International Journal of Reproductive BioMedicine
International Journal of Reproductive BioMedicine
IJRM
Medical Sciences
http://ijrm.ir
1
admin
2476-4108
2476-3772
10.29252/ijrm
en
jalali
1401
1
1
gregorian
2022
4
1
20
4
online
1
fulltext
en
لیزوفسفاتیدیک اسید تکوین فولیکولی را همراه با کاهش وقوع مرگ سلولی حمایت می کند: یک مطالعه تجربی
Lysophosphatidic acid supports the development of vitrified ovarian follicles by decreasing the incidence of cell death: An experimental study
Reproductive Biology
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:">لیزوفسفاتیدیک اسید (</span></span><span dir="LTR" style="font-size:10.0pt">LPA</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">) در فعال­سازی فولیکول، بلوغ و لقاح تخمک، لانه­گزینی جنین نقش دارد.</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<b> </b>مطالعه جهت بررسی اثرات </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">بر بهبود تکوین فولیکول</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های ایزوله شده از تخمدان</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های کامل منجمد شده و کشت شده موشی در محیط کشت طراحی گردید.</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:">مواد و روش ­ها: </span></span></b><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">در این مطالعه تجربی ابتدا تخمدان منجمد شده و نشده موش­های یک هفته ای ابتدا در محیط کشت حاوی 20 میلی­مول </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">به مدت یک هفته کشت شدند. سپس فولیکول</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های پره</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">انترال از آنها جدا شده و در حضور مقدار 40 میلی</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مول </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">دوباره به شکل انفرادی به مدت 12 روز کشت شدند و مطالعات بعدی بروی آنها انجام شد: تست زنده</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مانی با بکارگیری رنگ</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">آمیزی کلسئین </span></span><span dir="LTR" style="font-size:10.0pt">AM</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">، فلوسیتومتری با استفاده از آنکسین </span></span><span dir="LTR" style="font-size:10.0pt">v</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:"> و </span></span><span dir="LTR" style="font-size:10.0pt">Pi</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">و بررسی بروز ژن</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">ها با بکارگیری </span></span><span dir="LTR" style="font-size:10.0pt">real time-RT-PCR</span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">. میزان بلوغ تخمک</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">ها در گروه­های با هم مقایسه شدند و تعدادی از تخمک</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های </span></span><span dir="LTR" style="font-size:10.0pt">MII</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">برای لقاح بکارگرفته 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:">در تمام گروه</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های تحت تیمار با </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">میزان زنده</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مانی و تکوین فولیکول</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">ها بیشتر بود و همچنین وقوع مرگ سلولی و بروز ژن</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های پیش</span></span><span dir="LTR" style="font-size:11.0pt"><span arial="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">برنده اپوپتور در مقایسه با گروه</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های بدون تیمار کمتر بود (035/0 </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:">) و از حیث تکوین فولیکولی تفاوتی بین گروه</span></span><span lang="FA" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های انجمادی و غیرانجمادی دیده نشد اما از نظر بروز ژن </span></span><span dir="LTR" style="font-size:10.0pt">Bad</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">و ژن گیرنده</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">در گروه انجمادی تیمار شده با </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">تفاوت معنی</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">داری با گروه غیرانجمادی تیمار شده با </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">وجود داشت (028/0 </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></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 گیری:</span></span></b> <span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">باعث بهبود زنده</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مانی و تکوین فولیکول</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">های جدا شده شد. علی</span></span><span dir="LTR" style="font-size:11.0pt"><span arial="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">رغم تغییراتی که در بروز ژن­های مرتبط با آپوپتوز در گروه انجمادی رخ داده بود اما </span></span><span dir="LTR" style="font-size:10.0pt">LPA</span> <span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">اثرات مثبتی بر زنده</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">مانی و تکوین این فولیکول</span></span><span dir="LTR" style="font-size:11.0pt"><span cambria="" style="font-family:">­</span></span><span lang="FA" style="font-size:11.0pt"><span b="" mitra="" style="font-family:">ها داشت.</span></span></span></span></span></span></span></div>
<div style="text-align: justify;"><span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><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=""> Lysophosphatidic acid</span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> (LPA) </span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">contributes</span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times=""> to </span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">follicular activation, oocyte maturation, in vitro fertilization, and embryo implantation.</span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><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 was designed to evaluate the effects of LPA to improve the development of isolated follicles derived from whole mouse cultured vitrified ovaries. </span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><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=""> In this experimental study, first, the 1-wk-old mouse ovaries in the non-vitrified and vitrified groups were cultured in the presence of 20 µM of LPA for 1 wk. Then, their isolated preantral follicles were cultured individually for 12 days in the presence or absence of 40 µM of LPA. The following evaluations were done for the cultured follicles: a viability test using Calcein AM staining, flow cytometry using annexin V/Pi, and analysis of the expression of genes by real-time reverse transcription polymerase chain reaction. The maturation rates of the oocytes were compared </span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">among groups </span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">and some of the released metaphase II oocytes were subjected to in vitro fertilization. </span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><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=""> In all LPA treated groups, the rates of survival and follicular development were higher, and the incidence of cell death and expression of pro-apoptotic genes were lower, than in the non-LPA supplemented groups (p = 0.035). There was no significant difference between the vitrified and non-vitrified groups regarding follicular or oocyte development, but the expression of <i>Bad</i> and <i>LPA</i> receptors genes was significantly altered in the vitrified LPA supplemented group in </span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">comparison with the </span></span><span style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">non-vitrified LPA supplemented group (p = 0.028). </span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><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=""> LPA improved the survival and developmental potential of the isolated follicles. Despite some alterations in the expression of apoptosis-related genes in the vitrified ovaries, <i>LPA</i> had positive effects on the survival and development of these follicles.</span></span></span></span></span></span></div>
مرگ سلولی, بلوغ تخمک در شرایط آزمایشگاهی, اسید لیزوفسفاتیدیک, گیرنده های لیزوفسفاتیدیک اسید, فولیکول های تخمدان, انجماد.
Cell death, In vitro oocyte maturation, Lysophosphatidic acid, Lysophosphatidic acid receptors, Ovarian follicles, Vitrification.
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http://ijrm.ir/browse.php?a_code=A-10-158-3&slc_lang=en&sid=1
Neda
Abedpour
abedpour.n@umsu.ac.ir