Arginase inhibition mediates renal tissue protection in diabetic nephropathy by a nitric oxide synthase 3-dependent mechanism

Recently we showed that pharmacological blockade or genetic deficiency of arginase-2 confers kidney protection in diabetic mouse models. Here we tested whether the protective effect of arginase inhibition is nitric oxide synthase-3 (eNOS)-dependent in diabetic nephropathy. Experiments were conducted in eNOS knockout and their wild type littermate mice using multiple low doses of vehicle or streptozotocin and treated with continuous subcutaneous infusion of vehicle or the arginase inhibitor S-(2-Boronoethyl)-L-cysteine by an osmotic pump. Inhibition of arginases for 6 weeks in diabetic wild type mice significantly attenuated albuminuria, the increase in plasma creatinine and blood urea nitrogen, histopathological changes, kidney fibronectin and TNF-α expression, kidney macrophage recruitment, and oxidative stress compared to vehicle-treated diabetic wild type mice. Arginase inhibition in diabetic eNOS knockout mice failed to affect any of these parameters but reduced kidney macrophage recruitment and kidney TNF-α expression compared to vehicle-treated diabetic eNOS knockout mice. Furthermore, diabetic wild type and eNOS knockout mice exhibited increased kidney arginase-2 protein, arginase activity and ornithine levels. Thus, arginase inhibition mediates renal tissue protection in diabetic nephropathy by an eNOS-dependent mechanism and has an eNOS-independent effect on kidney macrophage recruitment.