补锌对耐力训练、力竭运动小鼠体内酶活性及结蛋白含量变化的影响
发布时间：2019-03-24 18:14
【摘要】：研究目的:本实验以雄性ICR小鼠为研究对象,通过补锌来探讨微量元素锌对耐力训练小鼠骨骼肌中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性变化,力竭运动小鼠骨骼肌中丙二醛(MDA)含量变化,GSH-Px、SOD、血清肌酸激酶(CK)活性变化,以及小鼠运动后骨骼肌中结蛋白(desmin)含量变化的影响。采用耐力测试、组织形态学、酶偶联免疫和免疫组化等技术,通过分析小鼠运动后骨骼肌中各种酶活性变化、细胞骨架蛋白desmin的分布情况、力竭运动时间及力竭运动后即刻骨骼肌的组织形态学变化,来探讨补锌对提高小鼠运动能力的影响。研究方法:研究对象为89只雄性ICR小鼠,适应饲养三天后进行一次游泳测试,剔除不会游泳的小鼠,然后将小鼠随机分为安静对照组、耐力训练组、耐力训练补锌组、力竭运动对照组以及力竭运动补锌组(安静对照组小鼠不做任何处理,力竭运动对照组仅进行一次力竭运动,力竭运动补锌组进行6周的补锌;以下耐力训练补锌组简称训练补锌组,力竭运动补锌组简称补锌力竭组,力竭运动对照组简称对照力竭组),耐力训练组和训练补锌组中又分别设2周组、4周组和6周组用来测各种酶活性的变化及运动后desmin的表达(各组n=10)。补锌方法为:训练补锌组和补锌力竭组于饮用水中加入Zn SO4·7H2O,[Zn]=225 mg/L,其余各组正常饮水。耐力训练组和训练补锌组分别进行2周、4周和6周的游泳耐力训练,每周游6 d,第一周每天游泳30 min,以后每周递加10 min,第六周时每天游泳90min。耐力训练组和训练补锌组于运动后24小时取材,对照力竭组和补锌力竭组于力竭运动后即刻取材。其中力竭判断的标准:小鼠沉入水中过10 s,放在平面上无法完成翻正反射。测试方法及指标:①力竭组对小鼠进行一次性力竭游泳运动并记录力竭运动时间,并以此来探讨补锌对提高机体抗疲劳能力的影响。②HE染色观察小鼠运动后骨骼肌组织形态学变化。③酶偶联等方法测定小鼠运动后骨骼肌中丙二醛(MDA)含量以及谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、血清肌酸激酶(CK)的活性。④免疫组化法检测力竭运动后即刻及耐力运动2周、4周、6周后小鼠骨骼肌中desmin的含量及分布。研究结果:(1)补锌能够显著延长小鼠游泳至力竭的时间,补锌力竭组小鼠游泳至力竭的时间长于对照力竭组,差异极显著(P0.01)。(2)训练补锌组小鼠运动2周、4周和6周后骨骼肌中SOD活性显著高于安静对照组和耐力训练组,与安静对照组相比P0.01,与耐力训练组相比P0.05;运动4周和6周后骨骼肌中谷胱甘肽过氧化物酶(GSH-Px)活性显著高于安静对照组和耐力训练组(P0.05)。力竭运动后即刻补锌力竭组小鼠骨骼肌中GSH-Px、SOD活性显著高于对照力竭组(P0.05)。(3)对照力竭组小鼠力竭运动后即刻血清中CK活性显著高于安静对照组和补锌力竭组,差异极显著;补锌力竭组小鼠运动后即刻血清中CK活性高于安静对照组(P0.05)。力竭运动后即刻补锌力竭组小鼠骨骼肌中丙二醛(MDA)含量显著低于对照力竭组(P0.05),对照力竭组小鼠力竭运动后即刻丙二醛含量高于安静对照组,差异极显著(P0.01)。(4)运动2周、4周、6周后训练补锌组小鼠骨骼肌中desmin免疫组化平均光密度值显著高于安静对照组(P0.05),耐力训练组小鼠骨骼肌中desmin免疫组化平均光密度值低于训练补锌组;力竭运动后即刻补锌力竭组小鼠骨骼肌中结蛋白平均光密度值高于对照力竭组,且具有显著性差异(P0.05)。研究结论:(1)补锌可以显著提高小鼠游泳至力竭的时间,增强小鼠骨骼肌中GSH-Px、SOD的活性,控制力竭运动后血清CK的升高,降低运动后肌肉中MDA的含量,延缓运动性疲劳的发生,对小鼠运动能力的提高具有促进作用。(2)耐力训练过程中补锌可以使小鼠的抗氧化酶活性得到提高,训练补锌组小鼠骨骼肌中GSH-Px、SOD活性显著高于安静对照组和耐力训练组,耐力训练组小鼠骨骼肌中GSH-Px、SOD活性高于安静对照组;补锌和耐力训练均可以提高小鼠的抗氧化能力,且具有协同作用,联合使用时可以显著提高小鼠的运动能力。(3)通过补锌,训练补锌组小鼠骨骼肌中desmin含量显著高于耐力训练组;补锌力竭组小鼠骨骼肌中的desmin含量显著高于对照力竭组,其原因可能是补锌可以促进小鼠骨骼肌纤维的再生,进而提高其抗疲劳能力和运动耐力。(4)一次性力竭运动会引发小鼠骨骼肌的微细损伤,desmin细胞骨架破坏是骨骼肌微损伤的形态学指标,前期补锌可以提高小鼠的运动耐力,降低力竭运动诱导的骨骼肌微细损伤程度。
[Abstract]:Objective: To study the changes of the activity of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in skeletal muscle of mice with endurance training by supplementing zinc to the study of male ICR mice. The changes of the content of malondialdehyde (MDA), GSH-Px, SOD and serum creatine kinase (CK) in the skeletal muscle of the mice after exhaustive exercise, and the influence of the content of desmin in the skeletal muscle following the exercise of the mice. By using the techniques of endurance test, tissue morphology, enzyme-coupled immunoassay, and immunohistochemistry, the changes of various enzyme activities in skeletal muscle, the distribution of skeletal protein desmin, the time of exhaustive exercise and the tissue morphological changes of the skeletal muscle after exhaustive exercise were analyzed by analyzing the changes of the activity of various enzymes in the skeletal muscle after the movement of the mouse. To study the effect of zinc-supplementing on the improvement of the mouse's ability to exercise. Methods:89 male ICR mice were studied. After three days of feeding, a swimming test was carried out to eliminate the mice that would not swim, and then the mice were randomly divided into a quiet control group, a endurance training group and a endurance training zinc-supplementing group. in the control group of exhaustive exercise and in the exercise of the exhaustive exercise, the mice in the control group did not do any treatment, and the exercise control group had only one exhaustive exercise, and the exercise of the exhaustive exercise of the zinc group was carried out for 6 weeks; the following endurance training zinc-supplementing group was simply a training zinc-supplementing group, 2-week,4-week and 6-week groups were used to measure the changes of various enzyme activities and the expression of desmin after exercise (n = 10 in each group). The zinc-supplementing method comprises the following steps of: training a zinc-supplementing group and a zinc-supplementing exhausted group in the drinking water, adding Zn SO4.7H2O,[Zn] = 225 mg/ L, and the rest groups of normal drinking water. The endurance training group and the training zinc-supplementing group were used for swimming endurance training for 2 weeks,4 weeks and 6 weeks, respectively. The endurance training group and the training zinc-supplementing group were used for 24 hours after the exercise. In which the standard of exhaustion judgment: the mouse is immersed in water for 10s, and the positive reflection can not be completed on the plane. Test method and index: the exhaustive swimming exercise of mice and the time of exhaustive exercise were carried out, and the effect of zinc-supplementing on the anti-fatigue ability of the body was discussed. The morphological changes of skeletal muscle after exercise in mice were observed with HHE staining. The content of malondialdehyde (MDA) in skeletal muscle and the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and serum creatine kinase (CK) were determined by enzyme-coupled method. The content and distribution of desmin in skeletal muscle of mice at 2 weeks,4 weeks and 6 weeks after exhaustive exercise were detected by immunohistochemistry. The results of the study: (1) The time of mouse swimming to exhaustion can be significantly prolonged by (1) zinc supplement, and the time of swimming to exhaustion of the mice in the exhaustion group is longer than that of the control force, and the difference is very significant (P0.01). (2) The activity of SOD in the skeletal muscle was significantly higher than that in the control group and the endurance training group after 2 weeks,4 weeks and 6 weeks after the exercise of the zinc-supplemented group. The activity of glutathione peroxidase (GSH-Px) in skeletal muscle after 4 and 6 weeks of exercise was significantly higher than that in the control group and the endurance training group (P0.05). The activity of GSH-Px and SOD in the skeletal muscle of the mice after exhaustive exercise was significantly higher than that of the control group (P0.05). (3) In the control group, the CK activity in the serum was significantly higher than that in the control group and the inexhaustible group after exhaustive exercise, and the CK activity in the serum was higher than that in the control group (P0.05). The content of malondialdehyde (MDA) in the skeletal muscle of the mice after exhaustive exercise was significantly lower than that of the control group (P0.05). (4) The average optical density of desmin in the skeletal muscle of the mice after 2 weeks,4 weeks and 6 weeks was significantly higher than that in the control group (P0.05). The average optical density of desmin in the skeletal muscle of the endurance training group was lower than that of the training zinc supplement group. The average optical density of the bound protein in the skeletal muscle of the mice after exhaustive exercise was higher than that of the control group, and there was a significant difference (P0.05). The results of the study: (1) The time of mouse swimming to exhaustion can be obviously improved, the activity of GSH-Px and SOD in the skeletal muscle of the mouse is enhanced, the increase of serum CK after exhaustive exercise is controlled, the content of MDA in the muscle after exercise is reduced, and the occurrence of exercise fatigue is delayed. And has the effect of promoting the improvement of the exercise capacity of the mouse. (2) The activity of GSH-Px and SOD in the skeletal muscle of the mice was significantly higher than that of the control group and the endurance training group, and the activity of GSH-Px and SOD in the skeletal muscle of the endurance training group was higher than that of the control group. The zinc-supplementing and the endurance training can improve the anti-oxidation ability of the mice and have the synergistic effect, and the exercise capacity of the mice can be obviously improved when the mice are used in combination. (3) The desmin content in the skeletal muscle of the mice was significantly higher than that of the endurance training group by supplementing the zinc, and the desmin content in the skeletal muscle of the zinc-supplemented group was significantly higher than that of the control group, and the reason may be that the zinc supplement can promote the regeneration of the skeletal muscle fiber of the mouse, So as to improve the anti-fatigue ability and the exercise endurance. (4) The single-time exhaustive sports meet the microdamage of the skeletal muscle of the mouse, and the desmin cytoskeleton damage is the morphological index of the microdamage of the skeletal muscle, and the early-phase zinc supplement can improve the exercise endurance of the mouse and reduce the degree of the fine injury of the skeletal muscle induced by the exhaustive exercise.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：G804.7
本文编号：2446566
[Abstract]:Objective: To study the changes of the activity of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in skeletal muscle of mice with endurance training by supplementing zinc to the study of male ICR mice. The changes of the content of malondialdehyde (MDA), GSH-Px, SOD and serum creatine kinase (CK) in the skeletal muscle of the mice after exhaustive exercise, and the influence of the content of desmin in the skeletal muscle following the exercise of the mice. By using the techniques of endurance test, tissue morphology, enzyme-coupled immunoassay, and immunohistochemistry, the changes of various enzyme activities in skeletal muscle, the distribution of skeletal protein desmin, the time of exhaustive exercise and the tissue morphological changes of the skeletal muscle after exhaustive exercise were analyzed by analyzing the changes of the activity of various enzymes in the skeletal muscle after the movement of the mouse. To study the effect of zinc-supplementing on the improvement of the mouse's ability to exercise. Methods:89 male ICR mice were studied. After three days of feeding, a swimming test was carried out to eliminate the mice that would not swim, and then the mice were randomly divided into a quiet control group, a endurance training group and a endurance training zinc-supplementing group. in the control group of exhaustive exercise and in the exercise of the exhaustive exercise, the mice in the control group did not do any treatment, and the exercise control group had only one exhaustive exercise, and the exercise of the exhaustive exercise of the zinc group was carried out for 6 weeks; the following endurance training zinc-supplementing group was simply a training zinc-supplementing group, 2-week,4-week and 6-week groups were used to measure the changes of various enzyme activities and the expression of desmin after exercise (n = 10 in each group). The zinc-supplementing method comprises the following steps of: training a zinc-supplementing group and a zinc-supplementing exhausted group in the drinking water, adding Zn SO4.7H2O,[Zn] = 225 mg/ L, and the rest groups of normal drinking water. The endurance training group and the training zinc-supplementing group were used for swimming endurance training for 2 weeks,4 weeks and 6 weeks, respectively. The endurance training group and the training zinc-supplementing group were used for 24 hours after the exercise. In which the standard of exhaustion judgment: the mouse is immersed in water for 10s, and the positive reflection can not be completed on the plane. Test method and index: the exhaustive swimming exercise of mice and the time of exhaustive exercise were carried out, and the effect of zinc-supplementing on the anti-fatigue ability of the body was discussed. The morphological changes of skeletal muscle after exercise in mice were observed with HHE staining. The content of malondialdehyde (MDA) in skeletal muscle and the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and serum creatine kinase (CK) were determined by enzyme-coupled method. The content and distribution of desmin in skeletal muscle of mice at 2 weeks,4 weeks and 6 weeks after exhaustive exercise were detected by immunohistochemistry. The results of the study: (1) The time of mouse swimming to exhaustion can be significantly prolonged by (1) zinc supplement, and the time of swimming to exhaustion of the mice in the exhaustion group is longer than that of the control force, and the difference is very significant (P0.01). (2) The activity of SOD in the skeletal muscle was significantly higher than that in the control group and the endurance training group after 2 weeks,4 weeks and 6 weeks after the exercise of the zinc-supplemented group. The activity of glutathione peroxidase (GSH-Px) in skeletal muscle after 4 and 6 weeks of exercise was significantly higher than that in the control group and the endurance training group (P0.05). The activity of GSH-Px and SOD in the skeletal muscle of the mice after exhaustive exercise was significantly higher than that of the control group (P0.05). (3) In the control group, the CK activity in the serum was significantly higher than that in the control group and the inexhaustible group after exhaustive exercise, and the CK activity in the serum was higher than that in the control group (P0.05). The content of malondialdehyde (MDA) in the skeletal muscle of the mice after exhaustive exercise was significantly lower than that of the control group (P0.05). (4) The average optical density of desmin in the skeletal muscle of the mice after 2 weeks,4 weeks and 6 weeks was significantly higher than that in the control group (P0.05). The average optical density of desmin in the skeletal muscle of the endurance training group was lower than that of the training zinc supplement group. The average optical density of the bound protein in the skeletal muscle of the mice after exhaustive exercise was higher than that of the control group, and there was a significant difference (P0.05). The results of the study: (1) The time of mouse swimming to exhaustion can be obviously improved, the activity of GSH-Px and SOD in the skeletal muscle of the mouse is enhanced, the increase of serum CK after exhaustive exercise is controlled, the content of MDA in the muscle after exercise is reduced, and the occurrence of exercise fatigue is delayed. And has the effect of promoting the improvement of the exercise capacity of the mouse. (2) The activity of GSH-Px and SOD in the skeletal muscle of the mice was significantly higher than that of the control group and the endurance training group, and the activity of GSH-Px and SOD in the skeletal muscle of the endurance training group was higher than that of the control group. The zinc-supplementing and the endurance training can improve the anti-oxidation ability of the mice and have the synergistic effect, and the exercise capacity of the mice can be obviously improved when the mice are used in combination. (3) The desmin content in the skeletal muscle of the mice was significantly higher than that of the endurance training group by supplementing the zinc, and the desmin content in the skeletal muscle of the zinc-supplemented group was significantly higher than that of the control group, and the reason may be that the zinc supplement can promote the regeneration of the skeletal muscle fiber of the mouse, So as to improve the anti-fatigue ability and the exercise endurance. (4) The single-time exhaustive sports meet the microdamage of the skeletal muscle of the mouse, and the desmin cytoskeleton damage is the morphological index of the microdamage of the skeletal muscle, and the early-phase zinc supplement can improve the exercise endurance of the mouse and reduce the degree of the fine injury of the skeletal muscle induced by the exhaustive exercise.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：G804.7
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