اثر عصاره برگ شبدر قرمز (Trifolium pretense L) بر بیان ژن‌های موثر در فولیکوژنز و تکثیر سلول‌های گرانولوزای تخمدان موش کوچک آزمایشگاهی نژاد NMRI

نوع مقاله: مقاله پژوهشی

نویسندگان

1 گروه زیست‌شناسی، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

2 گروه زیست شناسی و مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

چکیده

ناباروری، یکی از مشکلات عمده پزشکی در دنیای امروز است. امروزه برخی مطالعات به بیان اثرات ترکیبات فیتواستروژنی در درمان ناباروری پرداخته‌اند. شبدر قرمز یکی از گیاهانی است که حاوی ترکیبات فیتواستروژنی است. در پژوهش حاضر اثر عصاره برگ شبدر قرمز بر بیان ژن‌های موثر در فولیکوژنز و تکثیر سلول‌های گرانولوزا در موش ماده کوچک آزمایشگاهی نژاد NMRI بررسی شده است. در این پژوهش تجربی آزمایشگاهی تعداد 30 سر موش ماده نابالغ 20-18 روزه نژاد NMRI استفاده شد. جهت انجام تجربیات سلول‌های گرانولوزای فولیکولی جداسازی شدند، سلول‌های حاصل در محیط in vitro به 6 گروه به صورت تصادفی تقسیم بندی شدند. گروه شاهد (بدون تیمار) و گروه‌‌‌‌‌‌‌‌‌‌‌های تجربی با غلظت‌های (10، 15، 20، 40، 80  میکروگرم بر میلی‌لیتر) عصاره برگ شبدر قرمز تیمار و زیست ‌پذیری سلول‌ها از طریق رنگ­آمیزی تریپان بلو و رنگ سنجیMTT بررسی شد. همچنین تغییرات بیان ژن‌های BMP15 وFOXO1 به روش Real Time PCR ارزیابی گردید. نتایج حاصل از رنگ آمیزی تریپان بلو و آزمونMTT  نشان داد با افزایش غلظت و زمان تیمار با عصاره برگ شبدر میزان بقاء سلولی کاهش یافته است. همچنین بررسی سطح بیان ژن‌های BMP15 وFOXO1  در گروه تیماری 10 میکروگرم بر میلی‌لیتر با عصاره برگ شبدر قرمز در مقایسه با گروه شاهد به طور معناداری افزایش نشان داد (05/0 > p < /em>). با توجه به اثر عصاره برگ شبدر قرمز بر تکثیر سلول‌های گرانولوزا و بیان ژن‌های موثر در فولیکوژنز می‌توان این گیاه به عنوان کاندید مناسبی برای مطالعات بالینی درمان ناباروری مورد توجه بیشتری قرار گیرد.

کلیدواژه‌ها


عنوان مقاله [English]

Effect of Red Clover Leaf Extract on Expression of Genes Involved in Follicogenesis and Proliferation of Ovarian Granulosa Cells of NMRI Mice

نویسندگان [English]

  • Farzaneh Rangamiz 1
  • Javad Baharara 2
  • Khadijeh Shahrokhabadi 1
1 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
2 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
چکیده [English]

Infertility is one of the major medical problems in today’s world. Today, some studies have addressed the effects of phytoestrogenic compounds in the treatment of infertility. Red clover is one of the herbs containing phytoestrogenic compounds. In the present study, the effect of red clover leaf extract on expression of genes involved in follicogenesis and proliferation of granulosa cells in NMRI mice was investigated. In this experimental in vitro study, 30 NMRI 18-20 immature female rats were used. For experiments, follicular granulosa cells were isolated, and in vitro cells were randomly divided into 6 groups.Control (untreated) and experimental groups with concentrations (10, 15, 20, 40, 80 μg/ml) of treated red clover leaf extract and cell viability were evaluated by trypan blue staining and MTT staining. Expression changes of BMP15 and FOXO1 genes were evaluated by Real Time PCR. The results of trypan blue staining and MTT assay showed that cell viability decreased with increasing concentration and time of treatment with clover leaf extract. Moreover, the expression level of BMP15 and FOXO1 genes was significantly increased in the 10 μg/ml red clover leaf extract compared to the control group (p < /em> < 0.05). Due to the effect of red clover leaf extract on granulosa cell proliferation and expression of genes involved in follicogenesis, this plant can be considered as a suitable candidate for clinical studies in infertility treatment.
 

کلیدواژه‌ها [English]

  • Infertility
  • Red Clover
  • Follicogenesis
  • Granulosa Cell
  • BMP15 Genes
  • FOXO1 Genes
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