| 1.Research purposesIn this experiment,the UAS-Gal4 system was used to knock down the Drosophila Sima gene,and the Drosophila hypoxia device and exercise device were used to perform hypoxia and exercise intervention on Drosophila.To explore the effect of hypoxia combined exercise on the heart function of Drosophila,and to detect exercise capacity and sleep.Then,the effect of exercise intervention,hypoxia intervention and exercise and hypoxia intervention on the heart function of Drosophila was studied after knocking down the Sima gene.To provide a theoretical basis for studying the effect of sima gene-mediated hypoxia,exercise,and hypoxia combined exercise on cardiac function.2.research methodThe virgin flies with genotype v106187 were crossed with W1118 strain and Hand-GAL4 strain male flies respectively,and F1 generation virgin flies were obtained and randomly divided into 8 groups: quiet gene normal expression group(NC),hypoxia gene normal expression group(HC),exercise gene normal expression group(NE).,exercise and hypoxia gene normal expression group(HE),quiet Sima gene group(NCS),hypoxia SimaRNAi group(HCS),exercise SimaRNAi group(NES),exercise hypoxia SimaRNAi group(HES).in this case,the exercise group began to exercise on the 22 nd day of age,each training for1.5 hours,and continuous training for three weeks.The hypoxia group started hypoxia on the 22 nd day of age,hypoxia for 4 h,continuous hypoxia for three weeks.Index detection: 60 strains of Drosophila were randomly selected from Sima normal expression and Sima low expression for RT-PCR detection.The group randomly selected 30 fruit flies for M-mold cardiac function test,100 fruits is tested for climbing ability,32 Drosophila for sleep monitoring,3 research results3.1 RT-PCR test resultsW1118/v106187(gene normal group),Hand-Gal4/V106187(gene knockdown group)mRNA expression decreased Sima gene mRNA expression decreased,the down-regulation rate was 39.29%.The results proved F1 results The fly Sima gene knockdown line was successfully constructed.3.2 Drosophila heart function test results3.2.1 Drosophila heart rhythm index detection:3.2.1.1 Effect of hypoxia on rhythm of Drosophila heart Compared with the NC group,the heart rate of the HC group decreased,the heart rate index decreased,the HC cardiac cycle increased,and thediastolic interval increased(P<0.01).3.2.1.2 Effects of exercise on rhythm of Drosophila heart Compared with NC,the heart rate of the NE group was decreased,the heart rate index was decreased(P<0.01),the cardiac cycle was increased,and the diastolic interval was increased(P<0.05).3.2.1.3 Effect of exercise combined with hypoxia on rhythm of Drosophila heartThe HC heart rate is less than HCS,the NE heart rate is less than NES,and HE is less than HES.(P<0.01).The heart rate of NES was smaller than that of HES group(P<0.01).The NC arrhythmia index was smaller than NCS,the HC heart rate index was smaller than HCS,the NE heart rate index was smaller than NE group,and the HE heart rate index was smaller than the HES group(P<0.01).The NE cardiac cycle was greater than the NES group,and the HC was greater than the HCS group(P<0.01).HC was greater than HCS group(P<0.01),NE diastolic interval was greater than NES(P<0.05),HE diastolic interval was greater than HES group(P<0.01)Compared with the NC group,the heart rate of the HC,NE,and HE groups decreased,and the heart rate index of HC and NE decreased(P<0.01).HC(P<0.01)and NE cardiac cycle increased(P<0.05).The contraction interval of the HE group was increased(P<0.01).The diastolic interval was increased in the HC,NE,and HE groups(P<0.01).Compared with the NCS group,the heart rate of the HES group was decreased(P<0.01).The heart rate index of HCS,NES and HES decreased(P<0.01).The cardiac cycle of NCS and HES group increased(P<0.05).The diastolic interval of the HES group increased.3.2.2 Drosophila heart pumping blood function index detection3.2.2.1 Effect of hypoxia on blood function of DrosophilaCompared with NC,the HC shortening score was significantly increased(P<0.01),the HC contraction diameter was decreased,and(P<0.01)the HC diastolic diameter was decreased(P<0.01).Compared with NCS,the HCS shortening score increased significantly(P<0.01),and the HCS contraction diameter decreased(P<0.05).3.2.2.2 Effects of exercise on blood function of DrosophilaCompared with NC,NE shortened fraction increased(P<0.01)and NE diastolic diameter increased(P<0.05).Compared with NCS,NES shortening score increased(P<0.01)and NES diastolic diameter increased(P<0.05).3.2.2.3 Effects of exercise combined with hypoxia on blood function of DrosophilaThe NC shortening score was greater than NCS(P<0.01),the HC shortening score was greater than HCS,and the NE shortening score was greater than NES(P<0.01).The NC shrinkage diameter is smaller than NCS,the HC shrinkage diameter is smaller than HCS,the NE shrinkagediameter is smaller than NES,and the HE shrinkage diameter is smaller than HES(P<0.01).The NC diastolic diameter was significantly smaller than NCS(P<0.05).The HC diastolic diameter was significantly smaller than HCS(P<0.01),the NE diastolic diameter was smaller than NES(P<0.05),and the HE diastolic diameter was significantly smaller than HES(P<0.01).The NE and HC shortening scores were significantly increased compared with NC(P<0.01).HC and HE contraction diameter decreased(P<0.01).The diastolic diameter of Drosophila in the HC and HE groups decreased(P<0.01)and the NE diastolic diameter increased(P<0.01).Compared with NCS,the shortening scores of NES,HCS and HES increased(P<0.01).The HCS shrinkage diameter decreased(P<0.05).The NES diastolic diameter increased and the HES diastolic diameter increased(P<0.05).3.3 Drosophila climbing ability detection3.3.1 Impact of low oxygen on climbing abilityHC climbing ability was increased compared with NC group(P<0.01).Compared with NCS,HCS climbing ability increased(P<0.05).3.3.2 The impact of exercise on climbing ability Compared with NC,NE climbing ability increased(P<0.01).Compared with NCS,NES climbing ability increased(P<0.05)3.3.3 Impact of low oxygen combined exercise on climbing abilityHC is higher than HCS,NE is higher than NES,and HE is higher than HES(P<0.01).Compared with NC,the HC,NE,and HE climbing index increased.Compared with NCS,the HCS,NES,and HES climbing index increased(P<0.05).3.4 Drosophila sleep fragment number detection3.4.1 Effect of hypoxia on the number of sleep fragmentsCompared with NC,the number of NCS sleep fragments increased(P<0.01)3.4.2 The effect of exercise on the number of sleep segmentsCompared with NC,the number of NE sleep fragments decreased(P<0.01),and the number of NES sleep fragments decreased compared with NCS(P<0.05).3.4.3 Effect of exercise combined with hypoxia on the number of sleep segmentsThe number of NES sleep fragments was greater than NE(P<0.01),and the number of HE sleep fragments was reduced compared with NC,and the number of HES sleep fragments was reduced compared with NES.4 Conclusion4.1 Successfully constructed Sima gene knockdown fruit fly strain through UAS-GAL4,and found that Sima gene knockdown will lead toan increase in heart rate heart rate index,increase in contraction diameter and diastolic diameter,decrease in fractional score,and similar to dilated cardiomyopathy.symptom.And lead to a decline in exercise capacity and sleep quality.4.2.Sima gene knockdown of Drosophila and normal expression of Drosophila reduce heart rate index,delay aging-induced contraction,and reduce blood pumping function after hypoxia intervention.And have a positive effect on athletic ability.Also genetically normal fruit flies can reduce heart rate and improve cardiac cycle after hypoxia’s intervention.4.3 Sima gene knockdown of Drosophila and normal expression of Drosophila through exercise intervention can reduce heart rate,reduce heart rate index,improve cardiac cycle,delay the decline of Drosophi la relaxation,pumping function decline,and exercise capacity and sleep Have a positive effect.Also genetically normal fruit flies can reduce heart rate and improve cardiac cycle after exercise’s intervention. |
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