运动对PINK1 RNAi帕金森果蝇线粒体及其相关功能的影响

发布时间：2018-05-09 16:18

本文选题：运动 + PINK1　；参考：《广州体育学院》2017年硕士论文

【摘要】：目的:本文使用PINK1 RNAi帕金森果蝇作为研究对象,对果蝇进行3周运动训练,观察测量果蝇的爬行能力、寿命以及线粒体功能的变化,观察果蝇线粒体聚集情况、测定线粒体复合物I的活性,旨在证明运动能够对帕金森疾病产生积极的影响或作用,并寻找运动的作用机制,为今后对帕金森疾病的治疗或者预防提供理论依据。方法:将用GFP标记线粒体的果蝇进行分组,包括不患病的UAS-mtGFP/cyo;Mhc/Mhc果蝇,和患病的UAS-mtGFP/Cyo;Mhc-RNAi/TM2的果蝇两种基因型,对患病果蝇进行运动干预,运动持续3周,每周5天,每天运动时长不同,按时长分为4组,分为0分钟组、30分钟组、60分钟组、90分钟组,不患病的果蝇作为对照组,不进行运动训练。在运动训练过程中每周测量一次所有果蝇的爬行能力以及翅膀表型百分数(患病果蝇会出现翅膀的异常形态),并选取一部分果蝇进行解剖,取出胸部肌肉进行切片处理,使用激光共聚焦显微镜拍摄并观察计算线粒体聚合情况。三周运动结束后再对所有果蝇进行磨浆处理,提取线粒体,测定每组果蝇的线粒体复合物I的活性,。结果:1)翅膀表型:开始两周并没有发现对照组和不运动组翅膀表型百分数有显著性差异,在第三周的差异才存在显著性,尤其是30分钟组有着非常显著性差异(P0.01)。这说明运动三周对患病果蝇的翅膀表型还是存在着作用的。以每天30分钟运动量为佳。2)运动能力:第一周没有发现和不运动组存在的显著性差异,第二周的30分钟组和不运动组具有显著性差异,第三周30分钟组有非常显著性差异(P0.01)。这说明运动三周对患病果蝇的爬行能力有改善作用。以每天30分钟运动量为佳。3)寿命:不运动组的果蝇只能存活27天,而30分种组平均可以存活33天,60分钟和90分钟组平均可以存活30天左右,但都要低于正常的对照组的寿命。4)激光共聚焦显微镜下的线粒体聚集情况:同样也是30分钟组的线粒体聚集最少。而60分钟和90分钟组以及0分钟组聚集情况较为明显。5)线粒体复合物I的活性:有较为明显的变化,其中不运动组的活性较低,而30分钟组的活性较高。结论:1)适宜的运动能够在一定程度上改善患帕金森果蝇的翅膀表型,提高其运动能力和延长其寿命。2)运动训练能减少线粒体病态聚集以及提高线粒体复合物I的活性。
[Abstract]:Objective: to observe the changes of creeping ability, life span and mitochondrial function of Drosophila melanogaster, using PINK1 RNAi Parkinsonian fly as the research object, and observe the mitochondrial aggregation of Drosophila melanogaster after 3 weeks exercise training. The purpose of measuring the activity of mitochondrial complex I is to prove that exercise can have a positive effect or effect on Parkinson's disease, to find out the mechanism of exercise, and to provide a theoretical basis for the treatment or prevention of Parkinson's disease in the future. Methods: Drosophila melanogaster labeled with GFP was divided into two genotypes: UAS-mtGFP / Mhc- Mhc- Mhc / Mhc- Mhc / Mhc- Mhc / Mhc- Mhhc- Mhc / Mhc- Mhc- Mhhc- Mhhc- Mhhc- Mhhc / Mhhc- Mhhc- Mhc / Mhc- Mhhc- Mhhc- Mhhc- Mhhc@@ According to the time, the flies were divided into 4 groups, divided into 0 minute group, 30 minutes group, 60 minutes group and 90 minutes group. The fruit fly without disease was used as control group, and no exercise training was carried out. During exercise training, the crawling ability and the percentage of wing phenotypes of all fruit flies were measured once a week. The polymerization of mitochondria was observed and photographed by laser confocal microscope. After three weeks of exercise, all drosophila were treated with grinding plasma, mitochondria were extracted, and the activity of mitochondrial complex I in each group was determined. Results: wing phenotype: in the first two weeks, there was no significant difference in the percentage of wing phenotype between the control group and the non-exercise group, but there was a significant difference in the third week, especially in the 30-minute group. This suggests that three weeks exercise has an effect on the wing phenotype of the infected fruit fly. Exercise ability: there was no significant difference between the first week and the non-exercise group, there was significant difference between the 30 minute group and the non-exercise group in the second week, and there was a very significant difference between the 30 minute group and the non-exercise group in the third week (P 0.01). This shows that three weeks of exercise can improve the crawling ability of infected fruit flies. The average survival time of fruit flies in the non-exercise group was 27 days, while that in the 30 species group was 33 days, 60 minutes and 90 minutes, respectively. But they were lower than the normal control group. 4) the mitochondria aggregation under laser confocal microscope was also the lowest in the 30 minute group. However, the activity of mitochondrial complex I in 60 min, 90 min and 0 min groups was obvious. 5) the activity of mitochondrial complex I was significantly changed, among which the activity of no exercise group was lower, while that of 30 minute group was higher than that of non exercise group. Conclusion: (1) the suitable exercise can improve the wing phenotype of Drosophila parkinsonii to some extent, increase its motor ability and prolong its life span. 2) exercise training can reduce the pathological aggregation of mitochondria and increase the activity of mitochondrial complex I.
【学位授予单位】：广州体育学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：G804.2

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