Eukaryotic elongation factor 2 (eEF2) kinase (eEF2K) is a calmodulin-related protein kinase that inhibits protein translation via phosphorylating a specific substrate, eEF2. We previously demonstrated that eEF2K expression increases in mesenteric artery from spontaneously hypertensive rats (SHR) and that eEF2K mediates hypertension development in SHR. We recently revealed that A484954, a selective eEF2K inhibitor lowers blood pressure in SHR via vasorelaxation through the production of endothelium-derived relaxing factors. In this study, we examined whether A484954 induces diuretic effect. After male SHR and normotensive Wistar Kyoto rats (WKY) were given a single intraperitoneal injection of A484954 (2.5 mg/kg) or vehicle (0.5% carboxymethyl cellulose), urine was collected using metabolic cage. Isometric contraction of isolated renal arteries form SHR was measured. While A484954 did not induce diuretic effect in WKY, it increased urine output and water intake (6 and 9 hours after injection) in SHR. A484954 (0.01-10 μM) induced concentration-dependent relaxation in renal arteries, which was inhibited by pretreatment with a β blocker, propranolol. In summary, it was suggested that A484954 induces diuretic effect in SHR perhaps via renal vasorelaxation through β adrenergic receptor and subsequent increase in renal blood flow.