Inhibition of P2Y2 Attenuates Cisplatin-Induced AKI via Reduced Oxidative Stress, Inflammation and Cell Death.

Abstract

Introduction: Purinergic signaling has been recognized as important extracellular regulator in multiple physiological and pathophysiological conditions. Adenosine triphosphate-purinergic receptor P2Y2 signaling pathway is associated with glomerular nephritis (GN), diabetic nephropathy (DN), and chronic kidney disease. Recently, there has been evidence that global knockout of P2Y2 exacerbated bilateral ischemic reperfusion-induced acute kidney injury (AKI). However, its role in cisplatin-induced AKI (CIA) remains unknown. Cisplatin is a platinum-containing antineoplastic drug widely used in variety of solid malignant tumors. Nephrotoxicity is one of the major serious side effect that limit its clinical use. In the present study, we investigated whether inhibition of P2Y2 has an effect on CIA.

Methods: We used AR-C118925 (AR-C), a selective antagonist of P2Y2, and gene transfection for interruption of the P2Y2 pathway. Mice were pretreated with AR-C (10 mg/kg/day) and then challenged with cisplatin at a dose of 20 mg/kg. Seventy-two hours after cisplatin administration, all mice developed kidney failure. Knockdown and overexpression of P2Y2 in mice and mouse proximal tubular cells (mPTCs) were employed to validate that ARC acts through P2Y2 receptor.

Results: AR-C markedly ameliorated cisplatin-induced nephrotoxicity evidenced by improved renal function, renal morphology, and tubular injury marker expression. Further analysis of the mechanism revealed that AR-C significantly reduced kidney oxidative stress, inflammation, apoptosis, and necroptosis. Consistently, AR-C protects mPTCs from injury caused by cisplatin. To verify that AR-C acts through the P2Y2 receptor, we knocked down P2Y2 in mice or in mPTC cells. Both showed beneficial effects, while overexpression of P2Y2 promotes cisplatin-induced cell death.

Conclusion: Taken together, our study, for the first time revealed that P2Y2 plays an important role in CIA by regulating oxidative stress, inflammation, apoptosis, and necroptosis and its inhibitor, AR-C, is a potential drug for treating CIA.