مقایسه اثر اگزوزوم های مشتق از سلول های سرتولی با ویتامین C بر آسیب های ناشی از میدان الکترومغناطیس در سلول های بنیادی اسپرماتوگونی
محورهای موضوعی : مجله پلاسما و نشانگرهای زیستی
فرزانه سالک
1
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جواد بهارآرا
2
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خدیجه نژاد شاهرخ آبادی
3
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الهه امینی
4
1 - گروه زیست شناسی،دانشکده علوم،واحدمشهد دانشگاه آزاداسلامی ،مشهد،ایران.
2 - گروه زیست شناسی و مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهددانشگاه آزاد اسلامی ، مشهد.ایران
3 - گروه زیست شناسی،دانشکده علوم،واحدمشهد دانشگاه آزاداسلامی ،مشهد،ایران.
4 - دانشگاه خوارزمی، دانشکده علوم زیستی، گروه سلولی مولکولی، تهران، ایران
کلید واژه: میدان های الکترومغناطیسی, آپوپتوز, سرتولی, سلول های بنیادی اسپرماتوگونی, اگزوزوم,
چکیده مقاله :
زمینه و هدف: سلول های بنیادی اسپرماتوگونی( SSCs) به عنوان سلول های بنیادی بالغ برای اسپرماتوژنز ضروری هستند. میدان های الکترومغناطیسی(EMF) باعث اختلال در فعالیت زیستی این سلول ها و اسپرماتوژنز می شود. آنتی اکسیدان ها هم چون ویتامین C آسیب های ناشی از EMF را از طریق کاهش استرس اکسیداتیو بهبود می بخشند. هم چنین سیگنالینگ پاراکرین سلول های سرتولی در حفظ و تمایز SSCها نقشی اساسی دارد. در نتیجه به مقایسه اثر ویتامین C و اگزوزوم های مشتق از سلول های سرتولی بر آسیب های ناشی از EMF در SSCها پرداخته شد. روش کار: سلول های اسپرماتوگونی و سرتولی از بیضه موش های نر نابالغ جداسازی شد. فعالیت آلکالین فسفاتازی SSCها بررسی شد. SSCها در معرض میدان الکترومغناطیس 50 هرتز و شدت 5/2 میلی تسلا به مدت یک ساعت، پنج روز قرار گرفتند و توسط غلظت بهینه ویتامین C و غلظت های مختلف اگزوزوم تیمار شدند. سپس زیستایی، توانایی کلونی زایی و آپوپتوز این سلول ها بررسی گردید. یافته ها:EMF باعث کاهش زیستایی، کلونی زایی و تغییرات هسته SSCها شد. هم چنین نتایج به دست آمده با افزایش بیان ژن کاسپاز 9 و کاهش ژن SOD تأیید گردید. افزودن ویتامین C و اگزوزوم ها باعث بهبود تغییرات القائ شده توسط EMF در SSCها شد، با این وجود اگزوزوم ها دارای اثرات بهبود بخش بیشتری نسبت به ویتامین C بود نتیجه گیری: یافته ها اثربخشی اگزوزوم ها را به عنوان یک عامل درمانی جدید نشان داد که می تواند آسیب های ریز محیط SSCها را پس از قرار گرفتن در معرض EMF بازیابی کند.
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.
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