Effect Of Endurance Exercise On Cardiac Function And Sleep In Aging Sir2 Gene Silencing Drosophila

1 ObjectiveThis study uses UAS-Gal4 binary transgenic system to intervene in the Sir2 gene knockdown, using drosophila vertical rotation device to study the effects of endurance exercise on Sir2 gene knockdown in drosophila, as well as endurance exercise, Sir2 and heart function, sleep, life span, finding a way to improve the ability of drosophila, improve sleep quality and prolong the life span of drosophila.2 Materials and MethodsExperimental model construction:this experiment was conducted with three types,w[67c23] P{w[+mC]y[+mDint2]=EPgy2} Sirtl[EY22867]DnaJ-H[EY22867] （Sir2）,（y[1]）, W¹¹¹⁸ wild type and arm-Gal4 regulatory system.Methods:arm-Gal4 male and female control lines were W¹¹¹⁸ wild-type and sir2 female drosophila knockdown hybridization,F1 generations W¹¹¹⁸/arm-Gal4（control group） and sir2/arm-Gal4（exercise group）, two strains of virgin drosophila collected within 6-10 hours of eclosion of 3200 rats each, were randomly divided into 8 groups:2W-C-W¹¹¹⁸/arm-Gal4,2W-C-sir2/arm-Gal4, 2W-E-W¹¹¹⁸/arm-Gal4,2W-E-sir2/arm-Gal4,6W-C-W¹¹¹⁸/arm-Gal4, 6W-C-sir2/arm-Gal4,6W-E-W¹¹¹⁸/arm-Gal4,6W-E-sir2/arm-Gal4.each group are 400 drosophilas,20/tube.By use of the drosophila vertical rotational motion device, exercise group began to exercise from the second day of age, training two hours a day, exercising five days in a row,2 days off, movement for two weeks and six weeks respectively, device speed frequency set according to the actual situation of the drosophila movement do accordingly, （device frequency set to 280, at the beginning of time interval between 7s, after the movement around the adjustment to the time interval of 14s） from the build of the two strains were randomly selected.10 drosophilas for RT-PCR detection,16 drosophilas for DAMS,30 drosophilas for M-mold cardiac functions detection,100 drosophilas do climbing ability test,100 drosophilas to make lifespan statistics.3 Results3.1 The result of RT-PCRCompared with W¹¹¹⁸/arm-Gal4 group,sir2/arm-Gal4 group expression decreased, knockdown at a rate of 61.92%, suggesting that the Fl sir2 knockdown drosophila strains build is successful.3.2 The result of M-mold cardiac functionsCompared with 6W-C-W¹¹¹⁸/arm-Gal4 group, cardiac fibrillation decreased （P<0.05） in 6W-E-W¹¹¹⁸/arm-Gal4 group, suggesting that exercise can decrease the risk of wild type aging drosophila’s fibrillation, compared with 6W-C-W/arm-Gal4 group, arrhythmia index was significantly increased （P<0.01）, cardiac fibrillation increased significantly （P<0.01） in 6W-C-sir2/arm-Gal4 group, suggesting that the Sir2 gene knockdown will aggravate the arrhythmia and cardiac fibrillation occurrence in aging drosophila.Compared with 6W-C-sir2/arm-Gal4 group, arrhythmia index decreased （P<0.01）, cardiac fibrillation decreased（P<0.05）, shortening fraction increased （P<0.05） in 6W-E-Sir2/arm-Gal4 group, suggesting that exercise can improve heart function of Sir2 gene knockdown type aging drosophila and reduce the risk of cardiac rhythm abnormal and cardiac fibrillation.3.3 Circadian rhythm of sleep6W-C-W¹¹¹⁸/armGal4 group,6 W-C-Sir2/arm-Gal4 group, 6W-E-W¹¹¹⁸/arm-Gal4,6W-E-Sir2/arm-Gal4 group’s daytime activity, sleep time had no significant difference in the day, and the aging drosophila exercise group’s activity is less than the quiet group,and the night sleep time, total sleep time are more than the quiet group, 6W-C-Sir2/arm-Gal4 group activity at night is more than 6W-C-W¹¹¹⁸/arm-Gal4 group（P<0.05）. suggesting that the Sir2 gene knock down would increase the amount of night activity in the aging drosophila, and exercise could reduce the amount of night activity in aging drosophila.3.4 Exercise ability testCompared with 6W-C-W¹¹¹⁸/arm-Gal4 group,6W-E-W¹¹¹⁸/arm-Gal4 group index climbing was obviously increased（P<0.01）, 6W-C-Sir2/arm-Gal4 group index climbing was obviously decreased（P<0.01）, suggesting long-term exercise training can obviously improve the athleticism of the wild type aging drosophila, Sir2 gene knockdown can reduce the movement of aging drosophila. compared with 6W-C-Sir2/arm-Gal4 group,6W-E-Sir2/arm-Gal4 group index climbing was obviously increased（P<0.01）, suggesting exercise can strengthen the movement of Sir2 gene knockdown aging drosophila.3.5 Tests of the lifespan statisticsCompared with C-W¹¹¹⁸/arm-Gal4 group, C-Sir2/arm-Gal4 group’s lifespan shortened（P<0.01）, suggesting Sir2 gene knockdown will shorten the average lifespan of drosophila. Compared with C-W¹¹¹⁸/arm-Gal4 group,2C-E-W¹¹¹⁸/arm-Gal4 group longevity（P<0.01）,compared with W-E-W¹¹¹⁸/arm-Gal4 group longevity（P<0.01）, compared with C-Sir2/arm-Gal4 group,2W-E-Sir2/arm-Gal4 group longevity（P<0.01）, 6W-E-Sir2/arm-Gal4 group increased（P<0.01）,suggesting either only in adolescent period or from adolescent period to aging period,exercise can prolong lifespan inwild type and Sir2 gene Knockdown drosophila.4 Results4.1 Through the UAS-Gal4 binary transgenic system successfully achieved the Sir2 gene knockdown, Sir2 gene knockdown can increase the risk of cardiac dysfunction in drosophila, exercise can effectively reduce the occurrence of aging drosophila heart arrhythmia and fibrous tremor.4.2 Drosophila’s exercise capacity in age of recession, the Sir2 gene knockdown has superimposition effect with the increase of age, further reducing the drosophila’s movement ability, through endurance exercise intervention can slow aging drosophila’s exercise decline.4.3 Sir2 gene knockdown has no influence of young drosophila’s sleep-wake patterns, the influence of aging drosophila’s sleep-wake patterns which is active at night, it will lead to increased with fragmented sleep at night and the night of sleep time reduced. Implementation of endurance sports after the intervention, aging drosophila with fragmented sleep significantly reduced, night sleep time increased significantly.4.4 Sir2 gene knockdown will shorten the lifespan of drosophila, both in drosophila young period of the implementation of endurance exercise intervention or exercise intervention period from young to aging,it can improve the lifespan of drosophila decreased, there is no obvious difference between the two types, visibling exercise in young age can effect with retention.