急性耐力性离心运动对大鼠肌肉特异性microRNAs表达水平的影响

发布时间：2018-03-24 13:28

本文选题：myomiRs　切入点：血浆　出处：《南京体育学院》2017年硕士论文

【摘要】：研究目的:长期有规律的运动能够促使人体产生生物适应,促进和维持身体健康。离心运动(eccentric exercise,ECC)指肌肉在离心工作时承受额外负荷,产生抗重力和制动的动作,发挥制动作用。由于ECC生理学和机械学上的诸多特性,例如众所周知的ECC能够产生更大的肌力以及ECC特异性的心肺和代谢反应。ECC是改善健康人和运动员的肌肉力量和运动能力的经典运动方式,同时也在康复锻炼和临床上也越来越多地被采用。此外,长期周期性训练计划中包含更多的ECC训练可能是提高竞技能力的一个很有前景的训练手段和策略。但是目前对于ECC作用的生理学适应机制仍然不清楚。micro RNAs(miRNAs)是一类长度约为22nt的小分子非编码RNA,在转录后水平的基因调控中发挥重要的生理学作用,在骨骼肌生成、增殖、分化、凋亡等过程中发挥调控作用,作为一类新的调控分子,可能参与ECC诱导的生物学适应,提供新的角度理解ECC给机体带来的影响。因此,本课题选定7种肌肉特异性的miRNAs,目的是从miRNAs的角度研究ECC的诸多好处,从分子层面理解ECC,并且探索miRNAs可潜在的作为ECC的生物标志物。研究方法:选用8周龄健康雄性Sprague-Dawley(SD)大鼠80只,体重280±15g,随机分为安静组(rest)、运动后即刻组(0h)、1小时组(1h)、24小时组(24h)和48小时组(48h)。安静组不参加运动,0h、1h、24h和48h组进行急性下坡跑运动。跑台的坡度为-16°,速度为20m/min,时间90min。在各个时间点麻醉大鼠,腹腔取血5ml,后取右侧腿腓肠肌、比目鱼肌、股四头肌和心肌。利用实时荧光定量逆转录聚合酶链式反应(q RT-PCR)检测选定的几种肌肉特异性miRNAs(myomiRs)(miR-1、miR-133a、miR-133b、miR-206、miR-208a、miR-208b和miR-499)水平在血浆、血浆外泌体(exosome)和肌肉中变化情况。研究结果:在ECC后,血浆中几种肌肉特异性miRNAs都有不同程度的应答,而在选定的四种肌肉组织中,只有腓肠肌中miR-1和miR-133b水平发生变化,同时,血浆exosome中也发现了5种miRNAs对运动有不同程度的应答,运动后miR-133a水平在血浆中和血浆exosome中变化趋势相似,ECC后24h腓肠肌miR-133b水平较安静组显著上升,而血浆和血浆exosome中却呈现相反趋势。结论:ECC后miR-1在血浆、exosome和腓肠肌中变化趋势一致;miR-133a水平在四种组织中没有变化,但在血浆和exosome中都在ECC后升高;腓肠肌中与血浆中miR-133b水平变化呈现相反趋势,但腓肠肌中miR-133b与exosome中变化趋势相似,都在运动后24h达到最高。这些都提示miR-1、miR-133a和miR-133b作为ECC潜在生物标志物,在ECC带来的各种生物适应中发挥重要作用。
[Abstract]:Research objective: Long-term regular exercise can promote biological adaptation, promote and maintain good health. Centrifugal exercise exercise is the action of resisting gravity and braking when muscles bear extra load during centrifugal work. Play a role in braking. Because of the physiological and mechanical properties of ECC, For example, ECC is known to produce greater muscle strength and ECC specific cardiopulmonary and metabolic responses. ECC is a classic exercise way to improve muscle strength and exercise ability in healthy people and athletes. At the same time, it is also used more and more in rehabilitation exercise and clinical practice. In addition, More ECC training in long-term periodic training plan may be a promising training method and strategy to improve competitive ability. However, the physiological adaptation mechanism of ECC effect is still unclear. Micro RN AsmiRNs is a kind of training method and strategy. Small non-coding RNAs, which are about the length of 22nt, play an important physiological role in gene regulation at post-transcriptional level. It plays a regulatory role in skeletal muscle formation, proliferation, differentiation and apoptosis. As a new class of regulatory molecules, it may participate in the biological adaptation induced by ECC and provide a new perspective to understand the effects of ECC on the body. In this paper, seven muscle-specific miRNAss were selected to study the benefits of ECC from the point of view of miRNAs. To understand Ecc at the molecular level and to explore the potential biomarker of miRNAs as a biomarker of ECC, 80 healthy male Sprague-Dawley SD rats aged 8 weeks were selected. The body weight was 280 卤15g, which was randomly divided into quiet group (0 h / h) and 48 hour group (24 h) and 48 h group respectively. The rest group did not take part in the exercise for 24 h and 48 h for acute downhill running. The slope of the platform was -16 掳, the speed was 20 m / min, and the time was 90 min. Anesthetized rats at a point in time, 5 ml of blood was taken from abdominal cavity, then from right leg gastrocnemius muscle, soleus muscle, quadriceps femoris muscle and myocardium. The specific miR-1 miR-133amiR-133bmiR-206miR-208amiR-208b and miR-499b were detected in plasma by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (qRT-PCRR). Changes in plasma exosome and muscle. Results: after ECC, several muscle-specific miRNAs in plasma had varying degrees of response, while in the four selected muscle tissues, only miR-1 and miR-133b levels in gastrocnemius muscle changed, and at the same time, It was also found in plasma exosome that five miRNAs had different degree of response to exercise. The changes of miR-133a level in plasma and exosome were similar after exercise. The miR-133b level of gastrocnemius muscle was significantly higher than that of quiet group at 24 h after exercise. Conclusion the change trend of miR-1 in plasma exosome and gastrocnemius muscle is the same in plasma and gastrocnemius muscle. The level of miR-133a does not change in four tissues, but it is increased in both plasma and exosome after ECC. The change of miR-133b level in gastrocnemius muscle and plasma showed the opposite trend, but the change trend of miR-133b in gastrocnemius muscle was similar to that in exosome, and reached the highest at 24 hours after exercise. All these suggested that miR-1 miR-133a and miR-133b were potential biomarkers of ECC. Play an important role in various biological adaptation brought about by ECC.
【学位授予单位】：南京体育学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：G804.7

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