Abstract
Chronic metabolic acidosis occurs commonly in chronic renal failure (CRF). The proximal renal tubular cell is the site in the kidney of high oxidative metabolic activity and in CRF is associated with adaptive hypertrophy and hypermetabolism. We hypothesised that chronic acidosis may lead to increased generation of reactive oxygen species due to increased oxidative activity. We developed a novel model of chronic acidosis in LLC-PK1 cells and measured markers of oxidative stress and metabolism. Acidosis led to a reduction in cellular total glutathione and protein thiol content and an increase in glutathione peroxidase activity and NH3 generation. The expression of constitutively expressed heat stress protein (HSP) HSC70 and HSP60 increased at pH 7.0.
Journal Section:
Original Paper
References
1.
El-Nahas AM: Mechanisms of experimental and clinical renal scarring; in Davison AM, Stewart JS, Grunfeld J-P, Kerr DNS, Ritz E, Winearls CG (eds): Oxford Textbook of Clinical Nephrology, ed 2. Oxford, Oxford University Press, 1998, vol 3, pp 1749–1789.
2.
Bohle A, Mackensen-Haen S, Von Gise H, Grund KE, Wehrmann M, Batz CH, Bogenschutz O, Schmitt H, Nagy J, Muller C, Muller G: The consequences of tubulo-interstitial changes for renal function in glomerulopathies. Pathol Res Pract 1990;186:135–144.
[PubMed]
3.
Rustom R, Grime JS, Sells RA, Amara A, Jackson MJ, Shenkin A, Maltby P, Smith L, Hammad A, Brown M, Bone JM: Renal tubular peptide catabolism in chronic vascular rejection. Ren Fail 2001;23:517–531.
4.
Rustom R, Maltby P, Grime JS, Hughes A, Costigan M, Shenkin A, Critchley M, Bone JM: Tubular peptide hypermetabolism and urinary ammonia in chronic renal failure in man: A maladaptive response? Nephron 1998;79:306–311.
5.
Camargo MJF, Sumpio BE, Maack T: Renal hydrolysis of absorbed protein: Influence of load and lysosomal pH. Am J Physiol 1984;247:F856–F864.
6.
Nath KA, Hostetter MK, Hostetter TM: Pathophysiology of chronic tubulo-interstitial disease in rats. Interaction of dietary acid load, ammonia, and complement. J Clin Invest 1985;79:667–675.
7.
Rustom R, Grime S, Costigan M, Maltby P, Hughes A, Taylor W, Shenkin A, Critchley M, Bone JM: Na HCO3 reduces proximal renal tubular peptide catabolism, ammoniagenesis and tubular damage in renal patients. Ren Fail 1998;20:371–382.
8.
Maack T, Hyung P, Camargo MJF: Renal filtration, transport, and metabolism of proteins; in Seldin DW, Giebisch G (eds): The Kidney: Physiology and Pathophysiology. New York, Raven Press, 1985, pp 1773–1803.
9.
Nagami GT: Ammonia production and secretion by the proximal tubule. Am J Kidney Dis 1989;14:258–261.
[PubMed]
10.
Weeson LG: Compensatory growth and other growth responses of the kidney. Nephron 1989;51:149–184.
[PubMed]
11.
Harris DCH, Chan L, Schrier RW: Remnant kidney hypermetabolism and progression of chronic renal failure. Am J Physiol 1988;254:F267–F276.
12.
Preuss H, Baird K, Goldin H: Oxygen consumption and ammoniagenesis in isolated dog renal tubules. J Lab Clin Med 1974;83:937–946.
13.
Schoolwerth AC, Sandler RS, Hoffman PM, Klahr S: Effects of nephron reduction and dietary protein content on renal ammoniagenesis in the rat. Kidney Int 1975;7:397–404.
[PubMed]
14.
Andreoli S, Mc Ateer JA, Mallett C: Reactive oxygen molecule-mediated injury in endothelial and renal tubular cells in vitro. Kidney Int 1990;38:795–794.
15.
Nath KA, Fischereder M, Hostetter TH: The role of oxidants in progressive renal injury. Kidney Int 1994;45:S111–S115.
16.
Shah SV: The role of reactive oxygen metabolites in glomerular disease. Annu Rev Physiol 1995;57:245–262.
[PubMed]
17.
Andreoli SP, Mallett C, McAteer JA, Williams LV: Antioxidant defence mechanisms of endothelial cells and renal tubular epithelial cells in vitro: Role of the glutathione redox cycle and catalase. Pediatr Res 1992;32:360–365.
18.
Hagen TM, Aw TY, Jones DP: Glutathione uptake and protection against oxidative injury in isolated kidney cells. Kidney Int 1988;34:74–81.
[PubMed]
19.
Cole LA, Scheid JM, Tannen RL: Induction of mitochondrial metabolism and pH-modulated ammoniagenesis by rocking LLC-PK1 cells. Am J Physiol 1986;251:C293–C298.
20.
Igarshi P, Freed MI, Ganz MB, Reilly RF: Effects of chronic acidosis on Na(+)-H+ exchangers in LLC-PK1 renal epithelial cells. Am J Physiol 1992;263:F83–F88.
21.
Solzenbach F: Lactic dehydrogenase (crystalline); in Wood WA (ed): Methods in Enzymology. New York, Academic Press, 1966, vol 9, pp 278–288.
22.
Anderson M: Determination of glutathione and glutathione disulphide. Methods Enzymol 1998;113:548–555.
23.
McArdle A, Van der Meulen JH, Catapano M, Symons MCR, Faulkner JA, Jackson MJ: Free radical activity following contraction-induced injury to the extensor digitorum longus muscle of rats. Free Radic Biol Med 1999;26:1085–1091.
24.
Di Monte D, Rose D, Bellomo G, Eklow L, Orrenius S: Alterations in intracellular thiol homeostasis during the metabolism of menadione by isolated rat hepatocytes. Arch Biochem Biophys 1984;235:334–342.
[PubMed]
25.
Fukunaga K, Suzuki T, Takama K: Highly sensitised high-performance liquid chromatography for the measurement of malondialdeyde in biological samples. J Chromatogr 1993;621:77–81.
26.
Claiborne Al: Catalase activity; in Greenwald RA (ed): CRC Handbook of Methods for Oxygen Radical Research. Boca Raton, CRC Press, 1985, pp 283–284.
27.
Flohe L, Gunzler WA: Assay of glutathione peroxidase. Methods Enzymol 1984;105:114–121.
[PubMed]
28.
Crapo JD, McCord JM, Fridovich I: Preparation and assay of superoxide dismutase. Methods Enzymol 1978;3:382–393.
29.
McArdle A, Patwell D, Vasilaki A, Griffiths D, Jackson MJ: Contractile activity-induced oxidative stress: Cellular origin and adaptive responses. Am J Physiol Cell Physiol 2001;280:C621–C627.
30.
Khassaf M, Child RB, McArdle A, Brodie DA, Esanu C, Griffiths RD, Jackson MJ: Time course of responses of human skeletal muscle to exercise-induced oxidative stress. Appl Physiol 2001;90:1031–1036.
31.
Hul RN, Cherry WR, Weaver GW: The origin and characterization of a pig kidney cell strain, LLC-PK1. In Vitro 1976;12:670–677.
[PubMed]
32.
Gstraunthaler G, Landauer F, Pfaller W: Ammoniagenesis in LLC-PK1 cultures: Role of transamination. Am J Physiol 1992;263:C47–C54.
33.
Gottschalk CW, Lassiter WE, Mylle M: Localisation of urine acidification in the mammalian kidney. Am J Physiol 1960;198:581–585.
34.
Tannen RL, Sahai A: Response of ammonia metabolism to acute acidosis: Insights from cultured renal epithelium. Am J Kidney Dis 1989;14:281–284.
35.
Nissim I, Sahal A, Sandler RS, Tannen RL: The intensity of acidosis differentially alters the pathways of ammoniagenesis in LLC-PK1 cells. Kidney Int 1994;45:1014–1019.
36.
McCoy RN, Hill KE, Ayon MA, Stein JH, Burk RF: Oxidant stress following renal ischaemia: Changes in the glutathione redox ratio. Kidney Int 1988;33:812–817.
[PubMed]
37.
Beck FX, Neuhofer W, Muller E: Molecular chaperones in the kidney: Distribution, putative roles, and regulation. Am J Physiol 2000;279:F203–F215.
38.
Muller E, Neuhofer W, Ohno A, Rucker S, Thurau K, Beck FX: Heat shock proteins HSP25, HSP60, HSP72, HSP73 in isoosmotic cortex and hyperosmotic medulla of rat kidney. Pflugers Arch 1996;431:608–617.
39.
Somji S, Todd JH, Sens MA, Garrett SH, Sens DA: Expression of heat shock protein 60 in human proximal tubule exposed to heat, sodium arsenite and CdCl(2). Toxicol Lett 2000;19:115:127–136.
40.
Sawczuk IS, Hoke G, Olsson CA, Connor J, Buttyan R: Gene expression in response to acute unilateral ureteral obstruction. Kidney Int 1989;35:1315–1319.
[PubMed]
41.
Emami A, Schwartz JH, Borkan SC: Transient ischaemia or heat stress induces a cytoprotection protein in rat kidney. Am J Physiol 1991;260:F479–F485.
42.
Razzaque MS, Taguchi T: Role of glomerular epithelial cell-derived heat shock protein 47 in experimental lipid nephropathy. Kidney Int 1999;56:S256–S259.
43.
Liu D, Razzaque MS, Cheng M, Taguchi T: The renal expression of heat shock protein 47 and collagens in acute and chronic experimental diabetes in rats. Histochem J 2001;33:621–628.
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