The Combined Effect Of ATG5 And ATG7 On Autophagy And Ers Via PERK Signaling,and The Effect Of D-EAK On Mouse Osteoarthritis

ObjectiveTo explore the biological function of autophagy-related gene 5(ATG5)and autophagy-related gene 7(ATG7)in C28I2 cells,and further determine its relationship with endoplasmic reticulum stress.Forthermore,to investigate the effect of D-EAK on mouse osteoarthritis.MethodsUsing adenovirus construction and packaging technology,Ad-ATG5and Ad-ATG7 were successfully packaged.The expression of ATG5 and ATG7 in C28I2 cells was detected by RT-PCR,q PCR and Western blot methods.By performing different virus groups,detecting the combined effect of ATG5 and ATG7 on autophagy,ERS,proliferation and apoptosis in C28I2 chondrocytes,and further exploring the relationship between ATG5 and ATG7-induced biological function and endoplasmic reticulum stress.In addition,D-EAK oligopeptide was injected into the joint cavity of mouse with osteoarthritis to investigate the therapeutic effect of D-EAK.ResultsThe recombinant adenovirus Ad-ATG5(4×10~8pfu/ml)and Ad-ATG7(6×10~8pfu/ml)were successfully contructed.The combined effect of Ad-ATG5 and Ad-ATG7 could further promote autophagy and proliferation(P<0.05),and inhibit apoptosis and endoplasmic reticulum stress(P<0.05).However,inhibition of PERK pathway can significantly reverse the effect of Ad-ATG5 and Ad-ATG7.It demonstrated that ATG5and ATG7 regulate autophagy and ERS through the PERK signaling.In addition,the usage of D-EAK oligopeptide can effectively treat mouse osteoarthritis.ConclusionThe combined effect of ATG5 and ATG7 on autophagy and ERS via PERK signaling.The usage of D-EAK oligopeptide can effectively treat mouse osteoarthritis.