بررسی تأثیر استروژن بر میزان بیان نشانگرهای عصبی در طی تمایز نوروژنیک سلول‌های بنیادی مشتق از چربی

نوع مقاله : Original Article(s)

نویسندگان

1 استاد، گروه علوم تشریحی و بیولوژی مولکولی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

2 کارشناس ارشد، گروه زیست‌شناسی، دانشگاه آزاد اسلامی، واحد علوم و تحقیقات تهران، تهران، ایران

3 دانشجوی دکتری، گروه ژنتیک، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

4 دانشجوی پزشکی، دانشکده‌ی پزشکی و کمیته‌‌ی تحقیقات دانشجویی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

چکیده

مقدمه: با توجه به سهولت دستیابی به سلول‌های بنیادی حاصل از بافت چربی نسبت به سایر منابع سلول‌های بنیادی در صورتی که بتوان از عوامل رشد مانند استروژن جهت افزایش تمایز نرونی استفاده نمود، نرون‌های تمایز یافته، در پیوند اتولوگ بیماری‌های عصبی دژنراتیو خاص مانند بیماری پارکینسون و ضایعات نخاعی استفاده‌ی وسیعی دارد. هدف از این مطالعه، ارزیابی تأثیر استروژن بر تمایز عصبی سلول‌های بنیادی مشتق از بافت چربی از طریق ارزیابی بیان نشانگر نورون بالغ (2MAP یا 2Microtubule-associated protein)، نشانگر سلول‌های گلیال (Glial fibrillary acidic protein یا GFAP) و نشانگر سلول‌های پیش‌ساز عصبی (Nestin) بود.روش‌ها: پس از جداسازی سلول‌های بنیادی از بافت چربی، القای عصبی سلول‌های بنیادی از طریق روش تشکیل نروسفر انجام شد. پس از یک هفته، تجمعات سلولی نروسفر تشکیل شده جهت تمایز نهایی، به محیط کشت القای عصب منتقل شد و در گروه تیمار استروژن با دوز 10 نانومول تا انتهای دوره‌ی تمایز روزانه به محیط کشت القای عصب اضافه گردید. سپس ارزیابی بیان نشانگرهای Nestin، 2MAP و GFAP در سلول‌های تمایز یافته از طریق تکنیک RT-PCR (Reverse transcription-polymerase chain reaction) صورت گرفت. به علاوه، آزمایش MTT [diphenyl tetrazolium bromide assay 2,5 (yl-2- dimethyl thiazol-4,5)3] جهت ارزیابی تکثیر سلولی انجام شد.یافته‌ها: میانگین بیان نشانگرهای Nestin و GFAP در گروه شاهد نسبت به مورد بیشتر بود و این اختلاف میانگین، از لحاظ آماری معنی‌دار بود (050/0 > P)؛ در حالی که میزان بقای سلولی در دو گروه، اختلاف معنی‌داری را نشان نداد.نتیجه‌گیری: در مجموع، می‌توان دریافت استروژن سبب کاهش بیان نشانگرهای عصبی می‌شود. با این حال، جهت تعیین تأثیر استروژن بر تمایز عصبی سلول‌های بنیادی، پیشنهاد می‌شود این مطالعه به صورت گسترده‌تر و با استفاده از تکنیک‌های حساس‌تری بررسی گردد.

کلیدواژه‌ها

عنوان مقاله [English]

Effects of Estrogen on the Expression of Neural Markers in Differentiated Adipose-Derived Stem Cells

نویسندگان [English]

  • Shahnaz Razavi 1
  • Nafiseh Ahmadi 2
  • Mohammad Kazemi 3
  • Hamid Reza Sadeghian 4
1 Professor, Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
2 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
3 PhD Student, Department of Genetics, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
4 Student of Medicine, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: Over the past decade, researchers have used the existing knowledge about pathways messaging protocols to successfully stimulate stem cells to generate neurons. Due to the ease of access to adipose tissue-obtained stem cells rather than other sources, whereas estrogen factor could be used to improve neural differentiation, antilogous transplantation of differentiated neurons would be widely used for certain degenerative neurological diseases such as Parkinson's disease and spinal cord injuries. The purpose of this study was to evaluate the effect of estrogen on expression of microtubule-associated protein-2 (MAP2), glial fibrillary acidic protein (GFAP) and Nestin markers.Methods: After the isolation of stem cells from adipose tissue, the neural induction was carried out through neurosphere construction; then, final differentiation of the cells was performed. Neurosphere-singed cell were transferred to neural induction medium (control group). In the estrogen-treated group, estrogen was added to the culture medium until the end of the day of distinction. Then, evaluation of the expression of neural markers, was performed using reverse transcription-polymerase chain reaction (RT-PCR) technique. In addition, MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] was performed to assess cell viability.Findings: The mean expression of GFAP and Nestin markers were down regulated in treated group compared to controls (P < 0.05). The difference between the mean of MAP2 expression was not significant between the two groups. In addition, the difference between the mean of cell viability was not significant between two groups, too.Conclusion: In this study, we found that estrogen can decrease the expression of neuronal markers. However, to determine the effect of estrogen on neurogenic differentiation of stem cells, next studies should be done broader using other precise techniques.Keywords: Estrogen, Neurogenesis, Adipose-derived stem cell, Reverse transcription-polymerase chain reaction (RT-PCR)

کلیدواژه‌ها [English]

  • Estrogen, Neurogenesis
  • Adipose-derived stem cell
  • Reverse transcription-Polymerase chain reaction (RT-PCR)

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مجله دانشکده پزشکی اصفهان
دوره 33، شماره 345 - شماره پیاپی 345
مهر و آبان 1394
صفحه 1239-1249

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  • تاریخ دریافت: 20 اسفند 1393
  • تاریخ بازنگری: 11 آبان 1403
  • تاریخ پذیرش: 17 فروردین 1401

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