【作者】 王海燕；

【导师】 丁树哲；

【作者基本信息】 华东师范大学 ， 运动人体科学， 2010， 硕士

【摘要】 目前,心血管疾病居全球成人死因的首位；全球有2.46亿人被确诊为糖尿病,并预测到2025年全球糖尿病人数将超过3.8亿；癌症的每年新增病例数约为800万,死亡数约为620万；心血管疾病、癌症、糖尿病等正成为威胁人类健康的“头号杀手”。很多学者一直致力于这些疾病的发病机理和防治工作的研究,随着研究的深入,发现核受体不仅对机体的生长、发育、正常生理功能的维持和代谢过程起着重要的调节作用,而且与多种疾病如乳腺癌、前列腺癌、糖尿病及肥胖的发生、发展有直接关联。核受体成为疾病病理研究这一领域的一个重点。鸡卵白蛋白上游启动子转录因子家族(COUP-TFs)是属于核受体中孤核受体家族的一员,有COUP-TFⅠ和COUP-TFⅡ两个亚型,它们在组织中广泛表达,与多种基因的转录相关,在器官形成、神经发生、细胞分化等方面有重要的作用。COUP-TFs的突变或异常表达与人类的先天性心脏病、糖尿病和癌症等疾病相关。特别是COUP-TFⅡ基因在骨骼、肌肉、脂肪形成与代谢平衡中起调节作用。目的：比较研究一次性急性运动、高强度间歇性冲刺训练、耐力训练对SD大鼠及耐力训练对GK大鼠骨骼肌中COUP-TFⅠ和COUP-TFⅡ基因表达的的影响,探讨机体运动适应中糖代谢与COUP-TFⅠ、COUP-TFⅡ基因表达的变化关系。方法：将40只雄性SD大鼠,4周龄,体重100±5g；12只GK大鼠,8周龄,体重250±5g,SD大鼠随机分成SD对照组(C)、SD急性耐力训练组(D)、SD耐力训练组(E)、SD间歇性冲训练组(S),每组10只；GK大鼠随机分成GK对照组(H)、GK耐力训练组(R),每组6只。急性耐力训练：大鼠1-6周与安静组饲养方式相,处死前进行一次急性耐力训练,起始跑速为13m/min,最大速度不超过16.7m/min,训练时间60min；间歇性冲刺训练：每天3组,每组3次,每次10s极量强度(≥50m/min)的跑台运动,每次间歇时间30-60s,组间间歇时间3min；耐力训练：持续6周的跑台训练,起始跑速为13m/min,最大速度不超过16.7m/min,前2周每次训练40分钟,后4周每次训练60min,每周训练6天。所有大鼠6周饲养后处死,酶标仪检测血糖、血浆胰岛素、血浆脂联素、糖化血红蛋白和糖化血清蛋白。采用实时荧光定量PCR法检测COUP-TFⅠ和COUP-TFⅡ基因转录水平,Western Blotting检测COUP-TFⅠ蛋白表达水平。结果：(1)三种不同运动方式对SD大鼠体重增长及耐力训练对GK大鼠体重增长无显著影响。(2)耐力训练能够显著降低SD大鼠安静时的血糖水平,但对胰岛素抵抗指数无显著影响；耐力训练能显著降低GK大鼠血糖及胰岛素抵抗指数。(3)三种不同的运动方式能显著降低SD大鼠糖化血清蛋白含量；耐力训练能极显著降低GK大鼠糖化血清蛋白。(4)耐力训练能够显著升高SD大鼠与GK大鼠血脂联素浓度。(5)一次性急性运动与耐力训练能显著上调SD大鼠骨骼肌中COUP-TFⅠmRNA;耐力训练能显著下调GK大鼠骨骼肌中COUP-TFⅠmRNA表达。(6)一次性急性运动、耐力训练、高强度间歇性冲刺训练对SD大鼠COUP-TFⅠ蛋白表达无显著影响；耐力训练能极显著降低GK大鼠COUP-TFⅠ蛋白表达。(7)一次性急性运动、耐力训练、高强度间歇性冲刺训练能显著下调SD大鼠骨骼肌中COUP-TFⅡmRNA表达；耐力训练能显著下调GK大鼠COUP-TFⅡmRNA水平结论：(1)耐力训练有利于GK大鼠血糖降低水平,改善胰岛素抵抗。(2)急性运动、耐力训练、冲刺训练可使导致SD大鼠骨骼肌中COUP-TFⅠmRNA水平上升,COUP-TFⅡmRNA表达水平下降,而这三种运动对COUP-TFⅠ蛋白表达水平无显著影响,表现出与COUP-TFⅠmRNA水平变化不一致。说明COUP-TFⅠ和COUP-TFⅡ在基因转录水平参与了骨骼肌对急性耐力运动的快速应答,也参与了骨骼肌对长期耐力训练、间歇性大强度训练的慢性适应。运动可能倾向于激活骨骼肌COUP-TFⅠ基因表达,并抑制COUP-TFⅡ基因表达。(3)长期耐力训练改善GK大鼠胰岛素抵抗可能与骨骼肌COUP-TFⅠ基因、蛋白表达以及COUP-TFⅡ基因表达下调有关。(4)在正常生理和糖尿病病理条件下,耐力运动对骨骼肌COUP-TFⅠ基因表达有着决然相反的影响；考虑到GK大鼠与SD大鼠的品系、年龄、饮食差异,本实验不能排除以上因素对耐力运动效应的影响。(5)与COUP-TF相比,在正常生理和糖尿病病理条件下,COUP-TFⅡ基因表达对耐力运动的适应特征完全一致,表明耐力运动对COUP-TFⅡ基因表达的抑制在胰岛素抵抗的预防和改善中可能发挥更重要的作用。更多还原

【Abstract】 Currently, the most people die of cardiovascular disease in the world; the number of new cases of cancer each year is about 800 million, about 6.2 million of them die; 246 million people worldwide have been diagnosed with diabetes, and predicted that by 2025 The number of Diabetes will be more than 380 million. Cardiovascular disease,cancer,diabetes becoming a threat to human health.The pathogenesis of these diseases and control were studied by many scholars.With the in-depth study, scholars found that nuclear receptors not only play an important regulatory role for the body’s growth, development, maintenance of normal physiological functions and metabolic processes, but also are directly related to the pathological process a variety of diseases, development, such as breast cancer, prostate cancer, diabetes, diseases and obesity.The study of nuclear receptor become a major focus in this area.COUP-TFs belong to the nuclear hormone receptor family,they play an very important role in differentiation of organ and nerve cells. In recent years, studies have confirmed that COUP-TFs have two subtypes, they are COUP-TFⅠand COUP-TFⅡ.And they are widely expressed in various tissues and related to transcription. They also play an important role in the embryonic development and organ formation. And COUP-TFs mutation or abnormal expression of these two genes can cause heart disease, diabetes and cancer diseases. In particularly, the COUP-TFⅡgene is related to the formation and metabolism in skeletal muscle.COUP-TFⅡhas a role in reglating balance of the bone, muscle, fat.Objective:The aim of this study was to investigate the effects of an acute exercise,6-week high intensity interval training and endurance training on COUP-TFⅠand COUP-TFⅡgene expression of SD rats and the effect of endurance training on COUP-TFⅠand COUP-TFⅡgene expression of GK rats. We wanted to find out the relationship of COUP-TFⅠand COUP-TFⅡgene expression with glucose metabolism in the adaptation of exercise. Method:40 male SD rats, about 4 weeks old, weighing 100±5g; 12 GK rats, about 8 weeks old, weighing 250±5g, SD rats were randomly divided into SD control group (C, n=10), SD of acute group (D, n=10), SD endurance group (E, n=10), SD sprint group (S, n=10); GK rats were randomly divided into GK control group (H, n=6), GK endurance group (R, n =6). Endurance training in rats for 6 weeks of treadmill training, training six days a week, once a day for the Paosu starting 13m/min, a maximum speed of not more than 16.7m/min, day before the four weeks training in 40 minutes, After four weeks training,1 hour a day. After 6 weeks feeding, all rats were killed after the microplate reader detection of blood glucose, plasma insulin, plasma adiponectin and glycosylated hemoglobin and glycosylated serum protein. Real-time fluorescent quantitative PCR detection of COUP-TFⅠand COUP-TFⅡgene transcription level. Western Blotting detection of COUP-TFⅠgene expression level.Results:(1)Three types training had no apparent effect on the growth curve of SD rats; Endurance training had no apparent effect on the growth curve of GK rats.(2)Endurance training significantly ruduced blood glucose of SD rats and had no apparent effect on IRS; Endurance training significantly ruduced blood glucose and IRS of GK rat.(3)Three type exercises significantly ruduced GSP of SD rat; Endurance training significantly ruduced GSP of GK rats.(4) Endurance training significantly increased the ADP concentration of GK and SD rats.(5)Acute exercise and endurance training significantly increased COUP-TFⅠmRNA transcription of SD rats except high intensity interval training; Endurance training significantly reduced COUP-TFⅠmRNA transcription of GK rats.(6)Three type exercises had no apparent effect on COUP-TFⅠprotein expression of SD rats; Endurance training significantly reduced COUP-TFⅠprotein expression of GK rats.(7)Three type exercises significantly reduced COUP-TFⅡmRNA transcription of SD rats; Endurance training significantly reduced COUP-TFⅡmRNA transcription of GK rats.Conclusion:(1)Endurance training was beneficial to reduce the level of blood glucose in GK rats, which may mitigate IR.(2)An acute exercise,6-week high intensity interval training and endurance training increased COUP-TFⅠmRNA expression and reduced COUP-TFⅡmRNA expression, but had no apparent effect on the gene transcription level of COUP-TFⅠ. The transcription level of COUP-TFⅠand COUP-TFⅡmay involve in the skeletal muscle rapid response to an acute exercise, but also in the chronic adaptation to the high intensity interval training and endurance training. Exercise may tend to activate COUP-TFⅠgene expression, and inhibition of COUP-TFⅡgene expression in the skeletal muscle.(3)The effect of Long-term endurance training on improving insulin resistance in GK rats may be relevant to the expression, transcription of COUP-TFⅠand down down regulation expression of COUP-TFⅡ.(4)The effect of endurance exercise on skeletal muscle gene expression in COUP-TFⅠhas decidedly the opposite effect in normal physiological and diabetes conditions, which we should consider the GK rats and SD rat strains, age, diet differences,where this study can not rule out these factors on the effect of endurance exercise.(5)Compared with COUP-TFⅠ, COUP-TFⅡgene expression to the adaptations to endurance exercise exactly the same, indicating that the effect of endurance exercise inhibition COUP-TFⅡgene expression may play an important role in the prevention and improvement of insulin resistance.更多还原