Abstract
Metabolic and oxidative stresses have been implicated in ageing and the pathogenesis of chronic kidney disease. In this study, we investigated the glutathione (GSH), thiobarbituric acid reactive substances (TBARS) and lactate concentrations in different kidney regions under control conditions and after exposure to oxidative stress invoked by 0.2 mM H2O2. Slices of superficial cortex, outer or inner medulla were dissected from kidneys of male Wistar rats of 5-, 12-, 36- and 60-week old. Samples were incubated for 30 min ± 0.2 mM H2O2 prior to homogenisation and centrifugation. The concentrations of GSH, TBARS and lactate were measured by colorimetry. Each metabolite showed a distinctive pattern. For GSH, this was 12 weeks > 36 weeks > 60 weeks and 5 weeks with the highest concentration measured in the superficial cortex at 12 weeks. For TBARS and lactate, the pattern was for the lowest concentration at 12 weeks and the highest at 60 and 5 weeks. The highest lactate and TBARS concentrations were measured under oxidative stress conditions, particularly at 5 and 60 weeks. These results suggest that GSH in different kidney regions peaks at maturity and then reduces with increasing age.
Similar content being viewed by others
Abbreviations
- GSH:
-
Glutathione
- TBARS:
-
Thiobarbituric acid reactive species
- ROS:
-
Reactive oxygen species
- \({\text{O}}_{2}^{ - \cdot}\) :
-
Superoxide
- OH· :
-
Hydroxyl radical
- \({\text{O}}_{2}^{1}\) :
-
Singlet oxygen
- GSSG:
-
Oxidised glutathione
- MDA:
-
Malondialdehyde
- PBS:
-
Phosphate buffered saline
- BSA:
-
Bovine serum albumin
References
Sonntag C (1987) The chemical basis of radiation biology. Taylor & Francis, London
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic oxidative stress tolerance in crop plants. Plant Physiol Biochem 48:909–930
Halliwell B (1994) Free radicals, antioxidants, and human disease: curiosity, cause, or consequence? Lancet 344:721–724
Dalle-Donne I, Giustarini D, Colombo R, Rossi Milzano A (2003) Protein carbonylation in human diseases. Trends Mol Med 9:169–176
Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O (2012) Oxidative oxidative stress and antioxidant defense. WAO J 5:9–19
Anathy V, Roberson LC, Guala AS, Godburn KE, Budd RC, Janssen-Heininger YMW (2012) Redox-based regulation of apoptosis: S-glutathionylation as a regulatory mechanism to control cell death. Antioxid Redox Signal 16:496–505
Meister A, Anderson ME (1983) Glutathione. Annu Rev Biochem 52:711–760
Mates JM, Segura JA, Alonso FJ, Marrquez J (2012) Sulphur-containing non enzymatic antioxidants: therapeutic tools against cancer. Front Biosci 4:722–748
Simic MG, Nygaard OF (1983) Radioprotectors and anticarcinogens. Academic Press, New York
Reed DJ (1990) Glutathione: toxicological implications. Annu Rev Pharmacol Toxicol 30:603–631
Janero DR (1990) Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 9:515–540
Delfino RJ, Staimer N, Vaziri ND (2011) Air pollution and circulating biomarkers of oxidative oxidative stress. Air Qual Atmos Health 4:37–52
Lunec J (1989) Oxygen radicals: their measurement in vivo. Anal Proc 26:130–131
Pryor WA (1989) On the detection of lipid hydroperoxides in biological samples. Free Radic Biol Med 7:177–178
Halliwell B, Whiteman M (2004) Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean? Br J Pharmacol 142:231–255
Mizock BA, Falk JL (1992) Lactic acidosis in critical illness. Crit Care Med 20:80–93
Bellomo R (2002) Bench-to-bedside review: lactate and the kidney. Crit Care 6:322
Höhmann B, Frohnert PP, Kinne R, Bauman K (1974) Proximal tubular lactate transport in rat kidney: a micropuncture study. Kidney Int 5:261–270
Buege JA (1978) Microsomal lipid peroxidation. Methods Enzymol 52:302–310
Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358
Williams H, King N, Griffiths EJ, Suleiman M-S (2001) Glutamate-loading stimulates metabolic flux and improves cell recovery following chemical hypoxia in isolated cardiomyocyte. J Mol Cell Cardiol 33:2109–2119
Schulz JB, Lindenau J, Seyfried J, Dichgans J (2000) Glutathione, oxidative oxidative stress and neurodegeneration. FEBS J 267:4904–4911
Farooqui MY, Day MW, Zamorano DM (1987) Gluthathione and lipid peroxidation in the aging rat. Comp Biochem Physiol Part B 88:177–180
Currais A, Maher P (2013) Functional consequences of age-dependent changes in glutathione status in the brain. Antioxid Redox Signal 19:813–822
Jozwiak Z, Jasnowska B (1985) Changes in oxygen-metabolizing enzymes and lipid peroxidation in human erythrocytes as a function of age of donor. Mech Ageing Dev 32:77–83
Imanishi H, Nakai T, Abe T, Takino T (1985) Glutathione metabolism in red cell aging. Mech Ageing Dev 32:57–62
Louboutin J-P, Strayer D (2014) Role of oxidative oxidative stress in HIV-1-associated neurocognitive disorder and protection by gene delivery of antioxidant enzymes. Antiox 3:770–797
Jain A, Martensson J, Stole E, Auld PA, Meister A (1991) Glutathione deficiency leads to mitochondrial damage in brain. Proc Natl Acad Sci 88:1913–1917
Rook D, van Goudoever JB (2014) Oxidative stress and glutathione synthesis rates in early postnatal life. In: Perinatal and prenatal disorders. Springer, New York, pp 255–269
Subramanian MV, James T (2010) Age-related protective effect of deprenyl on changes in the levels of diagnostic marker enzymes and antioxidant defense enzymes activities in cerebellar tissue in Wistar rats. Cell Ox Stress Chaperones 15:743–751
Anandan R, Ganesan B, Obolesu T, Matthew S, Asha KK, Lakshmanan PT, Zynudheen AA (2013) Antiaging effect of dietary chitosan supplementation on glutathione-dependent antioxidant system in young and aged rats. Cell Stress Chaperones 18:121–125
Rana S, Allen T, Singh R (2002) Inevitable glutathione, then and now. Indian J Exp Biol 40:706–716
Baird M, Samis H (1971) Regulation of catalase activity in mice of different ages. Gerontol 17:105–115
Hazelton GA, Lang CA (1985) Glutathione peroxidase and reductase activities in the aging mouse. Mech Ageing Dev 29:71–81
Ito Y, Kajkenova O, Feuers RJ, Udupa KB, Desai VG, Epstein J, Hart RW, Lipschitz DA (1998) Impaired glutathione peroxidase activity accounts for the age-related accumulation of hydrogen peroxide in activated human neutrophils. J Gerontol Ser A 53:M169–M175
Marí M, Morales A, Collel A, Garcia-Ruiz C, Kaplowitz N, Fernandes-Checa JC (2013) Mitochondrial glutathione: features, regulation and role in disease. Biochim et Biophys Acta 1830:3317–3328
Martensson J, Meister A, Martensson J (1991) Glutathione deficiency decreases tissue ascorbate levels in newborn rats: ascorbate spares glutathione and protects. Proc Natl Acad Sci 88:4656–4660
Gecit İ, Kavak S, Meral I, Pirincci N, Gunes M, Demir H, Cengiz N, Ceylan K (2011) Effects of shock waves on oxidative oxidative stress, antioxidant enzyme and element levels in kidney of rats. Biol Trace Elem Res 144:1069–1076
Chen Y-F, Cowley AW Jr, Zou A-P (2003) Increased H2O2 counteracts the vasodilator and natriuretic effects of superoxide dismutation by tempol in renal medulla. Am J Physiol 285:R827–R833
Zou C-G, Agar NS, Jones GL (2001) Oxidative insult to human red blood cells induced by free radical initiator AAPH and its inhibition by a commercial antioxidant mixture. Life Sci 69:75–86
Halliwell B, Gutteridge J (1989) Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol 186:1–85
Choi J-H, Yu BP (1990) Unsuitability of TBA test as a lipid peroxidation marker due to prostaglandin synthesis in the aging kidney. Age 13:61–64
Boaz M, Matas Z, Biro A, Katzir Z, Green M, Fainaru M, Smetana S (1999) Serum malondialdehyde and prevalent cardiovascular disease in hemodialysis. Kidney Int 56:1078–1083
Sheng B, He D, Zhou J, Chen X, Nan X (2011) The protective effects of the traditional Chinese herbs against renal damage induced by extracorporeal shock wave lithotripsy: a clinical study. Urol Res 39:89–97
Shih P-H, Yen G-C (2007) Differential expressions of antioxidant status in aging rats: the role of transcriptional factor Nrf2 and MAPK signaling pathway. Biogerontology 8:71–80
Rao G, Xia E, Richardson A (1990) Effect of age on the expression of antioxidant enzymes in male Fischer F344 rats. Mech Ageing Dev 53:49–60
Acknowledgements
This research was conducted at the University of New England, Australia and supported by a grant from the School of Science and Technology.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that there are no potential conflicts of interest involved in the publication of this work
Animal ethics
All experiments were approved by the animal ethics committee of the UNE (AEC09/152, EC10/036 and AEC11/100).
Rights and permissions
About this article
Cite this article
Thiab, N.R., King, N. & Jones, G.L. Effects of ageing on metabolite and oxidant concentrations in different regions of rat kidney under normal and stress conditions. Mol Cell Biochem 408, 55–61 (2015). https://doi.org/10.1007/s11010-015-2482-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-015-2482-3