POSSIBLE PROTECTIVE MECHANISMS OF ALPHA-KETOGLUTARATE ON FRUIT FLY DROSOPHILA MELANOGASTER CANTON S UNDER EXPOSURE TO DIFFERENT STRESSORS

Authors

  • M.P. Lylyk Кафедра біохімії та біотехнології, ДВНЗ «Прикарпатський національний університет імені Василя Стефаника»
  • M.M. Bayliak Кафедра біохімії та біотехнології, ДВНЗ «Прикарпатський національний університет імені Василя Стефаника»

Abstract

Alpha-ketoglutaric acid is an important intermediate in Krebs cycle and in metabolism of amino acids. In recent years, the role of alpha-ketoglutaric acid as a powerful natural detoxifying agent was demonstrated. This work aimed to study the ability of the sodium salt of alpha-ketoglutaric acid (AKG) to alleviate the toxic effects of sodium nitroprusside on the larvae development and to increase the resistance of D. melanogaster Canton S flies to cold stress. Sodium nitroprusside (SNP) is widely used as a donor of nitric oxide in medicine but may be toxic for many organisms at high concentrations due to the induction of oxidative/nitrosative stress. This study shows, that food supplementation with AKG alleviates the inhibitory effect of SNP on D. melanogaster Сanton S pupation. The number of larvae reached the pupa stage on diet with 10 mM AKG and 1 mM SNP was 20% higher than on diet with 1 mM SNP. The reduced food intake of larvae grown on medium with SNP was not changed by AKG supplementation, and AKG also did not affect the levels of total iron and nitrite ions released from SNP. SNP decomposition is also accompanied by the release of highly toxic cyanide moieties, and the ability of AKG to interact with cyanide ions was reported earlier. It seems that the cyanide-binding capability may be important in prevention of the toxic effects of SNP on D. melanogaster. Two-day-old adult Сanton S flies reared on 10 mM AKG were more resistant to cold stress treatment and faster recovered from chill coma (0°C for 15 min) than control ones. Flies reared on AKG had the higher catalase activity and the higher levels of high- and low-molecular mass thiols comparing with the controls. Proline level was 1.4-fold higher in AKG-reared females in comparison with control females. The obtained results suggest, that dietary AKG provides cold stress resistance of two-day-old D. melanogaster flies due to the enhancement of antioxidant system capacity and synthesis of cryoprotective amino acids, such as proline.

Key words: alpha-ketoglutarate, Drosophila melanogaster, pupation, sodium nitroprusside, cold stress, antioxidant defense, proline.

References

Aebi H. Catalase in vitro. Meth. Enzymol. 1984; 105: 121-126.

Babich H., Zuckerbraun H. L., Ricklis A. S., Blau L. In vitro toxicity of sodium nitroprusside to human endothelial ECV304 cells. Environ. Toxicol. Pharmacol. 1998; 5(2): 135-144.

Bergman I., Loxley R. New spectrophotometric method for the determination of proline in tissue hydrolyzates. Anal. Chem. 1970; 42(7): 702-706.

Bhattacharya R., Satpute R., Hariharakrishnan J. et al. Acute toxicity of some synthetic cyanogens in rats and their response to oral treatment with alpha-ketoglutarate. Food Chem. Toxicol. 2009; 47: 2314-2320.

Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976; 72(1-2): 248-254.

David R. J., Gibert P., Pla E. et al. Cold stress tolerance in Drosophila: analysis of chill coma recovery in D. melanogaster. J. Therm. Biol. 1998; 23(5): 291-299.

Ellman G. L. Tissue sulfhydryl groups. Arch. Biochem. Biophys. 1959; 82: 70-77.

Harrison A. P., Pierzynowski S. G. Biological effects of 2-oxoglutarate with particular emphasis on the regulation of protein, mineral and lipid absorption/metabolism, muscle performance, kidney function, bone formation and cancerogenesis, all viewed from a healthy ageing perspective state of the art-review article. J. Physiol. Pharmacol. 2008; 59(Suppl. 1): 91-106.

Lee T. S., Kolthoff I. M., Leussing D. L. Reaction of ferrous and ferric iron with 1,10-phenanthroline. I. dissociation constants of ferrous and ferric phenanthroline. J. Am. Chem. Soc. 1948; 70(7): 2348–2352.

Lozinsky O., Lushchak O., Storey J., Storey K., Lushchak V. Sodium nitroprusside toxicity in Drosophila melanogaster: delayed pupation, reduced adult emergence, and induced oxidative stress. Archives of insects biochemistry and physiology. 2012; 80: 166-185. doi: 10.1002/arch.21033

Lylyk M., Shmihel H., Kozachok O., Bayliak M. Alpha-ketoglutarate modifies toxic action of sodium nitroprusside and ethanol on Drosophila melanogaster. Ukr. Biochem. J., Supplement 2 “Materials of XI Ukrainian Biochemical congress (Kyiv, October 6-10, 2014); 86(5): 249-250.

Lushchak V. I. Adaptive response to oxidative stress: bacteria, fungi, plants and animals. Comp. Biochem. Physiol. C. Toxicol. Pharmacol. 2011; 153: 175-190. doi: 10.1016/j.cbpc.2010.10.004

Lushchak, O. V., Rovenko, B. M., Gospodaryov, D. V., Lushchak, V. I. Drosophila melanogaster larvae fed by glucose and fructose demonstrate difference in oxidative stress markers and antioxidant enzymes of adult flies. Comp. Biochem. Physiol. A Mol. Integr. Physiol. 2011; 160(1): 27-34. doi: 10.1016/j.cbpa.2011.04.019

Misener S. R., Chen C., Walker V. K. Cold tolerance and proline metabolic gene expression in Drosophila melanogaster. J. Insect. Physiol. 2001; 47(4-5): 393-400.

Mitchell B., Bhandari R., Bebarta V. et al. Toxicokinetic profiles of α-ketoglutarate cyanohydrin, a cyanide detoxification product, following exposure to potassium cyanide. Toxicol. Lett. 2013; 222(1): 83-89. doi: 10.1016/j.toxlet.2013.07.008

Niemiec T., Sikorska J., Harrison A. et al. Alpha-ketoglutarate stabilizes redox homeostasis and improves arterial elasticity in aged mice. Physiol. Pharmacol. 2011; 62: 37-43.

Privat C., Lantoine F., Bedioui F. et al. Nitric oxide production by endothelial cells: comparison of three methods of quantification. Life Sci. 1997; 61(12): 1195-1202.

Whillier S., Garcia B., Chapman B. Glutamine and α-ketoglutarate as glutamate sources for glutathione synthesis in human erythrocytes. FEBS J. 2011; 278: 3152-3163. doi: 10.1111/j.1742-4658.2011.08241.x

Published

2015-06-30

Issue

Section

SHORT COMMUNICATIONS