Previous studies have indicated that inhibitors of the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE) protect against ischemia reperfusion injury (IRI) in the brain and heart. However, the role of endogenous 20-HETE in renal IRI has not been well characterized. The present study compared the susceptibility to IRI in Dahl salt-sensitive (S) rats in which the renal formation of 20-HETE is impaired versus that seen in normal Sprague-Dawley (SD) rats and in SS.5LEW 4A+ (4A+) congenic and SS.BN5 (BN5) consomic strains in which the CYP4A region of chromosome 5 was transferred from the Lewis or Brown Norway rats into the S genetic background. S, 4A+ and BN5 rats were fed a low salt diet and SD rats were fed a standard diet. IRI was induced by bilateral renal ischemia for 30 minutes followed by reperfusion. The 20-HETE production was measured using liquid chromatography-mass spectrometry (LC/MS). The renal production level of 20-HETE was significantly greater in SD, 4A+ and BN5 rats as compared to S rats and renal 20-HETE levels rose to a greater extent in these strains than in S rats following renal ischemia reperfusion. Plasma creatinine concentration rose to 3.7 ± 0.1 (n=6), 2.0 ± 0.4 (n=6), 1.9 ± 0.3 (n=6) and 2.2 ± 0.3 (n=6) mg/dl 24 hours following renal ischemia in S, 4A+, BN5 and SD rats, respectively. The kidney of S rats following IRI exhibited renal tubular necrosis and protein casts. The % of necrotic tubules in the corticomedullary regions of S rats (42.5 ± 3.5 %) was significantly higher than that seen in 4A+ (10.8 ± 3.8 %) or BN5 (10.2 ± 1.5 %) rats. The number of apoptotic tubular cells in the corticomedullary regions of S rats (125.3 ± 8.2 cells/field) was significantly higher than that seen in BN5 rats (46.8 ± 5.1 cells/field). Administration of the 20-HETE antagonist (6, 15-20-hydroxyeicosadecanoic acid) or the 20-HETE synthesis inhibitor (HET0016) abolished the resistance of 4A+ and BN5 rats to renal IRI and plasma creatinine levels rose to 4.0 ± 0.4 (n=4) and 3.8 ± 0.1 (n=4) mg/dl, respectively. The % of necrotic tubules respectively increased to 33.6 ± 6.9 % and 38.5 ± 5.5 % in 4A+ and BN5 rats and the number of apoptotic tubular cells increased to 119.2 ± 10.7 cells/field in BN5 rats. These data indicate that endogenously released 20-HETE plays a protective role in renal IRI.