Research article

Age-related protein and mRNA expression of glutathione peroxidases (GPx) and Hsp-70 in different regions of rat kidney with and without stressor

  • Received: 14 December 2015 Accepted: 14 April 2016 Published: 18 April 2016
  • Small molecular weight oxygen free radical species (ROS) involved in oxidative stress can cause damage to cellular macromolecules including proteins, DNA and lipids. One of the most important enzymes involved in ROS detoxification is glutathione peroxidase (GPx). Here we study the age-related expression of GPx isoenzymes in various parts of the rat kidney with and without exposure to external oxidative stress. These results are correlated to the age dependent changes in the expression of the chaperone, Hsp-70.
    Protein and mRNA expression of GPx1 and GPx4 was studied in different regions of the kidney in ageing rats in the presence and absence of the external stressor 0.2 mM H2O2. Protein levels were examined by Western blot analysis following detection with appropriate antibodies and mRNA levels were analysed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) using appropriate primer sequences. mRNA expression for the chaperone Hsp70 was investigated in parallel.
    After reaching a peak at maturity (12 weeks), GPx1 protein and mRNA levels decreased with age under both control and stress conditions, and were higher in the cortex than in the outer and inner medulla. GPx4 protein and mRNA levels showed few comparable age-related changes. By contrast with the observed age-related decrease in GPx1 expression, chaperone Hsp-70 mRNA expression greatly increased with age.
    These findings suggest that the age-related decline in GPx1 expression in the cortex may be partly offset by a reciprocal change in Hsp-70 expression. These results are consistent with the oxidative stress theory of ageing.

    Citation: Noor Riyadh Thiab, Nicola King, Mary McMillan, Amer Almashhadany, Graham L Jones. Age-related protein and mRNA expression of glutathione peroxidases (GPx) and Hsp-70 in different regions of rat kidney with and without stressor[J]. AIMS Molecular Science, 2016, 3(2): 125-137. doi: 10.3934/molsci.2016.2.125

    Related Papers:

  • Small molecular weight oxygen free radical species (ROS) involved in oxidative stress can cause damage to cellular macromolecules including proteins, DNA and lipids. One of the most important enzymes involved in ROS detoxification is glutathione peroxidase (GPx). Here we study the age-related expression of GPx isoenzymes in various parts of the rat kidney with and without exposure to external oxidative stress. These results are correlated to the age dependent changes in the expression of the chaperone, Hsp-70.
    Protein and mRNA expression of GPx1 and GPx4 was studied in different regions of the kidney in ageing rats in the presence and absence of the external stressor 0.2 mM H2O2. Protein levels were examined by Western blot analysis following detection with appropriate antibodies and mRNA levels were analysed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) using appropriate primer sequences. mRNA expression for the chaperone Hsp70 was investigated in parallel.
    After reaching a peak at maturity (12 weeks), GPx1 protein and mRNA levels decreased with age under both control and stress conditions, and were higher in the cortex than in the outer and inner medulla. GPx4 protein and mRNA levels showed few comparable age-related changes. By contrast with the observed age-related decrease in GPx1 expression, chaperone Hsp-70 mRNA expression greatly increased with age.
    These findings suggest that the age-related decline in GPx1 expression in the cortex may be partly offset by a reciprocal change in Hsp-70 expression. These results are consistent with the oxidative stress theory of ageing.


    加载中
    [1] Miyoshi N, Oubrahim H, Chock PB, et al. (2006) Age-dependent cell death and the role of ATP in hydrogen peroxide-induced apoptosis and necrosis. Proc Natl Acad Sci USA 103: 1727-1731. doi: 10.1073/pnas.0510346103
    [2] Gomes P, Simao S, Silva E, et al. (2009) Aging increases oxidative stress and renal expression of oxidant and antioxidant enzymes that are associated with an increased trend in systolic blood pressure. Ox Med Cell Longevity 2: 138-145. doi: 10.4161/oxim.2.3.8819
    [3] Clark B (2000) Biology of renal aging in humans. Adv renal replacement therapy 7: 11-21.
    [4] Davies I, Fotheringham AP, Faragher BE (1989) Age-associated changes in the kidney of the laboratory mouse. Age Ageing 18: 127-133. doi: 10.1093/ageing/18.2.127
    [5] Epstein M (1996) Aging and the kidney. J Am Soc Nephrol 7: 1106-1122.
    [6] Hirokawa K (1975) Characterization of age-associated kidney disease in Wistar rats. Mech Age Devel 4: 301-316. doi: 10.1016/0047-6374(75)90031-7
    [7] Zhou XJ, Rakheja D, Yu XQ, et al. (2008) The aging kidney. Kidney int 74: 710-720.
    [8] Maurer G, Dieterle F, Perentes E, et al. (2013) Use of β-2-microglobulin to assess glomerular alterations and damage in the kidney. Google Patents.
    [9] Lim BJ, Yang HC, Fogo AB (2014) Animal models of regression/progression of kidney disease. Drug Discovery Today: Disease Models 11: 45-51. doi: 10.1016/j.ddmod.2014.06.003
    [10] Gecit İ, Kavak S, Meral I, et al. (2011) Effects of shock waves on oxidative stress, antioxidant enzyme and element levels in kidney of rats. Biol Trace Element Res 144: 1069-1076. doi: 10.1007/s12011-011-9124-8
    [11] 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 Stress Chaperones 15: 743-751. doi: 10.1007/s12192-010-0177-y
    [12] Carrillo MC, Alvarez MDL, Parody J, et al. (2012) Reactive oxygen species act as signaling molecules in liver carcinogenesis. http://dx.doi.org/10.5772/45966.
    [13] Shigenaga MK, Hagen TM, Ames BN (1994) Oxidative damage and mitochondrial decay in aging. Proc Natl Acad Sci USA 91: 10771-10778. doi: 10.1073/pnas.91.23.10771
    [14] Fraga CG, Shigenaga MK, Park JW, et al. (1990) Oxidative damage to DNA during aging: 8-hydroxy-2’-deoxyguanosine in rat organ DNA and urine. Proc Natl Acad Sci USA 87: 4533-4537. doi: 10.1073/pnas.87.12.4533
    [15] Stadtman ER (1992) Protein oxidation and aging. Sci 257: 1220-1224. doi: 10.1126/science.1355616
    [16] El‐Sawalhi MM, Darwish HA, Mausouf MN, et al. (2013) Modulation of age‐related changes in oxidative stress markers and energy status in the rat heart and hippocampus: a significant role for ozone therapy. Cell Biochem Function 31: 518-525. doi: 10.1002/cbf.2930
    [17] Harman D (1998) Aging and oxidative stress. J Int Fed Clin Chem 10: 24-27.
    [18] Valavanidis A, Vlachogianni T, Fiotakis K, et al. (2013) Pulmonary oxidative stress, inflammation and cancer: respirable particulate matter, fibrous dusts and ozone as major causes of lung carcinogenesis through reactive oxygen species mechanisms. Int J Environ Res Public Health 10: 3886-3907. doi: 10.3390/ijerph10093886
    [19] Wang CH, Wu SB, Wu YT, et al. (2013) Oxidative stress response elicited by mitochondrial dysfunction: implication in the pathophysiology of aging. Exp Biol Med 238: 450-460.
    [20] Semsei I, Rao G, Richardson A (1991) Expression of supreoxide dismutase and catalase in rat brain as a function of age. Mech Ageing Develop 58: 13-19. doi: 10.1016/0047-6374(91)90116-H
    [21] Wang P, Li CG, Qi Z, et al. (2015) Acute Exercise Induced Mitochondrial H2O2 Production in Mouse Skeletal Muscle: Association with p 66Shc and FOXO3a Signaling and Antioxidant Enzymes. Ox Med Cell Longevity 2015: 536456.
    [22] Sohal R, Sohal BH, Brunk UT (1990) Relationship between antioxidant defenses and longevity in different mammalian species. Mech Ageing Devel 53: 217-227. doi: 10.1016/0047-6374(90)90040-M
    [23] Holmström KM, Finkel T (2014) Cellular mechanisms and physiological consequences of redox-dependent signalling. Nature Reviews Mol Cell Biol 15: 411-421. doi: 10.1038/nrm3801
    [24] Pollack M, Leeuwenburgh C (1999) Molecular mechanisms of oxidative stress in aging: free radicals, aging, antioxidants and disease. Handbook of Oxidants and Antioxidants in Exercise. Amsterdam: Elsevier Science BV, 881-923.
    [25] Vida C, Gonzalez EM, De la Fuente M (2014) Increase of oxidation and inflammation in nervous and immune systems with aging and anxiety. Curr Pharmaceut Design 20: 4656-4678. doi: 10.2174/1381612820666140130201734
    [26] Thiab NR, King N, Jones GL (2015) Effect of ageing and oxidative stress on antioxidant enzyme activity in different regions of the rat kidney. Mol Cell Biochem 408: 253-260. doi: 10.1007/s11010-015-2503-2
    [27] Feder ME, Hofmann GE (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Ann Rev Physiol 61: 243-282. doi: 10.1146/annurev.physiol.61.1.243
    [28] Fehrenbach E, Niess A (1998) Role of heat shock proteins in the exercise response. Exercise Immunol Rev 5: 57-77.
    [29] Jolly C, Morimoto RI (2000) Role of the heat shock response and molecular chaperones in oncogenesis and cell death. J Natl Cancer Inst 92: 1564-1572. doi: 10.1093/jnci/92.19.1564
    [30] Neuer A, Spandorfer SD, Giraldo P, et al. (2000) The role of heat shock proteins in reproduction. Human Reproduction Update 6: 149-159. doi: 10.1093/humupd/6.2.149
    [31] Gabai VL, Meriin A, Yaglom JA, et al. (1998) Role of Hsp70 in regulation of stress-kinase JNK: implications in apoptosis and aging. FEBS Lett 438: 1-4. doi: 10.1016/S0014-5793(98)01242-3
    [32] Sőti C, Csermely P (2000) Molecular chaperones and the aging process. Biogeront 1: 225-233. doi: 10.1023/A:1010082129022
    [33] Shipp CK, Watson K, Jones GL (2011) Associations of HSP90 client proteins in human breast cancer. Anticancer Res 31: 2095-2101.
    [34] Boswell-Casteel RC, Johnson JM, Duggan KD, et al. (2015) Overproduction and biophysical characterization of human HSP70 proteins. Protein Expres Purif 106: 57-65. doi: 10.1016/j.pep.2014.09.013
    [35] Chin JH, Okazaki M, Hu ZW, et al. (1996) Activation of heat shock protein (hsp) 70 and proto-oncogene expression by alpha1 adrenergic agonist in rat aorta with age. J Clin Invest 97: 2316-2323. doi: 10.1172/JCI118674
    [36] Sharma S, Singh R, Kaur M, et al. (2010) Late-onset dietary restriction compensates for age-related increase in oxidative stress and alterations of HSP 70 and synapsin1 protein levels in male Wistar rats. Biogeront 11: 197-209. doi: 10.1007/s10522-009-9240-4
    [37] Beckman KB, Ames BN (1998) The free radical theory of aging matures. Physiol Rev 78: 547-581.
    [38] Harman D (1992) Free radical theory of aging. Mutation Research/DNAging 275: 257-266. doi: 10.1016/0921-8734(92)90030-S
    [39] Xiong Q, Xie P, Li H, et al. (2010) Acute effects of microcystins exposure on the transcription of antioxidant enzyme genes in three organs (liver, kidney, and testis) of male Wistar rats. J Biochem Mol Toxicol 24: 361-367. doi: 10.1002/jbt.20347
    [40] Sadi G, Eryilmaz N, Tütüncüoğlu E, et al. (2012) Changes in expression profiles of antioxidant enzymes in diabetic rat kidneys. Diabetes/Metabolism Research and Rev 28: 228-235. doi: 10.1002/dmrr.1302
    [41] Livak KJ, Schmittgen TD (2001) Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2ΔΔCT Method. Met 25: 402-408.
    [42] Jung K, Henke W (1996) Developmental changes of antioxidant enzymes in kidney and liver from rats. Free Rad Biol Med 20: 613-617. doi: 10.1016/0891-5849(95)02090-X
    [43] Rikans LE, Hornbrook KR (1997) Lipid peroxidation, antioxidant protection and aging. Biochim Biophys Acta 1362: 116-127. doi: 10.1016/S0925-4439(97)00067-7
    [44] Cand F, Verdetti J (1989) Superoxide dismutase, glutathione peroxidase, catalase, and lipid peroxidation in the major organs of the aging rats. Free Rad Biol Med 7: 59-63. doi: 10.1016/0891-5849(89)90101-9
    [45] Sanz N, Diez-Fernández C, Andrés D, et al. (2002) Hepatotoxicity and aging: endogenous antioxidant systems in hepatocytes from 2-, 6-, 12-, 18-and 30-month-old rats following a necrogenic dose of thioacetamide. Biochim Biophys Acta 1587: 12-20. doi: 10.1016/S0925-4439(02)00048-0
    [46] Shih PH, Yen GC (2007) Differential expressions of antioxidant status in aging rats: the role of transcriptional factor Nrf2 and MAPK signaling pathway. Biogeront 8: 71-80. doi: 10.1007/s10522-006-9033-y
    [47] Chen YF, Cowley Jr AW, Zou AP (2003) Increased H2O2 counteracts the vasodilator and natriuretic effects of superoxide dismutation by tempol in renal medulla. Am J Physiol 285: R827-R833.
    [48] Cao C, Leng YM, Huang W, et al. (2003) Glutathione peroxidase 1 is regulated by the c-Abl and Arg tyrosine kinases. J Biol Chem 278: 39609-39614. doi: 10.1074/jbc.M305770200
    [49] Zou A-P, Li N, Cowley AW (2001) Production and actions of superoxide in the renal medulla. Hypertension 37: 547-553. doi: 10.1161/01.HYP.37.2.547
    [50] Rao G, Xia E, Richardson A (1990) Effect of age on the expression of antioxidant enzymes in male Fischer F344 rats. Mech Ageing Devel 53: 49-60. doi: 10.1016/0047-6374(90)90033-C
    [51] Verbeke P, Fonager J, Clark BFC, et al. (2001) Heat shock response and ageing: mechanisms and applications. Cell Biol Int 25: 845-857. doi: 10.1006/cbir.2001.0789
    [52] Lee YK, Manalo D, Liu AC (1996) Heat shock response, heat shock transcription factor and cell aging. Neurosignals 5: 180-191. doi: 10.1159/000109187
    [53] Borkan SC, Emami A, Schwartz JH (1993) Heat stress protein-associated cytoprotection of inner medullary collecting duct cells from rat kidney. Am J Physiol 265: F333-F341.
    [54] Liu AYC, Lee YK, Manalo D, et al. (1996) Attenuated heat shock transcriptional response in aging: molecular mechanism and implication in the biology of aging, in Stress-Inducible Cellular Responses. Springer, 393-408.
  • Reader Comments
  • © 2016 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Metrics

Article views(5362) PDF downloads(1184) Cited by(1)

Article outline

Figures and Tables

Figures(6)  /  Tables(2)

/

DownLoad:  Full-Size Img  PowerPoint
Return
Return

Catalog