H,K-ATPase and carbonic anhydrase response to chronic systemic rat gastric hypoxia

  • Ulfah Lutfiah Department Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Sri W.A. Jusman Department Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Mochammad Sadikin Department Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
DOI: https://doi.org/10.13181/mji.v24i3.1066
Keywords: carbonic anhydrase, gastric ulcer, H, K-ATPase, hypoxia
Abstract viewed: 1117 times
PDF downloaded: 813 times

Abstract

Background: Hypoxia may induce gastric ulcer associated with excessive hidrogen chloride (HCl) secretion. Synthesis of HCl involves 2 enzymes, H,K-ATPase and carbonic anhydrase (CA). This study aimed to clarify the underlying cause of gastric ulcer in chronic hypoxic condition, by investigating the H,K-ATPase and CA9 response in rats.

Methods: This study was an in vivo experiment, to know the relationship between hypoxia to expression of H,K-ATPase and CA9 mRNA, and H,K-ATPase and total CA specific activity of chronic systemic rat gastric hypoxia. The result was compared to control. Data was analyzed by SPSS. If the data distribution was normal and homogeneous, ANOVA and LSD post-hoc test were used. However, if the distribution was not normal and not homogeneous, and still as such after transformation, data was treated in non-parametric using Kruskal-Wallis and Mann Whitney test. Twenty five male Sprague-Dawley rats were divided into 5 groups: rats undergoing hypoxia for 1, 3, 5, and 7 days placed in hypoxia chamber (10% O2, 90% N2), and one control group. Following this treatment, stomach of the rats was extracted and homogenized. Expression of H,K-ATPase and CA9 mRNA was measured using real time RT-PCR. Specific activity of H,K-ATPase was measured using phosphate standard solution, and specific activity of total CA was measured using p-nitrophenol solution.

Results: The expression of H,K-ATPase mRNA was higher in the first day (2.159), and drastically lowered from the third to seventh day (0.289; 0.108; 0.062). Specific activities of H,K-ATPase was slightly higher in the first day (0.765), then was lowered in the third (0.685) and fifth day (0.655), and was higher in the seventh day (0.884). The expression of CA9 mRNA was lowered progressively from the first to seventh day (0.84; 0.766; 0.736; 0.343). Specific activities of total CA was low in the first day (0.083), and was higher from the third to seventh day (0.111; 0.136; 0.144).

Conclusion: In hypoxia condition, expression of H,K-ATPase and CA9 mRNA were decreased, but the specific activity of H,K-ATPase and total CA were increased.

Author Biography

Ulfah Lutfiah, Department Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
Department Biochemistry and Biology Moleculer, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia

References

  1. Gilany K, Vafakhah M. Hypoxia: a review. J Paramed Sci. 2010;1(2):43-60.

  2. Taylor CT. Mitochondria and cellular oxygen sensing in the HIF pathway. Biochem J. 2008;409(1):19-26. http://dx.doi.org/10.1042/BJ20071249

  3. Semenza GL. Oxygen-dependent regulation of mitochondrial respiration by hypoxia-inducible factor 1. Biochem J. 2007;405(1):1-9. http://dx.doi.org/10.1042/BJ20070389

  4. Alberti KG. The biochemical consequences of hypoxia. J Clin Pathol Suppl (R Coll Pathol). 1977;11:14-20. http://dx.doi.org/10.1136/jcp.s3-11.1.14

  5. Syam AF, Simadibrata M, Wanandi SI, Hernowo BS, Sadikin M, Rani AA. Gastric ulcers induced by systemic hypoxia. Acta Med Indones.2011;43(4):243-8.

  6. Shin JM, Munson K, Vagin O, Sachs G. The gastric HK-ATPase: structure, function, and inhibition. Pflugers Arch. 2009;457(3):609-22. http://dx.doi.org/10.1007/s00424-008-0495-4

  7. Munson K, Lambrecht N, Shin JM, Sachs G. Analysis of the membrane domain of the gastric H+/K+-ATPase. J Exp Biol. 2000;203:161-70.

  8. Harper HA. Review of physiological chemistry. 15th ed. Los Altos: Lange Medical publication; 1975. http://dx.doi.org/10.1007/978-3-662-09766-3

  9. ncbi.nlm.nih.gov [Internet]. Primer BLAST tools [cited 2013 Mar 8]. Available from: http://www.ncbi.nlm.nih.gov/tools/primer-blast/.

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

  11. Smolka AJ, Goldenring JR, Gupta S, Hammond CE. Inhibition of gastric H,K-ATPase activity and gastric epithelial cell IL-8 secretion by the pyrrolizine derivative ML 3000. BMC Gastroenterol. 2004;4(4):1-11. doi:10.1186/1471-230X-4-4

  12. Özdemir H, Küfrevioğlu O, Çetinkaya R. Effects of glycation on erythrocyte anhydrase-I and II patients with diabetes mellitus. Turk J Med Sci. 2000;30:135-41.

  13. Wood EJ. An introduction to centrifugation by T C Ford and J M Graham. Biochem Edu. 1991;19(3):157. http://dx.doi.org/10.1016/0307-4412(91)90066-H

  14. Healton EB, Savage DG, Brust JC, Garrett TJ, Lindenbaum J. Neurologic aspects of cobalamin deficiency. Medicine (Baltimore). 1991;70(4):229-45. http://dx.doi.org/10.1097/00005792-199107000-00001

  15. Mayer A, Höckel M, Vaupe P. Carbonic anhydrase IX expression and tumor oxygenation status do not correlate at the microregional level in locally advanced cancers of the uterine cervic. Clin Cancer Res. 2005,11(20):7220-5. http://dx.doi.org/10.1158/1078-0432.CCR-05-0869

  16. Pastorekova S, Zavada J. Carbonic anhydrase IX (CA IX) as a potential target for cancer therapy. Cancer Ther. 2004;2:245-62.

  17. Leppilampi M, Saarnio J, Karttunen TJ, Kivelä J, Patoreková S, Pastorek J, et al. Carbonic anhydrase isozymes IX and XII in gastric tumors. World J Gastroenterol. 2003;9(7):1398-403. http://dx.doi.org/10.3748/wjg.v9.i7.1398

  18. Hilvo M, Baranauskiene L, Salzano AM, Scaloni A, Matulis D, Innocenti A, et al. Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. J Biol Chem. 2008;283(41):27799-809. http://dx.doi.org/10.1074/jbc.M800938200

  19. Allen A, Garner A. Mucus and bicarbonate secretion in the stomach and their possible role in mucosal protection. Gut. 1980;21(3):249-62. http://dx.doi.org/10.1136/gut.21.3.249

Published
2015-11-09
How to Cite
1.
Lutfiah U, Jusman SW, Sadikin M. H,K-ATPase and carbonic anhydrase response to chronic systemic rat gastric hypoxia. Med J Indones [Internet]. 2015Nov.9 [cited 2024Apr.20];24(3):133-8. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/1066
Issue
Vol. 24 No. 3 (2015): September
Section
Basic Medical Research

Copyright (c) 2015 Ulfah Lutfiah, Sri W.A. Jusman, Mochammad Sadikin

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Authors who publish with Medical Journal of Indonesia agree to the following terms:

  1. Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
  2. Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.