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대한생식의학회지 제31권 제2호 2010년
GnRH-agonist에 의한 인간 과립-황체화 세포의 세포사멸과 PBR 단백질의 발현
연세대학교 의과대학 산부인과학교실, 한양대학교 자연과학대학 생명과학과, 을지의과대학교 생명과학연구소
김세광, 염윤희, 윤정미, 배상욱, 양현원, 조동제, 윤용달, 송찬호
Apoptosis and Peripheral Benzodiazepin Receptor (PBR) Expression in Human Granulosa-Luteal Cells by GnRH-agonist
Sei Kwang Kim,Yun-Hee Youm, Jeong-Mi Yoon, Sang Wook Bai, Hyunwon Yang, Dong Jae Cho, Yong-Dal Yoon, Chan Ho Song
Department of Obstetrics and Gynecology, College of Medicine, Yonsei University, Department of Life Sciences, College of Natural Sciences, Hanyang University Life Science Institute, Eulji University School of Medicine
Objective: To investigate whether GnRH-agonist (GnRH-Ag) using in IVF-ET affects apoptosis of human granulosa-luteal cells and expression of peripheral benzodiazepine receptor (PBR) protein involved in the apoptosis of the cells. Methods: Granulosa-luteal cells obtained during oocyte retrieval were cultured and treated with 10-5 M GnRH-Ag. Apoptosis of the cells by the treatment was confirmed using DNA fragmentation analysis 24 h after culture. The presence of PBR protein within the cells was examined by immunofluorescence staining and the expression of the protein was analyzed by Western blotting. In addition, it was measured for progesterone and nitric oxide (NO) produced by granulosa-luteal cells after GnRH-Ag treatment. To evaluate the relationship between NO production and PBR expression, sodium nitroprusside (SNP) as a NO donor was added in media and investigated the expression of PBR protein by Western blotting. Results: Apoptosis increased in the granulosa-luteal cells 24 h after GnRH-Ag treatment, whereas the expression of PBR protein significantly decreased. Furthermore, the production of progesterone and nitric oxide (NO) by the cells significantly fell from 12 h after the treatment. In the results of Western blotting after SNP treatment, the expression of PBR protein increased in the treatment with SNP alone to the granulosa-luteal cells, but was suppressed in the treatment with GnRH-Ag and SNP. Additionally, the staining result of PBR protein in the cells showed the even distribution of it through the cell. Conclusion: These results demonstrate that GnRH-Ag treatment induces apoptosis, decreasing expression of PBR protein and NO production in human granulosa-luteal cells. The present study suggests that one of the apoptosis mechanism of human granulosa-luteal cells by GnRH-Ag might be a signal transduction pathway via NO and PBR.
키워드 : Apoptosis, GnRH-agonist, Granulosa-luteal cells, Nitric Oxide (NO), Peripheral benzodiazepine receptor (PBR)
교신저자 : skkim@yumc.yonsei.ac.kr
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