Protective effect of crocin on electromagnetic field-induced testicular damage and heat shock protein A2 expression in male BALB/c mice

Document Type : Original Article

Authors

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

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

Abstract

Objective(s): Exposure to electromagnetic fields (EMF) emitted from mobile phones may cause a deleterious effect on human health and may affect the male reproductive system. Crocin, a carotenoid isolated from Crocus Sativus L. (Saffron), is a phar‌macologically active component of saffron. So, this study was conducted to investigate the protective effect of crocin on the male reproductive system of 60 day old mice after EMF exposure.
Materials and Methods: Twenty-four male Balb/c mice were randomly divided into 4 groups: 1. Em group (2100 MHZ); 2. Cr group (50 mg/kg); 3. Em+Cr group (2100 MHZ+50 mg/kg), and 4. Control group. Sperm parameters (count, and abnormal percent), testis weight index, testis volume, seminiferous tubule diam‌eter, germinal epithelium thickness, LH, FSH and testosterone serum level, testicular Heat shock protein A2 (HspA2) immunoreactivity, and apoptosis were evaluated.
Results: HspA2 immunoreactivity, apoptosis in the germinal epithelium and abnormal sperm were increased in Em group compared with the control group (P<0.05). Sperm count, LH, and testosterone serum level were decreased in the Em group compared with the control group (P<0.05). These parameters were improved in the Em+Cr group compared with Em group significantly (P<0.05).
Conclusion: our findings revealed that EMF exposure leads to harmful impressions on the male reproductive system, while crocin can attenuate EMF-induced destructive effects.

Keywords


1. Agarwal A, Singh A, Hamada A, Kesari K. Cell phones and male infertility: a review of recent innovations in technology and consequences. Int Braz J Urol 2011;37:432-54.
2. Mausset AL, de Seze R, Montpeyroux F, Privat A. Effects of radiofrequency exposure on the GABAergic system in the rat cerebellum: clues from semi-quantitative immunohistochemistry. Brain Res 2001;912:33-46.
3. Kesari KK, Kumar S, Nirala J, Siddiqui MH, Behari J. Biophysical evaluation of radiofrequency electromagnetic field effects on male reproductive pattern. Cell Biochem Biophys 2013;65:85-96.
4. Kesari KK, Kumar S, Behari J. Mobile phone usage and male infertility in Wistar rats. Indian J Exp Biol 2010;48:987-992.
5. Kesari KK, Kumar S, Behari J. Effects of radiofrequency electromagnetic wave exposure from cellular phones on the reproductive pattern in male Wistar rats. Appl Biochem Biotechnol 2011;164:546-559.
6. Deepinder F, Makker K, Agarwal A. Cell phones and male infertility: dissecting the relationship. Reprod Biomed Online 2007;15:266-270.
7. La Vignera S, Condorelli RA, Vicari E, D’Agata R, Calogero AE. Effects of the exposure to mobile phones on male reproduction: a review of the literature. J Androl. 2012;33:350-356.
8. Kang N, Shang XJ, Huang YF. [Impact of cell phone radiation on male reproduction]. Zhonghua Nan Ke Xue 2010;16:1027-1030.
9. Scieglinska D, Piglowski W, Mazurek A, Malusecka E, Zebracka J, Filipczak P, et al. The HspA2 protein localizes in nucleoli and centrosomes of heat shocked cancer cells. J Cell Biochem 2008;104:2193-2206.
10. Mayer MP, Bukau B. Hsp70 chaperones: cellular functions and molecular mechanism. Cell Mol Life Sci 2005;62:670-684.
11. Beere HM. “The stress of dying”: the role of heat shock proteins in the regulation of apoptosis. J Cell Sci 2004;117:2641-2651.
12. Huszar G, Willetts M, Corrales M. Hyaluronic acid (Sperm Select) improves retention of sperm motility and velocity in normospermic and oligospermic specimens. Fertil Steril 1990;54:1127-1134.
13. Huszar G, Stone K, Dix D, Vigue L. Putative creatine kinase M-isoform in human sperm is identifiedas the 70-kilodalton heat shock protein HspA2. Biol Reprod 2000;63:925-932.
14. Jalili C, Salahshoor MR, Moradi S, Pourmotabbed A, Motaghi M. The therapeutic effect of the aqueous extract of boswellia serrata on the learning deficit in kindled rats. Int J Prev Med 2014;5:563-568.
15. Assimopoulou AN, Sinakos Z, Papageorgiou VP. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytother Res 2005;19:997-1000.
16. Mohamadpour AH, Ayati Z, Parizadeh MR, Rajbai O, Hosseinzadeh H. Safety Evaluation of Crocin (a constituent of saffron) Tablets in Healthy Volunteers. Iran J Basic Med Sci 2013;16:39-46.
17. Asadi MH, Zafari F, Sarveazad A, Abbasi M, Safa M, Koruji M, et al. Saffron improves epididymal sperm parameters in rats exposed to cadmium. Nephrourol Mon 2014;6:1-6.
18. Wang Y, Han T, Zhu Y, Zheng CJ, Ming QL, Rahman K, et al. Antidepressant properties of bioactive fractions from the extract of Crocus sativus L. J Nat Med 2010;64:24-30.
19. Nam KN, Park YM, Jung HJ, Lee JY, Min BD, Park SU, et al. Anti-inflammatory effects of crocin and crocetin in rat brain microglial cells. Eur J Pharmacol 2010;648:110-116.
20. Aung HH, Wang CZ, Ni M, Fishbein A, Mehendale SR, Xie JT, et al. Crocin from Crocus sativus possesses significant anti-proliferation effects on human colorectal cancer cells. Exp Oncol 2007;29:175-180.
21. Zheng YQ, Liu JX, Wang JN, Xu L. Effects of crocin on reperfusion-induced oxidative/nitrative injury to cerebral microvessels after global cerebral ischemia. Brain Res 2007;1138:86-94.
22. Hosseinzadeh H, Abootorabi A, Sadeghnia HR. Protective effect of Crocus sativus stigma extract and crocin (trans-crocin 4) on methyl methanesulfonate-induced DNA damage in mice organs. DNA Cell Biol 2008;27:657-664.
23. Hosseinzadeh H, Sadeghnia HR, Ghaeni FA, Motamedshariaty VS, Mohajeri SA. Effects of saffron (Crocus sativus L.) and its active constituent, crocin, on recognition and spatial memory after chronic cerebral hypoperfusion in rats. Phytother Res 2012;26:381-386.
24. Salahshoor MR, Khazaei M, Jalili C, Keivan M. Crocin improves damage induced by nicotine on a number of reproductive parameters in male mice. Int J Fertil Steril 2016;10:71-78.
25. Jalili C, Salahshoor MR, Pourmotabbed A, Moradi S, Roshankhah S, Darehdori AS, et al. The effects of aqueous extract of Boswellia Serrata on hippocampal region CA1 and learning deficit in kindled rats. Res Pharm Sci 2014;9:351-358.
26. Sadoughi S. Effects of crocin on ovarian follicle and serum sex hormone in letrozole-induced polycystic ovarian syndrome in rat model. JAUMS 2017;17:198-210.
27. Durgun M, Dasdag S, Erbatur S, Yegin K, Durgun SO, Uzun C, et al. Effect of 2100 MHz mobile phone radiation on healing of mandibular fractures: an experimental study in rabbits. Biotechnology  Biotechnological Equipment 2016;30:112-120.
28. Fani M, Mohammadipour A, Ebrahimzadeh-Bideskan AR. 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. Iran J Reprod BioMed (Yazd) 2018;16:519-528.
29. Jalilvand N, Hosseini MM, Beheshti F, Ebrahimzadeh-Bideskan A. Protective effect of PPARγ agonist pioglitazone, on testicular tissue and sperm parameters in hypothyroid rats 2019;38:1-10.
30. Bakhtiary Z, Shahrooz R, Ahmadi A, Zarei L. Evaluation of antioxidant effects of crocin on sperm quality in cyclophosphamide treated adult mice. Vet Res Forum 2014;5:213-218.
31. Sehitoglu I, Tumkaya L, Kalkan Y, Bedir R, Cure MC, Zorba OU, et al. Biochemical and histopathological effects on the rat testis after exposure to electromagnetic field during fetal period. Arch Esp Urol 2015;68:562-568.
32. Ebrahimzadeh-Bideskan AR, Mansouri S, Ataei ML, Jahanshahi M, Hosseini M. The effects of soy and tamoxifen on apoptosis in the hippocampus and dentate gyrus in a pentylenetetrazole-induced seizure model of ovariectomized rats. Anat Sci Int 2018;93:218-230.
33. Baghishani F, Mohammadipour A, Hosseinzadeh H, Hosseini M, Ebrahimzadeh-Bideskan AR. The effects of tramadol administration on hippocampal cell apoptosis, learning and memory in adult rats and neuroprotective effects of crocin. Metab Brain Dis 2018;33:907-916.
34. Hosseinian S, Ebrahimzadeh-Bideskan AR, Shafei MN, Sadeghnia HR, Soukhtanloo M, Shahraki S, et al. Nigella sativa extract is a potent therapeutic agent for renal inflammation, apoptosis, and oxidative stress in a rat model of unilateral ureteral obstruction. Phytother Res 2018;32:2290-2298.
35. Mobaraki F, Seghatoleslam M, Fazel A, Ebrahimzadeh-Bideskan AR. Effects of MDMA (ecstasy) on apoptosis and heat shock protein (HSP70) expression in adult rat testis. Toxicol Mech Methods 2018;28:219-229.
36. Rajabzadeh A, Ebrahimzadeh-Bideskan AR, Fazel A, Sankian M, Rafatpanah H, Haghir H. The effect of PTZ-induced epileptic seizures on hippocampal expression of PSA-NCAM in offspring born to kindled rats. J Biomed Sci 2012;19:56-57.
37. Jalayeri-Darbandi Z, Rajabzadeh A, Hosseini M, Beheshti F, Ebrahimzadeh-Bideskan AR. The effect of methamphetamine exposure during pregnancy and lactation on hippocampal doublecortin expression, learning and memory of rat offspring. Anat Sci Int 2018;93:351-363.
38. Haeri P, Mohammadipour A, Heidari Z, Ebrahimzadeh-Bideskan A. Neuroprotective effect of crocin on substantia nigra in MPTP-induced Parkinson’s disease model of mice. Anat Sci Int. 2018;94:119-127.
39. Merhi ZO. Challenging cell phone impact on reproduction: a review. J Assist Reprod Genet 2012;29:293-297.
40. Lee SK, Park S, Gimm YM, Kim YW. Extremely low frequency magnetic fields induce spermatogenic germ cell apoptosis: possible mechanism. Biomed Res Int. 2014;2014:1-8.
41. Hanci H, Odaci E, Kaya H, Aliyazicioglu Y, Turan I, Demir S, et al. The effect of prenatal exposure to 900-MHz electromagnetic field on the 21-old-day rat testicle. Reprod Toxicol 2013;42:203-209.
42. Ozguner M, Koyu A, Cesur G, Ural M, Ozguner F, Gokcimen A, et al. Biological and morphological effects on the reproductive organ of rats after exposure to electromagnetic field. Saudi Med J 2005;26:405-410.
43. Tas M, Dasdag S, Akdag MZ, Cirit U, Yegin K, Seker U, et al. Long-term effects of 900 MHz radiofrequency radiation emitted from mobile phone on testicular tissue and epididymal semen quality. Electromagn Biol Med 2014;33:216-222.
44. Saygin M, Caliskan S, Karahan N, Koyu A, Gumral N, Uguz A. Testicular apoptosis and histopathological changes induced by a 2.45 GHz electromagnetic field. Toxicol Ind Health 2011;27:455-463.
45. Lee HJ, Pack JK, Kim TH, Kim N, Choi SY, Lee JS, et al. The lack of histological changes of CDMA cellular phone-based radio frequency on rat testis. Bioelectromagnetics 2010;31:528-534.
46. Hosseinzadeh H, Shamsaie F, Mehri S. Antioxidant activity of aqueous and ethanolic extracts of Crocus sativus  L. stigma and its bioactive constituents, crocin and safranal. Pharmacognosy Mag 2009;5:419-424.
47. Mousavi M, Baharara J, Zafar-Balanezhad S, Shaheokh-Abadi K. The effect of saffron aqua extract on angiogenesis in chick chorioalantoic membrane. ZJRMS 2014;16:55-58.
48. Al-Akhras MA, Elbetieha A, Hasan MK, Al-Omari I, Darmani H, Albiss B. Effects of extremely low frequency magnetic field on fertility of adult male and female rats. Bioelectromagnetics 2001;22:340-344.
49. Kim YW, Kim HS, Lee JS, Kim YJ, Lee SK, Seo JN, et al. Effects of 60 Hz 14 microT magnetic field on the apoptosis of testicular germ cell in mice. Bioelectromagnetics 2009;30:66-72.
50. Lee JS, Ahn SS, Jung KC, Kim YW, Lee SK. Effects of 60 Hz electromagnetic field exposure on testicular germ cell apoptosis in mice. Asian J Androl 2004;6:29-34.
51. Ramadan LA, Abd-Allah AR, Aly HA, Saad-el-Din AA. Testicular toxicity effects of magnetic field exposure and prophylactic role of coenzyme Q10 and L-carnitine in mice. Pharmacol Res 2002;46:363-370.
52. Kim HS, Park BJ, Jang HJ, Ipper NS, Kim SH, Kim YJ, et al. Continuous exposure to 60 Hz magnetic fields induces duration- and dose-dependent apoptosis of testicular germ cells. Bioelectromagnetics 2014;35:100-107.
53. Whitley D, Goldberg SP, Jordan WD. Heat shock proteins: a review of the molecular chaperones. J Vasc Surg 1999;29:748-751.
54. Rockett JC, Mapp FL, Garges JB, Luft JC, Mori C, Dix DJ. Effects of hyperthermia on spermatogenesis, apoptosis, gene expression, and fertility in adult male mice. Biol Reprod 2001;65:229-239.
55. Beere HM, Wolf BB, Cain K, Mosser DD, Mahboubi A, Kuwana T, et al. Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat Cell Biol 2000;2:469-475.
56. Gaucher J, Reynoird N, Montellier E, Boussouar F, Rousseaux S, Khochbin S. From meiosis to postmeiotic events: the secrets of histone disappearance. FEBS J 2010; 277:599-604.
57. Redgrove KA, Nixon B, Baker MA, Hetherington L, Baker G, Liu DY, et al. The molecular chaperone HSPA2 plays a key role in regulating the expression of sperm surface receptors that mediate sperm-egg recognition. PLoS One 2012;7:1-16.