ﻣﻘﺎﯾﺴﻪ ﮐﯿﻔﯿﺖ ﺗﺨﻤﮏﻫﺎي ﺣﺎﺻﻞ از ﻣﻮش ﻃﺒﯿﻌﯽ و ﻣﻮش اﻟﻘﺎﯾﯽ ﺳﻨﺪرم ﺗﺨﻤﺪان ﭘﻠﯽﮐﯿﺴﺘﯿﮏ ﺑﻪ دﻧﺒﺎل ﺑﺮرﺳﯽ ژن 9-GDF

Comparison of oocyte quality from normal mouse and mouse with polycystic ovarian syndrome following evaluation of GDF-9 gene

ﺳﻨﺪرومﺗﺨﻤﺪانﭘﻠﯽﮐﯿﺴﺘﯿﮏ )Polycystic ovarian syndrome: PCOS( ﯾﮑﯽ از ﺷﺎﯾﻊﺗﺮﯾﻦ ﻋﻠﻞ ﻧﺎﺑﺎروري اﺳﺖ. ﯾﮑﯽ از روشﻫﺎي ﮐﻤﮏ ﺑﺎروري ﺑﺮاي ﺑﯿﻤﺎران PCOS اﺳﺘﻔﺎده از روش ﺑﻠﻮغ آزﻣﺎﯾﺸﮕﺎﻫﯽ ) in-vitro maturation: IVM( اﺳﺖ. از ﻧﺸﺎﻧﻪﻫﺎي ﮐﯿﻔﯿﺖ ﺗﺨﻤﮏ ﺑﯿﺎن ﻣﻨﺎﺳﺐ ژن 9-GDF اﺳﺖ. اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺑﻪ ﺑﺮرﺳﯽ ﺗﻔﺎوت ﮐﯿﻔﯿﺖ ﺗﺨﻤﮏ/ﻓﻮﻟﯿﮑﻮلﻫﺎيPCOS ﺑﺎ ﺗﺨﻤﮏ/ﻓﻮﻟﯿﮑﻮلﻫﺎي ﻃﺒﯿﻌﯽ در ﺟﺮﯾﺎن IVM ﻣﯽﭘﺮدازد. اﻟﻘﺎء PCOS در ﻣﻮشﻫﺎي ﻧﮋاد NMRI )ﺗﻌﺪاد= 10( از ﻃﺮﯾﻖ ﺗﺰرﯾﻖ 4 ﻣﯿﻠﯽﮔﺮمﺑﺮﮐﯿﻠﻮﮔﺮم اﺳﺘﺮادﯾﻮل واﻟﺮات ﺣﻞ ﺷﺪه در 0/2 ﻣﯿﻠﯽﮔﺮم روﻏﻦ ﮐﻨﺠﺪ ﺑﻪ ﺻﻮرت ﯾﮏ ﺑﺎر در روز و ﺑﻪ ﻣﺪت 60 روز اﻧﺠﺎم ﺷﺪ. ﺗﺨﻤﮏﻫﺎ و ﻓﻮﻟﯿﮑﻮلﻫﺎ در ﮔﺮوهﻫﺎي ﮐﻨﺘﺮل و PCOS ﺟﻤﻊآوري ﺷﺪﻧﺪ و در ﻣﺤﯿﻂMEM-α ﻏﻨﯽ ﺷﺪه ﺑﺎ 10% ﺳﺮم ﺟﻨﯿﻦ ﮔﺎوي ﺑﻪ ﻣﺪت 24-48 ﺳﺎﻋﺖ ﮐﺸﺖ داده ﺷﺪﻧﺪ. ارزﯾﺎﺑﯽ ﻣﯿﺰان ﺑﻠﻮغ آزﻣﺎﯾﺸﮕﺎﻫﯽ و ﺑﯿﺎن ژن 9-GDF )روشReal-Time PCR ( در ﮔﺮوهﻫﺎي ﻣﺨﺘﻠﻒ اﻧﺠﺎم ﺷﺪ. آﻧﺎﻟﯿﺰ آﻣﺎري ﺑﺎ روش ANOVA ﯾﮏﻃﺮﻓﻪ اﻧﺠﺎم ﺷﺪ. در ﮔﺮوه PCOS، ﮐﺎﻫﺶ ﻣﻌﻨﯽداري در ﺗﻌﺪاد ﻓﻮﻟﯿﮑﻮلﻫﺎي اوﻟﯿﻪ، ﭘﺮيآﻧﺘﺮال، آﻧﺘﺮال و ﺟﺴﻢ زرد ﻣﺸﺎﻫﺪه ﺷﺪ. ﻫﻤﭽﻨﯿﻦ ﻓﻮﻟﯿﮑﻮلﻫﺎي ﮐﯿﺴﺘﯿﮏ و ﻓﻮﻟﯿﮑﻮلﻫﺎﯾﯽ ﺑﺎ ﻻﯾﻪ ﮔﺮاﻧﻮﻟﻮزاي دژﻧﺮهﺷﺪه ﺑﺎﻻ ﻣﺸﺎﻫﺪه ﺷﺪ. 52٪ ﺗﺨﻤﻚﻫﺎ و 50 % ﻓﻮﻟﯿﮑﻮلﻫﺎ در ﮔﺮوه PCOS ﺑﺎﻟﻎ ﺷﺪﻧﺪ. در ﮔﺮوه ﮐﻨﺘﺮل، 83/33 درﺻﺪ ﺗﺨﻤﮏﻫﺎ و 75 درﺻﺪ ﻓﻮﻟﯿﮑﻮلﻫﺎ ﺑﺎﻟﻎ ﺷﺪﻧﺪ )0.05

Polycystic ovarian syndrome (PCOS) is one of the most common causes of infertility. One of assisted reproductive methods for PCOS patients is using of in-vitro maturation (IVM). Normal GDF-9 gene expression is one of the symptoms of good oocyte quality. This study examines the differences between quality of PCOS oocytes/follicles with normal oocytes/follicles during IVM. Materials and Methods: Induction of PCOS in NMRI mice (n=10) was performed by injection of 4mg/kg estradiol valerate dissolved in 0.2mg sesame oil once daily for 60 days. Oocytes and follicles were collected from control and PCOS groups, and cultured in the MEM-α medium supplemented with 10% FBS for 24-48 hrs. The evaluation of IVM rate and GDF-9 gene expression (Real-Time PCR method) was performed in different groups. Statistical analysis was performed using one-Way ANOVA test. Results: In the PCOS group, a significant decrease was observed in the number of primary follicles, preantral, antral, and corpus luteum. Increased cystic follicles and follicles with degenerated granulosa layer were also observed. 52% and 50% of PCOS oocytes and follicles was matured, respectively. In the control group, 83.33% of oocytes and 75% of follicles matured (P<0.05). GDF-9 gene expression was 1.00±0.08 and 1.24±0.2 in normal oocytes and follicles (P<0.01) and 0.66±0.02 and 0.37±0.02 in PCOS oocytes and follicles, respectively (P<0.001). Conclusion: Quality of PCOS oocytes, and especially, follicles (GDF-9 gene expression) was decreased following IVM. Therefore, using of PCOS oocytes was suggested for IVM