Epidermal growth factor (EGF) is a potent mitogen implicated in gastroprotection and ulcer healing, but its possible interaction with nitric oxide (NO) and sensory nerves on healing after acute gastric damage has not been assessed. We examined the effects of topical application of a small dose (0.5 mg/kg) of capsaicin to stimulate sensory nerves and a larger parenteral dose of capsaicin (125 mg/kg s.c.) to deactivate these neurons or the effect of systemic administration of NG-nitro-L-arginine methyl ester (L-NAME) (20 mg/kg i.v.) to suppress NO synthase on healing of gastric lesions induced by 3.5 h of water immersion and restraint stress (WRS) in rats without or with EGF administration. Rats were sacrificed at 0, 6, 12, or 24 h after WRS and the gastric blood flow (GBF) was measured by the H2 gas clearance technique. Exposure to WRS produced many gastric lesions, with a marked decrease in GBF, but at 12 h these lesions started to heal and the lesion number was reduced by 75% after 24 h. This was accompanied by progressive increase in the GBF and an increase in expression of EGF mRNA in gastric mucosa, as detected by RT-PCR. Pretreatment with L-NAME or functional ablation of sensory nerves by capsaicin significantly delayed the healing of WRS lesions and accompanying hyperemia. In contrast, pretreatment with EGF (100 micrograms/kg s.c.) or glyceryl trinitrate (10 mg/kg i.g.), a donor of NO, or stimulation of sensory nerves by topical capsaicin significantly enhanced the healing of these lesions and increased the GBF. The acceleration of the healing and accompanying hyperemia induced by EGF at 12 h after WRS were completely reversed in rats pretreated with L-NAME or in those with capsaicin denervation. Addition of L-arginine but not D-arginine to L-NAME restored the healing of stress lesions and gastric hyperemia induced by this peptide. Removal of salivary glands, which reduced luminal content of EGF and DNA synthesis by about fourfold compared to rats with intact glands, produced a significant delay in healing, and this was further aggravated by capsaicin denervation. We conclude that EGF, sensory nerves, and NO play an important role in the healing of gastric mucosa from lesions induced by stress and that sensory nerves and NO appear to interact with EGF in the mechanism of mucosal recovery from stress lesions.