Evaluation of the anti-oxidant effect of ascorbic acid on apoptosis and proliferation of germinal epithelium cells of rat testis following malathion-induced toxicity

Document Type : Original Article

Authors

1 Department of Anatomy and Cell Biology, Mashhad University of Medical Sciences, Mashhad, Iran

2 Microanatomy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): The aim of this study was to determine the protective role of ascorbic acid on apoptosis and proliferation of spermatogonia and primary spermatocyte cells after malathion administration as an organophosphate pesticide in rat testis.
Materials and Methods: Thirty male Wistar rats were randomly divided into five groups of 6 rats each, including control (no intervention), sham (normal saline 0.09%), malathion (50 mg/kg), malathion plus ascorbic acid (50 mg/kg and 200 mg/kg, respectively), and ascorbic acid (200 mg/kg) groups. Malathion and ascorbic acid were administrated via intraperitoneal injection once per day and seven times per week. After 6 weeks, animals were sacrificed, and testis tissue was used for evaluation of apoptosis and proliferation of germinal epithelium cells using the TUNEL and PCNA staining techniques.
Results: The results of TUNEL staining showed that the numbers of apoptotic cells in spermatogonia and primary spermatocyte cells were significantly increased in the malathion 50 mg/kg group vs control group (P<0.001). Co-administration of malathion 50 mg/kg and ascorbic acid 200 mg/kg significantly decreased the apoptotic cells in both cell types in comparison with malathion 50 mg/kg group (P<0.001). The results of PCNA staining revealed that the proliferation of these cells was significantly decreased in malathion 50 mg/kg group vs control group (P<0.001), and malathion 50 mg/kg plus ascorbic acid 200 mg/kg administration increased the proliferation of cells compared with malathion 50 mg/kg group (P<0.001).
Conclusion: The results provide evidence that ascorbic acid showed preventive effects on malathion-induced toxicity in male rat testis.

Keywords

References

1. Espinoza-Navarro O, Bustos-Obregón E. Effects of malathion on cellularity and sperm differentiation in testis and epididymis of adult rats. Int J Morphol 2014; 32:119-124.
2. Tchounwou PB, Patlolla AK, Yedjou CG, Moore PD. Environmental exposure and health effects associated with Malathion toxicity. Toxicity and Hazard of Agrochemicals: InTech; 2015.
3. Ahmadi S, Jafari M, Asgari A, Salehi M. Acute effect of diazinon on lipid peroxidation level and activities of antioxidant enzymes in rat spleen. J Kermanshah Univ Med Sci 2012; 16:1-9.
4. Bonner MR, Coble J, Blair A, Beane Freeman LE, Hoppin JA, Sandler DP, et al. Malathion exposure and the incidence of cancer in the agricultural health study. Am J Epidemiol 2007; 166:1023-1034.
5. Al-Attar AM. Physiological and histopathological investigations on the effects of alpha-lipoic acid in rats exposed to malathion. J Biomed Biotechnol 2010; 2010:1-8.
6. Croucher L, Jewess P, Roberts MC. Metabolic Pathways of Agrochemicals: Part 2: Insecticides and Fungicides: Royal Society of Chemistry; 2007.
7. Costa LG. Toxic effects of pesticides. Casarett and Doull’s Toxicology: The Basic Science of Poisons 2008; 8:883-930.
8. Abdollahi M, Mostafalou S, Pournourmohammadi S, Shadnia S. Oxidative stress and cholinesterase inhibition in saliva and plasma of rats following subchronic exposure to malathion. Comp Biochem Physiol C Toxicol Pharmacol 2004; 137:29-34.
9. Cerretani D, Riezzo I, Fiaschi AI, Centini F, Giorgi G, D’Errico S, et al. Cardiac oxidative stress determination and myocardial morphology after a single ecstasy (MDMA) administration in a rat model. Int J Legal Med 2008; 122:461-469.
10. Marefati N, Eftekhar N, Kaveh M, Boskabadi J, Beheshti F, Boskabady MH. The effect of Allium cepa extract on lung oxidant, antioxidant, and immunological biomarkers in ovalbumin-sensitized rats. Med Princ Pract 2018; 27:122-128.
11. Hengartner MO. The biochemistry of apoptosis. Nature 2000; 407:770-776.
12. Dadhich RK, Real FM, Zurita F, Barrionuevo FJ, Burgos M, Jiménez R. Role of apoptosis and cell proliferation in the testicular dynamics of seasonal breeding mammals: a study in the Iberian mole, Talpa occidentalis. Biol Reprod 2010; 83:83-91.
13. Oral B, Guney M, Demirin H, Ozguner M, Giray SG, Take G, et al. Endometrial damage and apoptosis in rats induced by dichlorvos and ameliorating effect of antioxidant vitamins E and C. Reprod Toxicol 2006; 22:783-790.
14. Slimen S, Saloua el F, Najoua G. Oxidative stress and cytotoxic potential of anticholinesterase insecticide, malathion in reproductive toxicology of male adolescent mice after acute exposure. Iran J Basic Med Sci 2014; 17:522-530.
15. Sánchez-Peña L, Reyes B, López-Carrillo L, Recio R, Morán-Martínez J, Cebrián M, et al. Changes on sperm chromatin structure in organophosphorus agricultural workers. Toxicol Appl Pharmacol 2004; 196:108-113.
16. Geng X, Shao H, Zhang Z, Ng JC, Peng C. Malathion-induced testicular toxicity is associated with spermatogenic apoptosis and alterations in testicular enzymes and hormone levels in male Wistar rats. Environ Toxicol Pharmacol 2015; 39:659-667.
17. Kelman Z. PCNA: structure, functions and interactions. Oncogene 1997; 14:629.
18. Hirshfield AN. Patterns of [3H] thymidine incorporation differ in immature rats and mature, cycling rats. Biol Reprod 1986; 34:229-235.
19. Chang C-D, Ottavio L, Travali S, Lipson K, Baserga R. Transcriptional and posttranscriptional regulation of the proliferating cell nuclear antigen gene. Mol Cell Biol 1990; 10:3289-3296.
20. Unek G, Ozmen A, Kipmen-Korgun D, Korgun ET. Immunolocalization of PCNA, Ki67, p27 and p57 in normal and dexamethasone-induced intrauterine growth restriction placental development in rat. Acta histochemica 2012; 114:31-40.
21. Jaskulski D, Gatti C, Travali S, Calabretta B, Baserga R. Regulation of the proliferating cell nuclear antigen cyclin and thymidine kinase mRNA levels by growth factors. J Biol Chem 1988; 263:10175-10179.
22. Chieffi P, Franco R, Fulgione D, Staibano S. PCNA in the testis of the frog, Rana esculenta: a molecular marker of the mitotic testicular epithelium proliferation. Gen Comp Endocrinol 2000; 119:11-16.
23. Aluigi M, Guida C, Falugi C. Apoptosis as a specific biomarker of diazinon toxicity in NTera2-D1 cells. Chem Biol Interact 2010; 187:299-303.
24. Agarwal A, Aponte-Mellado A, Premkumar BJ, Shaman A, Gupta S. The effects of oxidative stress on female reproduction: a review. Reprod Biol Endocrinol 2012; 10:49.
25. Siddique YH, Beg T, Afzal M. Antigenotoxic effects of ascorbic acid against megestrol acetate-induced genotoxicity in mice. Hum Exp Toxicol 2005; 24:121-127.
26. du Toit R, Volsteedt Y, Apostolides Z. Comparison of the antioxidant content of fruits, vegetables and teas measured as vitamin C equivalents. Toxicology 2001; 166:63-69.
27.Handelman GJ. Vitamin C deficiency in dialysis patients—are we perceiving the tip of an iceberg? : Oxford University Press; 2006.
28. Uzun FG, Kalender S, Durak D, Demir F, Kalender Y. Malathion-induced testicular toxicity in male rats and the protective effect of vitamins C and E. Food Chem Toxicol 2009; 47:1903-1908.
29. Ramanathan K, Balakumar B, Panneerselvam C. Effects of ascorbic acid and a-tocopherol on arsenic-induced oxidative stress. Hum Exp Toxicol 2002; 21:675-680.
30. Anbarkeh FR, Nikravesh MR, Jalali M, Sadeghnia HR, Sargazi Z, Mohammdzadeh L. Single dose effect of diazinon on biochemical parameters in testis tissue of adult rats and the protective effect of vitamin E. Iran J Reprod Med 2014; 12:731-736.
31. Kalender S, Uzun FG, Durak D, Demir F, Kalender Y. Malathion-induced hepatotoxicity in rats: the effects of vitamins C and E. Food Chem Toxicol 2010; 48:633-638. 32. Possamai FP, Fortunato JJ, Feier G, Agostinho FR, Quevedo J, Wilhelm Filho D, et al. Oxidative stress after acute and sub-chronic malathion intoxication in Wistar rats. Environ Toxicol Pharmacol 2007; 23:198-204. 33. Fortunato JJ, Agostinho FR, Reus GZ, Petronilho FC, Dal-Pizzol F, Quevedo J. Lipid peroxidative damage on malathion exposure in rats. Neurotox Res 2006; 9:23-28. 34. Choudhary N, Goyal R, Joshi SC. Effect of malathion on reproductive system of male rats. J Environ Biol 2008; 29:259-262.
35. Amirkabirian N, Teimouri F, Esmaily H, Mohammadirad A, Aliahmadi A, Abdollahi M. Protection by pentoxifylline of diazinon-induced toxic stress in rat liver and muscle. Toxicol Mech Methods 2007; 17:215-221.
36. Uzunhisarcikli M, Kalender Y, Dirican K, Kalender S, Ogutcu A, Buyukkomurcu F. Acute, subacute and subchronic administration of methyl parathion-induced testicular damage in male rats and protective role of vitamins C and E. Pest Biochem Physiol 2007; 87:115-122.
37. Ozmen O. Cardiotoxicity and apoptotic activity in subacute endosulfan toxicity and the protective effect of vitamin C in rabbits: a pathological study. J Environ Pathol Toxicol Oncol 2013; 32.
38. Sargazi Z, Nikravesh MR, Jalali M, Sadeghnia HR, Anbarkeh FR. Apoptotic effect of organophosphorus insecticide diazinon on rat ovary and protective effect of vitamin E. Iran J Toxicol  2016; 10:37-44.
39. Mobaraki F, Seghatoleslam M, Fazel A, Ebrahimzadeh-Bideskan A. Effects of MDMA (ecstasy) on apoptosis and heat shock protein (HSP70) expression in adult rat testis. Toxicol Mech Methods 2018; 28:219-229.
40. Khordad E, Fazel A, Ebrahimzadeh Bideskan A. The effect of ascorbic acid and garlic administration on lead-induced apoptosis in rat offspring,s eye retina. Iran Biomed J 2013; 17:206-213.
41. Hall P, Levison D, Woods A, Yu C, Kellock D, Watkins J, et al. PCNA immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol 1990; 162:285-294.
42. Bravo R, Macdonald-Bravo H. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol 1987; 105:1549-1554.
43. Rajabzadeh AA, Bideskan AR, Haghir H, Fazel AR. Morphometrical study of polysialylated neural cell adhesion molecule positive cells in rat pups hippocampus following induction of seizure during pregnancy. Iran Biomed J 2011; 15:157-163.
44. Penna-Videau S, Bustos-Obregón E, Cermeño-Vivas J, Chirino D. Malathion Affects Spermatogenic Proliferation in Mouse. Int J Morphol 2012; 30:1399-1407.
45. Güney M, Oral B, Demirin H, Özgüner M, Take G, Mungan T, et al. Evaluation of caspase-dependent apoptosis during methyl parathion-induced endometrial damage in rats: Ameliorating effect of Vitamins E and C. Environ Toxicol Pharmacol 2007; 23:221-227.
46. Yu F, Wang Z, Ju B, Wang Y, Wang J, Bai D. Apoptotic effect of organophosphorus insecticide chlorpyrifos on mouse retina in vivo via oxidative stress and protection of combination of vitamins C and E. Exp Toxicol Pathol 2008; 59:415-423.
47. Fani M, Mohammadipour A, Ebrahimzadeh-Bideskan A. The effect of crocin on testicular tissue and sperm parameters of mice offspring from mothers exposed to atrazine during pregnancy and lactation periods: An experimental study. Int J Reprod Biomed (Yazd) 2018; 16:519-528.
48. Jia Z, Misra HP. Reactive oxygen species in in vitro pesticide-induced neuronal cell (SH-SY5Y) cytotoxicity: Role of NFκB and caspase-3. Free Radic Biol Med 2007; 42:288-298.
49. Carlson K, Ehrich M. Distribution of SH‐SY5Y human neuroblastoma cells in the cell cycle following exposure to organophosphorus compounds. J Biochem Mol Toxicol 2008; 22:187-201.
50. Sarabia L, Maurer I, Bustos-Obregon E. Melatonin prevents damage elicited by the organophosphorous pesticide diazinon on mouse sperm DNA. Ecotoxicol Environ Saf 2009; 72:663-668.
Iranian Journal of Basic Medical Sciences
Volume 23, Issue 5
May 2020
Pages 569-575

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