en اثرات ضد آپوپتوزی محیط مشروط مشتق از سلول بنیادی مزانشیمی مغز استخوان بر آسیب بافت بیضه در رت های دریافت کننده سیکلوفسفامید: یک مطالعه آزمایشگاهی Reproductive Oncology Original Article Original Article <div style="text-align: justify;"><span style="font-size:12pt"><span style="text-justify:inter-ideograph"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مقدمه: </span></span></b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">سیکلوفسفامید (</span></span><span dir="LTR" style="font-size:10.0pt">CP</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) اثرات منفی بر سیستم تولید&shy;مثل دارد. سلول&shy;های بنیادی و متابولیت&shy;های آنها جهت افزایش باروری پس از شیمی درمانی مورد استفاده قرار گرفته&shy;اند. </span></span></span></span></span></span></span><br> <span style="font-size:12pt"><span style="text-justify:inter-ideograph"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">هدف: </span></span></b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">این مطالعه با هدف بررسی تأثیر محیط مشروط (</span></span><span dir="LTR" style="font-size:10.0pt">Conditioned medium: CM</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) مشتق شده از سلول&shy;های بنیادی مزانشیمی مشتق از مغز استخوان بر اثرات سمی </span></span><span dir="LTR" style="font-size:10.0pt">CP</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">بر بیضه</span></span><span dir="LTR" style="font-size:11.0pt"><span arial="" style="font-family:">&shy;</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">ها انجام شد.</span></span><b><span dir="LTR" style="font-size:11.0pt"><span style="font-family:"Arial","sans-serif""></span></span></b></span></span></span></span></span><br> <span style="font-size:12pt"><span style="text-justify:inter-ideograph"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مواد و روش&shy; ها: </span></span></b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">سلول‌های بنیادی مزانشیمی مغز استخوان استخراج، محیط مشروط جمع‌آوری و 25 برابر تغلیظ شد. 24 سر موش&shy;های صحرایی نر نژاد ویستار (8 هفته، 250-200 گرم) به صورت تصادفی به 4 گروه تقسیم شدند: کنترل، </span></span><span dir="LTR" style="font-size:10.0pt">CP</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، </span></span><span dir="LTR" style="font-size:10.0pt">CP+DMEM</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، </span></span><span dir="LTR" style="font-size:10.0pt">CP+CM</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> .</span></span><span dir="LTR" style="font-size:10.0pt">CP</span><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> به میزان 100 میلی&shy;گرم بر کیلوگرم به شکل تک دوز تزریق شد. دو هفته پس از تجویز سیکلوفسفامید، </span></span><span dir="LTR" style="font-size:10.0pt">CM</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">از طریق مجرای وابران بیضه تزریق شد. پارامترهای اسپرم، هیستوپاتولوژی بیضه و سطح تستوسترون دو ماه پس از درمان بررسی شد. بیان ژن‌های </span></span><i><span dir="LTR" style="font-size:10.0pt">Bax</span></i><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> و </span></span><i><span dir="LTR" style="font-size:10.0pt">Bcl2</span></i> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">توسط </span></span><span dir="LTR" style="font-size:10.0pt">real-time polymerase chain reaction</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> مورد ارزیابی قرار گرفت.</span></span></span></span></span></span></span><br> <span style="font-size:12pt"><span style="text-justify:inter-ideograph"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">نتایج: </span></span></b><span dir="LTR" style="font-size:10.0pt">CP</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> بر بافت‌شناسی بیضه (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) و کیفیت اسپرم (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) تأثیر منفی داشت. </span></span><span dir="LTR" style="font-size:10.0pt">CP</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> بیان ژن&shy;های مرتبط با آپوپتوز را تغییر داد (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">). درمان با </span></span><span dir="LTR" style="font-size:10.0pt">CM</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> بیان </span></span><i><span dir="LTR" style="font-size:10.0pt">Bax</span></i><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> را صورت معنی</span></span><span dir="LTR" style="font-size:11.0pt"><span arial="" style="font-family:">&shy;</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">داری کاهش داد (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">)، در حالی که بیان </span></span><i><span dir="LTR" style="font-size:10.0pt">Bcl2</span></i><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> را به طور معنی</span></span><span dir="LTR" style="font-size:11.0pt"><span arial="" style="font-family:">&shy;</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">داری افزایش نشان داد (01/0 </span></span><span lang="FA" style="font-size:10.0pt">= </span><span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">). </span></span><span dir="LTR" style="font-size:10.0pt">CP</span><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> تعداد اسپرم (03/0 </span></span><span dir="LTR" style="font-size:10.0pt">=</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">)، زنده&shy;مانی (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) تحرک (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">)، تعداد اسپرماتوگونی (001/0 </span></span><span dir="LTR" style="font-size:10.0pt"><</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) و ضخامت اپیتلیال لوله‌های بیضه (02/0 </span></span><span dir="LTR" style="font-size:10.0pt">=</span> <span dir="LTR" style="font-size:10.0pt">p</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) را بهبود داد.</span></span><b><span dir="LTR" style="font-size:11.0pt"><span style="font-family:"Arial","sans-serif""></span></span></b></span></span></span></span></span><br> <span style="font-size:12pt"><span style="text-justify:inter-ideograph"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">نتیجه&shy; گیری: </span></span></b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">این یافته‌ها نشان می‌دهد که </span></span><span dir="LTR" style="font-size:10.0pt">CM</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">تولید شده از سلول</span></span><span dir="LTR" style="font-size:11.0pt"><span arial="" style="font-family:">&shy;</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های بنیادی مزانشیمی مشتق از مغز استخوان ممکن است در درمان&shy;های ناباروری ارزشمند باشد و عوارض جانبی </span></span><span dir="LTR" style="font-size:10.0pt">CP</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">را کاهش دهد.</span></span><b><span dir="LTR" style="font-size:11.0pt"><span style="font-family:"Arial","sans-serif""></span></span></b></span></span></span></span></span><br> <br> <b><i><span dir="RTL" lang="FA" style="font-size:10.0pt"><span b="" mitra="" style="font-family:">کلمات کلیدی: </span></span></i></b><i><span dir="RTL" lang="FA" style="font-size:10.0pt"><span b="" mitra="" style="font-family:">سلول&shy;های بنیادی مزانشیمی مغز استخوان<b>، </b>سیکلوفسفامید، محیط شرطی شده، آپوپتوز، اسپرماتوژنز</span></span></i></div> <p style="text-align: justify;"><span style="font-family:Times New Roman;"><span style="font-size:12pt"><span new="" roman="" times=""><b>Abstract </b></span></span><br> <span style="font-size:12pt"><span new="" roman="" times=""><b>Background:</b> Cyclophosphamide (CP) has some negative effects on the reproductive system. Stem cells and their metabolites are being utilized to enhance fertility after chemotherapy.</span></span><br> <span style="font-size:12pt"><span new="" roman="" times=""><b>Objective:</b> This study aimed to investigate the impact of conditioned medium (CM) derived from bone marrow mesenchymal stromal stem cells (BM-MSCs) on the toxic effects of CP on testicles.</span></span><br> <span style="font-size:12pt"><span new="" roman="" times=""><b>Materials and Methods: </b>BM-MSCs were isolated, a CM was collected and 25-fold concentrated. 24 male Wistar rats (8 wk, 200-250 gr) were randomly divided into following groups: control, CP, CP+Dulbecco&rsquo;s Modified Eagle Medium (DMEM), CP+CM. CP was given at a single dose of 100 mg/kg. 2 wk after the CP administration, CM was injected into the testicular efferent duct. Sperm parameters, testicular histopathology, and the level of testosterone were analyzed 2 months after treatment. The expression of B-cell lymphoma 2 (<i>Bcl2</i>) and <i>Bcl2</i>-associated X protein (<i>Bax</i>) genes were evaluated by real-time polymerase chain reaction.</span></span><br> <span style="font-size:12pt"><span new="" roman="" times=""><b>Results:</b> CP had a negative effect on testis histology (p < 0.001) and sperm quality (p < 0.001). It changed the expression of genes associated with apoptosis (p < 0.001). Treatment with CM reduced the expression of <i>Bax</i> (p < 0.001), while significantly increasing the expression of <i>Bcl2</i> (p = 0.01). It improved sperm count (p = 0.03), viability (p < 0.001), motility (p < 0.001), spermatogonial count (p < 0.001), and epithelial thickness of testicular tubules (p = 0.02).</span></span><br> <span style="font-size:12pt"><span new="" roman="" times=""><b>Conclusion:</b> These findings suggest that CM produced from BM-MSCs may be valuable for therapeutic approaches in reproductive medicine and may lessen the side effects of CP.</span></span></span></p> سلول­های بنیادی مزانشیمی مغز استخوان, سیکلوفسفامید, محیط شرطی شده, آپوپتوز, اسپرماتوژنز Bone marrow mesenchymal stem cells, Cyclophosphamide, Conditioned medium, Apoptosis, Spermatogenesis. 89 100 http://ijrm.ir/browse.php?a_code=A-10-2038-3&slc_lang=en&sid=1 Zeynab Esmailpour zeynabes265@gmail.com 0000-0003-3450-316X No Students Research Committee, Arak University of Medical Sciences, Arak, Iran. Soheila Madadi soheila.madadi@yahoo.com 0000-0003-4746-6652 No Department of Anatomy, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran. Maryam Baazm dr.baazm@arakmu.ac.ir 0000-0002-1248-6314 Yes Department of Anatomy, School of Medicine, Arak University of Medical Sciences, Arak, Iran. Molecular and Medicine Research Center, School of Medicine, Arak University of Medical Sciences, Arak, Iran.