Aerobic Combined Resistance Exercise Regulates MiRNA-92a By Activating AKT Pathway To Promote Endothelial Progenitor Function In Type 2 Diabetic Mice

ObjectiveTo explore the effects of aerobic combined resistance exercise on the expression of angiogenic function,AKT and miRNA-92 a in endothelial progenitor cells(EPCs)of type 2 diabetic mice.Identify the effects of PI3K/AKT pathway inhibitors on Caveolin1,PI3 K,AKT and miRNA-92 a in EPCs.To elucidate the effect of aerobic combined resistance exercise on the angiogenesis function of EPCs in type 2 diabetic mice by activating AKT pathway.MethodsEight-week-old db/db(C57BL/KS.db background)male mice(blood glucose 300 mg/dl or more)were used as a mouse model of type 2 diabetes.Based on the random numbers generated by the computer-generated random number table(Research Randomizer,2008),the mice were randomly assigned to the control group and the mixed exercise group(aerobic + resistance exercise)(6 in each group).Different exercise interventions were used in each group.After 8 weeks of exercise training(6 days a week),changes in body weight and blood glucose before and after exercise intervention were measured.EPCs were isolated from mouse bone marrow by density gradient centrifugation,immunofluorescence DilAcLDL and FITC-UEA-1 double positive staining and flow cytometry were used to detect the expression of CD133 surface markers.EPCs identification.The PI3K/AKT pathway inhibitor LY294002 was added to observe the in vitro angiogenic ability of EPCs and the expression of Caveolin1,AKT,PI3 K and miRNA-92 a.The BD Matrigel ? matrix method detects the in vitro angiogenic capacity of EPCs.Real-time PCR and Western Blot were used to detect the expression of Caveolin1,AKT,PI3 K and miRNA-92 a.Statistical analysis was performed using SPSS 22.0 statistical software.ResultsThe protein expression levels of Phospho-AKT and AKT in EPCs in the aerobic combined resistance exercise group were significantly higher than those in the control group(P<0.05).The mRNA expression levels of AKT,Caveolin1 and PI3 K in EPCs of the aerobic combined resistance exercise group were higher than those of the control group(P<0.05).The expressions of miR-92a-1-5p,miR-92a-2-5p and miR-92a-3p in EPCs of aerobic combined anti-resistance group were significantly lower than those of the control group(P<0.05).Compared with the control group,the in vitro neovascularization of EPCs in the aerobic combined resistance exercise group had longer trunk length and more nodes,and the difference was statistically significant(P<0.05).After adding PI3K/AKT pathway inhibitor LY294002,the expression of AKT protein in EPCs of aerobic combined resistance exercise group decreased by 48.33% and that of Phospho-AKT decreased by 49.72%,the difference was statistically significant(P<0.05).After adding PI3K/AKT inhibitor LY294002,the mRNA expression of AKT in aerobic combined resistance exercise group decreased by 56.38% and PI3 K decreased by 95.16%,the difference was statistically significant(P<0.05).After adding PI3K/AKT pathway inhibitor LY294002,the expression level of Caveolin1 mRNA in aerobic combined resistance exercise group increased by 56.61%,the difference was statistically significant(P<0.05).After the addition of the PI3K/AKT pathway inhibitor,the expression level of miR-92a-1-5p in the aerobic combined anti-resistance group increased by 44.83%,miR-92a-2-5p increased by 38.56%,and miR-92a-3p increased by 37.14%.The difference was statistically significant(P<0.05).PI3K/AKT inhibitors inhibited the angiogenesis function of EPCs in vitro,resulting in shortened trunk length and node reduction in vitro,and the difference was statistically significant(P<0.05).The microscopic view of Figure 23 A directly observed sparse and short vessels..Conclusions1.Aerobic combined resistance exercise can effectively improve the angiogenic function of EPCs in type 2 diabetic mice,and increase AKT and decrease the expression of miRNA-92 a.2.Adding PI3K/AKT pathway inhibitor can effectively inhibit the expression of AKT and PI3 K in EPCs of aerobic combined resistance exercise group,and up-regulate the expression of miRNA-92 a,thereby inhibiting the angiogenesis function of EPCs.3.Aerobic combined resistance exercise may down-regulate the expression of miRNA-92 a by activating the AKT pathway and promote the angiogenesis of EPCs in type 2 diabetic mice.