运动对SD大鼠与GK大鼠骨骼肌中COUP-TF Ⅰ与COUP-TFⅡ基因表达的影响

发布时间：2018-07-06 08:58

本文选题：孤核受体 + 运动　；参考：《华东师范大学》2010年硕士论文

【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。
[Abstract]:At present, cardiovascular disease ranks first in the world's cause of death; 246 million people worldwide have been diagnosed with diabetes, and it is predicted that the number of global diabetes will exceed 380 million by 2025; the number of new cases of cancer is about 8 million a year, and the number of deaths is about 6 million 200 thousand; cardiovascular disease, cancer, diabetes and so on are becoming the "No. 1" killing of human health. Hand.
Many scholars have been working on the pathogenesis and prevention and treatment of these diseases. With the further research, it is found that the nuclear receptor not only plays an important role in the growth and development of the body, the maintenance of normal physiological function and the metabolic process, but also develops and develops with a variety of diseases such as breast cancer, prostate cancer, diabetes and obesity. There is a direct correlation. Nuclear receptors have become a focus in the field of pathological research.
The chicken oocyte albumin upstream promoter transcription factor family (COUP-TFs) is a member of the nucleus receptor family of the nuclear receptor. There are two subtypes of COUP-TF I and COUP-TF II. They are widely expressed in the tissues, related to the transcription of a variety of genes, and have an important role in the mutation of.COUP-TFs in organogenesis, neurogenesis, and cell differentiation. Abnormal expression is associated with human congenital heart disease, diabetes and cancer. In particular, the COUP-TF II gene plays a regulatory role in bone, muscle, fat formation and metabolic balance.
Objective: To compare the effects of SD rats and endurance training on the expression of COUP-TF I and COUP-TF II gene in skeletal muscles of GK rats and the relationship between the metabolism of glucose and the expression of COUP-TF I and COUP-TF II in the exercise adaptation.
Methods: 40 male SD rats, 4 weeks old, weight 100 5g, 12 GK rats, 8 weeks old and 250 + 5g, were randomly divided into SD control group (C), SD acute endurance training group (D), SD endurance training group (E), 10 rats in each group, 6 rats in each group. Practice: 1-6 weeks of rats with the quiet group feeding mode, before the death of an acute endurance training, the starting speed is 13m/min, the maximum speed is not more than 16.7m/min, training time 60min, intermittent sprint training: 3 groups of 3 times each day, each time 10s maximum intensity (> 50m/min) of the run, each interval time 30-60s, intermission time of 3 Min; endurance training: a 6 week running platform training with a starting speed of 13m/min, the maximum speed of not exceeding 16.7m/min, the first 2 weeks of training of 40 minutes, each training 60min for the first 4 weeks and 6 days of training every week. All rats were sacrificed after 6 weeks of feeding, and the enzyme was used to detect blood glucose, plasma insulin, plasma adiponectin, glycated hemoglobin and glycosylated serum protein. The transcriptional levels of COUP-TF I and COUP-TF II genes were detected by real-time fluorescence quantitative PCR. The expression level of COUP-TF I protein was detected by Western Blotting.
Result:
(1) three different exercises had no significant effect on weight gain and endurance training of SD rats on weight gain of GK rats.
(2) endurance training could significantly reduce the level of blood glucose in SD rats, but had no significant effect on the insulin resistance index, and endurance training could significantly reduce the blood glucose and insulin resistance index in GK rats.
(3) three different ways of exercise can significantly reduce the content of glycated serum protein in SD rats; endurance training can significantly reduce glycated serum protein in GK rats.
(4) endurance training can significantly increase the level of adiponectin in SD rats and GK rats.
(5) one time acute exercise and endurance training can significantly increase the COUP-TF I mRNA in the skeletal muscles of SD rats, and endurance training can significantly reduce the expression of COUP-TF I mRNA in the skeletal muscles of GK rats.
(6) one time acute exercise, endurance training, and high intensity intermittent sprint training had no significant effect on the expression of COUP-TF I protein in SD rats; endurance training could significantly reduce the expression of COUP-TF I protein in GK rats.
(7) one time acute exercise, endurance training, and high intensity intermittent sprint training can significantly reduce the expression of COUP-TF II mRNA in the skeletal muscle of SD rats; endurance training can significantly reduce the level of COUP-TF II mRNA in GK rats
Conclusion:
(1) endurance training is beneficial to lowering blood glucose level and improving insulin resistance in GK rats.
(2) acute exercise, endurance training and sprint training could increase the level of COUP-TF I mRNA in the skeletal muscles of SD rats and decrease the expression level of COUP-TF II mRNA, and the three kinds of exercise had no significant influence on the expression level of COUP-TF I protein, which showed that the level of COUP-TF I mRNA was not consistent with the level of COUP-TF I mRNA. With the rapid response of skeletal muscle to acute endurance exercise, it also participates in the chronic adaptation of skeletal muscle to long-term endurance training and intermittent large intensity training. Exercise may tend to activate the expression of COUP-TF I gene in skeletal muscle and inhibit the expression of COUP-TF II gene.
(3) long-term endurance training can improve insulin resistance in GK rats, which may be related to the down regulation of COUP-TF I gene, protein expression and COUP-TF II gene expression in skeletal muscle.
(4) in normal physiological and diabetic pathological conditions, endurance exercise has the opposite effect on the expression of COUP-TF I gene in skeletal muscle. Considering the strain, age, and dietary differences of GK rats and SD rats, this experiment can not exclude the effects of the above factors on the endurance exercise effect.
(5) compared with COUP-TF, the adaptation characteristics of COUP-TF II gene expression to endurance exercise in normal physiological and diabetic pathological conditions show that the inhibition of COUP-TF II gene expression by endurance exercise may play a more important role in the prevention and improvement of insulin resistance.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

【参考文献】