Comparison of the effect of exosomes derived from Sertoli cells with vitamin C on damage induced by electromagnetic field (50 Hz) in spermatogonial stem cells

Subject Areas :

Farzaneh Salek ¹ (Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran)
Javad Baharara ² (Department of Biology & Research Center for Animal Development Applied Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran.)
Khadijeh Nejad Shahrokhabadi ³ (Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran)
Elaheh Amini ⁴ (Department of Cellular & Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran)

Received: 2020-09-13 Accepted : 2020-09-20 Published : 2020-09-22

Keywords: Electromagnetic field, Apoptosis, Sertoli cells, Exosomes, Spermatogonial Stem Cells,

Abstract :

Introduction & Objective: Spermatogonial stem cells (SSCs) as adult stem cells are crucial for spermatogenesis. Electromagnetic fields (EMF) leads to biological activity disruption of these cells and spermatogenesis. Antioxidants like vitamin C can reduce the damage caused by EMF through oxidative stress reduction. Recent studies also reported the key role of Sertoli cell paracrine signaling in regulating the maintenance and differentiation of SSCs. Thus, we examined and compared the effect of vitamin C and exosomes derived from Sertoli cells on damage induced by EMF in SSCs. Material and Methods: SSCs and Sertoli cells were isolated from the testes of immature male mice. The alkaline phosphatase activity of SSCs was investigated. SSCs were exposed to 50 Hz EMF intensity of 2.5 mT for one hour/five days and were treated with the optimal concentration of vitamin C and various concentrations of exosome. Then the rate of viability, colonization capacity, and apoptosis of these cells were examined. Results: Our results showed the destructive effect of EMF by reducing viability, colonization rate and alteration of SSCs nuclei. Also, these results were confirmed by increasing expression level of Caspase 9 as apoptotic gene and down-regulation of SOD as antioxidant gene. The addition of vitamin C and exosomes improved the alterations induced by EMF in SSCs; however exosomes had more ameliorative effect in comparison with vitamin C on these alterations. Conclusion: These findings demonstrated the capacity and effectiveness of exosomes as a new therapeutic agent that can restore SSCs microenvironment damaged caused by EMF exposure.

References:

1. Abumadighem, A., Solomon, R., Stepanovsky, A., Kapelushnik, J., Shi, Q., Meese, E. (2018). Development of spermatogenesis in vitro in three-dimensional culture from spermatogonial cells of busulfan-treated immature mice. Int J Mol Sci, 19(12); 3804.

2.Azab, AE., Khalat, AM., Ebrahim, SA. (2018). Electromagnetic fields and its harmful effects on the male reproductive electromagnetic fields and its harmful effects on the male reproductive system. Biosci Bioeng, 4; 1–13.

3.Azizi, H., Hamidabadi, HG., Skutella, T. (2018). Differential proliferation effects after short-term cultivation of mouse spermatogonial stem cells on different feeder layers. Cell J, 21;186–193.

4.Bahaodini, A., Owjfard, M., Tamadon, A., Jafari, SM. (2015). Low frequency electromagnetic fields long-term exposure effects on testicular histology, sperm quality and testosterone levels of male rats. Asian Pacific J Reprod, 4;195–200.

5.Baharara, J., Amini, E., Salek-abdollahi, F., Nikdel, N., Asadi-Samani, M. (2015). Protective effect of date palm pollen ( Phoenix dactylifera ) on sperm parameters and sexual hormones in male NMRI mice exposed to low frequency electromagnetic field( 50 Hz ). J HerbMed Pharmacol, 4; 75–80.

6.Baharara, J., Hosseini, N., Farzin, TR. (2016). Extremely low frequency electromagnetic field sensitizes cisplatin-resistant human ovarian adenocarcinoma cells via P53 activation. Cytotechnology, 68; 1403–1413.

7.David, S., Orwig, KE. (2020). Spermatogonial stem cell culture in oncofertility. Urol Clin North Am, 47; 227–244.

8.Ding, Z., Li, J., Li, F., Mephryar, MM., Wu, S., Zhang, C. (2017). Vitamin C and vitamin E protected B95-8 and Balb/c-3T3 cells from apoptosis induced by intermittent 50Hz ELF-EMF radiation. Iran J Public Health, 46; 23–34.

9.Duan, Y., Wang, Z., Zhang, H., He, Y., Lu, R., Zhang, R. (2013). The preventive effect of lotus seedpod procyanidins on cognitive impairment and oxidative damage induced by extremely low frequency electromagnetic field exposure. Food Funct, 4; 1252–1262.

10.Fathi, E., Farahzadi, R., Rahbarghazi, R., Kafil, HS., Yolmeh, R. (2017). Rat adipose-derived mesenchymal stem cells aging reduction by zinc sulfate under extremely low frequency electromagnetic field exposure is associated with increased telomerase reverse transcriptase gene expression. Vet Res Forum an Int Q J, 8; 89–96.

11.Fereshteh, Z., Schmidt, SA., Al-Dossary, AA., Accerbi, M., Arighi, C., Cowart, J. (2018). Murine oviductosomes (OVS) microRNA profiling during the estrous cycle: Delivery of OVS-borne microRNAs to sperm where miR-34c-5p localizes at the centrosome. Sci Rep, 8; 16094.

12.Fu, C., Rojas, T., Chin, AC., Cheng, W., Bernstein, IA., Albacarys, LK. (2018). Multiple aspects of male germ cell development and interactions with Sertoli cells require inositol hexakisphosphate kinase-1. Sci Rep, 8; 1–13.

13.Gassei, K., Orwig, KE. (2016). Experimental methods to preserve male fertility and treat male factor infertility. Fertil Steril, 105; 256–266.

14.Górski, R., Kotwicka, M., Skibińska, I., Jendraszak, M., Wosiński, S. (2020). Effect of low-frequency electric field screening on motility of human sperm. Ann Agric Environ Med, 27; 427–434.

15.Guney, M., Ozguner, F., Oral, B., Karahan, N., Mungan, T. (2007). 900 MHz radiofrequency-induced histopathologic changes and oxidative stress in rat endometrium: Protection by vitamins E and C. Toxicol Ind Health, 23; 411–420.

16.Gurunathan, S., Kang, M., Jeyaraj, M., Qasim, M., Kim, J. (2019). Review of the isolation, characterization, biological function, and multifarious therapeutic approaches of exosomes. Cells, 8; 307.

17.Huang, B., Lu, J., Ding, C., Zou, Q., Wang, W., Li, H. (2018). Exosomes derived from human adipose mesenchymal stem cells improve ovary function of premature ovarian insufficiency by targeting SMAD. Stem Cell Res Ther, 9; 1–12.

18.Javeed, N., Mukhopadhyay, D. (2017). Exosomes and their role in the micro-/macro-environment: A comprehensive review. J Biomed Res, 31; 386–394.

19.Kaur, G., Vadala, S., Dufour, JM. (2017). An overview of a Sertoli cell transplantation model to study testis morphogenesis and the role of the Sertoli cells in immune privilege. Environ Epigenetics, 3; 1–10.

20.Kesari, KK., Agarwal, A., Henkel, R. (2018). Radiations and male fertility. Reprod Biol Endocrinol, 16; 118.

21.Koziorowska, A., Kozioł, K., Gniady, S., Romerowicz-Misielak, M. (2018). An electromagnetic field with a frequency of 50 Hz and a magnetic induction of 2.5 mT affects spermatogonia mouse cells(GC-1spg line). Prz Elektrotechniczny, 94; 132–135.

22.Liu, C., Duan, W., Xu, S., Chen, C., He, M., Zhang, L. (2013). Exposure to 1800 MHz radiofrequency electromagnetic radiation induces oxidative DNA base damage in a mouse spermatocyte-derived cell line. Toxicol Lett, 218; 2–9.

23.Mancuso, F., Calvitti, M., Milardi, D., Grande, G., Falabella, G., Arato, I. (2018). Testosterone and FSH modulate Sertoli cell extracellular secretion: Proteomic analysis. Mol Cell Endocrinol, 476; 1–7.

24.Marjault, HB., Allemand, I. (2016). Consequences of irradiation on adult spermatogenesis: Between infertility and hereditary risk. Mutat Res - Rev Mutat Res, 770; 340–348.

25.Masaki, K., Sakai, M., Kuroki, S., Jo, JI., Hoshina, K., Fujimori, Y., Oka, K. (2018). FGF2 has distinct molecular functions from gdnf in the mouse germline niche. Stem Cell Reports, 10; 1782–1792.

26.Navid, S., Rastegar, T., Baazm, M., Alizadeh, R., Talebi, A., Gholami, K. (2017). In vitro effects of melatonin on colonization of neonate mouse spermatogonial stem cells. Syst Biol Reprod Med, 63; 370–381.

27.Pandey, N., Giri, S. (2018). Melatonin attenuates radiofrequency radiation(900 MHz)-induced oxidative stress, DNA damage and cell cycle arrest in germ cells of male Swiss albino mice. Toxicol Ind Health, 34; 315–327.

28.Salek, F., Baharara, J., Nejad Shahrokhabadi, K., Amini, E. (2015). The guardians of germ cells; the role of Sertoli-derived exosomes on electromagnetic field-induced oxidative stress in mouse spermatogonial stem cells in vitro. Int J Radiat Biol Under review.

29.Saygin, M., Ozmen, O., Erol, O., Ellidag, HY., Ilhan, I., Aslankoc, R. (2018). The impact of electromagnetic radiation(2.45 GHz, Wi-Fi) on the female reproductive system: The role of vitamin C. Toxicol Ind Health, 34; 620–630.

30.Sharma, G., Sisodia, R., Meghnani, E. (2015). Radiation induced testicular injury and its amelioration by prunus domestica in Swiss albino mice. Iran J Radiat Res, 13; 45–54.

31.Solek, P., Majchrowicz, L., Bloniarz, D., Krotoszynska, E., Koziorowski, M. (2017). Pulsed or continuous electromagnetic field induce p53/p21-mediated apoptotic signaling pathway in mouse spermatogenic cells in vitro and thus may affect male fertility. Toxicology, 382; 84–92.

32.Solek, P., Majchrowicz, L., Koziorowski, M. (2018). Aloe arborescens juice prevents EMF-induced oxidative stress and thus protects from pathophysiology in the male reproductive system in vitro. Environ Res, 166; 141–149.

33.Sun, L., Li, D., Song, K., Wei, J., Yao, S., Li, Z. (2017). Exosomes derived from human umbilical cord mesenchymal stem cells protect against cisplatin-induced ovarian granulosa cell stress and apoptosis in vitro. Sci Rep, 7; 1–13.

34.Wang, J., Cao, H., Xue, X., Fan, C., Fang, F., Zhou, J. (2014). Effect of vitamin C on growth of caprine spermatogonial stem cells invitro. Theriogenology, 81; 545–555.

35.Yahyazadeh, A., Deniz, ÖG., Kaplan, AA., Altun, G., Yurt, KK., Davis, D. (2018). The genomic effects of cell phone exposure on the reproductive system. Environ Res, 167; 684–693.

36.Yang, C., Yao, C., Tian, R., Zhu, Z., Zhao, L., Li, P. (2019). miR-202-3p Regulates sertoli cell proliferation, synthesis function, and apoptosis by targeting LRP6 and cyclin D1 of Wnt/β-catenin signaling. Mol Ther - Nucleic Acids, 14; 1–19.

37.Yin, K., Wang, S., Zhao, RC. (2019). Exosomes from mesenchymal stem/stromal cells: A new therapeutic paradigm. Biomark Res, 7; 1–8.

38.Zhang, W., Yang, C., Guo, W., Guo, X., Liu, C. (2018). Protective effect of bone marrow mesenchymal stem cells-derived exosomes against testicular ischemia-reperfusion injury in rats. J South Med Univ, 38; 910–916.

39.Zhang, XF., Choi, YJ., Han, JW., Kim, E., Park, JH., Gurunathan, S. (2015). Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells. Int J Nanomedicine, 10; 1335–1357