Management of heat stress in Drosophila melanogaster with Abhrak bhasma and ascorbic acid as antioxidant supplements

Rambhadur P. Subedi; Rekha R. Vartak; Purushottam G. Kale

doi:10.7324/JABB.2018.60204

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Abstract

Global warming and climate change are the most serious consequences of anthropogenic activities. Although global warming poses threat to all living organisms, poikilotherms are most vulnerable among them. In the current study, management of heat stress in insects that constitute the largest group of poikilotherms has been studied using Drosophila melanogaster as a model. Various catalytic and non-catalytic oxidative parameters, at physiological as well as molecular level, were analyzed after exposing the Drosophila flies to high temperature. The efficacy of Abhrak bhasma and ascorbic acid in ameliorating the heat stress was also investigated. It was observed that heat stress alters various parameters indicating oxidative stress. Heat stress influences the activity of superoxide dismutase and catalase enzymes and also has an effect on total reduced glutathione (GSH) content as well as GSH: oxidized form of GSH (GSSG) ratio. Dietary supplement of Abhrak bhasma and ascorbic acid was found to alter the changes in antioxidant parameters induced due to heat stress. These flies also showed an increase in expression of cap “n” collar C and heat shock protein 70 genes, which play a crucial role in the management of stressful conditions. Overall, supplementation of diet with Abhrak bhasma and ascorbic acid was found to boost the capacity of Drosophila to counter the effects of heat stress.

Keyword: Heat stress Oxidative stress Drosophila Supplement Ascorbic acid.

Citation:

Subedi RP, Vartak RR, Kale PG. Management of heat stress in Drosophila melanogaster with Abhrak bhasma and ascorbic acid as antioxidant supplements. J App Biol Biotech. 2018;6(2):20-26. DOI: 10.7324/JABB.2018.60204

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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Reference

1. Solomon S, Plattner GK, Knutti R, and Friedlingstein P. Irreversible climate change due to carbon dioxide emissions. Proc Natl Acad Sci 2009; 106:1704–1709. https://doi.org/10.1073/pnas.0812721106
2. Altan O, Pabuçcuoglu A, Altan A, Konyalioglu S, and Bayraktar H. Effect of heat stress on oxidative stress, lipid peroxidation and some stress parameters in broilers. Br Poult Sci 2003; 44:545-550. https://doi.org/10.1080/00071660310001618334
3. Bhatia B, Kale PG, Daoo JV, and Meshram P. Testicular oxidative stress protective effects of abhraka bhasma in male wistar rats after heat exposure. Int J Pharm Pharm Sci 2013; 5:472-477.
4. Keil G, Cummings E, and Magalhaes JP. Being cool: how body temperature influences ageing and longevity. Biogerontology 2015; 16:383–397. https://doi.org/10.1007/s10522-015-9571-2
5. Watson J. Oxidants, antioxidants and the current incurability of metastatic cancers. Open Biol 2013; 3:120144. https://doi.org/10.1098/rsob.120144
6. Rahal A, Kumar A, Singh V, Yadav B, Tiwari R, Chakraborty S et al. Oxidative stress, prooxidants, and antioxidants: The interplay. BioMed Res Int 2014; vol. 2014, Article ID 761264, 19 pages.
7. Khaliq A, Javed M, Sohail M, and Sagheer M. Environmental effects on insects and their population dynamics. J Entomol Zool Stud 2014; 2:1-7.
8. Brewer MS. Natural antioxidants: sources, compounds, mechanisms of action, and potential applications. Compr Rev Food Sci Food Saf 2011; 10:221–247. https://doi.org/10.1111/j.1541-4337.2011.00156.x
9. Chand K, Rajeshwari, Hiremathad A, Singh M, Santos MA, and Keri RS. A review on antioxidant potential of bioactive heterocycle benzofuran: Natural and synthetic derivatives. Pharmacol Rep 2017; 69:281-295. https://doi.org/10.1016/j.pharep.2016.11.007
10. Pandey KB and Rizvi SI. Plant polyphenols as dietary antioxidants in human health and disease. Oxid Med Cell Longev 2009; 2:270–278. https://doi.org/10.4161/oxim.2.5.9498
11. Subedi RP, Vartak RR, and Kale PG. Effect of abhrak bhasma on the physiology and behaviour of Drosophila melanogaster. Int J Adv Res 2017; 5:1629-1641. https://doi.org/10.21474/IJAR01/3991
12. Bradford MM. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248-254. https://doi.org/10.1016/0003-2697(76)90527-3
13. Aebi HE. Method of Enzymatic Analysis. Vol. 3. Germany-Deerfield, FL: VCH, Weinheim; 1983. Catalase; pp. 273–286.
14. Marklund S and Marklund G. Involvement of the superoxide anion radical in the autooxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974; 47:469-474. https://doi.org/10.1111/j.1432-1033.1974.tb03714.x
15. Rahman I, Kode A, and Biswas SK. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat protoc 2006; 1:3159-3165. https://doi.org/10.1038/nprot.2006.378
16. Brand-williams W, Cuvelier ME, and Berset C. Use of free radical method to evaluate antioxidant activity. LWT-Food Science and Technology 1995; 28:25-30. https://doi.org/10.1016/S0023-6438(95)80008-5
17. Zeb A and Ullah F. A simple spectrophotometric method for the determination of thiobarbituric acid reactive substances in fried fast foods. J Anal Methods Chem 2016; vol. 2016, Article ID 9412767:5 pages.
18. Devasagayam TP, Boloor KK, and Ramasarma T. Methods for estimating lipid peroxidation: an analysis of merits and demerits. Indian J Biochem Biophys 2003; 40:300-8.
19. Bourg EL. Oxidative stress, aging and longevity in Drosophila melanogaster. FEBS Lett 2001; 498:183–186. https://doi.org/10.1016/S0014-5793(01)02457-7
20. Fleming J.E., Niedzwiecki A., Reveillaud I. Stress induced expression of CuZn superoxide dismutase and catalase in senescent Drosophila melanogaster. In: Packer L., Cadenas E., editors. New Strategies in Prevention and Therapy: Biological Oxidants and Antioxidants. Stuttgart: Hippokrates Verlag; 1994; pp. 181–192.
21. Winterbourn CC. Toxicity of iron and hydrogen peroxide: the Fenton reaction. Toxicol Lett 1995; 82–83:969-974. https://doi.org/10.1016/0378-4274(95)03532-X
22. Deutsch JC. Ascorbic acid oxidation by hydrogen peroxide. Anal Biochem 1998; 255:1-7. https://doi.org/10.1006/abio.1997.2293
23. Noctor G and Foyer CH. Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Mol Biol 1998; 49:249–279. https://doi.org/10.1146/annurev.arplant.49.1.249
24. Sestini EA, Carlson JC, and Allsopp R. The effects of ambient temperature on life span, lipid peroxidation, superoxide dismutase, and phospholipase A2 activity in Drosophila melanogaster. Exp Gerontol 1991; 26:385-395. https://doi.org/10.1016/0531-5565(91)90050-V
25. Feder ME and Hofmann GE. Heat-shock proteins, molecular chaperones, and the stress response: Evolutionary and Ecological Physiology. Annu Rev Physiol 1999, 61:243–282. https://doi.org/10.1146/annurev.physiol.61.1.243
26. Sykiotis GP and Bohmann D. Stress-Activated Cap'n'collar Transcription Factors in Aging and Human Disease. Sci Signal 2010; 3:re3. https://doi.org/10.1126/scisignal.3112re3
27. Mosser DD, Caron AW, Bourget L, Meriin AB, Sherman MY, Morimoto RI et al. The chaperone function of hsp70 is required for protection against stress-induced apoptosis. Mol Cell Biol 2000; 20:7146–7159. https://doi.org/10.1128/MCB.20.19.7146-7159.2000

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1. Solomon S, Plattner GK, Knutti R, and Friedlingstein P. Irreversible climate change due to carbon dioxide emissions. Proc Natl Acad Sci 2009; 106:1704–1709. https://doi.org/10.1073/pnas.0812721106

2. Altan O, Pabuçcuoglu A, Altan A, Konyalioglu S, and Bayraktar H. Effect of heat stress on oxidative stress, lipid peroxidation and some stress parameters in broilers. Br Poult Sci 2003; 44:545-550. https://doi.org/10.1080/00071660310001618334

3. Bhatia B, Kale PG, Daoo JV, and Meshram P. Testicular oxidative stress protective effects of abhraka bhasma in male wistar rats after heat exposure. Int J Pharm Pharm Sci 2013; 5:472-477.

4. Keil G, Cummings E, and Magalhaes JP. Being cool: how body temperature influences ageing and longevity. Biogerontology 2015; 16:383–397. https://doi.org/10.1007/s10522-015-9571-2

5. Watson J. Oxidants, antioxidants and the current incurability of metastatic cancers. Open Biol 2013; 3:120144. https://doi.org/10.1098/rsob.120144

6. Rahal A, Kumar A, Singh V, Yadav B, Tiwari R, Chakraborty S et al. Oxidative stress, prooxidants, and antioxidants: The interplay. BioMed Res Int 2014; vol. 2014, Article ID 761264, 19 pages.

7. Khaliq A, Javed M, Sohail M, and Sagheer M. Environmental effects on insects and their population dynamics. J Entomol Zool Stud 2014; 2:1-7.

8. Brewer MS. Natural antioxidants: sources, compounds, mechanisms of action, and potential applications. Compr Rev Food Sci Food Saf 2011; 10:221–247. https://doi.org/10.1111/j.1541-4337.2011.00156.x

9. Chand K, Rajeshwari, Hiremathad A, Singh M, Santos MA, and Keri RS. A review on antioxidant potential of bioactive heterocycle benzofuran: Natural and synthetic derivatives. Pharmacol Rep 2017; 69:281-295. https://doi.org/10.1016/j.pharep.2016.11.007

10. Pandey KB and Rizvi SI. Plant polyphenols as dietary antioxidants in human health and disease. Oxid Med Cell Longev 2009; 2:270–278. https://doi.org/10.4161/oxim.2.5.9498

11. Subedi RP, Vartak RR, and Kale PG. Effect of abhrak bhasma on the physiology and behaviour of Drosophila melanogaster. Int J Adv Res 2017; 5:1629-1641. https://doi.org/10.21474/IJAR01/3991

12. Bradford MM. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248-254. https://doi.org/10.1016/0003-2697(76)90527-3

13. Aebi HE. Method of Enzymatic Analysis. Vol. 3. Germany-Deerfield, FL: VCH, Weinheim; 1983. Catalase; pp. 273–286.

14. Marklund S and Marklund G. Involvement of the superoxide anion radical in the autooxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974; 47:469-474. https://doi.org/10.1111/j.1432-1033.1974.tb03714.x

15. Rahman I, Kode A, and Biswas SK. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat protoc 2006; 1:3159-3165. https://doi.org/10.1038/nprot.2006.378

16. Brand-williams W, Cuvelier ME, and Berset C. Use of free radical method to evaluate antioxidant activity. LWT-Food Science and Technology 1995; 28:25-30. https://doi.org/10.1016/S0023-6438(95)80008-5

17. Zeb A and Ullah F. A simple spectrophotometric method for the determination of thiobarbituric acid reactive substances in fried fast foods. J Anal Methods Chem 2016; vol. 2016, Article ID 9412767:5 pages.

18. Devasagayam TP, Boloor KK, and Ramasarma T. Methods for estimating lipid peroxidation: an analysis of merits and demerits. Indian J Biochem Biophys 2003; 40:300-8.

19. Bourg EL. Oxidative stress, aging and longevity in Drosophila melanogaster. FEBS Lett 2001; 498:183–186. https://doi.org/10.1016/S0014-5793(01)02457-7

20. Fleming J.E., Niedzwiecki A., Reveillaud I. Stress induced expression of CuZn superoxide dismutase and catalase in senescent Drosophila melanogaster. In: Packer L., Cadenas E., editors. New Strategies in Prevention and Therapy: Biological Oxidants and Antioxidants. Stuttgart: Hippokrates Verlag; 1994; pp. 181–192.

21. Winterbourn CC. Toxicity of iron and hydrogen peroxide: the Fenton reaction. Toxicol Lett 1995; 82–83:969-974. https://doi.org/10.1016/0378-4274(95)03532-X

22. Deutsch JC. Ascorbic acid oxidation by hydrogen peroxide. Anal Biochem 1998; 255:1-7. https://doi.org/10.1006/abio.1997.2293

23. Noctor G and Foyer CH. Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Mol Biol 1998; 49:249–279. https://doi.org/10.1146/annurev.arplant.49.1.249

24. Sestini EA, Carlson JC, and Allsopp R. The effects of ambient temperature on life span, lipid peroxidation, superoxide dismutase, and phospholipase A2 activity in Drosophila melanogaster. Exp Gerontol 1991; 26:385-395. https://doi.org/10.1016/0531-5565(91)90050-V

25. Feder ME and Hofmann GE. Heat-shock proteins, molecular chaperones, and the stress response: Evolutionary and Ecological Physiology. Annu Rev Physiol 1999, 61:243–282. https://doi.org/10.1146/annurev.physiol.61.1.243

26. Sykiotis GP and Bohmann D. Stress-Activated Cap'n'collar Transcription Factors in Aging and Human Disease. Sci Signal 2010; 3:re3. https://doi.org/10.1126/scisignal.3112re3

27. Mosser DD, Caron AW, Bourget L, Meriin AB, Sherman MY, Morimoto RI et al. The chaperone function of hsp70 is required for protection against stress-induced apoptosis. Mol Cell Biol 2000; 20:7146–7159. https://doi.org/10.1128/MCB.20.19.7146-7159.2000