Glutathione (GSH) protects against oxidative damage in lots of tissues, including retinal pigment epithelium (RPE). Oxidative stress-mediated senescence and dying of RPE and subsequent dying of photoreceptors happen to be noticed in age-related macular degeneration (AMD). Even though the effects of GSH depletion happen to be described formerly, questions remain concerning the molecular mechanisms. We herein examined the downstream results of GSH depletion on stress-caused premature senescence (SIPS) and cell dying in human RPE cells. Briefly, cultured ARPE-19 cells were depleted of GSH using: (1) incubation in cystine (Cys2)-free culture medium (2) treatment with buthionine sulphoximine (BSO, 1000 |¨¬M) to bar de novo GSH synthesis for twenty-four-48 h or (3) treatment with erastin (10 |¨¬M for 12-24 h) to hinder Cys2/glutamate antiporter (system xc-). These treatments decreased cell viability and elevated both soluble and fat reactive oxygen species (ROS) generation but didn’t affect mitochondrial ROS or mitochondrial mass. Western blot analysis revealed decreased expression of ferroptotic modulator glutathione peroxidase 4 (GPX4). Elevated autophagy was apparent, as reflected by elevated LC3 expression, autophagic vacuoles, and autophagic flux. Additionally, GSH depletion caused SIPS, as evidenced by elevated number of the senescence-connected |?-galactosidase-positive cells, elevated senescence-connected heterochromatin foci (SAHF), in addition to cell cycle arrest in the G1 phase. GSH depletion-dependent cell dying was avoided by selective ferroptosis inhibitors (8 |¨¬M Fer-1 and 600 nM Lip-1), iron chelator DFO (80 |¨¬M), in addition to autophagic inhibitors Baf-A1 (75 nM) and three-MA (10 mM). Inhibiting autophagy with Baf-A1 (75 nM) or 3-MA (10 mM) promoted SIPS. In comparison, inducing autophagy with rapamycin (100 nM) attenuated SIPS. Our findings claim that GSH depletion induces ferroptosis, autophagy, and SIPS. Additionally, we discovered that autophagy is activated while ferroptosis and reduces SIPS, suggesting an important role of autophagy in ferroptosis and SIPS.