| Objective This study was based on animal experiments to investigate the effect of combined exercise intervention on body weight and blood glucose improvement in type 2 diabetic mice through 12 weeks of exercise intervention in type 2 diabetic mice,and to explore combine exercise for type 2 diabetes.Effect of the function of mouse endothelial cells,and further study of Caveolin-1/PI3K/Akt related protein and expression of endothelial progenitor cell function.Scientific evaluation of combine campaigns lays the foundation for a clear role mechanism and provides scientific evidence and guiding significance for effective care interventions fordiabetic patients.Methods(1)Eight-week-old db/db(C57BL/KS.db background)male mice(blood glucose 300 mg/dl or more)were used as a model of type 2 diabetes mellitus.The mice were divided into control group,combined exercise group,resistance exercise group and aerobic exercise group.Mice were randomly assigned to aerobic exercise group,resistance exercise group,combined exercise,and control group(6 per group)at five time points of1 week,2 weeks,4 weeks,8 weeks,and 12 weeks.Each group took differentexercise intervention programs.After 1 week,2 weeks,4 weeks,8 weeks,and12 weeks of exercise training(6 days per week),the body weight and blood glucose changes before and after exercise intervention were measured at different time points.(2)Four groups of db/db mice were sacrificed at 1 week,2weeks,4 weeks,8 weeks,and 12 weeks after intervention.In vitro,db/db mouse bone marrow derived endothelial progenitor cells(EPCs)were induced to culture in vitro.The morphology of EPCs was observed by cell morphology;the positive rates of DiI-ac-LDL and FITC-UEA-I double staining and the CD133 phenotype detected by CD133 were used to identify EPCs;EPCs cell function test: CCK-8 assay was used to determine the proliferation and cells.Adhesion experiments to observe the adhesion,cell scratches experimental observation of migration ability,matrigel culture observed angiogenesis.(3)Western-blot.Protein expression levels of Caveolin-1,p-PI3 K p85,and p-Akt were detected.Results(1)After exercise for 12 weeks,the three exercise interventions of the combine exercise group,the resistance exercise group,and the aerobic exercise group were able to reduce body weight and blood glucose levels(P<0.05),and the combined exercise group was superior to the aerobic exercise group and the resistance.Resistance exercise group(P<0.05).(2)EPCs were cultured,identified and functionally tested: EPCs adhered slowly and adhered to the wall in human fibronectin-coated dishes for 24 h.The adherence to the wall was almost complete in 4 days.In the whole bone marrow culture method,the adherent cells were seen in the 4th day,and they were round,short shuttle-shaped,and some were long shuttle-shaped;the adherent cells in the 6th to 8th days were mostly spindle-shaped and round;With the prolongation of culture time,the number of round cells decreased,the number of spindle cells gradually increased,and cell colonies appeared.10 days later,EPCs proliferatedin large numbers,and spindle-shaped adherent cells continued to form on their edges.The cells were overgrown in 14 days.At the bottom,there may be "paving stone"-like changes,and the cells after passaging may be arranged end to end;The positive rates of DiI-ac-LDL uptake and FITC-UEA-I double staining in whole bone marrow adherent culture EPCs were(73.50±7.47)%.The detection rate of CD133 with phenotype was 63.29%;The proliferative capacity of EPCs: There were significant differences between the combined exercise group,the resistance exercise group,and the aerobic exercise group compared with the control group(P<0.05),there was no statistical difference between the groups(P>0.05);EPCs adhesion,migration,blood vessels Ability to generate:The difference between the combined exercise group,the resistance exercise group,and the aerobic exercise group was statistically significant(P<0.05).The comparison between the combined exercise group,the resistance exercise group,and the aerobic exercise group were statistically significant(P<0.05).The combined exercise group was superior to the resistance exercise group and the aerobic exercise group.(3)Western-blot: The protein expression levels of Caveolin-1,p-PI3 K p85,and p-Akt in EPCs in the combined exercise group,the resistance exercise group,the aerobic exercise group,and the control group were significantly different(P<0.05);There was a statistically significant difference between the combined exercise group,the resistance exercise group,and the aerobic exercise group(P<0.05).The joint exercise group was higher than the resistance exercise group and the aerobic exercise group.Conclusion(1)Combine exercise,resistance exercise and aerobic exercise can improve the body weight and blood glucose level of db/db mice,and enhance the proliferation,adhesion,migration and angiogenesis of EPCs.The effect may be through the Caveolin-1 protein.The PI3K/Akt pathway mediates up-regulationof the target gene,which in turn enhances the function of EPCs.(2)The combine exercise is optimal for the three types of exercise interventions. |
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