GnRH-agonist에 의한 인간 과립-황체화 세포의 세포사멸과 PBR 단백질의 발현 = Apoptosis and Peripheral Benzodiazepin Receptor (PBR) Expression in Human Granulosa-Luteal Cells by GnRH-agonist

다국어 초록 (Multilingual Abstract)

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.

참고문헌 (Reference)

• 1 "directly induces apoptotic cell death in the rat ovary biochemical and in situ detection of deoxyribonucleic acid fragmentation in granulosa cells" 245-52, endocrinology1994
• 2 "Tumor selective G2/M cell cycle arrest and apoptosis of epithelial and hematological malignancies by BBL22 a benzazepine" 97 : 7494-7499, 2000
• 3 "The use of gonadotrophin releasing-hormone analogues some clinical and experimental inve- stigations of a direct effect on the human ovary" 7 39-1 47,
• 4 "The role of nitric oxide in oocyte meiotic maturation and ovulation:meiotic abnormalities of endothelial nitric oxide synthase knock-out mouse oocytes" 139 : 2944-2954, 1998
• 5 "The role of nitric oxide in oocyte meiotic maturation and ovulation:meiotic abnormalities of endothelial nitric oxide synthase knock-out mouse oocytes" 139 : 2944-2954, 1998
• 6 "The ovarian androgen producing cells:a review of structure function relationships" 6 : 371-99, 1985
• 7 "The involvement of nitric oxide in the ovulatory process in the rat" 135 : 2287-90, 1994
• 8 "Pregnancy rate and ovarian hyperstimu- lation after luteal human chorionic gonadotropin in in vitro fertilization stimulated with gonadotropin？releasing hormone analog and menotropins" 53 : 92-6, 1990
• 9 "Peripheral benzodiazepine receptor agonists exhibit potent anti-apoptotic activities" 265 : 457-461, 1999
• 10 "Peripheral benzodiaze- pine receptor in cholesterol transport and steroido- genesis" 62 : 21-28, 1997
• 11 "Nonsupple- mented luteal phase characteristics after the admi- nistration of recombinant human chorionic gonado- tropin, recombinant luteinizing hormone, or gona- dotropin-releasing hormone (GnRH) agonist to in- duce final oocyte maturation in in vitro fertilization patients after ovarian stimulation with recombinant follicle-stimulating hormone and GnRH antagonist cotreatment." 88 : 4186-4192, 2003
• 12 "Nitric oxide:physiology,pathophysiology,and pharmacology" 43 : 109-42, 1991
• 13 "Nitric oxide:an autocrine regulator of human granulosa-luteal cell steroidogenesis" 135 : 1799-806, 1994
• 14 "Nitric oxide modu- lates human chorionic gonadotropin-induced ovu- lation in the rabbit" 67 : 548-552, 1997
• 15 "Nitric oxide decreases estradiol synthesis in rat luteal cells in vitro:possible role for nitric oxide in functional luteal regression" 137 : 3531-3539, 1996
• 16 "Nitric oxide as a bifunctional regulator of apoptosis" 84 : 253-256, 1999
• 17 "Nitiric oxide and the control of reproduction" 65 : 1-16, 2001
• 18 "Mitochondrial peripheral-type benzodiazepine receptor expression: correlation with gonadotropin-releasing hormone (GnRH) agonist？induced apoptosis in the corpus luteum." 58 (58): 1389-1393, 1999
• 19 "Mitochondria as regulators of apoptosis:doubt no more" 1366 : 151-165, 1998
• 20 "Leung PCK Expression and regulation of gonadotropin-releasing hormone and GnRH receptor messenger ribonucleic acids in human granulosa luteal cells" 1740-6, endocrinology1994
• 21 "Isolation,characterization,and culture of cell sub- populations forming the pregnant rat corpus luteum" 130 : 954-66, 1992
• 22 "Isolation of the mitochondrial benzodiazepine receptor:association with the voltage-dependent anion channel and the adenine dinucleotide carrier" 89 : 3170-4, 1992
• 23 "Is nitric oxide an autocrine modulator of bovine granulosa cell function" 10 : 471-478, 1998
• 24 "Interleukin-1? suppresses apoptosis in rat ovarian follicles by increasing nitric oxide production" 136 : 3120-3127, 1995
• 25 "Inhibition of hCG-induced ovarian and uterine weight augmentation in the im- mature rat by analogs of GnRH" 152 : 432-6, 1976
• 26 "Induction of apoptosis by a gonadotropin-releasing hormone agonist during early pregnancy in the rat" 3 : 51-57, 1998
• 27 "Hormonal regulation of granu- losa cell inhibin biosyuthesis" 114 : 2711-9, 1986
• 28 "Hormonal regulation of apo- ptosis in early antral follicle:follicle-stimulating hormone as a major survival factor" 137 : 1447-1456, 1996
• 29 "Gonadotropin-releasing hormone-agonist inhibits synthesis of nitric oxide and steroidogenesis by luteal cells in the pregnant rat" 68 : 2222-2231, 2003
• 30 "Gonadotropin-releasing hormone receptor gene expression in human ovary and granulosa-lutein cells" 80 : 430-434, 1995
• 31 "Gonadotropin-relea- sing hormone agonist affects rat ovarian follicle development by interfering with FSH and growth factors on the prevention of apoptosis" 60 : 241-247, 2001
• 32 "GnRH agonist action on granulosa cells at varying follicular stages" 202 : 155-164, 2003
• 33 "GnRH action on luteal steroidogenesis during pregnancy" 64 : 618-623, 1999
• 34 "Extrapituitary actions of gonado- tropin-releasing hormone:stimulation of insulin-like growth factor-binding protein-4 and aterisa" 134 : 1365-72, 1994
• 35 "Expression and regulation of gonadotropin-releasing hormone(GnRH)and GnRH receptor messenger ribonucleic acids in human granulosa-luteal cells" 135 : 1740-6, 1994
• 36 "Evidence for an inverse relati- onship between apoptosis and inducible nitric oxide synthesis expression in rat granulosa cells:a possi- ble role of nitric oxide in ovarian follicle atresia" 45 : 745-751, 1998
• 37 "Enigma of the peripheral benzodiazepine receptor" 51 : 629-650, 1999
• 38 "Changes in rat luteal ultrastructure and P450scc mRNA and protein content after in vivo treatment with a gonadotropin-releasing hormone agonist" 44 : 382-91, 1991
• 39 "Bcl-2 resistant mitochondrial toxicity mediated by the isoquinoline carboxamide PK 11195 involves de novo generation of reactive oxygen species" 84 : 1397-1404, 2001
• 40 "Assisted reproduction-in vitro fertilization success is improved by ovarian stimulation with exogenous gonadotropins and pi- tuitary suppression with gonadotropin-releasing hor- mone analogues" 20 : 249-252, 1999