Background and Objective: Licochalcone A (Lico A) has anti-inflammatory, oxidative and bacterial activities. However, the effect of Lico A on the oxidative damage of RGC in glaucoma has not been reported. This study investigated the mechanism of Lico A-mediated regulation of Hydrogen Peroxide (H2O2)-induced oxidative damage of RGC in glaucoma via p53/AMPK/mTOR pathway. Materials and Methods: The Lico A was extracted from the licoherbs residues. Oxidative damage of glaucoma retinal ganglion cells model (Mod group) was constructed using H2O2-induced RGC-5 cells and effects of 5, 10 and 20 μmol/L Lico A on cell proliferation, apoptosis, oxidative stress and expression of p53/AMPK/mTOR pathway-related proteins were analyzed. Results: The Lico A was successfully extracted and with the increase of Lico A concentration, the clearance of DPPH, ABTS, O2– and H2O2 also increased. As against Mod, the proliferation activity of RGC-5 was increased, the AR was decreased, the avidity of SOD and GSH-Px had an increase, the degree of MDA and the expression of p53, AMPK and LC3II/LC3I was decreased and the expression of mTOR was visibly raised in the 5, 10 and 20 μmol/L Lico A treatment groups. Conclusion: The Lico A has a strong antioxidant effect. In addition, Lico A can inhibit H2O2-induced apoptosis and autophagy in glaucomatous RGC through p53/AMPK/mTOR signaling pathway and improve OS injury.