補(bǔ)鋅對(duì)耐力訓(xùn)練、力竭運(yùn)動(dòng)小鼠體內(nèi)酶活性及結(jié)蛋白含量變化的影響
發(fā)布時(shí)間：2019-03-24 18:14
【摘要】：研究目的:本實(shí)驗(yàn)以雄性ICR小鼠為研究對(duì)象,通過補(bǔ)鋅來(lái)探討微量元素鋅對(duì)耐力訓(xùn)練小鼠骨骼肌中谷胱甘肽過氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性變化,力竭運(yùn)動(dòng)小鼠骨骼肌中丙二醛(MDA)含量變化,GSH-Px、SOD、血清肌酸激酶(CK)活性變化,以及小鼠運(yùn)動(dòng)后骨骼肌中結(jié)蛋白(desmin)含量變化的影響。采用耐力測(cè)試、組織形態(tài)學(xué)、酶偶聯(lián)免疫和免疫組化等技術(shù),通過分析小鼠運(yùn)動(dòng)后骨骼肌中各種酶活性變化、細(xì)胞骨架蛋白desmin的分布情況、力竭運(yùn)動(dòng)時(shí)間及力竭運(yùn)動(dòng)后即刻骨骼肌的組織形態(tài)學(xué)變化,來(lái)探討補(bǔ)鋅對(duì)提高小鼠運(yùn)動(dòng)能力的影響。研究方法:研究對(duì)象為89只雄性ICR小鼠,適應(yīng)飼養(yǎng)三天后進(jìn)行一次游泳測(cè)試,剔除不會(huì)游泳的小鼠,然后將小鼠隨機(jī)分為安靜對(duì)照組、耐力訓(xùn)練組、耐力訓(xùn)練補(bǔ)鋅組、力竭運(yùn)動(dòng)對(duì)照組以及力竭運(yùn)動(dòng)補(bǔ)鋅組(安靜對(duì)照組小鼠不做任何處理,力竭運(yùn)動(dòng)對(duì)照組僅進(jìn)行一次力竭運(yùn)動(dòng),力竭運(yùn)動(dòng)補(bǔ)鋅組進(jìn)行6周的補(bǔ)鋅;以下耐力訓(xùn)練補(bǔ)鋅組簡(jiǎn)稱訓(xùn)練補(bǔ)鋅組,力竭運(yùn)動(dòng)補(bǔ)鋅組簡(jiǎn)稱補(bǔ)鋅力竭組,力竭運(yùn)動(dòng)對(duì)照組簡(jiǎn)稱對(duì)照力竭組),耐力訓(xùn)練組和訓(xùn)練補(bǔ)鋅組中又分別設(shè)2周組、4周組和6周組用來(lái)測(cè)各種酶活性的變化及運(yùn)動(dòng)后desmin的表達(dá)(各組n=10)。補(bǔ)鋅方法為:訓(xùn)練補(bǔ)鋅組和補(bǔ)鋅力竭組于飲用水中加入Zn SO4·7H2O,[Zn]=225 mg/L,其余各組正常飲水。耐力訓(xùn)練組和訓(xùn)練補(bǔ)鋅組分別進(jìn)行2周、4周和6周的游泳耐力訓(xùn)練,每周游6 d,第一周每天游泳30 min,以后每周遞加10 min,第六周時(shí)每天游泳90min。耐力訓(xùn)練組和訓(xùn)練補(bǔ)鋅組于運(yùn)動(dòng)后24小時(shí)取材,對(duì)照力竭組和補(bǔ)鋅力竭組于力竭運(yùn)動(dòng)后即刻取材。其中力竭判斷的標(biāo)準(zhǔn):小鼠沉入水中過10 s,放在平面上無(wú)法完成翻正反射。測(cè)試方法及指標(biāo):①力竭組對(duì)小鼠進(jìn)行一次性力竭游泳運(yùn)動(dòng)并記錄力竭運(yùn)動(dòng)時(shí)間,并以此來(lái)探討補(bǔ)鋅對(duì)提高機(jī)體抗疲勞能力的影響。②HE染色觀察小鼠運(yùn)動(dòng)后骨骼肌組織形態(tài)學(xué)變化。③酶偶聯(lián)等方法測(cè)定小鼠運(yùn)動(dòng)后骨骼肌中丙二醛(MDA)含量以及谷胱甘肽過氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、血清肌酸激酶(CK)的活性。④免疫組化法檢測(cè)力竭運(yùn)動(dòng)后即刻及耐力運(yùn)動(dòng)2周、4周、6周后小鼠骨骼肌中desmin的含量及分布。研究結(jié)果:(1)補(bǔ)鋅能夠顯著延長(zhǎng)小鼠游泳至力竭的時(shí)間,補(bǔ)鋅力竭組小鼠游泳至力竭的時(shí)間長(zhǎng)于對(duì)照力竭組,差異極顯著(P0.01)。(2)訓(xùn)練補(bǔ)鋅組小鼠運(yùn)動(dòng)2周、4周和6周后骨骼肌中SOD活性顯著高于安靜對(duì)照組和耐力訓(xùn)練組,與安靜對(duì)照組相比P0.01,與耐力訓(xùn)練組相比P0.05;運(yùn)動(dòng)4周和6周后骨骼肌中谷胱甘肽過氧化物酶(GSH-Px)活性顯著高于安靜對(duì)照組和耐力訓(xùn)練組(P0.05)。力竭運(yùn)動(dòng)后即刻補(bǔ)鋅力竭組小鼠骨骼肌中GSH-Px、SOD活性顯著高于對(duì)照力竭組(P0.05)。(3)對(duì)照力竭組小鼠力竭運(yùn)動(dòng)后即刻血清中CK活性顯著高于安靜對(duì)照組和補(bǔ)鋅力竭組,差異極顯著;補(bǔ)鋅力竭組小鼠運(yùn)動(dòng)后即刻血清中CK活性高于安靜對(duì)照組(P0.05)。力竭運(yùn)動(dòng)后即刻補(bǔ)鋅力竭組小鼠骨骼肌中丙二醛(MDA)含量顯著低于對(duì)照力竭組(P0.05),對(duì)照力竭組小鼠力竭運(yùn)動(dòng)后即刻丙二醛含量高于安靜對(duì)照組,差異極顯著(P0.01)。(4)運(yùn)動(dòng)2周、4周、6周后訓(xùn)練補(bǔ)鋅組小鼠骨骼肌中desmin免疫組化平均光密度值顯著高于安靜對(duì)照組(P0.05),耐力訓(xùn)練組小鼠骨骼肌中desmin免疫組化平均光密度值低于訓(xùn)練補(bǔ)鋅組;力竭運(yùn)動(dòng)后即刻補(bǔ)鋅力竭組小鼠骨骼肌中結(jié)蛋白平均光密度值高于對(duì)照力竭組,且具有顯著性差異(P0.05)。研究結(jié)論:(1)補(bǔ)鋅可以顯著提高小鼠游泳至力竭的時(shí)間,增強(qiáng)小鼠骨骼肌中GSH-Px、SOD的活性,控制力竭運(yùn)動(dòng)后血清CK的升高,降低運(yùn)動(dòng)后肌肉中MDA的含量,延緩運(yùn)動(dòng)性疲勞的發(fā)生,對(duì)小鼠運(yùn)動(dòng)能力的提高具有促進(jìn)作用。(2)耐力訓(xùn)練過程中補(bǔ)鋅可以使小鼠的抗氧化酶活性得到提高,訓(xùn)練補(bǔ)鋅組小鼠骨骼肌中GSH-Px、SOD活性顯著高于安靜對(duì)照組和耐力訓(xùn)練組,耐力訓(xùn)練組小鼠骨骼肌中GSH-Px、SOD活性高于安靜對(duì)照組;補(bǔ)鋅和耐力訓(xùn)練均可以提高小鼠的抗氧化能力,且具有協(xié)同作用,聯(lián)合使用時(shí)可以顯著提高小鼠的運(yùn)動(dòng)能力。(3)通過補(bǔ)鋅,訓(xùn)練補(bǔ)鋅組小鼠骨骼肌中desmin含量顯著高于耐力訓(xùn)練組;補(bǔ)鋅力竭組小鼠骨骼肌中的desmin含量顯著高于對(duì)照力竭組,其原因可能是補(bǔ)鋅可以促進(jìn)小鼠骨骼肌纖維的再生,進(jìn)而提高其抗疲勞能力和運(yùn)動(dòng)耐力。(4)一次性力竭運(yùn)動(dòng)會(huì)引發(fā)小鼠骨骼肌的微細(xì)損傷,desmin細(xì)胞骨架破壞是骨骼肌微損傷的形態(tài)學(xué)指標(biāo),前期補(bǔ)鋅可以提高小鼠的運(yùn)動(dòng)耐力,降低力竭運(yùn)動(dòng)誘導(dǎo)的骨骼肌微細(xì)損傷程度。
[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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：G804.7
本文編號(hào)：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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：G804.7
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