en بررسی تأثیر ترکیب ملاتونین و ال-کارنیتین بر بلوغ آزمایشگاهی در تخمک موش: یک مطالعه تجربی Assisted Reproductive Technologies Original Article Original Article <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:">ملاتونین و ال-کارنیتین عوامل مهارکننده رادیکال آزاد، آنتی&shy;اکسیدان و ضد آپوپتوز هستند که به بهبودی رشد تخمک کمک می&shy;کنند.</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 تعیین<b> </b>اثرات احتمالی ترکیب دو عامل آنتی&shy;اکسیدان ملاتونین و ال-کارنیتین بر مورفولوژی، بلوغ، آپوپتوز و بیان ژن&shy;های </span></span><i><span dir="LTR" style="font-size:10.0pt">BMP-15</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">GDF-9</span></i><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> در تخمک موش مورد بررسی قرار گرفت.</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:">تحریک تخمک&shy;گذاری با تزریق 5/7 واحد بین المللی </span></span><span dir="LTR" style="font-size:10.0pt">PMSG</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> به صورت داخل صفاقی در 60 موش ماده نژاد </span></span><span dir="LTR" style="font-size:10.0pt">NMRI</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> انجام شد. در روز دوم پس از تزریق، 70 کمپلکس کومولوس-اووسیت از هر موش جمع&shy;آوری گردید. تخمک&shy;های برداشت شده به طور تصادفی به 4 گروه شامل کنترل و 3 گروه درمان ملاتونین و ال-کارنیتین به تنهایی و ترکیبی از ملاتونین و ال-کارنیتین تقسیم شدند. مورفولوژی، میزان بلوغ، آپوپتوز و بیان ژن&shy;های </span></span><i><span dir="LTR" style="font-size:10.0pt">BMP-15</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">GDF-9</span></i><span style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> تخمک&shy;ها با </span></span><span dir="LTR" style="font-size:10.0pt">real-time PCR</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">ارزیابی شدند.</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 lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">درمان ترکیبی ملاتونین و ال-کارنیتین در مقایسه با گروه کنترل باعث افزایش قطر تخمک&shy;ها گردید (05/0 </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:">). نتایج آزمون </span></span><span dir="LTR" style="font-size:10.0pt">TUNEL</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> نشان داد که میزان آپوپتوز در گروه ملاتونین + ال-کارنیتین کمترین میزان بود. علاوه بر این نشان داده شد که تعداد تخمک&shy;های بالغ شده در تمام گروه&shy;های مورد مطالعه نسبت به گروه کنترل افزایش داشته و بیشترین میزان بلوغ در گروه ملاتونین + ال کارنیتین مشاهده شد. همچنین سطح بیان ژن&shy;های </span></span><i><span dir="LTR" style="font-size:10.0pt">BMP-15</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">GDF-9</span></i><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> نیز در گروه&shy;های درمان در مقایسه با گروه کنترل به طور قابل توجهی افزایش داشته است.</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:">نتایج ما پیشنهاد می&shy;کند که تجویز ترکیبی ملاتونین و ال-کارنیتین به عنوان انتخاب مؤثرتری برای افزایش بلوغ تخمک در شرایط آزمایشگاهی می&shy;باشد.</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> &nbsp; <div style="text-align: justify;"><span style="font-size:11pt"><span style="line-height:normal"><span sans-serif="" style="font-family:Calibri,"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Background:</span></span></b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> Melatonin and L-carnitine are free radical scavengers with antiapoptotic and antioxidant properties that improve oocyte development</span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">. </span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span sans-serif="" style="font-family:Calibri,"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Objective:</span></span></b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> This study aimed to find the possible effect of combining 2 antioxidant agents of melatonin and L-carnitine on oocyte morphology, maturation, apoptosis, and expression of bone morphogenetic protein 15 (<i>BMP-15</i>) and growth differentiation factor 9 (<i>GDF-9</i>) genes in a mice model.&nbsp; </span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span sans-serif="" style="font-family:Calibri,"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Materials and Methods:</span></span></b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> To overstimulation, 60 female NMRI mice were injected intraperitoneally using mare serum gonadotropin. On day 2 post-injection, 70 cumulus-oocyte complexes were collected from each mouse. The collected oocytes randomly were then divided into 4 groups including, the control, melatonin, L-carnitine, melatonin + L-carnitine groups. The morphology and maturation rate of the oocytes was evaluated using a light microscope. Apoptosis was identified by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay and the expression of <i>BMP-15</i> and growth and differentiation factor <i>GDF-9</i> genes was also evaluated by real-time polymerase chain reaction.</span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span sans-serif="" style="font-family:Calibri,"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Results:</span></span></b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> Oocyte diameter significantly was increased in combination treatment of L-carnitine and melatonin compared to other groups (p < 0.05). L-carnitine group showed the highest mean percentage of oocyte cytoplasmic pattern. Results of the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling indicated that the lowest apoptosis rate belonged to the melatonin + L-carnitine group. Moreover, the combination groups showed the highest number of oocytes and maturation rate. The <i>BMP-15</i> and <i>GDF-9</i> genes were significantly upregulated in all treatment groups compared to the control group.</span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span sans-serif="" style="font-family:Calibri,"><b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">Conclusion:</span></span></b><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> Our results suggested a combination of melatonin + L-carnitine administration as a more effective choice for in vitro promotion of oocyte maturation. </span></span></span></span></span></div> بلوغ آمایشگاهی تخمک, ملاتونین, ال-کارنتین. In vitro oocyte maturation, Melatonin, L-carnitine. 527 538 http://ijrm.ir/browse.php?a_code=A-10-2634-1&slc_lang=en&sid=1 Raziye Chegini Raziyechegini1369@gmail.com 0009-0003-6675-9052 No Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Morteza Sadeghi Ms.sadeghi@yahoo.com 0000-0003-3435-0010 No Human Genetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. Sadegh Shirian shirian@shirazu.ac.ir 0000-0003-4137-9056 No Department of Pathology, School of Veterinary Medicine, Shahrekord University, Shahrekord, Iran. Fatemeh Sabbaghziarani f.sabbagh@qums.ac.ir 0000-0002-5167-3569 No Cellular and Molecular Research Center, Research Institute for Prevention of Non-communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran. Ehsan Aali E.aali@qums.ac.ir 0000-0002-2231-118X No Cellular and Molecular Research Center, Research Institute for Prevention of Non-communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran. Pouriya Soleimani pouriya.soleimani@gmail.com 0000-0003-1307-7966 No Department of Nursing, Faculty of Nursing and Midwifery, Qazvin University of Medical Sciences, Qazvin, Iran. Mohammad Reza Ashtari Majelan mohammadashtarimajelan17@gmail.com 0000-0002-1166-8490 No Cellular and Molecular Research Center, Research Institute for Prevention of Non-communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran. Fariba Zafari f.zafari@qums.ac.ir 0000-0001-6673-278X Yes Cellular and Molecular Research Center, Research Institute for Prevention of Non-communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran. Shahram Darabi shdarabi@qums.ac.ir 0000-0002-9405-6640 No Cellular and Molecular Research Center, Research Institute for Prevention of Non-communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran.