تأثیر محافظتی آلفا لیپوئیک اسید در برابر انجماد سلول های بنیادی اسپرماتوگونی موشی بر الگوی بیان ژن های دخیل در آپوپتوز

نوع مقاله : مقاله پژوهشی

نویسندگان
کاظم نوروزی 1
حسن نظری 2
ابراهیم احمدی 3
ناصر شمس اسفندآبادی 4
آزیتا افضلی 5
1 کیلومتر 2 جاده سامان دانشگاه شهرکرد
2 پژوهشکده فناوری جنین دام، دانشگاه شهرکرد، شهرکرد، ایران
3 پژوهشکده فناوری جنین دام، دانشگاه شهرکرد
4 گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه شهرکرد، شهرکرد- ایران.
5 گروه علوم تشریح، دانشکده پزشکی، دانشگاه علوم پزشکی سمنان، سمنان- ایران.
10.22034/ijvcs.2026.15030.1100
چکیده
هدف از این مطالعه بررسی اثر محافظتی آلفا لیپوئیک اسید (ALA) بر آسیب ناشی از انجماد-ذوب سلول‌های بنیادی اسپرماتوگونی (SSCs) موش نابالغ از طریق ارزیابی بیان ژن‌های دخیل در آپوپتوز بود. سلول‌های بنیادی اسپرماتوگونی از بیضه موش‌های ۴-۸ روزه نژاد C57 جدا شده و پس از تأیید هویت با روش‌های ایمونوسیتوشیمی (نشانگر PLZF) و سنجش فعالیت آلکالین فسفاتاز، به مدت سه هفته کشت داده شدند. سپس سلول‌ها در چهار گروه انجمادی شامل کنترل (بدون آنتی‌اکسیدان) و سه دوز 2/0، 1 و 5 میکرومولار آلفالیپوئیک اسید منجمد شدند. پس از ذوب، بیان ژن‌های Bax، Bcl2، Casp3 و Casp9 با روش Real-time PCR بررسی گردید. نتایج نشان داد که غلظت 1 میکرومولار ALA به‌طور معنی‌داری بیان ژن پروآپوپتوتیک Bax و نیز ژن‌های Casp3 و Casp9 را کاهش و بیان ژن ضد آپوپتوتیک BCL2 را افزایش می‌دهد (05/0>P). غلظت 2/0 میکرومولار نیز کاهش معنی‌داری در بیان Bax و Casp3 ایجاد کرد. بالاترین دوز (5 میکرومولار) اثر معنی‌داری نداشت. میتوان نتیجه گرفت که آلفا لیپوئیک اسید، به ویژه در غلظت 1 میکرومولار، با تعدیل بیان ژن‌های کلیدی آپوپتوز می‌تواند از سلول‌های بنیادی اسپرماتوگونی موش در برابر آسیب ناشی از فرآیند انجماد-ذوب محافظت کند و به عنوان یک افزودنی مؤثر در محیط انجماد این سلول‌ها پیشنهاد می‌گردد.
کلیدواژه‌ها
سلول‌های بنیادی اسپرماتوگونی
آلفا لیپوئیک اسید
انجماد
آپوپتوز
بیان ژن
موش
موضوعات

عنوان مقاله English

Protective Role of Alpha-Lipoic Acid During Freezing on the Expression of Apoptosis-Related Genes in Mouse Spermatogonial Stem Cells

نویسندگان English

Kazam Norouzi 1
Hassan Nazari 2
Ebrahim Ahmadi 3
Naser Shams Esfandabadi 4
Azita Afzali 5
1 Faculty of Veterinary Medicine, Shahrekord University
2 Research Institute of Animal Embryo Technology
3 Research Institute of Animal Embryo Technology, Shahrekord university
4 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord- Iran.
5 Department of Anatomical Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan- Iran.
چکیده English

This study aimed to examine the protective role of alpha-lipoic acid (ALA) against damage caused by freeze-thawing in immature mouse spermatogonial stem cells (SSCs) by analyzing the expression of genes related to apoptosis. SSCs were extracted from the testes of 4–8-day-old C57 mice and, after verification through immunocytochemistry using the PLZF marker and an alkaline phosphatase activity test, were cultured for three weeks. The cells were then frozen in four groups: a control group without antioxidants and three groups treated with different ALA concentrations (0.2, 1, and 5 µM). Following thawing, the expression levels of BAX, BCL2, Caspase-3, and Caspase-9 were measured using Real-time PCR. Findings revealed that 1 µM ALA significantly lowered the expression of the pro-apoptotic genes BAX, Caspase-3, and Caspase-9, while significantly increasing the anti-apoptotic gene BCL2 (P<0.05). The 0.2 µM dose also notably reduced BAX and Caspase-3 expression. However, the highest concentration (5 µM) showed no significant impact. The study concludes that alpha-lipoic acid, especially at 1 µM, can safeguard mouse spermatogonial stem cells from freeze-thaw damage by regulating key apoptosis-related gene expression and may serve as a beneficial additive in the cryopreservation medium for these cells.

کلیدواژه‌ها English

Spermatogonial stem cells
Alpha-lipoic acid
Cryopreservation
Apoptosis
Gene expression
Mouse
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علوم درمانگاهی دامپزشکی ایران
دوره 19، شماره 2 - شماره پیاپی 33
پاییز و زمستان 1404
اسفند 1404
صفحه 47-60