Gene expression and enzyme activities of carbonic anhydrase and glutaminase in rat kidneys induced by chronic systemic hypoxia

Authors

  • Andi N.K. Syarifin Faculty of Medicine, Universitas Halu Oleo, Kendari
  • Sri W.A. Jusman Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Mohamad Sadikin Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta

DOI:

https://doi.org/10.13181/mji.v24i3.1190

Keywords:

CA9, chronic systemic hypoxia, GLS1, HIF-1α, kidney
Abstract viewed: 1867 times
PDF downloaded: 800 times

Abstract

Background: Hypoxia can cause acidosis. Kidney plays an essential role in maintaining acid-base balance, which involves the activities of carbonic anhydrase (CA) and glutaminase (GLS). This study is aimed to determine the expression and activities of the CA9 and GLS1 enzymes in relation to hypoxia inducible factor-1α (HIF-1α), a transcription factor protein which is a marker of hypoxia.

Methods: This study was an in vivo experimental study with coupled paralel design. used 25 male Sprague-Dawley rats weighing 150-200 g. Rats were divided into 5 groups: the control group (normoxic condition) and 4 treatment groups. The latter were kept in a hypoxic chamber (10% O2: 90% N2) for 1, 3, 5 and 7 days. All rats were euthanized after treatment, kidneys excised, tissues homogenized and investigated for gene expression of CA9, GLS1 and HIF-1α. On protein level, total enzymatic activities of CA and GLS and protein of HIF-1α were also investigated. Data were analyzed statistically using ANOVA for significance, and as its alternative, used Mann-Whitney and Kruskal-Wallis test.

Results: Results showed that HIF-1α mRNA increased during hypoxia, but not HIF-1α protein. It seemed that acidosis occurs in kidney tissue, indicated by increased CA9 and GLS1 mRNA expression and specific activity of total CA and GLS1. Expression of CA9 and GLS1 mRNA both showed strong positive correlation with HIF-1α mRNA, but not with HIF-1α protein.

Conclusion: It is suggested that during chronic systemic hypoxia, gene expression of CA9 and GLS1 and their enzyme activities were increased as a response to acidosis and related with the expression of HIF-1α mRNA.

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References

Alterio V, Hilvo M, Di Fiore A, Supuran CT, Pan P, Parkkila S, et al. Crystal structure of catalytic domain of the tumor-associated human carbonic anhydrase IX. Proc Natl Acad Sci U S A. 2009;106(38):16233-8. http://dx.doi.org/10.1073/pnas.0908301106

Pastorekova S, Ratcliffe PJ, Pastorek J. Molecular mechanisms of carbonic anhydrase IX-mediated pH regulation under hypoxia. BJU Int. 2008;101(Suppl4):8-15. http://dx.doi.org/10.1111/j.1464-410X.2008.07642.x

Phypers B, Pierce JMT. Lactate physiology in health and disease. Anes Crit Pain. 2006;6(3):128-32. http://dx.doi.org/10.1093/bjaceaccp/mkl018

Kovavevic Z, McGivan JD. Mitochondrial metabolism of glutamine and glutamate and its physiological signifinance. Physiol Rev. 1983;63(2):547-605.

Sing KW, Sofian-azirun M, Tayyab S. Protein analysis of Chrysomya megacephala maggot meal. Anim Nutr Food Tech. 2012;12:35-46.

Özdemir H, Küfrevio?lu OI, Çetinkay R. Effects of glycation on erythrocyte carbonic anhydrase-I and II patients with diabetes mellitus. Turk J Med Sci. 2000;30:135-41.

Imada A, Igarasi S, Nakaham K, Isono M. Asparaginase and glutaminase activities of micro-organisms. J Gen Microbiol. 1973;76(1):85-99. http://dx.doi.org/10.1099/00221287-76-1-85

Nathiya K, Nath SS, Angayarkanni J, Palaniswamy M. Optimised production of L-glutaminase: a tumour inhibitor from Aspergillus flavus cultured on agro-industrial residues. Afr J Biotechnol. 2011;10(63):13887-94. DOI: 10.5897/AJB11.1251

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Method. 2001;25(4):402-8. http://dx.doi.org/10.1006/meth.2001.1262

Prijanti AR. Peran hypoxia inducable factor -1a dalam pengaturan ekspresi renin [dissertation]. Jakarta: Universitas Indonesia; 2010. Indonesian.

Gunaratnam L, Bonventre JV. HIF in kidney disease and development. J Am Soc Nephrol. 2009;20(9):1877-87. http://dx.doi.org/10.1681/ASN.2008070804

Haase VH. Hypoxia-inducible factors in the kidney. Am J Renal Physiol. 2006;291(2):F271-81. http://dx.doi.org/10.1152/ajprenal.00071.2006

Semenza GL. Hypoxia-inducible factor-1: oxygen homeostasis and disease pathophysiology. Trends Mol Med. 2001;7(8):345-50. http://dx.doi.org/10.1016/S1471-4914(01)02090-1

Maxwel P. HIF-1: an oxygen response system with special relevance to the kidney. J Am Soc Nephrol. 2003;14(11):2712-22. http://dx.doi.org/10.1097/01.ASN.0000092792.97122.E0

Stroka DM, Burkhardt T, Desbaillets I, Wenger RH, Neil DA, Bauer C, et al. HIF-1 is expressed in normoxic tissue and displays an organ-specific regulation under systemic hypoxia. FASEB J. 2001;15(13):2445-53. doi:10.1096/fj.01-0125com

Ziel KA, Campbell CC, Wilson GL, Gillespie MN. Ref-1/Ape is critical for formation of the hypoxia-inducible transcriptional complex on hypoxic response element of rat pulmonary artery endothelial cell VEGF gene. FASEB J. 2004;18(9):986-8. http://dx.doi.org/10.1096/fj.03-1160fje

Eckardt KU, Bernhardt WM, Weidemann A, Warnecke C, Rosenberger C, Wiesener MS. Role of hypoxia in pathogenesis of renal disease. Kidney Int Suppl. 2005;99:S46-51. http://dx.doi.org/10.1111/j.1523-1755.2005.09909.x

Sherwood L. Human physiology: from cells to systems. 8th ed. USA: Brooks/Cole Cenage Learning; 2010.

Hilvo M, Rafajová M, Pastoreková S, Pastorek J, Parkkila S. Expression of carbonic anhydrase IX in mouse tissues. J Histochem Cytochem. 2004;52(10):1313-21. http://dx.doi.org/10.1177/002215540405201007

Harper HA. Review of physiological chemistry. 15th ed. California: Lange Medical Publications. 1973.

Thangalevu K, Pan CQ, Karlberg T, Balaji G, Uttamchandani M, Suresh V, et al. Structural basis for the allosteric inhibitory mechanism of human kidney-type glutaminase (KGA) and its regulation by Ref-Mek-Erk signaling in cancer cell metabolism. PNAS. 2012;15(109):7705-10. doi: 10.1073/pnas.1116573109

Chun YS, Kim MS, Park JW. Oxygen-dependent and -independent regulation of HIF-1alpha. J Korean Med Sci. 2002;17(5):581-8. http://dx.doi.org/10.3346/jkms.2002.17.5.581

Edwards JR. Amino acid metabolism: The urea cycle and urea cycle disorders. In Fulfillment for the requirements of Advanced Nutrition I. 2012.

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Published

2015-11-09

How to Cite

1.
Syarifin AN, Jusman SW, Sadikin M. Gene expression and enzyme activities of carbonic anhydrase and glutaminase in rat kidneys induced by chronic systemic hypoxia. Med J Indones [Internet]. 2015Nov.9 [cited 2024Dec.25];24(3):139-45. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/1190

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Section

Basic Medical Research
Abstract viewed = 1867 times
PDF downloaded = 800 times

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