بررسي نقش آگونيست و آنتاگونيست گيرنده آدرنرژيك بتا-2 در بيان ژن هاي اتصال شكافي(دو كانكسين 37/43) در سلول هاي كومولوس زنان با پاسخ ضعيف تخمدان

Evaluation role of beta-2 adrenoceptor agonist and antagonist in the expression of gap junction genes (connexin 37/43) in cumulus cells of women with poor ovarian response

كانكسن ها (Connexin) پروتيين غشايي در ساختار اتصال شكافي يا گپ جانكشن ها هستند كه عهده دار انتقال يون ها و مولكول هاي پيام رسان به تخمك اند. هدف اين مطالعه بررسي نقش گيرنده بتا-دو آدرنرژيك در روند رشد فوليكول براساس بيان ژن دو كانكسن (Connexin)، 37 و 43 گپ جانكشن هاست كه نقش اوليه را در از سرگيري مجدد روند ميوتيك و بلوغ اووسيت دارند. روش بررسي اين مطالعه موردي-شاهدي از فروردين 98 تا آبان 1399 در مركز تحقيقات بهداشت باروري بيمارستان امام خميني تهران در دو گروه مطالعه (زنان با پاسخ ضعيف تخمدان) و كنترل (زنان اهدا كننده تخمك) انجام شد. هر گروه با معيار بولونيا (Bologna) و ضوابط ورود اندكس توده بدني زير m2/kg 28 و محدوده سني45-20 سال و ضوابط خروج عدم مصرف دارو به جز تحريك كننده هاي تخمدان و عدم ابتلا به بيماري وارد مطالعه شدند. سيكل تحريك تخمك گذاري اجرا و كومولوس ها پس از پانكچر جدا و در محيط كشت سلولي قرار گرفتند. ايزوپروترنول (آگونيست) و پروپرانولول (آنتاگونيست) گيرنده بتا-دو با غلظتnM 100 به محيط اضافه و استخراج RNA انجام و cDNA سنتز گرديد. بيان ژن توسط تكنيك Real-time PCR تعيين شد. يافته ها بيان ژن هر دو كانكسن (Connexin) در گروه مطالعه بدون دارو (0/001

Connexon is a membrane structural protein in the gap junctions. These cellular connections are responsible for transporting ions and messenger molecules to the oocyte. This study aimed to investigate the role of beta-2-adrenoceptors in the process of follicle growth based on the expression of the two connexins 37/43 in the gap junctions that have a primary role in the mitotic resumption and oocyte maturation. Methods This case-control study was conducted from April 2019 to November 2020 at the Reproductive Health Research Center of Tehran Imam Khomeini Hospital was performed on women with the poor ovarian response (POR) and control (women donate eggs) groups. Both groups had entry criteria with a body mass index under 28 m2/kg and 20-45 years old. Exit criteria were including no drug use except ovarian stimulants and no illness. The diagnosis was made agreeing to the Bologna model criterion. Ovulation stimulation cycle was performed, and then after the puncture, cumulus cells were isolated by enzyme and were freezing in -80 centigrade until the time of inserting into the cell culture medium. Isoproterenol (agonist) and propranolol (antagonist) at a concentration of 100 nM were added to the culture medium as the beta-2 adrenoceptors selective drugs. After culture, RNA extraction was performed and the concentration was read by Nanodrop, and then cDNA was synthesized. Gene expression was determined by real-time PCR. Results The findings of connexin expression in the three study groups: without the drug (P<0.001), propranolol (P<0.001), and isoproterenol (P<0.001) were significant compared to the control group. Isoproterenol decreased expression but propranolol increased it (P<0.001). Conclusion These findings confirm the important role of connexins 37 and 43 in cumulus cleft junctions that propranolol was able to increase its expression. Therefore, we suggest firstly these two connexins can be an effective target for oocyte growth and maturation. Secondly, propranolol could be a new treatment for women with POR and be effective in assisted reproductive technology (ART).