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Apoptosis of human luteinized granulosa cell via change of mitochondria

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dc.contributor.author김, 미란-
dc.contributor.author박, 동욱-
dc.contributor.author김, 영아-
dc.contributor.author조, 태섭-
dc.contributor.author황, 경주-
dc.contributor.author임, 윤경-
dc.contributor.author민, 철기-
dc.date.accessioned2012-01-17T04:23:41Z-
dc.date.available2012-01-17T04:23:41Z-
dc.date.issued2001-
dc.identifier.issn0494-4755-
dc.identifier.urihttp://repository.ajou.ac.kr/handle/201003/5283-
dc.description.abstractObjective : To determine the effects of extracellular ATP on mitochondrial function and apoptosis during human luteinized granulosa cell cultures. Methods : The addition of various concentrations of ATP (0, 0.1, 0.25, 0.5, 0.75 mM) to luteinized granulosa cells obtained during In vitro fertilization ovum pickup procedures. After culture for 24 hours, purinoceptor activity and functional changes in mitochondria were measured by patch clamp, flow cytometry, and confocal microscopy. Results : Measurement by patch clamp of the granulosa cell membrane potential after ATP addition to cultured granulosa cells showed that both the inward and outward currents were expressed. After treatment of the granulosa cells with JC-1, measurement of the mitochondrial activity by confocal microscopy showed that the with increasing concentrations of ATP the relative ratio of undamaged mitochondria (red/green ratio) tended to decrease (P=0.027). After double staining of the cultured granulosa cells with Annexin V and Propidium Iodide, quantitative flow cytometry analysis showed that apoptosis increased with increasing concentrations of ATP (7.88%, 8.44%, 11.40%, 13.52%, 18.57%). Conclusion : The results of this study shows that apoptosis of granulosa cells increases with increasing extracellular ATP concentrations in cultured human luteinized granulosa cells. This is observed to be a consequence of cell membrane purinoceptor activity and functional changes in the mitochondria. It is therefore thought that remodelling processes of the ovarian tissue is regulated by neuroendocrine factors of the extracellular ATP.en
dc.description.abstract목적 : 세포외 ATP가 인간 황체화 과립세포 배양시 미토콘드리아의 기능적 변화와 세포자멸사 과정에 미치는 영향을 알아보고자 하였다.
연구방법 : 체외수정술을 시행 받은 환자들을 대상으로 난자채취시 획득한 황체화 과립세포에 각기 다른 농도의 ATP (0, 0.1, 0.25, 0.5, 0.75 mM)를 첨가하여 24시간 배양한 후 patch clamp, flow cytometry, confocal microscope을 통하여 urinoceptor의 활성 및 미토콘드리아의 기능적 변화를 측정하였다.
결과 : 배양된 과립세포에 ATP를 첨가한 후 patch clamp 기법을 이용하여 과립 세포막 전위차를 측정한 결과 inward current와 outward current가 모두 발현되었다. JC-1을 과립세포에 처리한 후 공촛점 현미경을 이용하여 미토콘드리아의 활성도를 측정한 결과 ATP의 농도가 높아질수록 손상 받지 않은 미토콘드리아의 상대적 빈도 (red/green ratio)가 감소하는 양상을 보였다 (P=0.027). 배양된 과립세포를 Annexin V와 Propidium Iodide로 이중 염색한 후 flow cytometry로 정량 분석한 결과 ATP의 농도가 증가할수록 세포자멸사의 정도가 증가함을 보여주었다 (7.88%, 8.44%, 11.40%, 13.52%, 18.57%).
결론 : 인간 황체화 과립세포의 배양에 있어서 세포외 ATP는 농도에 비례하여 과립세포의 세포자멸사를 유도하는 것으로 나타났으며 이러한 과정은 세포막의 purinoceptor의 활성 및 미토콘드리아의 기능학적 변화를 통하여 이루어짐을 관찰함으로써 난소의 재형성과정이 세포외 ATP에 의한 신경내분비학적 조절을 받고 있으리라 사료된다.
ko
dc.formatapplication/pdf-
dc.language.isoko-
dc.titleApoptosis of human luteinized granulosa cell via change of mitochondria-
dc.title.alternativeATP에 의해 유도된 인간 난소 황체화 과립세포의 세포자멸사에 관한 연구-
dc.typeArticle-
dc.identifier.urlhttps://www.ogscience.org/journal/view.php?number=3637-
dc.subject.keywordPurinoceptor-
dc.subject.keywordApoptosis-
dc.subject.keywordLuteinized granulosa cell-
dc.subject.keyword퓨린수용체-
dc.subject.keyword세포자멸사-
dc.subject.keyword황체화 과립세포-
dc.contributor.affiliatedAuthor김, 미란-
dc.contributor.affiliatedAuthor황, 경주-
dc.type.localJournal Papers-
dc.citation.titleKorean journal of obstetrics and gynecology-
dc.citation.volume44-
dc.citation.number11-
dc.citation.date2001-
dc.citation.startPage2091-
dc.citation.endPage2097-
dc.identifier.bibliographicCitationKorean journal of obstetrics and gynecology, 44(11). : 2091-2097, 2001-
dc.relation.journalidJ004944755-
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Obstetrics & Gynecology
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