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Pyruvate Protects Giardia Trophozoites from Cysteine-Ascorbate Deprived Medium Induced Cytotoxicity

  • Raj, Dibyendu (Vivekananda College) ;
  • Chowdhury, Punam (Division of Parasitology, National Institute of Cholera and Enteric Diseases) ;
  • Sarkar, Rituparna (Division of Parasitology, National Institute of Cholera and Enteric Diseases) ;
  • Saito-Nakano, Yumiko (Department of Parasitology, National Institute of Infectious Diseases) ;
  • Okamoto, Keinosuke (Collaborative Research Center of Okayama University for Infectious Diseases in India) ;
  • Dutta, Shanta (Division of Parasitology, National Institute of Cholera and Enteric Diseases) ;
  • Nozaki, Tomoyoshi (Department of Biomedical Chemistry, School of International Health, Graduate School of Medicine, The University of Tokyo) ;
  • Ganguly, Sandipan (Division of Parasitology, National Institute of Cholera and Enteric Diseases)
  • Received : 2017.05.08
  • Accepted : 2017.10.27
  • Published : 2018.02.28

Abstract

Giardia lamblia, an anaerobic, amitochondriate protozoan parasite causes parasitic infection giardiasis in children and young adults. It produces pyruvate, a major metabolic product for its fermentative metabolism. The current study was undertaken to explore the effects of pyruvate as a physiological antioxidant during oxidative stress in Giardia by cysteine-ascorbate deprivation and further investigation upon the hypothesis that oxidative stress due to metabolism was the reason behind the cytotoxicity. We have estimated intracellular reactive oxygen species generation due to cysteine-ascorbate deprivation in Giardia. In the present study, we have examined the effects of extracellular addition of pyruvate, during oxidative stress generated from cysteine-ascorbate deprivation in culture media on DNA damage in Giardia. The intracellular pyruvate concentrations at several time points were measured in the trophozoites during stress. Trophozoites viability under cysteine-ascorbate deprived (CAD) medium in presence and absence of extracellular pyruvate has also been measured. The exogenous addition of a physiologically relevant concentration of pyruvate to trophozoites suspension was shown to attenuate the rate of ROS generation. We have demonstrated that Giardia protects itself from destructive consequences of ROS by maintaining the intracellular pyruvate concentration. Pyruvate recovers Giardia trophozoites from oxidative stress by decreasing the number of DNA breaks that might favor DNA repair.

Keywords

References

  1. Adam RD. The biology of Giardia spp. Microbiol Rev 1991; 55: 706-732.
  2. Adam RD. Biology of Giardia lamblia. Clin Microbiol Rev 2001; 14: 447-475. https://doi.org/10.1128/CMR.14.3.447-475.2001
  3. Lane S, Lloyd D. Current trends in research into the waterborne parasite Giardia. Crit Rev Microbiol 2002; 28: 123-147. https://doi.org/10.1080/1040-840291046713
  4. Tak V, Mirdha BR, Yadav P, Vyas P, Makharia GK, Bhatnagar S. Molecular Characterization of Giardia intestinalis assemblages from human isolates at a tertiary care center of India. Ind J Med Micro 2014; 32: 19-25. https://doi.org/10.4103/0255-0857.124290
  5. Farthing MJ. The molecular pathogenesis of giardiasis. J Pediatr Gastroenterol Nutr 1997; 24: 79-88. https://doi.org/10.1097/00005176-199701000-00018
  6. Atkinson HJ. Respiration in Nematodes. In Zuckerman BM ed, Nematodes as Biological Models, Vol. 2. London, UK. Academic Press. 1980, p 101-138.
  7. Brown DM, Upcroft JA, Upcroft P. Free radical detoxification in Giardia duodenalis. Mol Biochem Parasitol 1995; 72: 47-56. https://doi.org/10.1016/0166-6851(95)00065-9
  8. Lujan HD, Byrd LG, Mowatt MR, Nash TE. Serum Cohn fraction IV-1 supports the growth of Giardia lamblia in vitro. Infect Immun 1994; 62: 4664-4666.
  9. Gillin FD, Reiner DS, McCann PP. Inhibition of growth of Giardia lamblia by difluoromethylornithine, a specific inhibitor of polyamine biosynthesis. J Protozool 1984; 31: 161-613. https://doi.org/10.1111/j.1550-7408.1984.tb04308.x
  10. Brown DM, Upcroft JA, Upcroft P. Cysteine is the major lowmolecular weight thiol in Giardia duodenalis. Mol Biochem Parasitol 1993; 61: 155-158. https://doi.org/10.1016/0166-6851(93)90169-X
  11. Tekwani BL, Mehlotra RK. Molecular basis of defence against oxidative stress in Entamoeba histolytica and Giardia lamblia. Microbes Infect 1999; 1: 385-394. https://doi.org/10.1016/S1286-4579(99)80055-0
  12. Ghosh E, Ghosh A, Ghosh AN, Nozaki T, Ganguly S. Oxidative stress-induced cell cycle blockage and a protease-independent programmed cell death in microaerophilic Giardia lamblia. Drug Des Devel Ther 2009; 3: 103-110.
  13. Raj D, Ghosh E, Mukherjee AK, Nozaki T, Ganguly S. Differential gene expression in Giardia lamblia under oxidative stress: significance in eukaryotic evolution. Gene 2014; 535: 131-139. https://doi.org/10.1016/j.gene.2013.11.048
  14. Paget TA, Kelly ML, Jarrol EL, Lindmark DG, Lloyd D. The effects of oxygen on fermentation in Giardia lamblia. Mol Biochem Parasitol 1993; 57: 65-72. https://doi.org/10.1016/0166-6851(93)90244-R
  15. Mead JF. Free Radical Mechanisms of Lipid Damage and Consequences for Cellular Membranes. In Pryor WA ed, Free Radicals in Biology. New York, USA. Academic Press. 1976, p 51-68.
  16. Schuessel K, Frey C, Jourdan C, Keil U, Weber CC, Muller-Spahn F, Muller WE, Eckert A. Aging sensitizes toward ROS formation and lipid peroxidation in PSIM146L transgenic mice. Free Radic Biol Med 2006; 40: 850-862. https://doi.org/10.1016/j.freeradbiomed.2005.10.041
  17. Bar-Or D, Rael LT, Lau EP, Rao NK, Thomas GW, Winkler JV, Yukl RL, Kingston RG, Curtis CG. An analog of the human albumin N-terminus (Asp-AlaHis-Lys) prevents formation of copperinduced reactive oxygen species. Biochem Biophys Res Commun 2001; 284: 856-862. https://doi.org/10.1006/bbrc.2001.5042
  18. Evans PJ, Gallesi D, Mathieu C, Hernandez MJ, Felipe M, Halliwell B, Puppo A. Oxidative stress occurs during soybean nodule senescence. Planta 1999; 208: 73-79. https://doi.org/10.1007/s004250050536
  19. Rael LT, Thomas GW, Craun ML, Curtis CG, Bar-Or R, Bar-Or D. Lipid peroxidation and the thiobarbituric acid assay: standardization of the assay when using saturated and unsaturated fatty acids. J Biochem Mol Biol 2004; 37: 749-752.
  20. Nandi N, Sen A, Banerjee R, Kumar S, Kumar V, Ghosh AN, Das P. Hydrogen peroxide induces apoptosis-like death in Entamoeba histolytica trophozoites. Microbiology 2010; 156: 1926-1941. https://doi.org/10.1099/mic.0.034066-0
  21. Andrae U, Singh J, Ziegler-Skylakakis K. Pyruvate and related alpha-ketoacids protect mammalian cells in culture against hydrogen peroxide-induced cytotoxicity. Toxicol Lett 1985; 28: 93-98. https://doi.org/10.1016/0378-4274(85)90015-3
  22. Crestanello JA, Kamelgard J, Whitman GJ. The cumulative nature of pyruvate's dual mechanism for myocardial protection. J Surg Res 1995; 59: 198-204. https://doi.org/10.1006/jsre.1995.1154
  23. Holleman AF. Notice sur l'action de l'eau oxyge'ne'e sur les acides ace'toniques et sur les dice'tones 1.2. Recl Trav Chim Pays Bas 1904; 23: 169-171 (in French).
  24. Biagini GA, Park JH, Lloyd D, Edwards MR. The antioxidant potential of pyruvate in the amitochondriate diplomonads Giardia intestinalis and Hexamita inflata. Microbiology 2001; 147: 3359-3365. https://doi.org/10.1099/00221287-147-12-3359
  25. Herbener GH. A morphometric study of age-dependent changes in mitochondrial population of mouse liver and heart. J Gerontol 1976; 31: 8-12. https://doi.org/10.1093/geronj/31.1.8
  26. Vorbeck ML, Martin AP, Park JK, Townsend JF. Aging related decrease in hepatic cytochrome oxidase of the Fischer 344 rat. Arch Biochem Biophys 1982; 214: 67-79. https://doi.org/10.1016/0003-9861(82)90009-1
  27. Ma'ayeh SY, Knorr L, Svard SG. Transcriptional profiling of Giardia intestinalis in response to oxidative stress. Int J Parasitol 2015; 45: 925-938. https://doi.org/10.1016/j.ijpara.2015.07.005
  28. Ganguly S, Raj D. Giardia and Giardiasis: an Overview of Recent Developments. The Handbook of Microbial Bioresources. In Gupta VK, Sharma GD, Tuohy MG, Gaur R eds, The Handbook of Microbial Bioresources. London, UK. CABI Wallingford. 2016, p 211-227.
  29. Raj D, Saini P, Nozaki T, Ganguly S. Involvement of pyruvate on oxidative stress management in the microaerophilic protozoan parasite Giardia lamblia. Int J Adv Res 2015; 4: 1148-1166.

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