Advertisement

Journal of Biosciences

, Volume 29, Issue 2, pp 179–187 | Cite as

Effects of anisotonicity on pentose-phosphate pathway, oxidized glutathione release and t-butylhydroperoxide-induced oxidative stress in the perfused liver of air-breathing catfish,Clarias batrachus

  • Nirmalendu Saha
  • Carina Goswami
Article

Abstract

Both hypotonic exposure (185 mOsmol/l) and infusion of glutamine plus glycine (2 mmol/l each) along with the isotonic medium caused a significant increase of14CO2 production from [1-14C]glucose by 110 and 70%, respectively, from the basal level of 18.4 ± 1.2 nmol/g liver/min from the perfused liver ofClarias batrachus. Conversely, hypertonic exposure (345 mOsmol/l) caused significant decrease of14CO2 production from [1-14C]glucose by 34%.14CO2 production from [6-14C]glucose was largely unaffected by anisotonicity. The steady-state release of oxidized glutathione (GSSG) into bile was 1.18 ±0.09 nmol/g liver/min, which was reduced significantly by 36% and 34%, respectively, during hypotonic exposure and amino acid-induced cell swelling, and increased by 34% during hypertonic exposure. The effects of anisotonicity on14CO2 production from [1-14C]glucose and biliary GSSG release were also observed in the presence of t-butylhydroperoxide (50 (Amol/1). The oxidative stress-induced cell injury, caused due to infusion of t-butylhydroperoxide, was measured as the amount of lactate dehydrogenase (LDH) leakage into the effluent from the perfused liver; this was found to be affected by anisotonicity. Hypotonic exposure caused significant decrease of LDH release and hypertonic exposure caused significant increase of LDH release from the perfused liver. The data suggest that hypotonically-induced as well as amino acid-induced cell swelling stimulates flux through the pentose-phosphate pathway and decreases loss of GSSG under condition of mild oxidative stress; hypotonically swollen cells are less prone to hydroperoxide-induced LDH release than hypertonically shrunken cells, thus suggesting that cell swelling may exert beneficial effects during early stages of oxidative cell injury probably due to swelling-induced alterations in hepatic metabolism.

Keywords

Anisotonicity cell injury cell volume Clarias batrachus GSSG oxidative stress pentose-phosphate pathway perfused liver t-butylhydroperoxide 

Abbreviations Used

GSH

Glutathione

GSSG

oxidized glutathione

LDH

lactate dehydrogenase

RVD

regulatory volume decrease

RVI

regulatory volume increase

t-BOOH

t-butylhydroperoxide

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Akerboom T P M and Sies H 1981 Assay of glutathione, glutathione disulfide and glutathione mixed disulfides in biological samples;Methods Enzymol. 77 373–382PubMedCrossRefGoogle Scholar
  2. Akerboom T P M, Bilzer M and Sies H 1982 The relationship of biliary glutathione disulphide content in the perfused rat liver;J. Biol. Chem. 257 4248–4252PubMedGoogle Scholar
  3. Bianchini L, Fossat B, Porthe-Nibelle J, Ellory J C and Lahlou B 1988 Effects of hyposmotic shock on ion fluxes in isolated trout hepatocytes;J. Exp. Biol. 137 303–318PubMedGoogle Scholar
  4. Brigelius R 1983 Mixed disulphides: biological functions and increase in oxidative stress; inOxidative stress (ed.) H Sies (London: Academic Press) pp 303–318Google Scholar
  5. Goswami C and Saha N 1998 Glucose, pyruvate and lactate efflux by the perfused liver of a teleost,Clarias batrachus during aniso-osmotic exposure;Comp. Biochem. Physiol. A119 999–1007Google Scholar
  6. Hallbrucker C, Ritter M, Lang F, Gerok W and Häussinger D 1993 Hydroperoxide metabolism in rat liver: K+ channel activation, cell volume changes eicosanoid formation;Eur. J. Biochem. 211 449–458PubMedCrossRefGoogle Scholar
  7. Häussinger D 1996 The role of cellular hydration of cell function;Biochem. J. 321 697–710Google Scholar
  8. Häussinger D, Hallbrucker C, Saha N, Lang F and Gerok W 1992 Cell volume and bile acid excretion;Biochem. J. 288 681–689PubMedGoogle Scholar
  9. Häussinger D and Lang F 1991 Cell volume in regulation of hepatic function: a new mechanism for metabolic control;Biochim. Biophys. Acta 1071 331–350PubMedGoogle Scholar
  10. Häussinger D, Lang F, Bauers K and Gerok W 1990 Control of hepatic nitrogen metabolism and glutathione release by cell volume-regulatory mechanisms;Eur. J. Biochem. 193 891–898CrossRefGoogle Scholar
  11. Häussinger D, Roth E, Lang F and Gerok W 1993 Cellular hydration state: an important determinant of protein catabolism in health and disease;Lancet 341 1330–1332PubMedCrossRefGoogle Scholar
  12. Hoffmann E K and Simonsen L O 1989 Membrane mechanism in volume and pH regulation in vertebrate cells;Physiol. Rev. 69 315–382PubMedGoogle Scholar
  13. Jensen F B 1995 Regulatory volume decrease in carp red blood cells. Mechanisms and oxygenation-dependency of volume-activated potassium and amino acid transport;J. Exp. Biol. 198 155–165PubMedGoogle Scholar
  14. Livingstone D R, Garcia Martinez P, Michel X, Narbonne J F, O’Hara S, Ribera D and Winston G 1990 Oxyradical production as a pollution-mediated mechanism of toxicity in the common musselMytilus edulis L;Aquatic Toxicol. 15 231–236Google Scholar
  15. Nelson D L and Cox M M 2000Lehninger principles of biochemistry 3rd edition (New York: Worth Publishers, Macmillan Press Ltd)Google Scholar
  16. Okada Y and Maeno E 2001 Apoptosis, cell volume regulation and volume-regulatory chloride channels;Comp. Biochem. Physiol. 130 377–383CrossRefGoogle Scholar
  17. Perlman D F, Mush M W and Goldstein L 1996 Band 3 in cell volume regulation in fish erythrocytes;Cell. Mol. Biol. 42 975–984PubMedGoogle Scholar
  18. Saha N, Dkhar J and Ratha B K 1995 Induction of ureogenesis in the perfused liver of a freshwater teleostHeteropneustes fossilis, infused with different concentrations of ammonium chloride;Comp. Biochem. Physiol. B112 733–741Google Scholar
  19. Saha N and Ratha B K 1998 Ureogenesis in Indian air-breathing teleosts: Adaptation to environmental constraints;Comp. Biochem. Physiol. 120 195–208CrossRefGoogle Scholar
  20. Saha N, Schreiber R, von Dahl S, Lang F, Gerok W and Häussinger D 1993 Endogenous hydroperoxide formation, cell volume and cellular K+ balance in the perfused rat liver;Biochem. J. 296 701–707PubMedGoogle Scholar
  21. Saha N, Stoll B, Lang F and Häussinger D 1992 Effect of anisotonic cell volume modulation on glutathione-S-conjugate release, t-butylhydroperoxide metabolism and the pentosephosphate shunt in the perfused rat liver;Eur. J. Biochem. 209 437–444PubMedCrossRefGoogle Scholar
  22. Sies H 1985 Hydroperoxides and thiol oxidants in the study of oxidative stress in intact cells; inOxidative stress (ed.) H Sies (London: Academic Press) pp 73–90Google Scholar
  23. Stoll B and Häussinger D 1989 Functional hepatocyte heterogeneity vascular 2-oxoglutarate is almost exclusively taken up by perivenous glutamine-synthetase containing hepatocytes;Eur. J. Biochem. 181 709–716PubMedCrossRefGoogle Scholar
  24. Winston G W and Di Giulio R T 1991 Prooxidant and antioxidant mechanisms in aquatic organisms;Aquatic Toxicol. 19 137–161CrossRefGoogle Scholar
  25. Vorhaben J E and Campbell J W 1972 Glutamine synthetase: A mitochondrial enzyme in uricotelic species;J. Biol. Chem. 247 2763–2767PubMedGoogle Scholar

Copyright information

© Indian Academy of Sciences 2004

Authors and Affiliations

  1. 1.Biochemical Adaptation Laboratory, Department of ZoologyNorth-Eastern Hill UniversityShillongIndia

Personalised recommendations