Effects Of Regular Exercise On Heart Function Of Drosophila With High Sugar Diet Carrying Gene Knockout Of Sugar Transporters

1 ObjectiveThe UAS/GAL4 system was used to regulate the expression of the sugar transporter gene and high-sugar diet intervention in drosophila,The flies were measured for body weight,triglyceride levels,blood sugar levels,heart function,climbing ability,to investigate the effects of high-sugar-diet combined with glucose transporter gene knockout on triglyceride and glucose levels,heart function and climbing ability in drosophila,Screening out sugar transporter regulatory genes associated with heart function in drosophila on a high-sugar-diet.To further study the relationship between sugar transporters and heart dysfunction,and to study the positive effects of exercise on glucose metabolism disorders in drosophila and the heart.2 MethodsUAS/GAL4 system was used to regulate the expression of glucose transporter gene in drosophila,drosophila of UAS-RNAi strain?Glut1 and Glut3 genes and Tret1-1 and Tret1-2 genes?,a sugar transporter regulatory gene,were crossed with drosophila of arm-gal4 strain and W¹¹¹⁸ strain respectively,To realize gene knockout and normal expression of sugar transporter in drosophila,They were divided into normal expression group and knockout group.Two groups of virgin flies bred in F1 within 8 hours were collected and randomly cultured on Normal Diet?ND?and High Sugar Diet?HSD?.They were divided into blank control group?W-ND?,gene knockout group?ARM-ND?,high-sugar-diet group?W-HSD?,gene knockout and high-sugar diet group?ARM-HSD?,400 in each group.The normal diet group was cultured in ordinary medium for 7days,while the high sugar diet group was cultured in high sugar for 7 days.The normal medium was adapted on the first day,the samples were taken after the 7 days of high sugar.Test indicators:Randomly selected from four groups of drosophila strains,20 files were weighed,15 were measured for triglyceride and blood sugar levels,30 for heart function and 100 for climbing ability.3 Results3.1 Test results of triglyceride and glucose levels in drsophilaGlut3:The triglyceride level of the W-HSD group was higher than that of the W-ND and ARM-ND groups?P<0.01?.The level of triglyceride in the ARM-HSD group was higher than that in the W-ND and ARM-ND groups?P<0.01?.The glucose level of the W-HSD group was higher than that of the W-ND group and the ARM-HSD group?P<0.01?,the level of triglyceride in ARM-HSD group was higher than ARM-ND group?P<0.01?.Glut1:The levels of triglyceride and glucose levels in the W-HSD group were higher than that in the W-ND group?P<0.01?.The levels of triglyceride and glucose in the ARM-HSD group were higher than those in the ARM-ND group?P<0.01?.The triglyceride level of ARM-ND group was higher than that of W-ND group?P<0.01?.Tret1-1:The triglyceride level in the W-ND group was higher than that in the W-HSD and ARM-ND groups?P<0.05?.The triglyceride level in the ARM-HSD group was higher than that in the W-HSD group?P<0.05?.The glucose level of the W-ND group was higher than that of the ARM-ND and ARM-HSD groups?P<0.01,P<0.05?.Tret1-2:The triglyceride level of ARM-HSD group was higher than that of W-HSD group?P<0.01?.Triglyceride and glucose levels in the ARM-HSD group were higher than those in the W-ND group?P<0.01?and P<0.05).Triglyceride and glucose levels in the ARM-HSD group were higher than those in the ARM-ND group?P<0.01,P<0.05?.The triglyceride level of the W-ND group was higher than that of the W-HSD group?P<0.01?.The triglyceride level of ARM-ND group was higher than that W-ND group?P<0.01?.3.2 Results of drosophila heart M-mode detectionGlut3:The arrhythmia index of the W-ND group was higher than that of the ARM-ND group and ARM-HSD group?P<0.01,P<0.05?.Glut1:W-ND group fibrillation was lower than ARM-ND group?P<0.05?.The diastolic diameter of the W-ND group was higher than that of the ARM-ND and W-HSD groups?P<0.01?.Arrhythmia index of ARM-ND group was higher than that of W-ND group and ARM-HSD group?P<0.01?.Tret1-1:The diastolic diameter of ARM-ND group was higher than that of W-HSD group?P<0.05?.The diastolic diameter and arrhythmia index of the ARM-HSD group were higher than that of the W-HSD group?P<0.05?.Arrhythmia index of ARM-HSD group was higher than that of W-ND group?P<0.05?.Tret1-2:Arrhythmia index of ARM-ND group was higher than that of W-ND group,W-HSD group and ARM-HSD group?P<0.01?.3.3 Drosophila climbing index resultsGlut3:The climbing index of the W-HSD group was higher than that of the W-ND and ARM-ND groups?P<0.05,P<0.01?.The climbing level of W-ND was higher than that of ARM-ND group?P<0.01?.The climbing level of ARM-HSD was higher than that of W-ND and ARM-ND?P<0.01?.Glut1:The climbing index of the W-HSD group was higher than that of the ARM-HSD group,ARM-ND group?P<0.05?and the W-ND group?P<0.01?.Tret1-1:The climbing index of the W-ND group was higher than that of the ARM-ND group?P<0.05?.The climbing index of the W-HSD group was higher than that of the ARM-ND and ARM-HSD groups?P<0.01,P<0.05?.Tret1-2:The climbing index of the W-HSD group was higher than that of the W-ND group?P<0.05?.The climbing index of ARM-HSD group was higher than that of W-ND group?P<0.01?.3.4 qRT-PCR detection resultsGlut3 mRNA gene expression rate in ARM-ND group was73.5%.Glut3 mRNA gene expression rate of ARM-HSD group was68.9%.The Glut3 gene knockout strain was successfully constructed.Glut1 mRNA gene expression rate in Glut1 group ARM-ND group was 72%.The mRNA gene expression rate of Glut1 in ARM-HSD group was 65%,indicating that the Glut1 gene knockout strain was successfully constructed.Tret1-1 mRNA gene expression rate in ARM-ND group was54.9%.Tret1-1 mRNA gene expression rate in ARM-HSD group was56.7%.Tret1-1 knockout strain was successfully constructed.Tret1-2 mRNA gene expression rate in tret1-2 ARM-ND group was69.9%.Tret1-2 mRNA gene expression rate in ARM-HSD group was70%.Tret1-2 knockout strain was successfully constructed4 Conclusion.4 Conclusion?1?HSD can cause the metabolism disorder of triglyceride and glucose in drosophila,resulting in its small size.?2?The effects of HSD and gene knockout of glucose transferase on the cardiac function of drosophila were as follows:diastolic diameter decreased,fibrillation and arrhythmia index increased.?3?Glut1 can be used as a candidate gene to study the effects of regular exercise on cardiac function decline and glucose metabolism disorder induced by HSD in drosophila.HSD can improve the climbing characteristics of one-week-old drosophila.1 ObjectiveBased on the screening results,the experiment further explored the effects of regular exercise on glycometabolism and cardiac function of flies in combination with HSD and sugar transporter gene knockout.The UAS/GAL4 system was used to regulate the expression of the screened glucose transporter gene in drosophila,intervention with regular exercise and high-sugar-diet,Within 24 hours of the end of the exercise and diet intervention,test the heart function,climbing ability,triglycerides and blood sugar levels,The effects of regular exercise on heart function decline and glucose metabolism disorder of flies induced by HSD were also discussed.2 MethodsTo further investigate the effect of regular exercise on glucose metabolism disorder in flies caused by sugar transporter gene knockout combined with HSD.By using the UAS/GAL4 system,the sugar transporter regulatory gene UAS-RNAi line of drosophila?Glut1?was hybridized with arm-gal4 line and W¹¹¹⁸ respectively.To realize gene knockout and normal expression of sugar transporter in drosophila,Two groups of virgin drsophila that were bred in F1 within 8 hours were collected for HSD and regular exercise intervention,Divided into normal diet group?W-ND?,High-sugar-diet group?W-HSD?,Gene knockout group?ARM-ND?,Gene knockout high-sugar-diet group?ARM-HSD?,exercise group?W-ND-E?,exercise and HSD group?W-HSD-E?,The exercise gene knockout group?ARM-ND-E?,exercise gene knockout high-sugar-diet group?ARM-HSD-E?,total of 8 groups,Each group of 400,The 7 day exercise with high-sugar regimen ended after sampling.Using the drosophila anti-gravity climbing property,The third-generation drosophila movement device was used for regular exercise training,exercise speed control is 0.18r/s,1.5 hours persession,Starting at16:00 every day,test tube diameter d=2.80cm,7 consecutive days of training.Test indicators:8 groups of flies were randomly selected,20 flies were weighed,15 flies measured their triglyceride levels as well as their glucose levels,30 were tested for heart function and 100 for climbing ability.3 Results3.1 Test results of triglyceride and glucose levels in drosophilaIn the exercise group,the triglyceride level of flies in the W-ND-E group was lower than the W-HSD-E,ARM-ND-E and ARM-HSD-E groups,There was a significant difference?P<0.05?.The level of triglyceride in ARM-ND-E group was lower than that in W-HSD-E group and ARM-HSD-E group,with significant difference?P<0.01?.Compared to the control group,the level of ARM-ND group triglyceride in the exercise group was higher than that in ARM-HSD-E group,There was a significant difference?P<0.05?.In the exercise group,The glucose level of the W-ND-E group was lower than that of the W-HSD-E and ARM-ND-E groups,with significant differences?P<0.05,P<0.01?,The glucose level of the ARM-ND-E group was lower than that of the W-HSD-E and ARM-HSD-E groups,and the difference was extremely significant?P<0.01?.Compared with the control group,the glucose level of the exercise group was all decreased,but there was no significant difference?P>0.05?.3.2 Results of drosophila heart M-mode detectionArrhythmia index in ARM-ND group was significantly higher than that in ARM-HSD,W-ND and W-HSD groups?P<0.01?.Arrhythmia index of ARM-ND group was significantly higher than that of W-ND-E,W-HSD-E,ARM-ND-E,ARM-HSD-E group?P<0.01?.The fibrillations of the W-ND-E group was significantly lower than the ARM-ND group,the fibrillations of the W-HSD-E group was significantly lower than the W-HSD group,and the fibrous tremor of the ARM-HSD-E group was significantly lower than that of the ARM-HSD group,all of which had extremely significant differences?P<0.01?.The diastolic diameter of the W-HSD-E group was significantly higher than that of the ARM-ND-E group and ARM-HSD-E group?P<0.01?.Compared with the control group,the diastolic diameter of the W-HSD-E group was higher than that of the W-HSD group,The diastolic diameter of the ARM-ND-E group was higher than that of the ARM-ND group,The diastolic diameter of the ARM-HSD-E group was higher than that of the ARM-HSD group,There were significant differences?P<0.01?.3.3 Drosophila climbing index resultsIn the exercise group,the climbing index of the W-ND-E group was lower than that of the W-HSD-E group?P<0.01?,The climbing index of the W-ND-E group was higher than that of the ARM-ND-E group,and the difference was significant?P<0.01?.The climbing index of ARM-ND-E group was lower than that of W-HSD-E and ARM-HSD-E group,and the difference was very significant?P<0.01?.Compared with the control group,the climbing index of the W-ND group was lower than that of the W-ND-E group?P<0.01?,the climbing index of the W-HSD group was lower than that of the W-HSD-E group?P<0.01?,and the climbing index of the ARM-HSD group was lower than that of the ARM-HSD-E group with significant difference?P<0.01?.3.4 q RT-PCR detection resultsCompared with the W-ND group,the expression rate of Glut1 mRNA gene in the W-HSD-E group was 97.26%.The mRNA expression rate of Glut1 in ARM-ND-E group was 82.1%.The mRNA gene expression rate of Glut1 in ARM-HSD-E group was 71.3%,indicating that the Glut1gene knockout strain was successfully constructed.4 Conclusion?1?A high-sugar diet combined and Glut1 gene knockout can affect heart function in drosophila,Its main expression is diastolic function drops and fibrillations,arrhythmia index increases.?2?Regular exercise can effectively reduce the heart fibrillations and arrhythmia index of one-week old drosophila.Regular exercise improves the glycolipid metabolism of drosophila caused by HSD.?3?The exercise ability of drosophila is affected by HSD,and reduced by Glut1 gene knockdown.