抗阻訓練、耐力訓練對SD大鼠骨骼肌氧化還原及肌衛(wèi)星細胞增殖影響的研究
發(fā)布時間：2018-08-03 18:51
【摘要】：骨骼肌占機體總重量的40%左右,它既是我們人體首要的能量供應站,又是機體能量代謝的關鍵場所,維持骨骼肌正常的收縮狀態(tài)和生理功能是保證機體健康的重要前提。改善骨骼肌功能的主要方式就是進行合理的運動訓練,但其在生物學方面的機制目前還尚未完全明確。一般認為骨骼肌對運動訓練的適應與骨骼肌衛(wèi)星細胞有關,而肌衛(wèi)星細胞的激活則可能是由于運動訓練導致的局部骨骼肌微細損傷;也可能是由于運動訓練產生的刺激導致了炎性因子、細胞因子等的釋放增加。無論何種途徑,都是發(fā)生在骨骼肌氧化還原狀態(tài)產生陡然改變的前提下,運動會引起NAD+含量改變,而NAD+的含量發(fā)生變化又會對SIRT1的活性產生影響,SIRT1作為機體氧化還原狀態(tài)的感受器有著非常關鍵的作用。運動訓練是否通過引起骨骼肌氧化還原狀態(tài)的改變導致SIRT1下游細胞事件的適應性改變還需進一步研究。目的:本研究旨在探討抗阻和有氧運動訓練對骨骼肌衛(wèi)星細胞增殖的影響,進而探討骨骼肌氧化還原與肌衛(wèi)星細胞增殖的內在聯(lián)系,為探索有效促進骨骼肌重塑,改善骨骼肌穩(wěn)態(tài)的運動方式提供理論與數(shù)據(jù)支持。方法:購買4周齡清潔級雄性SD大鼠24只,體重178.66土 1.69(g),隨機分為3組:安靜對照組(C,N=8);有氧運動訓練組(A,N=8);抗阻運動訓練組(R,N=8)。A組大鼠在動物跑臺上進行有氧運動訓練,3次/周,30min/次,速度為9m/min;R組大鼠以尾部負重的方式進行爬梯訓練,3次/周,3組/次,3次/組,每次間歇30-60s,每組間歇2-3min;大鼠的訓練周期為6周,周六周日休息。訓練結束后對大鼠稱重并麻醉處死,迅速取下完整的腓腸肌。采用Elisa檢測腓腸肌組織中NADH、NAD 總含量;采用Real-time PCR技術檢測腓腸肌組織中Nampt、Sirt1、P53、P21、MyoD、Myf5、Myogenin、MSTN、Pax7 的 mRNA 水平;采用 Western blotting 技術檢測腓腸肌組織中 Nampt、Sirt1、P53、Acetyl-P53、P21、MyoD、MSTN、Pax7的蛋白水平。結果:(1)與C組相比,A組大鼠6周訓練后體重、腓腸肌SMI值均無顯著變化(p0.05),R組大鼠6周訓練后體重顯著增加(p0.05),腓腸肌SMI值無顯著變化(p0.05);與A組相比,R組大鼠6周訓練后體重極顯著增加(P0.01,腓腸肌SMI值無顯著變化(P0.05)。(2)與C組相比,R組大鼠腓腸肌內P53、P21的mRNA水平顯著下調(P0.05),MSTN的mRNA水平極顯著下調(P0.01);與A組相比,R組大鼠腓腸肌內MSTN的mRNA水平極顯著下調(P0.01)。(3)與C組相比,A組大鼠腓腸肌內Nampt的蛋白水平顯著上調(p0.05),R組大鼠腓腸肌內SIRT1的蛋白水平顯著上調(p0.05),P53、Acetyl-P53的蛋白水平顯著下調(p0.05),Pax7的蛋白水平極顯著下調(p0.01)。(4)與C組相比,A組大鼠腓腸肌內NAD總的相對含量極顯著升高(p0.01),而NAD+/NADH比值無顯著變化(p0.05),R組大鼠腓腸肌內NAD+/NADH比值顯著升高(p0.05),NAD+/NAD總比值極顯著升高(p0.01),而NAD+、NAD總的相對含量無顯著變化(p0.05);與A組相比,R組大鼠腓腸肌內NAD總的相對含量極顯著降低(p0.01,NAD+/NAD總比值顯著升高(p0.05)。結論:(1)SIRT1的活性主要是由NAD+/NADH來調控,而并非NAD+的含量。(2)抗阻運動訓練能夠有效激活骨骼肌衛(wèi)星細胞,促進其增殖、分化。(3)抗阻運動訓練通過改變肌細胞內的氧化還原水平,進而引起SIRT1調控的下游細胞事件發(fā)生適應性改變。
[Abstract]:Skeletal muscle accounts for about 40% of the total weight of the body. It is not only the primary energy supply station in our body, but also the key place for the body's energy metabolism. Maintaining the normal contraction and physiological functions of the skeletal muscle is an important prerequisite to ensure the health of the body. The main formula for improving the skeletal muscle function is to exercise reasonable exercise, but it is in biology. The mechanism of the study is not yet fully understood. It is generally believed that the adaptation of skeletal muscle to exercise training is related to the skeletal muscle satellite cells, and the activation of the muscle satellite cells may be due to the slight injury of the local skeletal muscle caused by exercise training; it may also be caused by the stimulation of exercise training resulting in inflammatory factors, cytokines, and so on. The release increases. In any way, the movement will cause a change in the NAD+ content on the premise of a sudden change in the redox state of the skeletal muscle, and the changes in the content of NAD+ will affect the activity of SIRT1. As a receptor for the redox state of the body, SIRT1 has a very critical role. The changes in the redox state of skeletal muscle cause the adaptation of the SIRT1 downstream cell events. Objective: This study aims to explore the effects of resistance and aerobic exercise on the proliferation of skeletal muscle satellite cells, and to explore the internal relationship between the redox and the proliferation of muscle satellite cells. Theory and data support were provided to promote skeletal muscle remodeling and improve skeletal muscle homeostasis. Methods: 24 clean male SD rats of 4 weeks old, 178.66 soil 1.69 (g), were randomly divided into 3 groups: C (N=8), aerobic exercise training group (A, N=8), and R (N=8) group.A rats on the animal platform Oxygen exercise training, 3 times per week, 30min/ times, was 9m/min. Group R rats were trained with tail load, 3 times / week, 3 groups / times, 3 times / groups, each interval 30-60s, each group was 2-3min. The rats' training cycle was 6 weeks and Saturday Sunday rest. After the training, the rats were weighed and executed, and the complete gastrocnemius muscle was quickly removed. Detect the total content of NADH and NAD in the gastrocnemius muscle tissue with Elisa, and detect the level of Nampt, Sirt1, P53, P21, MyoD, Myf5, Myogenin, and the level of protein in the gastrocnemius muscle tissue by Real-time PCR technique. The SMI value of the gastrocnemius muscle was not significantly changed after 6 weeks of training in the A group (P0.05). The weight of the gastrocnemius muscle was significantly increased after 6 weeks of training (P0.05) and the gastrocnemius SMI value was not significantly changed (P0.05). Compared with the group A, the body weight of the rats in the group of R group was significantly increased (P0.01, the SMI value of the gastrocnemius muscle was not significantly changed (P0.05). (2) the group of rat fibula was compared with that of the group. The mRNA level of P53 and P21 in the intestinal muscle decreased significantly (P0.05), and the mRNA level of MSTN was significantly down (P0.01). Compared with the group A, the mRNA level of MSTN in the gastrocnemius muscle of the group R was significantly down (P0.01). (3) the protein level of the gastrocnemius muscle of the rats was significantly up-regulated compared with those in the group. P0.05), the protein level of P53, Acetyl-P53 decreased significantly (P0.05), and the protein level of Pax7 decreased significantly (P0.01). (4) compared with the C group, the relative content of NAD in the gastrocnemius muscle of A group rats increased significantly (P0.01), but the NAD+/NADH ratio had no significant change (P0.05). The total relative content of NAD+ and NAD was not significantly changed (P0.05), and the total relative content of NAD in the gastrocnemius muscle of group R was significantly lower than that of A group (P0.01, the total NAD+/NAD ratio increased significantly (P0.05). (1) SIRT1 activity was mainly regulated by NAD+/NADH, but not the content. (2) resistance exercise training could be effective. Activation of skeletal muscle satellite cells to promote its proliferation and differentiation. (3) resistance exercise training by altering the level of redox in the muscle cells, thus causing adaptive changes in the downstream cell events regulated by SIRT1.
【學位授予單位】：華東師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：G804.2
[Abstract]:Skeletal muscle accounts for about 40% of the total weight of the body. It is not only the primary energy supply station in our body, but also the key place for the body's energy metabolism. Maintaining the normal contraction and physiological functions of the skeletal muscle is an important prerequisite to ensure the health of the body. The main formula for improving the skeletal muscle function is to exercise reasonable exercise, but it is in biology. The mechanism of the study is not yet fully understood. It is generally believed that the adaptation of skeletal muscle to exercise training is related to the skeletal muscle satellite cells, and the activation of the muscle satellite cells may be due to the slight injury of the local skeletal muscle caused by exercise training; it may also be caused by the stimulation of exercise training resulting in inflammatory factors, cytokines, and so on. The release increases. In any way, the movement will cause a change in the NAD+ content on the premise of a sudden change in the redox state of the skeletal muscle, and the changes in the content of NAD+ will affect the activity of SIRT1. As a receptor for the redox state of the body, SIRT1 has a very critical role. The changes in the redox state of skeletal muscle cause the adaptation of the SIRT1 downstream cell events. Objective: This study aims to explore the effects of resistance and aerobic exercise on the proliferation of skeletal muscle satellite cells, and to explore the internal relationship between the redox and the proliferation of muscle satellite cells. Theory and data support were provided to promote skeletal muscle remodeling and improve skeletal muscle homeostasis. Methods: 24 clean male SD rats of 4 weeks old, 178.66 soil 1.69 (g), were randomly divided into 3 groups: C (N=8), aerobic exercise training group (A, N=8), and R (N=8) group.A rats on the animal platform Oxygen exercise training, 3 times per week, 30min/ times, was 9m/min. Group R rats were trained with tail load, 3 times / week, 3 groups / times, 3 times / groups, each interval 30-60s, each group was 2-3min. The rats' training cycle was 6 weeks and Saturday Sunday rest. After the training, the rats were weighed and executed, and the complete gastrocnemius muscle was quickly removed. Detect the total content of NADH and NAD in the gastrocnemius muscle tissue with Elisa, and detect the level of Nampt, Sirt1, P53, P21, MyoD, Myf5, Myogenin, and the level of protein in the gastrocnemius muscle tissue by Real-time PCR technique. The SMI value of the gastrocnemius muscle was not significantly changed after 6 weeks of training in the A group (P0.05). The weight of the gastrocnemius muscle was significantly increased after 6 weeks of training (P0.05) and the gastrocnemius SMI value was not significantly changed (P0.05). Compared with the group A, the body weight of the rats in the group of R group was significantly increased (P0.01, the SMI value of the gastrocnemius muscle was not significantly changed (P0.05). (2) the group of rat fibula was compared with that of the group. The mRNA level of P53 and P21 in the intestinal muscle decreased significantly (P0.05), and the mRNA level of MSTN was significantly down (P0.01). Compared with the group A, the mRNA level of MSTN in the gastrocnemius muscle of the group R was significantly down (P0.01). (3) the protein level of the gastrocnemius muscle of the rats was significantly up-regulated compared with those in the group. P0.05), the protein level of P53, Acetyl-P53 decreased significantly (P0.05), and the protein level of Pax7 decreased significantly (P0.01). (4) compared with the C group, the relative content of NAD in the gastrocnemius muscle of A group rats increased significantly (P0.01), but the NAD+/NADH ratio had no significant change (P0.05). The total relative content of NAD+ and NAD was not significantly changed (P0.05), and the total relative content of NAD in the gastrocnemius muscle of group R was significantly lower than that of A group (P0.01, the total NAD+/NAD ratio increased significantly (P0.05). (1) SIRT1 activity was mainly regulated by NAD+/NADH, but not the content. (2) resistance exercise training could be effective. Activation of skeletal muscle satellite cells to promote its proliferation and differentiation. (3) resistance exercise training by altering the level of redox in the muscle cells, thus causing adaptive changes in the downstream cell events regulated by SIRT1.
【學位授予單位】：華東師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：G804.2
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