Alpha Lipoic Acid Impact on Apoptosis-Related Gene Expression in Mature Mouse Oocyte

محورهای موضوعی :

Zahrah Khezli ¹ , Saeed Zavareh ² , Mojdeh Salehnia ³

1 - Anatomy Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 - School of Biology, Damghan University, Damghan, Iran
3 - Anatomy Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

تاریخ دریافت : 1401/05/31 تاریخ پذیرش : 1402/08/17 تاریخ انتشار : 1402/12/11

کلید واژه: GV oocytes, Alpha-lipoic acid, Apoptotic genes,

چکیده مقاله :

Utilizing antioxidants offers a promising strategy for mitigating the effects of oxidative stress. This study was designed to assess the influence of alpha-lipoic acid (ALA) on the maturation of mouse oocytes and their apoptosis-related genes. Germinal vesicle oocytes, obtained from female mice aged 6-8 weeks, were subjected to in vitro maturation. Among these, 488 oocytes were exposed to 100 μM ALA, while 506 oocytes matured without ALA over a 16-hour duration. Subsequent evaluations were conducted to determine oocyte maturation rates. A portion of the mature oocytes at the metaphase II (MII) stage underwent in vitro fertilization, while the remaining oocytes were utilized to analyze the expression of Caspase 3, Bad, Bax, and Bcl2 through real-time RT-PCR. To quantify cell numbers, resulting blastocysts were stained with DAPI (4',6-diamidino-2-phenylindole). The group treated with ALA exhibited significantly higher rates of MII oocytes (77.30%) and a greater proportion of embryos developing into the blastocyst stage (33.87%) in comparison with the control group (55.81% and 25.06%, respectively; P<0.05). Moreover, the average cell count within blastocysts significantly increased in the ALA-treated group (82.37) compared to the control group (68.5; P<0.05). Furthermore, the pro-apoptotic genes expression decreased significantly, while the expression of the Bcl2 gene exhibited a significant increase in the ALA-treated group in comparison to the control group (P<0.05). In conclusion, the supplementation of the maturation medium for mouse oocytes with ALA resulted in improvements in oocyte development and overall embryo quality. This effect was attributed to the downregulation of pro-apoptotic genes and the upregulation of the anti-apoptotic gene.

 

چکیده انگلیسی:

Utilizing antioxidants offers a promising strategy for mitigating the effects of oxidative stress. This study was designed to assess the influence of alpha-lipoic acid (ALA) on the maturation of mouse oocytes and their apoptosis-related genes. Germinal vesicle oocytes, obtained from female mice aged 6-8 weeks, were subjected to in vitro maturation. Among these, 488 oocytes were exposed to 100 μM ALA, while 506 oocytes matured without ALA over a 16-hour duration. Subsequent evaluations were conducted to determine oocyte maturation rates. A portion of the mature oocytes at the metaphase II (MII) stage underwent in vitro fertilization, while the remaining oocytes were utilized to analyze the expression of Caspase 3, Bad, Bax, and Bcl2 through real-time RT-PCR. To quantify cell numbers, resulting blastocysts were stained with DAPI (4',6-diamidino-2-phenylindole). The group treated with ALA exhibited significantly higher rates of MII oocytes (77.30%) and a greater proportion of embryos developing into the blastocyst stage (33.87%) in comparison with the control group (55.81% and 25.06%, respectively; P<0.05). Moreover, the average cell count within blastocysts significantly increased in the ALA-treated group (82.37) compared to the control group (68.5; P<0.05). Furthermore, the pro-apoptotic genes expression decreased significantly, while the expression of the Bcl2 gene exhibited a significant increase in the ALA-treated group in comparison to the control group (P<0.05). In conclusion, the supplementation of the maturation medium for mouse oocytes with ALA resulted in improvements in oocyte development and overall embryo quality. This effect was attributed to the downregulation of pro-apoptotic genes and the upregulation of the anti-apoptotic gene.

 

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