一次性离心运动后低氧暴露对骨骼肌utrophin的影响

发布时间：2018-03-11 21:06

本文选题：dystrophin　切入点：utrophin　出处：《北京体育大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的：Utrophin是dystrophin的同源蛋白,在dystrophin蛋白不足时有替代dystrophin的潜力。本研究选用EIMD为切入点采用损伤特征明显的离心运动方式模拟“高住低训”模式,从utrophin蛋白的角度来研究低氧对EIMD膜损伤影响及可能的分子机制。方法：将70只八周龄健康的SD雄性大鼠(体重196.75±8.14g)随机分成：安静组(C)；常氧下运动,常氧组(N)；常氧下运动,低氧组(H)；后两组又随机分为3个亚组：24H组、48H组、72H组。共7组,每组10只。低氧环境的氧浓度为12.7%。每组各取2只腹腔注射荧光染料EBD (Evans blue dye,伊文氏蓝)来检测肌细胞膜通透性。运动方式采用坡度为-16度的大负荷间歇性下坡跑,运动速度为26.8米/分,运动5分钟×10组,组间歇1分钟。运动结束后,取大鼠小腿腓肠肌用Western blot、RT-PCR和荧光免疫组化方法进行dystrophin基因表达、utrophin蛋白含量和基因表达等指标的测定。结果：一次性离心运动后N24h和H24h组均出现EBD阳性纤维,常氧组EBD阳性细胞数先增加后减少,低氧组EBD阳性细胞数逐渐降低,均显著高于安静组(P0.05)。低氧组和常氧组dystrophin基因表达量随时间推移均逐渐下降,且均明显低于安静组(P0.05)。与安静组相比,H24h和H48h组utrophin蛋白含量显著增高(P0.05),72h组降低明显,恢复至接近安静组水平。随着时间推移,低氧各组utrophin蛋白含量持续升高,均明显高于安静组(P(0.05)。Utrophin各组基因表达与蛋白含量变化并不完全相符,说明该蛋白的表达除了由基因转录调节的控制外,还可能存在其他的调节机制。结论：1.一次性离心运动后低氧暴露不仅使骨骼肌细胞膜损伤提前,而且会延缓肌细胞膜的修复。2.一次性离心运动后低氧暴露使utrophin蛋白的表达增加。
[Abstract]:Objective: Utrophin is the homologous protein of dystrophin and has the potential to replace dystrophin when the dystrophin protein is insufficient. In this study, we used EIMD as the starting point to simulate the "high living and low training" model by centrifugal exercise with obvious damage characteristics. To study the effect of hypoxia on EIMD membrane damage and its possible molecular mechanism from the point of view of utrophin protein. Methods: 70 healthy SD rats (body weight 196.75 卤8.14 g) were randomly divided into three groups: quiet group, normoxic group, normoxic group, normoxic group, normoxic group, normoxic group, and normal oxygen group. The latter two groups were randomly divided into 3 subgroups, 1: 24 H group, 48 H group and 72 H group. 10 rats in each group were given oxygen concentration of 12.70.Two rats in each group were injected intraperitoneally with fluorescent dye EBD Evans blue dye (Yi Wen's blue) to detect the permeability of muscle cell membrane. The speed of exercise was 26.8 m / min, 5 minutes 脳 10 group, the interval of group was 1 minute. The content and expression of dystrophin gene in gastrocnemius muscle of rats were determined by Western blottir RT-PCR and fluorescence immunohistochemistry. Results: EBD positive fibers were found in N24 h and H24 h groups after single centrifugation. The number of EBD positive cells in normoxic group increased first and then decreased, and the number of EBD positive cells in hypoxic group decreased gradually, which was significantly higher than that in quiet group. The expression of dystrophin gene in hypoxia group and normoxic group decreased gradually with the passage of time. Compared with the rest group, the content of utrophin protein in H24 h and H48 h groups decreased significantly and recovered to the level of quiet group. With time, the content of utrophin protein in hypoxic groups continued to increase. The expression of Utrophin gene was not consistent with the change of protein content, which indicated that the expression of the protein was controlled by gene transcription. There may be other regulatory mechanisms. Conclusion 1. Hypoxic exposure after one time centrifugation can not only lead to early damage of skeletal muscle cell membrane, but also delay the repair of muscle cell membrane. 2. Hypoxia exposure after one time centrifuge exercise can increase the expression of utrophin protein.
【学位授予单位】：北京体育大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R87

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