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생식샘자극호르몬분비호르몬이 사람 과립-황체화 세포의 스테로이드 생성과 세포자연사에 미치는 영향

Effects of Gonadotropin Releasing Hormone on Steroidogenesis and Apoptosis of Human Granulosa-Luteal Cells

양현원|이효진
한국발생생물학회 (The Korea Society Of Developmental Biology)|2009.12|발생과 생식 (DEVELOPMENT & REPRODUCTION)|제13권 제4호|pp. 353-362

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참고문헌

주제분류생물학

초록

GnRH는 국부적으로 난소에서 합성되며 난소내 과립 및 황체세포에 직접적으로 작용하여 난소의 기능을 조절하는 것으로 알려져 있으며. 특히, GnRH는 난소내 과립-황체화 세포의 세포자연사를 유도하는 보고하고 있다. 그러나 GnRH에 의한 세포자연사가 FSH에 의해 회복될 수 있는지는 명확히 밝혀져 있지 않다. 따라서 본 실험에서 난자 채취시 획득한 사람 과립-황체화 세포를 배양한 후 5, 50, 100 ng/㎖ GnRH와 1 IU/㎖ FSH를 처리하고 세포의 세포자연사 여부와 분비된 progesterone(P4)과 estradiol (E2) 양의 변화를 조사하였다. DNA 분절화 분석과 TUNEL 방법으로 세포자연사를 평가한 결과, GnRH는 농도 의존적으로 과립-황체화 세포의 세포자연사를 증가시켰고, 특히 100 ng/㎖ GnRH을 처리한 군에서 유의한 차이를 보이며 세포자연사 비율이 증가하였다. 또한 GnRH에 의한 세포자연사의 증가는 FSH에 의해 억제되는 것을 확인할 수 있었다. 화학발광면역 측정법을 이용하여 배양내 P4와 E2의 양을 측정한 결과, GnRH을 처리한 후 E2의 양은 변화가 없었던 반면 P4의 양은 감소하였다. 이러한 GnRH의 P4 합성 억제 효과는 세포자연사 결과 마찬가지로 FSH에 의해 회복되는 것을 확인할 수 있었다. 이상의 결과는 체외수정 및 배아이식 시술시 사용되고 있는 GnRH 작용제가 난소의 기능을 억제시킬 수 있을 것으로 보이나, 다량으로 투여되는 FSH에 의해 회복될 수 있음을 보여주고 있다. 이러한 실험 결과는 난소에 대한 GnRH의 생리적 기전을 이해하고 향후 새로운 과배란 유도 방법을 개발하는데 필요한 기초 자료로 사용될 수 있을 것으로 사료된다.

Abstract

GnRH and its receptor are known to express locally in ovary and to regulate the ovarian function by effecting on granulosa and lutein cells. It has been reported that GnRH directly causes apoptosis in the granulosa and lutein cells of the ovary. However, whether the apoptosis of the cells by GnRH is recovered by FSH as an anti-apoptotic factor. is not yet known. In the this study, therefore, we evaluated the apoptosis and the production of progesterone (P4) and estradiol (E2) after treatment with 5, 50, and 100 ng/㎖ GnRH and 1 IU/ml FSH in the granulosa-lutein cells that are obtained during oocyte-retrieval for IVF-ET. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in the cells treated with 100 ng/㎖ GnRH. In addition we found that FSH suppresses the apoptosis of the cells induced by GnRH. In the results of chemiluminescence assay for P4 and E2, P4 production was decreased by GnRH treatment, whereas E2 production was not changed. We also got to know that FSH inhibits the suppressive effect of GnRH on P4 production as the result of apoptosis. The present results suggest that GnRH agonist using in ovarian hyperstimulation protocol might induce the dysfunction of ovary, but its function could be recovered by FSH. These results also will be expected to use as the basic data to elucidate the physiological role of GnRH and to develop new ovarian hyperstimulation protocols for IVF-ET.

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