Effects Of Recombinant Adenovirus-Mediated FOXO1and FOXO3a On Myocardial Cells In Hypoxic And Reoxygenated Mice

Objective:To investigate the effects of recombinant adenovirus-mediated FOXO1 and FOXO3 a on myocardial cells of hypoxic/reoxygenated mice.Methods: H9C2 mouse myocardial cells hypoxia 3 h/complex oxygen 6 h,and establish hypoxi a/complex oxygen injury model.H9C2 cells were randomly divided into 5 group s: group A(blank Control group/Control group)and group B(Model/anoxia and reoxygenation group),group C(FOXO1 + group B),group D(FOXO3a +grou p B),group E(FOXO1 + FOXO3 a +group B),CCK 8 method is used to dete ct the survival rate of mice cardiomyocytes after hypoxia reoxygenation;Brdu m ethod was used to detect cell proliferation activity.TUNEL method was used to detect apoptosis of cells.Transwell method and scratch test were used to detect cell migration.Real-time PCR method was used to detect the expression level of the transcription m RNA associated with FOXO1 and FOXO3 a genes in each cell.The expression of FOXO1 and FOXO3 a in cells was detected by Western blot.ELISA detection of oxidative stress related indicators(superoxide dismutase(SOD)activity and reactive oxygen species(ROS)activity and malondialdehyde(MDA)content,nitric oxide(NO)content and total antioxidant capacity(T-A OC)activity).Results: Compared with group B,C,D group,at the same time t ransfection FOXO1 and FOXO3 a in the group E,after FOXO1 and FOXO3 a pr otein expression significantly enhanced in H9C2 cells,cell proliferation activity,migration and invasion ability obviously increased(P < 0.05),apoptosis rate sig nificantly decreased,cell alpha MHC,Nkx2.5,GATA-4 mRNA expression leve l increased significantly,while beta MHC,cTn T mRNA level significantly decrea sed,anti oxidative stress indicators(SOD activity and T AOC activity)expression levels,The expression levels of oxidative stress(ROS activity,MDA content a nd NO content)were decreased.Conclusion: Recombinant adenovirus mediated F OXO1 and FOXO3 a can alleviate hypoxia/reoxygenation(H/R)caused by oxidati ve stress caused by the damage of myocardial cells,improve the myocardial cell proliferation and survival ability,for the gene therapy of ischemic heart disease heart failure provides a new train of thought.