The effects of hot water treatments on antioxidant responses in red sweet pepper (Capsicum annuum L.) fruit during cold storage were investigated. Red sweet pepper fruits were treated with hot water at 55 °C for 1 (HWT-1 min), 3 (HWT-3 min), and 5 min (HWT-5 min) and stored at 10 °C for 4 weeks. The results indicated that HWT-1 min fruit showed less development of chilling injury (CI), electrolyte leakage, and weight loss. Excessive hot water treatment (3 and 5 min) caused cellular damage. Moreover, HWT-1 min slowed the production of hydrogen peroxide and malondialdehyde and promoted the ascorbate and glutathione contents for the duration of cold storage as compared to HWT-3 min, HWT-5 min, and control. HWT-1 min enhanced the ascorbate-glutathione cycle associated with ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase, but it was less effective in simulating catalase activity. Thus, HWT-1 min could induce CI tolerance in red sweet pepper fruit by activating the ascorbate-glutathione cycle via the increased activity of related enzymes and the enhanced antioxidant level.