Objective:To investigate the effect of glycine on the ATP depletion-induced autophagy and mechanism underlying it. Methods:GFP-LC3 plasmid was transiently transfected into Madin-Darby canine kidney (MDCK) cells with lipofectamine. Autophagosome in cells was observed under an inverted fluorescence microscope. Expression of GFP-LC3-Ⅱ/LC3-Ⅰ and the phosphorylation activity of AMPK in the transfected cells were analyzed by western blot. Results:We found that the clear punctate fluorescence particles were dispersed in the cytoplasm of the transfected cells. When ATP was deprived off by addition of rotenone and antimycin into MDCK cells,autophagosome and GFP-LC3-Ⅱ/LC3-Ⅰratio were increased and AMPK was activated in cells. Treatment with glycine resulted in decrease in autophagosome and GFP-LC3-Ⅱ/LC3-Ⅰ expression in the ATP-depleted cells. Treatment with glycine also inhibited phosphorylation of AMPK in the ATP-depleted cells. Conclusion:ATP depletion induces an autophagic cell death,which could be inhibited by glycine. The inhibitory effect of glycine on the ATP depletion-induced autophagy may be involved in the depression of AMPK phosphorylation.