MiR-184 Targets FOXO1 Regulates Human Bronchial Epithelial Cell Oxidative Stress Induced By Chlamydia Psittaci Through Wnt/β-catenin Signaling Pathway

Objective: To explore the effect of miR-184 targeting FOXO1 on the oxidative stress in host cells infected with Chlamydia psittaci,and the regulatory mechanism of Wnt/β-catenin signaling pathway on this process,providing a new experimental basis for further understanding the pathogenesis of C.psittaci.Methods: Human bronchial epithelial cells(HBE)were infected with C.psittaci with a multiplicity of infection(MOI)of 3 for 44 h,and the levels of miR-184,FOXO1,ROS,MDA,SOD,and GSH were detected to explore the effects of C.psittaci infection on oxidative stress and the expression levels of miR-184 and FOXO1 in C.psittaci-infected HBE cells.Additionally,in vitro transfection of miR-184 inhibitor or miR-184 mimic or co-transfection with FOXO1 siRNA and miR-184 inhibitor,detection of miR-184 and FOXO1 levels after C.psittaci infection,to evaluate the correlation between miR-184 and FOXO1,and detection of ROS,MDA,SOD and GSH levels,to evaluate the effect of miR-184 or FOXO1 on oxidative stress induced by C.psittaci infection,and the levels of Wnt3 a,β-catenin,C-myc and Cyclin D1 were detected in order to evaluate the activation of Wnt/β-catenin signaling pathway by miR-184 or FOXO1 after C.psittaci infection.Finally,after transfection with miR-184 mimic or FOXO1 siRNA,HBE cells were treated with Wnt/β-catenin signaling pathway inhibitor ICG-001 for 2 h,and then HBE cells were infected with C.psittaci with MOI of 3 for 44 h.The function of Wnt/β-catenin signaling pathway in C.psittaci-induced oxidative stress was also elucidated by detecting the levels of Wnt3 a,β-catenin,C-myc,Cyclin D1,ROS,MDA,SOD,and GSH.Results: After HBE cells were infected with C.psittaci with MOI of3 for 44 h,the expression of miR-184 was up-regulated,while FOXO1 was down-regulated,and the levels of ROS,MDA were increased,and the levels of SOD,GSH were decreased.The expression of miR-184,Wnt3 a,β-catenin,C-myc,and Cyclin D1 were down-regulated in HBE cells infected with C.psittaci after transfected with miR-184 inhibitor,while FOXO1 was up-regulated,and the levels of ROS,MDA were decreased,and the levels of SOD,GSH were increased.However,after HBE cells were transfected with miR-184 mimic,the change trend was opposite.Compared with the cells only transfected with miR-184 inhibitor,the expression of Wnt3 a,β-catenin,C-myc,and Cyclin D1 were up-regulated in HBE cells co-transfected with FOXO1 siRNA and miR-184 inhibitor infected with C.psittaci,while FOXO1 was down-regulated,and the levels of ROS,MDA were increased,and the levels of SOD,GSH were decreased.After ICG-001 treatment,the expression of Wnt3 a,β-catenin,C-myc,and Cyclin D1 were down-regulated,and the levels of ROS,MDA were decreased,and the levels of SOD,GSH were increased in C.psittaci infected HBE cells.Conclusion: These data suggest that C.psittaci infection induces oxidative stress in HBE cells,and C.psittaci infection can up-regulate the expression of miR-184 and down-regulate the expression of FOXO1 in HBE cells.Furthermore,miR-184 targeting FOXO1 can regulate C.psittaci-induced oxidative stress in HBE cells by activating the Wnt/β-catenin signaling pathway.