Uric acid promotes apoptosis in human proximal tubule cells by oxidative stress and the activation of NADPH oxidase NOX 4
Daniela Verzola, Elena Ratto, Barbara Villaggio, Emanuele Luigi Parodi, Roberto Pontremoli, Giacomo Garibotto, Francesca Viazzi, Daniela Verzola, Elena Ratto, Barbara Villaggio, Emanuele Luigi Parodi, Roberto Pontremoli, Giacomo Garibotto, Francesca Viazzi
Abstract
Mild hyperuricemia has been linked to the development and progression of tubulointerstitial renal damage. However the mechanisms by which uric acid may cause these effects are poorly explored. We investigated the effect of uric acid on apoptosis and the underlying mechanisms in a human proximal tubule cell line (HK-2). Increased uric acid concentration decreased tubule cell viability and increased apoptotic cells in a dose dependent manner (up to a 7-fold increase, p<0.0001). Uric acid up-regulated Bax (+60% with respect to Ctrl; p<0.05) and down regulated X-linked inhibitor of apoptosis protein. Apoptosis was blunted by Caspase-9 but not Caspase-8 inhibition. Uric acid induced changes in the mitochondrial membrane, elevations in reactive oxygen species and a pronounced up-regulation of NOX 4 mRNA and protein (p<0.05). In addition, both reactive oxygen species production and apoptosis was prevented by the NADPH oxidase inhibitor DPI as well as by Nox 4 knockdown. URAT 1 transport inhibition by probenecid and losartan and its knock down by specific siRNA, blunted apoptosis, suggesting a URAT 1 dependent cell death. In summary, our data show that uric acid increases the permissiveness of proximal tubule kidney cells to apoptosis by triggering a pathway involving NADPH oxidase signalling and URAT 1 transport. These results might explain the chronic tubulointerstitial damage observed in hyperuricaemic states and suggest that uric acid transport in tubular cells is necessary for urate-induced effects.
Conflict of interest statement
Competing Interests: The authors have declared that no competing interests exist.
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References
- Obermayr RP, Temml C, Gutjahr G, Knechtelsdorfer M, Oberbauer R, et al. (2008) Elevated uric acid increases the risk for kidney disease. J Am Soc Nephrol 19:2407–2013.
- Weiner DE, Tighiouart H, Elsayed EF, Griffith JL, Salem DN, et al. (2008) Uric acid and incident kidney disease in the community. J Am Soc Nephrol 19:1204–1211.
- Iseki K, Oshiro S, Tozawa M, Iseki C, Ikemiya Y, et al. (2001) Significance of hyperuricemia on the early detection of renal failure in a cohort of screened subjects. Hypertens Res 24:691–697.
- Johnson RJ, Nakagawa T, Jalal D, Sánchez-Lozada LG, Kang DH, et al. (2013) Uric acid and chronic kidney disease: which is chasing which? Nephrol Dial Transplant 28:2221–2228.
- Viazzi F, Leoncini G, Ratto E, Falqui V, Parodi A, et al. (2007) Mild hyperuricemia and subclinical renal damage in untreated primary hypertension. Am J Hypertens 20:1276–1282.
- Jalal DI, Rivard CJ, Johnson RJ, Maahs DM, McFann K, et al. (2010) Serum uric acid levels predict the development of albuminuria over 6 years in patients with type 1 diabetes: findings from the Coronary Artery Calcification in Type 1 Diabetes study. Nephrol Dial Transplant 25:1865–1869.
- Zoppini G, Targher G, Chonchol M, Ortalda V, Abaterusso C, et al. (2012) Serum uric acid levels and incident chronic kidney disease in patients with type 2 diabetes and preserved kidney function. Diabetes Care 35:99–104.
- Ohno I, Hosoya T, Gomi H, Ichida K, Okabe H, et al. (2001) Serum uric acid and renal prognosis in patients with IgA nephropathy. Nephron 87:333–339.
- Syrjänen J, Mustonen J, Pasternack A (2001) Hypertriglyceridaemia and hyperuricaemia are risk factors for progression of IgA nephropathy. Nephrol Dial Transplant 15:34–42.
- Sánchez-Lozada LG, Lanaspa MA, Cristóbal-García M, García-Arroyo F, Soto V, et al. (2012) Uric acid-induced endothelial dysfunction is associated with mitochondrial alterations and decreased intracellular ATP concentrations. Nephron Exp Nephrol 121:e71–78.
- Sautin YY, Nakagawa T, Zharikov S, Johnson RJ (2007) Adverse effects of the classic antioxidant uric acid in adipocytes: NADPH oxidase-mediated oxidative/nitrosative stress. Am J Physiol Cell Physiol 293:C584–C596.
- Ray PD, Huang BW, Tsuji Y (2012) Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 24:981–990.
- Sautin YY, Johnson RJ (2008) Uric acid: the oxidant-antioxidant paradox. Nucleosides Nucleotides Nucleic Acids 27:608–661.
- Nieto FJ, Iribarren C, Gross MD, Comstock GW, Culter RG (2000) Uric acid and serum antioxidant capacity: a reaction to atherosclerosis? Atherosclerosis 148:131–139.
- Kanellis J, Kang DH (2005) Uric acid as a mediator of endothelial dysfunction, inflammation, and vascular disease. Semin Nephrol 25:39–42.
- Kanellis J, Watanabe S, Li JH, Kang DH, Li P, et al. (2003) Uric acid stimulates monocyte chemoattractant protein-1 production in vascular smooth muscle cells via mitogen-activated protein kinase and cyclooxygenase-2. Hypertension 41:1287–1293.
- Kang DH, Han L, Ouyang X, Kahn AM, Kanellis J, et al. (2005) Uric acid causes vascular smooth muscle cell proliferation by entering cells via a functional urate transporter. Am J Nephrol 25:425–433.
- Sánchez-Lozada LG, Tapia E, Santamaría J, Avila-Casado C, Soto V, et al. (2005) Mild hyperuricemia induces vasoconstriction and maintains glomerular hypertension in normal and remnant kidney rats. Kidney Int 67:237–247.
- Kang DH, Park SK, Lee IK, Johnson RJ (2005) Uric acid-induced C-reactive protein expression: implication on cell proliferation and nitric oxide production of human vascular cells. J Am Soc Nephrol 16:3553–3562.
- Feig DI, Mazzali M, Kang DH, Nakagawa T, Price K, et al. (2006) Serum uric acid: a risk factor and a target for treatment? J Am Soc Nephrol 17 Suppl 2: 4):S69–73.
- Cirillo P, Gersch MS, Mu W, Scherer PM, Kim KM, et al. (2009) Ketohexokinase-dependent metabolism of fructose induces proinflammatory mediators in proximal tubular cells. J Am Soc Nephrol 20:545–553.
- Quan H, Peng X, Liu S, Bo F, Yang L, et al. (2011) Differentially expressed protein profile of renal tubule cell stimulated by elevated uric acid using SILAC coupled to LC-MS. Cell Physiol Biochem 27:91–98.
- Gasse P, Riteau N, Charron S, Girre S, Fick L, et al. (2009) Uric acid is a danger signal activating NALP3 inflammasome in lung injury inflammation and fibrosis. Am J Respir Crit Care Med 179:903–913.
- Wang C, Pan Y, Zhang QY, Wang FM, Kong LD (2012) Quercetin and allopurinol ameliorate kidney injury in STZ-treated rats with regulation of renal NLRP3 inflammasome activation and lipid accumulation. PLoS One 7:e38285.
- Havasi A, Borkan SC (2011) Apoptosis and acute kidney injury. Kidney Int 80:29–40.
- Ortiz A (2000) Nephrology forum: apoptotic regulatory proteins in renal injury. Kidney Int 58:467–485.
- Lhotta K, Gruber J, Sgonc R, Fend F, König P (1998) Apoptosis of tubular epithelial cells in familial juvenile gouty nephropathy. Nephron 79:340–344.
- Capellino S, Montagna P, Villaggio B, Soldano S, Straub RH, et al. (2008) Hydroxylated estrogen metabolites influence the proliferation of cultured human monocytes: possible role in synovial tissue hyperplasia. Clin Exp Rheumatol 26:903–909.
- Nicholson DW (1996) ICE/CED-3-like proteases as therapeutic targets for the control of inappropriate apoptosis. Nat Biotech 14:297–301.
- Miyazaki H, Sekine T, Endou H (2004) The multispecific organic anion transporter family: properties and pharmacological significance. Trends Pharmacol Sci 25:654–662.
- Hamada T, Ichida K, Hosoyamada M, Mizuta E, Yanagihara K, et al. (2008) Uricosuric action of losartan via the inhibition of urate transporter 1 (URAT 1) in hypertensive patients. Am J Hypertens 21:1157–1162.
- Palm F, Nordquist L. (2011) Renal tubulointerstitial hypoxia: cause and consequence of kidney dysfunction. Clin Exp Pharmacol Physiol 38:474–480.
- Sedeek M, Callera G, Montezano A, Gutsol A, Heitz F, et al. (2010) Critical role of Nox4-based NADPH oxidase in glucose-induced oxidative stress in the kidney: implications in type 2 diabetic nephropathy. Am J Physiol Renal Physiol 299:F1348–1358.
- Gorin Y, Block K, Hernandez J, Bhandari B, Wagner B, et al. (2005) Nox4 NAD(P)H oxidase mediates hypertrophy and fibronectin expression in the diabetic kidney. J Biol Chem 280:39616–39626.
- Gill PS, Wilcox CS (2006) NADPH oxidases in the kidney. Antioxid Redox Signal 8:1597–1607.
- Schreck C, O'Connor PM (2011) NAD(P)H oxidase and renal epithelial ion transport. Am J Physiol Regul Integr Comp Physiol 300:R1023–1029.
- Barnes JL, Gorin Y (2011) Myofibroblast differentiation during fibrosis: role of NAD(P)H oxidases. Kidney Int 79:944–956.
- New DD, Block K, Bhandhari B, Gorin Y, Abboud HE (2011) IGF-I increases the expression of fibronectin by Nox4-dependent Akt phosphorylation in renal tubular epithelial cells. Am J Physiol Cell Physiol 302: C122–130, 2012.
- Susztak K, Raff AC, Schiffer M, Böttinger EP (2006) Glucose-induced reactive oxygen species cause apoptosis of podocytes and podocyte depletion at the onset of diabetic nephropathy. Diabetes : 225–233.
- Lodha S, Dani D, Mehta R, Bhaskaran M, Reddy K, et al. (2002) Angiotensin II-induced mesangial cell apoptosis: role of oxidative stress. Mol Med 8:830–840.
- Liu Y (2009) Advanced oxidation protein products: a causative link between oxidative stress and podocyte depletion. Kidney Int 76:1125–1127.
- Morais C, Westhuyzen J, Metharom P, Healy H (2005) High molecular weight plasma proteins induce apoptosis and Fas/FasL expression in human proximal tubular cells. Nephrol Dial Transplant 20:50–58.
- Tsuda H, Kawada N, Kaimori JY, Kitamura H, Moriyama T, et al. (2012) Febuxostat suppressed renal ischemia-reperfusion injury via reduced oxidative stress. Biochem Biophys Res Commun 427:266–272.
- Hansell P, Welch WJ, Blantz RC, Palm F (2013) Determinants of kidney oxygen consumption and their relationship to tissue oxygen tension in diabetes and hypertension. Clin Exp Pharmacol Physiol 40:123–137.
- Sanchez-Niño MD, Benito-Martin A, Ortiz A (2010) New paradigms in cell death in human diabetic nephropathy. Kidney Int 78:737–744.
- Devaraux QL (1998) IAPs block apoptotic events induced by Caspase 8 and cytochrome c by direct inhibition of distinct caspases. EMBO J 17:2215–2223.
- Schimmer AD, Dalili S, Batey RA, Riedl SJ (2006) Targeting XIAP for the treatment of malignancy. Cell Death Differ 13:179–188.
- Han HJ, Lim MJ, Lee YJ, Lee JH, Yang IS, et al. (2007) Uric acid inhibits renal proximal tubule cell proliferation via at least two signaling pathways involving PKC, MAPK, cPLA2, and NF-kappaB. Am J Physiol Renal Physiol 292:F373–381.
- Bose B, Badve SV, Hiremath SS, Boudville N, Brown FG, et al. (2014) Effects of uric acid-lowering therapy on renal outcomes: a systematic review and meta-analysis. Nephrol Dial Transplant 29:406–413.
- Sánchez-Lozada LG, Tapia E, Soto V, Avila-Casado C, Franco M, et al. (2008) Treatment with the xanthine oxidase inhibitor febuxostat lowers uric acid and alleviates systemic and glomerular hypertension in experimental hyperuricaemia. Nephrol Dial Transplant 23:1179–1185.
- Høieggen A, Alderman MH, Kjeldsen SE, Julius S, Devereux RB, et al. (2004)The impact of serum uric acid on cardiovascular outcomes in the LIFE study. Kidney Int 65:1041–1049.
- Miao Y, Ottenbros SA, Laverman GD, Brenner BM, Cooper ME, et al. (2011) Effect of a reduction in uric acid on renal outcomes during losartan treatment: a post hoc analysis of the reduction of endpoints in non-insulin-dependent diabetes mellitus with the Angiotensin II Antagonist Losartan Trial. Hypertension 58:2–7.
- Smink PA, Bakker SJ, Laverman GD, Berl T, Cooper ME, et al. (2012) An initial reduction in serum uric acid during angiotensin receptor blocker treatment is associated with cardiovascular protection: a post-hoc analysis of the RENAAL and IDNT trials. J Hypertens 30:1022–1028.
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