IN VIVO RADIOPROTECTIVE EFFECTS OF WHEATGRASS (TRITICUM AESTIVUM) EXTRACT AGAINST X-IRRADAITION-INDUCED OXIDATIVE STRESS AND APOPTOSIS IN PERIPHERAL BLOOD LYMPHOCYTES IN RATS
Â Objectives: The present study was undertaken to investigate the possible protective potential of wheatgrass extract against radiation-induced toxicity in peripheral lymphocytes of rats exposed to a fractionated dose of X-rays.
Methods: Effects of the X-irradiation with and without wheatgrass were studied on various parameters in peripheral lymphocytes including antioxidant defense system and apoptosis. Male Sprague-Dawley rats were divided into four different groups: Normal control group, X-ray-irradiated group (21 Gy over a span of 7 days), wheatgrass-treated group (80 mg/100 g bodyweight for 2 weeks), and X-rays-irradiated + wheatgrass-treated group. All the biochemical indices which included lipid peroxidation (LPO), reduced glutathione, reactive oxygen species (ROS), and activities of antioxidant enzymes were investigated in lymphocytes. Terminal deoxynucleotidyl transferase dUTP nick-end labeling assay was carried out to assess the apoptosis in lymphocytes following whole-body X-irradiation.
Results: Whole-body X-irradiation to rats resulted in significant increase in LPO with concomitant depression of antioxidant enzymes activities, namely, superoxide dismutase, catalase, and glutathione peroxidise (GPx) in lymphocytes. Further, the present study witnessed a significant increase in the number of apoptotic lymphocytes in the X-irradiated animals. However, wheatgrass supplementation lowered the LPO levels, restored cellular antioxidant status, and provided significant protection against radiation-induced apoptosis.
Conclusions: Based on these observations, the present study suggests that wheatgrass extract has the potential to be used as an effective radioprotectant against radiation-induced oxidative stress and apoptosis in peripheral lymphocytes of whole-body X-ray-exposed rats.
2. Goans RE, Holloway EC, Berger ME, Ricks RC. Early dose assessment following severe radiation accidents. Health Phys 1997;72:513-8.
3. Mauch P, Constine L, Greenberger J, Knospe W, Sullivan J, Liesveld JL, et al. Hematopoietic stem cell compartment: Acute and late effects of radiation therapy and chemotherapy. Int J Radiat Oncol Biol Phys 1995;31:1319-39.
4. Morgan WF. Radiation-induced genomic instability. Health Phys 2011;100:280-1.
5. Dainiak N. Hematologic consequences of exposure to ionizing radiation. Exp Hematol 2002;30:513-28.
6. Jagetia GC, Venkatesha VA. Mangiferin protects human peripheral blood lymphocytes against Î³-radiation-induced DNA strand breaks: A fluorescence analysis of DNA unwinding assay. Nutr Res 2006;26:303-11.
7. Ozasa K, Shimizu Y, Suyama A, Kasagi F, Soda M, Grant EJ, et al. Studies of the mortality of atomic bomb survivors. Report 14, 1950-2003. An overview of cancer and noncancer diseases. Radiat Res 2012;177:229-43.
8. Riley PA. Free radicals in biology: Oxidative stress and the effects of ionizing radiation. Int J Radiat Biol 1994;65:27-33.
9. Bhosle SM, Huilgol NG, Mishra KP. Enhancement of radiation-induced oxidative stress and cytotoxicity in tumor cells by ellagic acid. Clin Chim Acta 2005;359:89-100.
10. Von Sonntag C. The Chemical Basis of Radiation Biology. London: Taylor & Francis; 1987. p. 65-84.
11. Livesey JC, Reed DJ. Chemical protection against ionizing radiation. Adv Radiat Res 1987;13:285-353.
12. Mansour HH, Hafez HF, Fahmy NM, Hanafi N. Protective effect of N-acetylcysteine against radiation induced DNA damage and hepatic toxicity in rats. Biochem Pharmacol 2008;75:773-80.
13. Glowe PJ, Glick JH, Weiler C. Phase I trials of WR 2721 and Cisplatinum. Int J Radiat Oncol Biol Phys 1984;10:1781-7.
14. Aydos OS, Avci A, Ozkan T, Karadag A, Gurleyik E, Altinok B, et al. Antiproliferative, apoptotic and antioxidant activities of wheatgrass (Triticumaestivum L.) extract on CML (K562) cell line. Turk J Med Sci 2011;41:657-63.
15. Lee SH, Lee YM, Lee HS, Kim DK. Antioxidative and antihyperglycemia effects of Triticumaestivum wheat sprout water extracts on the Streptozotocin-induced diabetic mice. Korean J Pharmacogn 2009;40:408-14.
16. Peryt B, Szymczyk T, Lesca P. Mechanism of antimutagenicity of wheat sprout extracts. Mutat Res 1992;269:201-15.
17. Tandon S, Arora A, Singh S, Monga J, Arora S. Antioxidant profiling of Triticumaestivum(wheatgrass) and its antiproliferative activity in MCF-7 breast cancer cell line. J Pharm Res 2011;4:4601-4.
18. Yi B, Kasai H, Lee HS, Kang Y, Park JY, Yang M. Inhibition by wheat sprout (Triticumaestivum) juice of bisphenol A-induced oxidative stress in young women. Mutat Res 2011;724:64-8.
19. Calzuola I, Gianfranceschi GL, Marsili V. Comparative activity of antioxidants from wheat sprouts, Morinda citrifolia, fermented papaya and white tea. Int J Food Sci Nutr 2006;57:168-77.
20. Kulkarni SD, Acharya R, Nair AG, Rajurkar NS, Reddy AV. Determination of elemental concentration profiles in tender wheatgrass (Triticumaestivum L.) using instrumental neutron activation analysis. Food Chem 2006;95:699-707.
21. Benincasa P, Galieni A, Manetta AC, Pace R, Guiducci M, Pisante M, et al. Phenolic compounds in grains, sprouts and wheatgrass of hulled and non-hulled wheat species. J Sci Food Agric 2015;95:1795-803.
22. Tyl CE, Bunzel M. Antioxidant activity-guided fractionation of blue wheat (UC66049 Triticumaestivum L.). J Agric Food Chem 2012;60:731-9.
23. Shirude AA. Phytochemical and pharmacological screening of wheatgrass juice (Triticumaestivum L.). Int J Pharm Sci Rev Res 2011;9:159-64.
24. Singh K, Pannu MS, Singh P, Singh J. Effects of wheat grass tablets on the frequency of blood transfusions in Thalassemia Major. Indian J Pediatr 2010;77:90-1.
25. Sharma P, Singla N, Dhawan DK. Evidence of zinc in affording protection against X-ray-induced brain injury in rats. Biol Trace Elem Res 2017;179:247-58.
26. Kamboj JK, Rana SV, Dhawan DK, Vahiphei K. Role of wheatgrass in prevention of carbon tetrachloride induced hepatotoxicity in rats. J Clin Exp Hepatol 2011;1:S1.
27. Gupta BL, Bhat RM, Gomathy KR, Susheela B. Radiation chemistry of ferrous sulphate-benzoic and xylenol orange system. Radiat Res 1978;75:269-77.
28. Hudson L, Hay FC. Practical Immunology. 4th ed. Oxford: Blackwell Scientific Publications; 1991.
29. Wills ED. Mechanism of lipid peroxide formation in animal tissues. Biochem J 1966;99:667-76.
30. Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys 1959;82:70-7.
31. Catalase LH. In: Bergmeyer HU, editor. Methods of Enzymatic Analysis. New York: Academic Press; 1971. p. 885-93.
32. FlohÃ© L, GÃ¼nzler WA. Assays of glutathione peroxidase. In: Packer L, editor. Methods in Enzymology. New York: Academic; 1984. p. 114-21.
33. Kono Y. Generation of superoxide radical during auto oxidation of hydroxylamine and an assay for superoxide dismutase. Arch Biochem Biophys 1978;186:189-95.
34. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurementswith the Folin-phenol reagent. J Biol Chem 1951;193:265-75.
35. Driver AS, Kodavanti PR, Mundy WR. Age-related in reactive oxygen species production in rat brain homogenates. Neurotoxicol Teratol 2000;22:175-81.
36. Singh N, Verma P, Pandey BR. Therapeutic potential of organic TriticumaestivumLinn. (Wheatgrass) in prevention and treatment of chronic diseases: An Overview. Int J Pharm Sci Drug Res 2012;4:10-4.
37. Edwards JC, Chapman D, Cramp WA, Yatvin MB. The effects of ionizing radiation on biomembrane structure and function. Prog Biophys Mol Biol 1984;43:71-93.
38. Ianzini F, Guidoni L, Indovina PL, Viti V, Erriu G, Onnis S, et al. Gamma-irradiation effects on phosphatidylcholine multilayer liposomes: Calorimetric, NMR, and spectrofluorimetric studies. Radiat Res 1984;98:154-66.
39. Sheriff NH, Hawas AM, Abdallah WE, Saleh IA, Shams KA, Hammouda FM. Potential role of milk thistle seed and its oil extracts against heart and brain injuries induced by Î³-radiation exposure. Int J Pharm Pharm Sci 2017;9:52-8.
40. Pamplona R. Advanced lipoxidation end-products. Chem Biol Interact 2011;192:14-20.
41. GarcÃa-Gil FA, Albendea CD, LÃ³pez-PingarrÃ³n L, Royo-Dachary P, MartÃnez-GuillÃ©n J, Piedrafita E, et al. Altered cellular membrane fluidity levels and lipid peroxidation during experimental pancreas transplantation. J Bioenerg Biomembr 2012;44:571-7.
42. Srinivasan M, Sudheer AR, Rajasekaran KN, Menon VP. Effects of curcuminanalog on Î³-radiation induced cellular changes in primary culture of isolated rat hepatocytes in vitro. Chem Biol Interact 2008;176:1-8.
43. Goel A, Dani V, Dhawan DK. Protective effects of zinc on lipid peroxidation, antioxidant enzymes and hepatic histoarchitecture in chlorpyrifos-induced toxicity. Chem Biol Interact 2005;156:131-40.
44. PrÃ¡ D, Franke SI, Henriques JA, Fenech M. Iron and genome stability: An update. Mutat Res 2012;733:92-9.
45. Dani V, Dhawan DK. Zinc as an antiperoxidative agent following iodine-131 induced changes on the antioxidant system and on the morphology of red blood cells in rat. Hell J Nucl Med 2006;9:22-6.
46. Malhotra A, Dhawan DK. Zinc improves antioxidative enzymes in red blood cells and hematology in lithium-treated rats. Nutr Res 2008;28:43-50.
47. Younes M, Siegers CP. Mechanistic aspects of enhanced lipid peroxidation following glutathione dependent depletion in vivo. Chem Biol Interact 1981;34:257-66.
48. Cario E, Jung S, Harder Dâ€™Heureuse J, Schulte C, Sturm A, Wiedenmann B, et al. Effects of exogenous zinc supplementation on intestinal epithelial repair in vitro. Eur J Clin Invest 2000;30:419-28.
49. Manasaveena V, Akula KK, Sangram V. A comparative evaluation of enzymatic antioxidant levels in pre and post therapy patients with oral cancer. Int J Pharm Pharm Sci 2014;6:52-6.
50. Fraga CG. Relevance, essentiality and toxicity of trace elements in human health. Mol Aspects Med 2005;26:235-44.
51. MatÃ©s JM, SÃ¡nchez-JimÃ©nez F. Antioxidant enzymes and their implications in pathophysiologic processes. Front Biosci 1999;4:D339-45.
52. Kumari SN, Sanjeev G, Rao VC. Effect of ethanolic extract of Cyanotis cristata leaves applied topically on wound healing in wistar rats. Int J Pharm Pharm Sci 2015;7:63-7.
53. Eder E, Wacker M, Lutz U, Nair J, Fang X, Batsch H, et al. Oxidative stress related DNA adducts in the liver of female rats fed with sunflower, rapeseed, olive or coconut oil supplemented diets. Chem Biol Interact 2006;159:81-9.
54. Hartwig A. Role of magnesium in genomic stability. Mutat Res 2001;475:113-21.
55. Ho E. Zinc deficiency, DNA damage and cancer risk. J Nutr Biochem 2004;15:572-8.
56. Hsu CY, Yang CM, Chen CM, Chao PY, Hu SP. Effects of chlorophyll-related compounds on hydrogen peroxide induced DNA damage within human lymphocytes. J Agric Food Chem 2005;53:2746-50.
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