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Aims: To observe the protective effects of L-citrulline on the renal I/R injury and elucidate the mechanisms involved.Main methods: Forty-eight rats were randomized into eight groups: Group 1: sham operated; Group 2: I/R (45 min renal ischemia and 24 h reperfusion); Group 3: I/R + L-citrulline (300 mg/kg, i.g.); Group 4: I/R + L-citrulline (600 mg/kg, i.g.); Group 5: I/R + L-citrulline (900 mg/kg, i.g.); Group 6: I/R + normal saline (NS, i.g.); Group 7: I/R + N sup omega nitro-L-arginine ester (L-NAME, 20 mg/kg, i.p.); Group 8: I/R + L-citrulline (900 mg/kg, i.g.) + L-NAME (20 mg/kg, i.p.).At the end of the repeffusion period, serum was collected and the kidneys were removed for histological and biochemical examinations.Key findings: Our results showed that pretreatment with L-citrulline (300, 600, and 900 mg/kg) significantly ameliorated the renal injury caused by I/R.Moreover, L-citrulline prevented the production of lipid peroxidation and increased the activity of superoxide dismutase (SOD) and the levels of glutathione (GSH) and nitric oxide (NO).The I/R-induced decreases in total nitric oxide synthase (TNOS) activity, inducible NOS (iNOS) activity, constitutive NOS activity (cNOS) and endothelial NOS (eNOS) protein expression in the renal cortex were significantly prevented.However, the L-citrulline-mediated protection was significantly antagonized by coadministration of L-NAME.Significance: These results suggested that L-citrulline administration exhibited significant protection on renal I/R injury.The protective effect of L-citrulline on renal I/R injury, at least in part, via the up-regulation of the eNOS protein expression and cNOS activity, maintained the basal production of NO level.