Renal ischemia–reperfusion injury is a major cause of acute kidney injury. We previously found that renal A₁ adenosine receptor (A₁AR) activation attenuated multiple cell death pathways including necrosis, apoptosis, and inflammation. Here, we tested whether induction of cytoprotective sphingosine kinase (SK)-1 and sphingosine-1-phosphate (S1P) synthesis might be the mechanism of protection. A selective A₁AR agonist (CCPA) increased the synthesis of S1P and selectively induced SK1 in mouse kidney and HK-2 cells. This agonist failed to protect SK1-knockout but protected SK2-knockout mice against renal ischemia–reperfusion injury indicating a critical role of SK1 in A₁AR-mediated renal protection. Inhibition of SK prevented A₁AR-mediated defense against necrosis and apoptosis in HK-2 cells. A selective S1P₁R antagonist (W146) and global in vivo gene knockdown of S1P₁Rs with small interfering RNA completely abolished the renal protection provided by CCPA. Mice selectively deficient in renal proximal tubule S1P₁Rs (S1P₁R^f/f PEPCK^Cre/–) were not protected against renal ischemia–reperfusion injury by CCPA. Mechanistically, CCPA increased nuclear translocation of hypoxia-inducible factor-1α in HK-2 cells and selective hypoxia-inducible factor-1α inhibition blocked A₁AR-mediated induction of SK1. Thus, proximal tubule SK1 has a critical role in A₁AR-mediated protection against renal ischemia–reperfusion injury.