Aim βguanadinopropionic acid (βGPA), a creatine analog has been demonstrated to lead to the chronic activation of AMPK with increasing mitochondrial biogenesis.However, little is known about the effect ofβGPA on lifespan extension.In current study, we explore whether βGPA produces the lifespan extension in adult Drosophila and underlying mechanism.Methods The flies were fed in a normal chow diet containing different concentrations (300 mM,900 mM,2700 mM)of βGPA.Western blot was also used to investigate protein expressions.RNA interference was employed to examine effects of lifespan in Drosophila by βGPA.Results 900 mM and 2700 mM βGPA significant extended the lifespan and activated AMPK both in male and female Drosophila with elevated level of autophagy.We found that Atg8 was significantly upregulated and P62 was significantly downregulated by 900 mM βGPA, on the other hand, when the expression of Atg5 protein, an essential protein for autophagy, was reduced by RNAi, the effect of βGPA on lifespan extension was abolished.Inhibition of AMPK by either AMPKRNAi or compound C, significantly attenuated the expression of autophagyrelated proteins and lifespan extension in Drosophila.We found that the activation of AMPK induced by βGPA inhibited mTORC1 activity,implicating that mTORC1 may not prevent the activation of ULK1/Atgl by AMPK.Conclusion βGPA can extend the lifespan of Drosophila and this effect is mediated by AMPKAtgl dependent autophagy.