Studies on the Regulation of Ovarian Granulosa Cell Apoptosis by Gonadotropins and Nitric Oxide

이석자 한국발생생물학회 1997 발생과 생식 Vol.1 No.2

To study the regulation of porcine follicular cell apostosis by gonadotropin, steroid, and nitric oxide, we analyzed DNA fragmentation, the hallmark of apoptosis, and nitrite production of porcine granulosa cells. Dissected indiidual follicles from ovary were separated in size (small, 2-3 mm; medium, 5-6 mm; large, 7-8 mm) and isolated granulosa cells were classified morpholocally as atretic or nonatretic. Nitrite concentration was measured by mixing follicular fluids with an equal volume of Griess reagent. Follicular nitric oxide (NO) concentration of healthy follicles was higher than that of atretic follicles. Apoptotic DNA fragmentation was suppressed in non-apoptotic granulosa cells. Follicular apoptosis was induced by androgen but prevented by gonadotropin in vitro. Apoptosis was confined to the granulosa cells. But it was not clear whether apoptosis of granulosa cells were isolated, incubated with or without gonadotropin, androgen and sodium nitroprusside (SNP), respectively at for 24 hrs. Cultured granulosa cells were used to extract genomic DNA and culture media was asssayed for nitrite concentration. Nitrite production of culture media was increased, while apoptotic DNA fragmentation was suppressed in PMSG, hCG, testosterone+SNP and SNP treated groups. Nitrite concentration in culture media was decreased, but apoptotic DNA fragmentation was induced in testosterone treated group. These data suggest that NO production and apoptosis may be involved of granulosa cell apoptosis induced by testosterone.

Gonadotropin과 Steroid에 의한 난소 과립세포의 Apoptosis와 Nitric Oxide 합성에 대한 영향

이석자,강한승,김외리,오미숙,강성구 한국통합생물학회 1995 한국동물학회 뉴스레터 Vol.12 No.2

Nitric Oxide에 의한 난소 과립세포의 Apoptosis 조절에 대한 연구

강한승,이석자,김외리,이채관,신창숙,김성태,강성구 인제대학교기초과학연구소 1998 자연과학 Vol.2 No.-

여포의 퇴화가 진행되는 기작에서 apoptotic cell death 와 nitric oxide (NO)의 연관성에 관하여 실험하였다. 난소의 apoptosis는 GnRH에 의해 유도되고 스테로이드에 의해 억제된다. 여포에서 과립 세포의 apoptosis에 대한 호르몬의 영향과 NO 생산효과를 조사하기 위하여 중간크기 (직경 5-6 ㎜)의 여포에서 non-apoptotic 과립세포를 분리하여 스테로이드, GnRH agonist, GNRH agonist+SNP, 그리고 NO 공여체인 SNP를 처리하여 24시간동안 배양하였다. 배양된 과립세포에서 게놈 DNA를 추출하여 apoptotic DNA절편화 경향을 알아보았고, 배양액과 griess reagent를 반응시켜 nitrite 농도를 측정하였다. 스테로이드 호르몬을 처리하였을 때, 대조군에 비해 NO 생성이 다소 증가하였으며 apoptotic DNA 절편화도 억제되었다. GnRH agonist 처리군에서는 apoptotic DNA 절편화가 가속화 양상을 나타내었으며 NO 생성은 다소 감소하였다. 그러나 GnRH agonist+SNP 그리고 SNP 처리군에서는 apoptotic DNA 퇴화가 억제되었으며 NO 농도는 대조군에 비해 현저히 높았다. 위의 실험 결과로 보아 과립세포 분화와 여포성숙과 퇴화과정에서 호르몬과 NO가 밀접한 관련이 있는 것으로 사료된다. To study the regulation of porcine follicular cell apoptosis by steroids, GnRH agonist and nitric oxide, we have analyzed DNA fragmentation, the hallmark of apoptosis, and nitrite production in porcine granulosa cells. In, ovarian apoptosis is induced by GnRH but is prevented by steroids. Apoptosis is confined to the granulosa cells but it was not clear whether apoptosis of granulosa cell was regulated by NO in vitro. So non-apoptotic granulosa cells isolated, grown at 37℃, for 24 hrs, and treated with / without SNP, GnRH agonist, and sterioid, respectively. In cultured granulosa cells were used to extract genomic DNA and culture media was assayed for nitrite concentration. Nitrite production of culture media was increased and apoptotic DNA fragmentation was supressed in SNP treated groups, GnRH agonist+SNP, and estrogen, progesterone. Nitrite concentration in culture media was decreased and apoptotic DNA was induced treated with GnRH agonist. These data suggest that NO production and apoptosis may be involved of granulosa cell differentiation and ovarian follicular atresia by SNP, estrogen and progesterone. Moreover, NO can overcome the granulosa cell apoptosis induced by GnRH agonist.

어머니의 양육 스트레스와 유아의 사회적 유능감, 정서조절능력 간의 상호관계에 관한 연구

조효숙,이석자,박영숙 국제차세대융합기술학회 2021 차세대융합기술학회논문지 Vol.5 No.5

본 연구는 어머니의 양육 스트레스와 유아의 사회적 유능감, 정서조절 능력 간의 관련성을 살펴보고 어 머니의 양육 스트레스가 유아의 사회적 유능감과 정서조절능력에 미치는 영향을 알아보는 목적으로 경기도와 서 울의 어린이집 만 3, 4, 5세 유아 145명의 담임교사와 유아의 주 양육자를 대상으로 질문지를 배포하여 자료를 수 집하였다. 양육 스트레스는 어머니가 정서조절능력과 사회적 유능감은 담임교사가 체크하도록 하였으며 SPSS 25.0을 활용하여 Pearson 상관관계 및 중다회귀분석을 실시하였다. 연구결과 첫째, 어머니의 양육 스트레스, 유아 의 사회적 효능감, 정서조절 능력 인식수준은 모두 평균보다 높았으며 둘째, 어머니의 양육 스트레스와 유아의 사 회적 유능감, 정서조절 능력은 부적 상관관계가 나타났다. 셋째, 어머니의 양육 스트레스가 증가할수록 유아의 사 회적 유능감과 정서조절능력은 낮아져 어머니의 양육 스트레스는 유아의 사회적 유능감과 정서조절능력에 부정적 인 영향을 미치는 것으로 나타났다. 본 연구는 어머니의 양육 스트레스가 유아의 사회정서 발달에 영향을 끼침을 밝힘으로 어머니의 양육 스트레스의 감소와 긍정적 해소가 유아의 성장발달에 중요함을 인식시키고 주 양육자의 양육 스트레스를 감소시키고 해소시킬 수 있는 방법과 유아의 사회적 유능감, 정서조절능력을 높일 수 있는 프로 그램 개발을 제안하는데 연구의 의의와 시사점이있다. The purpose of this study is to examine the relationship between the parenting stress of the Mother's, the social competence of the infant, and the emotional control ability and to find out how the parenting stress affects the infant's social competence and emotional control ability. The subjects of the study were 145 homeroom teachers and Mother's, working at daycare centers in Seoul and Gyeonggi-do, The research method Infant mothers checked parenting stress, homeroom teachers checked infants' emotional control ability and social competence, and Pearson correlation and multiple regression analysis were conducted using SPSS 25.0. As a result of the study, first, the level of perception of the three variables are higher than average. Second, there was a negative correlation between the parenting stress of the Mother's the infant's social competence, and emotional control ability. Third, as the parenting stress of the Mother's increased, the infant's social competence and emotional control ability decreased, and the parenting stress of the Mother's negatively affected the infant's social competence and emotional control ability. Follow-up studies suggest recognizing the importance of reducing and positive relieving mother parenting stress in infant growth development by revealing that mother parenting stress affects infant social emotional development, and suggesting development of infant social competence and emotional control.

생식소 자극 호르몬과 스테로이드 호르몬에 의한 생쥐 여포의 bad와 bax 유전자 발현

강한승,김외리,이석자,이채관,신창숙,김성태,강성구 인제대학교 1998 仁濟論叢 Vol.14 No.1

Abstract The purpose of this study was to investigate the effects of gonadotropin and steroid hormone on the expression of mouse follicular bad and box genes that are known to induce apoptosis. Ovary of immature mice were obtained at 0, 24, 48 and 72 hours time intervals after Pregnant Mare's Serum Gonadotropins (PMSG, 5 I.U.) injection. Large and preovulatory follicles collected at 24hrs after PMSG injection were cultured with or without various chemicals such as gonadotropin, progesterone, estrogen, testosterone for 24hrs at 37℃. After 24hrs culture, the follicles was used for total RNA extracted, subjected to RT-PCR for the analysis of bad and bax expression. We found that expressions of bad and bax genes in ovary was markedly reduced before and after PMSG injections. When the preovulatory follicles were cultured for 24 hrs in culture media with progesterone and estrogen, the expression of bad and bax genes was decreased. However testosterone significantly induce the expressions of bad and bax genes in follicles. These data demonstrated that gonadotropin and sex steroid hormone may play an important roles in the regulation of mouse follicular differentiation. PMSG, progesterone and estrogen may prevent apoptosis via suppressing the expressions of bad and bax genes but testosterone may accelerate apoptosis via inducing the bad and bax gene expressions. Key words : Apoptosis, bad, bax, mouse follicle, RT-PCR