發(fā)布時(shí)間：2018-05-09 16:18

  本文選題：運(yùn)動(dòng) + PINK1��； 參考：《廣州體育學(xué)院》2017年碩士論文

【摘要】：目的:本文使用PINK1 RNAi帕金森果蠅作為研究對(duì)象,對(duì)果蠅進(jìn)行3周運(yùn)動(dòng)訓(xùn)練,觀察測(cè)量果蠅的爬行能力、壽命以及線粒體功能的變化,觀察果蠅線粒體聚集情況、測(cè)定線粒體復(fù)合物I的活性,旨在證明運(yùn)動(dòng)能夠?qū)ε两鹕膊‘a(chǎn)生積極的影響或作用,并尋找運(yùn)動(dòng)的作用機(jī)制,為今后對(duì)帕金森疾病的治療或者預(yù)防提供理論依據(jù)。方法:將用GFP標(biāo)記線粒體的果蠅進(jìn)行分組,包括不患病的UAS-mtGFP/cyo;Mhc/Mhc果蠅,和患病的UAS-mtGFP/Cyo;Mhc-RNAi/TM2的果蠅兩種基因型,對(duì)患病果蠅進(jìn)行運(yùn)動(dòng)干預(yù),運(yùn)動(dòng)持續(xù)3周,每周5天,每天運(yùn)動(dòng)時(shí)長(zhǎng)不同,按時(shí)長(zhǎng)分為4組,分為0分鐘組、30分鐘組、60分鐘組、90分鐘組,不患病的果蠅作為對(duì)照組,不進(jìn)行運(yùn)動(dòng)訓(xùn)練。在運(yùn)動(dòng)訓(xùn)練過(guò)程中每周測(cè)量一次所有果蠅的爬行能力以及翅膀表型百分?jǐn)?shù)(患病果蠅會(huì)出現(xiàn)翅膀的異常形態(tài)),并選取一部分果蠅進(jìn)行解剖,取出胸部肌肉進(jìn)行切片處理,使用激光共聚焦顯微鏡拍攝并觀察計(jì)算線粒體聚合情況。三周運(yùn)動(dòng)結(jié)束后再對(duì)所有果蠅進(jìn)行磨漿處理,提取線粒體,測(cè)定每組果蠅的線粒體復(fù)合物I的活性,。結(jié)果:1)翅膀表型:開始兩周并沒有發(fā)現(xiàn)對(duì)照組和不運(yùn)動(dòng)組翅膀表型百分?jǐn)?shù)有顯著性差異,在第三周的差異才存在顯著性,尤其是30分鐘組有著非常顯著性差異(P0.01)。這說(shuō)明運(yùn)動(dòng)三周對(duì)患病果蠅的翅膀表型還是存在著作用的。以每天30分鐘運(yùn)動(dòng)量為佳。2)運(yùn)動(dòng)能力:第一周沒有發(fā)現(xiàn)和不運(yùn)動(dòng)組存在的顯著性差異,第二周的30分鐘組和不運(yùn)動(dòng)組具有顯著性差異,第三周30分鐘組有非常顯著性差異(P0.01)。這說(shuō)明運(yùn)動(dòng)三周對(duì)患病果蠅的爬行能力有改善作用。以每天30分鐘運(yùn)動(dòng)量為佳。3)壽命:不運(yùn)動(dòng)組的果蠅只能存活27天,而30分種組平均可以存活33天,60分鐘和90分鐘組平均可以存活30天左右,但都要低于正常的對(duì)照組的壽命。4)激光共聚焦顯微鏡下的線粒體聚集情況:同樣也是30分鐘組的線粒體聚集最少。而60分鐘和90分鐘組以及0分鐘組聚集情況較為明顯。5)線粒體復(fù)合物I的活性:有較為明顯的變化,其中不運(yùn)動(dòng)組的活性較低,而30分鐘組的活性較高。結(jié)論:1)適宜的運(yùn)動(dòng)能夠在一定程度上改善患帕金森果蠅的翅膀表型,提高其運(yùn)動(dòng)能力和延長(zhǎng)其壽命。2)運(yùn)動(dòng)訓(xùn)練能減少線粒體病態(tài)聚集以及提高線粒體復(fù)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.
【學(xué)位授予單位】：廣州體育學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：G804.2

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