β‐Guanidinopropionic acid extends the lifespan of D rosophila melanogaster via an AMP ‐activated protein kinase‐dependent increase in autophagy

Summary Previous studies have demonstrated that AMP ‐activated protein kinase ( AMPK ) controls autophagy through the mammalian target of rapamycin (mTOR) and Unc‐51 like kinase 1 ( ULK 1/Atg1) signaling, which augments the quality of cellular housekeeping, and that β‐guanidinopropionic acid (β‐ GPA ), a creatine analog, leads to a chronic activation of AMPK . However, the relationship between β‐ GPA and aging remains elusive. In this study, we hypothesized that feeding β‐ GPA to adult Drosophila produces the lifespan extension via activation of AMPK ‐dependent autophagy. It was found that dietary administration of β‐ GPA at a concentration higher than 900 m m induced a significant extension of the lifespan of Drosophila melanogaster in repeated experiments. Furthermore, we found that Atg8 protein, the homolog of microtubule‐associated protein 1A/1B‐light chain 3 ( LC 3) and a biomarker of autophagy in Drosophila , was significantly upregulated by β‐ GPA treatment, indicating that autophagic activity plays a role in the effect of β‐ GPA . On the other hand, when the expression of Atg5 protein, an essential protein for autophagy, was reduced by RNA interference ( RNA i), the effect of β‐ GPA on lifespan extension was abolished. Moreover, we found that AMPK was also involved in this process. β‐ GPA treatment significantly elevated the expression of phospho‐T172‐ AMPK levels, while inhibition of AMPK by either AMPK ‐ RNA i or compound C significantly attenuated the expression of autophagy‐related proteins and lifespan extension in Drosophila . Taken together, our results suggest that β‐ GPA can induce an extension of the lifespan of Drosophila via AMPK ‐Atg1‐autophagy signaling pathway.