抗阻训练、耐力训练对SD大鼠骨骼肌氧化还原及肌卫星细胞增殖影响的研究
发布时间：2018-08-03 18:51
【摘要】：骨骼肌占机体总重量的40%左右,它既是我们人体首要的能量供应站,又是机体能量代谢的关键场所,维持骨骼肌正常的收缩状态和生理功能是保证机体健康的重要前提。改善骨骼肌功能的主要方式就是进行合理的运动训练,但其在生物学方面的机制目前还尚未完全明确。一般认为骨骼肌对运动训练的适应与骨骼肌卫星细胞有关,而肌卫星细胞的激活则可能是由于运动训练导致的局部骨骼肌微细损伤;也可能是由于运动训练产生的刺激导致了炎性因子、细胞因子等的释放增加。无论何种途径,都是发生在骨骼肌氧化还原状态产生陡然改变的前提下,运动会引起NAD+含量改变,而NAD+的含量发生变化又会对SIRT1的活性产生影响,SIRT1作为机体氧化还原状态的感受器有着非常关键的作用。运动训练是否通过引起骨骼肌氧化还原状态的改变导致SIRT1下游细胞事件的适应性改变还需进一步研究。目的:本研究旨在探讨抗阻和有氧运动训练对骨骼肌卫星细胞增殖的影响,进而探讨骨骼肌氧化还原与肌卫星细胞增殖的内在联系,为探索有效促进骨骼肌重塑,改善骨骼肌稳态的运动方式提供理论与数据支持。方法:购买4周龄清洁级雄性SD大鼠24只,体重178.66土 1.69(g),随机分为3组:安静对照组(C,N=8);有氧运动训练组(A,N=8);抗阻运动训练组(R,N=8)。A组大鼠在动物跑台上进行有氧运动训练,3次/周,30min/次,速度为9m/min;R组大鼠以尾部负重的方式进行爬梯训练,3次/周,3组/次,3次/组,每次间歇30-60s,每组间歇2-3min;大鼠的训练周期为6周,周六周日休息。训练结束后对大鼠称重并麻醉处死,迅速取下完整的腓肠肌。采用Elisa检测腓肠肌组织中NADH、NAD 总含量;采用Real-time PCR技术检测腓肠肌组织中Nampt、Sirt1、P53、P21、MyoD、Myf5、Myogenin、MSTN、Pax7 的 mRNA 水平;采用 Western blotting 技术检测腓肠肌组织中 Nampt、Sirt1、P53、Acetyl-P53、P21、MyoD、MSTN、Pax7的蛋白水平。结果:(1)与C组相比,A组大鼠6周训练后体重、腓肠肌SMI值均无显著变化(p0.05),R组大鼠6周训练后体重显著增加(p0.05),腓肠肌SMI值无显著变化(p0.05);与A组相比,R组大鼠6周训练后体重极显著增加(P0.01,腓肠肌SMI值无显著变化(P0.05)。(2)与C组相比,R组大鼠腓肠肌内P53、P21的mRNA水平显著下调(P0.05),MSTN的mRNA水平极显著下调(P0.01);与A组相比,R组大鼠腓肠肌内MSTN的mRNA水平极显著下调(P0.01)。(3)与C组相比,A组大鼠腓肠肌内Nampt的蛋白水平显著上调(p0.05),R组大鼠腓肠肌内SIRT1的蛋白水平显著上调(p0.05),P53、Acetyl-P53的蛋白水平显著下调(p0.05),Pax7的蛋白水平极显著下调(p0.01)。(4)与C组相比,A组大鼠腓肠肌内NAD总的相对含量极显著升高(p0.01),而NAD+/NADH比值无显著变化(p0.05),R组大鼠腓肠肌内NAD+/NADH比值显著升高(p0.05),NAD+/NAD总比值极显著升高(p0.01),而NAD+、NAD总的相对含量无显著变化(p0.05);与A组相比,R组大鼠腓肠肌内NAD总的相对含量极显著降低(p0.01,NAD+/NAD总比值显著升高(p0.05)。结论:(1)SIRT1的活性主要是由NAD+/NADH来调控,而并非NAD+的含量。(2)抗阻运动训练能够有效激活骨骼肌卫星细胞,促进其增殖、分化。(3)抗阻运动训练通过改变肌细胞内的氧化还原水平,进而引起SIRT1调控的下游细胞事件发生适应性改变。
[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.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：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.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：G804.2
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