Angiotensin II is a critical mediator of diabetic nephropathy. Pharmacologic inhibition of Ang II slows disease progression beyond what could be predicted by the blood pressure lowering effects alone, suggesting the importance of non-hemodynamic pathways of Ang II in mediating disease. Podocyte injury and loss are cardinal features of diabetic nephropathy. Mounting evidence suggests that the podocyte is a direct target of Ang II-mediated signaling in diabetic renal disease. We have tested the hypothesis that high glucose leads to the activation of a local angiotensin system in podocytes and delineated the underlying pathways involved. Cultured podocytes were exposed to standard glucose (5mM), high glucose (40 mM) or mannitol as an osmotic control. Angiotensin II levels in cell lysates were measured in the presence or absence of inhibitors of angiotensin converting enzyme (captopril), chymase (chymostatin), and renin (aliskiren) activity. The effects of glucose on renin and AT1R expression and protein levels were determined. Exposure to high glucose resulted in a 2.1-fold increase in Ang II levels mediated through increased renin activty, as exposure to high glucose increased renin levels and pre-incubation with Aliskiren abrogated glucose-induced Ang II production. Relevance to the in vivo setting was demonstrated by showing glomerular upregulation of the (pro)renin receptor in a podocyte distribution early in the course of experimental diabetic nephropathy. Furthermore, high glucose increased AT1R levels by immunofluorescence and western blot. Taken together, the resultant activation of a local renin angiotensin system by high glucose may promote progressive podocyte injury and loss in diabetic nephropathy.