exercising states, there were states of quiet、fixed time(90min’s swimming)、exhaustion instantly,and exhaustion recovery 24 hours later,at the end of each exercising states, taking some coagulant blood from eyeball to separate serum, and then disconnecting spondyle of the rats to take liver tissue,purging off blood with ice PBS,bloting up blood with filter paper. serum and livers were stord in -30℃to detect SOD、GSH-PX activities,and MDA content,expressing the results with average±stdev(x士s).
Research results
(1)state of quiet,compared with group A,there were descending trends of MDA contents in rats’serum and liver of groups B and D ,but not markedly(p﹥0.05);there were remarkably increase of GSH-PX,SOD activities in groups B and D(p﹤0.05).
(2)at the end of fixed time(90min’s swimming), compared with group A, there were remarkably descending of MDA contents in rats’serum and liver of groups B、C and D (p﹤0.05); there were remarkably increase of SOD activities in rats’serum and liver of groups B、C and D (p﹤0.05); there was remarkably increase of GSH-PX activity in rats’serum of group D (p﹤0.05);there were remarkably increase of GSH-PX activity in rats’liver of groups B、C and D (p﹤0.05).
(3)exhaustion instantly, compared with group A, there were remarkably descending of MDA contents and remarkably increase of SOD activities in rats’serum and liver of groups C and D (p﹤0.05),compared with group B,there were remarkably descending of MDA contents and remarkably increase of GSH-PX and SOD activities in group D(p﹤0.05); compared with group A,there was remarkably increase of GSH-PX activity in rats’serum and liver of group D(p﹤0.05),there was remarkably increase of GSH-PX activity in rats’liver of group C(p﹤0.05).
(4)exhaustion recovery 24 hours later, compared with group A, there were remarkably descending of MDA contents in rats’serum and liver of groups B、C and D(p﹤0.05),of which group D is much lower than that of group B(p﹤0.05);compared with group A, there were remarkably increase of SOD activities in rats’serum and liver of groups C and D(P﹤0.05),there were remarkably increase of GSH-PX activities in rats’liver of groups B、C and D(p﹤0.05);compared with group B there was also remarkably increase of GSH-PX activity in rats’liver of group D(p﹤0.05).
(5)compared with group D, the exhaustion time was remarkably prolonged in groups A、Band C(p﹤0.01); compared with group A,the exhaustion time was also remarkably prolonged in groups Band C(p﹤0.05).
Conclusions
(1)Sijunzi formula can cooperate with exercise training to prolong swimming time exhaustion of rats,there is a obvious effect of sijunzi formula on increasing exercise ability and resisting fatigue.
(2)Sijunzi formula can reduce MDA content in rats’serum and liver,restrain lipid oxygenation,and reduce free radical content efficiently.Sijunzi formula can enhance enzyme activities of SOD、GSH-PX in rats’serum and liver,reminding that,sijunzi formula can enhance antioxidation abilities in different training states.
(3)Exercise training can reduce MDA content in rats’serum and liver、restrain lipid peroxide、enhance enzyme activities of SOD、GSH-PX ,and boost up antioxidation abilities.
引文
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