Nitric Oxide가 인간 정자세포의 기능에 미치는 효과에 대한 연구

주보선,문화숙,박수진,문재연,조재동,김한도,Joo, Bo-Sun,Moon, Hwa-Sook,Park, Sue-Jin,Moon, Jae-Yeoun,Cho, Jae-Dong,Kim, Han-Do 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.1

This study was performed to determine the effects of nitric oxide on human sperm cell function. Semen samples were obtained from normal healthy volunteers. Motile spermatozoas collected by swim-up method were incubated up to 24 hours in Ham's F-10 medium supplemented with a various concentration of sodium nitroprusside (nitric oxide releasing agent). Sperm motility, hyperactivation, acrosome reaction rate, and acrosin activity were determined. The results are as follows; 1. 1mM of SNP resulted in a significant decrease in sperm motility ($44.8%{\pm}8.9%:78.1%{\pm}6.3%$, and hyperactivation $(10.4%{\pm}6.4%:47.7%:{\pm}9.5%)$ after incubation for 3 hours compared with the control group (Ham's F-10 alone), but had no effect on acrosome reaction. 2. At $100{\mu}M$ SNP, sperm motility was reduced after incubation for 6 hours $(54.8%{\pm}3.2%)$ compared with that of the control group $(82.7%{\pm}8.9%)$, but hyperactivation and acrosome reaction were not affected. 3. However, a lower concentration (less than $10{\mu}M$) of SNP had no effect on sperm motility and hyperactivation for 8 hours of incubation but significantly decreased them when incubation periods were increased up to 24 hours compared with the control group. On the other hand, $1{\mu}M$ and $10{\mu}M$ SNP significantly increased the acrosome reaction rate in both acrosomal status ($17.3%{\pm}5.2%$, $23.5%{\pm}4.7%$, respectively) and acrosin activity ($34.3{\mu}IU{\pm}10.5{\mu}IU,\;45.6{\mu}IU{\pm}5.6{\mu}IU$, respectively) as compared with the control group $(7.0%{\pm}4.0%,\;9.5{\mu}IU{\pm}3.4{\mu}IU)$. These results indicate that SNP, NO releasing agent, has a dose-dependent effects on the sperm cell function. Therefore it may positively affect the fertilization by promoting acrosomal reaction at a lower concentration (less than $10{\mu}M$).

Regular exercise increases reproductive outcomes of aged female in mouse model

주보선 ( Bo Sun Joo ),( Sun Hee Park ),( Su Kyung Lee ),( Sae Hee Park ),( Sang Chil Kwon ),( Hwa Sook Moon ) 대한산부인과학회 2007 대한산부인과학회 학술대회 Vol.93 No.-

2세포기로의 조기난할 배아 선발을 이용한 체외수정술의 임신율 증가

박세희,주보선,이수경,김경서,문화숙,Park, Sea Hee,Joo, Bo Sun,Lee, Su Kyung,Kim, Kyung Sue,Moon, Hwa Sook 대한생식의학회 2005 Clinical and Experimental Reproductive Medicine Vol.32 No.1

Objective: Evaluation of embryos using early cleavage to 2-cell stage has been proposed, but a critical time-point for selecting embryos is unclear. The aim of the present study is to provide a guideline including critical time-point in the selection of early cleaving embryo for the reduction of multiple pregnancies as well as the increase of pregnancy rate in human IVF. Methods: This prospective study was performed in 116 cycles from 85 patients who underwent conventional IVF or ICSI at the infertility clinic of Good Moonhwa Hospital from January 2002 to December 2003. Early cleavage (EC) of embryos to 2-cell stage was assessed at 25 h and 27 h postinsemination/microinjection. Embryos that had early cleaved at each time point were designated as EC-1 and EC-2, respectively, while others were designated as non-early cleavage (NEC). Results: At least one early cleavage embryo was observed in 54 (46.6%) for the EC-1 and 84 (72.4%) for the EC-2 of the 116 cycles assessed. Clinical pregnancy rates (PR) were significantly higher in the EC-1 group (66.7%) compared to the EC-2 group (53.6%) or the NEC group (31.2%) (p<0.05). Significant improvement of the pregnancy rate was found when at least two or more embryos were early cleaved at 25 h postinsemination or when the proportion of early cleavage embryo at 25 h postinsemination was higher than 20% (p<0.05). Conclusion: The critical time-point for the selection of early cleavage embryos with high implantation potential is more effective in 25 h postinsemination/microinjection compared to 27 h. The proportion as well as number of early cleavage embryos is also an important factor for the prediction of pregnancy outcome and the chance of multiple pregnancies. These results demonstrated that the evaluation of early cleavage embryos to 2-cell stage is an easy, simple, and objective method for the selection of good quality embryos suitable for embryo transfer.

성선자극호르몬 및 GnRH agonist을 동시에 중단하는 1$\sim$2일 단기 코스팅이 임신율을 포함한 난소과자극증후군 예방에 미치는 효과

이수경,주보선,박세희,이수경,김경서,문성은,문화숙,Lee, Soo-Kyung,Joo, Bo-Sun,Park, Sea-Hee,Lee, Su-Kyung,Kim, Kyung-Seo,Moon, Sung-Eun,Moon, Hwa-Sook 대한생식의학회 2007 Clinical and Experimental Reproductive Medicine Vol.34 No.1

목 적: 성선자극호르몬과 GnRH agonist (GnRH)를 동시에 중단하는 1$\sim$2 일간의 단기 coasting이 임신율을 포함한 난소과자극증후군 예방에 미치는 효과를 조사하고자 한다. 연구방법: 체외수정시술을 위한 과배란유도 시 15 mm 이상의 난포가 20 개 이상이고 혈중 E$_2$ 농도가 4,000 pg/ml 이상일 때 coasting을 시도한 37 명의 여성들을 대상으로 하였다. Coasting은 성선자극호르몬과 GnRH agonist를 동시에 중단하였으며, 초음파상 난포의 상태와 혈중 E$_2$ 농도 에 따라 1일 또는 2일 동안 시행하였다. 혈중 E$_2$ 농도, 채취된 난자 수, 수정율, 임신율 등을 후향적으로 비교 분석하였다. 결 과: 평균 혈중 E$_2$ 농도는 coasting 시작 당일 6,993 pg/ml 에서 hCG 투여일에 3,396 pg/ml로 감소하였다. 평균 채취 난자 수와 수정율은 15.7개와 70%였다. 15명 (40.6%)이 임신을 하였고 착상율은 15.2%였다. 26명 (70.3%)이 1일, 11명 (29.7%)이 2일 동안 coasting 하였다. 평균 혈중 농도의 감소율은 1일 coasting한 군에서 43%, 2일 costing한 군은 15% (첫날)와 81% (둘째날)이었다. 임신율은 두 군간 유사하였고, 중증도 이상의 OHSS는 발생하지 않았으며 3명 (8.1%)에서 경미한 OHSS가 나타났다. 결 론: 성선자극호르몬과 GnRH agonist의 통시 중단에 의한 1$\sim$2 일의 단기 coasting은 체외수정의 결과에 영향을 주지않고 OHSS 를 예방하는데 성공적으로 적용될 수 있을 것으로 사료된다. Objective: To evaluate the effect of short coasting, by withdrawing both gonadotropins and GnRH agonist (GnRHa), on the prevention in severe ovarian hyperstimulation syndrome(OHSS) without compromising pregnancy outcome. Method: Thirty-seven women who had been coasted during COH for IVF were coasted when $\geq$20 follicles > 15 mm with serum E$_2$ level of 4,000 pg/ml were detected. Coasting was initiated for one or two days depending on the status of follicle on ultrasound and serum E$_2$ level. Both gonadotropin and GnRHa were withheld for coasting. Retrospective study was carried and changes of serum E$_2$ levels, number of oocytes retrieved, fertilization rate, pregnancy rate were compared and analyzed. Results: The mean serum E$_2$ level fell from 6,993 pg/ml on the onset of coasting to 3,396 pg/ml on the day of hCG administration. The mean number of oocytes retrieved and fertilization rate were 15.7 and 70.0%, respectively. Fifteen patients were pregnant (40.6%) and implantation rate was 15.2%. Twenty-six (70.3%) patients were coasted for one day and 11 (29.7%) were coasted for two days. The mean decrease rate of serum E$_2$ level was 43% in one day coasting group and 15% (1$^{st}$ day) and 81% (2$^{nd}$ day) in two day coasting group. The pregnancy outcome was similar between the two groups. After coasting, no severe or moderate OHSS occurred in any patients and mild OHSS occurred in 3(8.1%) patients. Conclusions: Coasting for one or two days can be used successfully in the prevention of OHSS without compromising IVF cycle outcome.

공배양의 작용기전에 관한 연구

김미경,주보선,김미선,문화숙,이규섭,김한도,Kim, Mi-Kyoung,Joo, Bo-Sun,Kim, Mi-Sun,Moon, Hwa-Sook,Lee, Kyu-Sup,Kim, Han-Do 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.1

Objective: A number of studies to improve in vitro culture conditions have been tried over past ten years by using co-culture system with helper somatic cells. However, the mechanism of coculture is poorly understood. This study was designed to understand the mechanism for the mode of actual action of co-culture using co-culture system of ICR strain's 1-cell embryos with human oviduct epithelial cells by examining the effect of conditioned medium and contactless coculture using a cell culture insert on the embryo development and by measuring the level of superoxide anion from conditioned medium after co-culture. Methods: ICR strain's zygote embryos were cultured in medium alone (control), coculture, conditioned medium, or contactless coculture system for 6 days. Conditioned media (CM) were prepared as following 5 groups. All CM were collected after culturing oviduct cells for 2 days. CM-1 was stored at $-20^{\circ}C$ until use, and CM-2 was prepared just before use as a culture medium. CM-3 was cocultured with embryos and retrieved just before use. CM-4 and CM-5 were derives from the microfilteration of CM-2 and CM-3, respectively, using Microcon-10 (10 kDa molecular weight cut-off). The percentage of the embryos developed to hatched blastocyst stage and the level of superoxide anion in supernatant from medium alone culture (control), coculture, and contactless coculture were measured. Results: The rates of embryo development to the hatched blastocyst stage were significantly higher in coculture (43%) than in control (0%) (p<0.05). The CM-1 group had no embryo development since 2-cell embryonic stage, whereas the CM-2, CM-3, CM-4 and CM-5 groups had the improved development to 4 or 8 cell embryo stage, but the similar rate of development to hatched blastocyst compared to control. The effect of coculture on embryo development was disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in coculture group compared to control. Conclusion: It is concluded that the present coculture system overcomes the 2-cell block in vitro and improves the embryo development. This beneficial effect may be due to the direct cell-cell contact between embryo and helper cells or the removal of deleterious components from medium rather than the embryotrophic factors.

Dose-Dependent Inhibitory Effect of Nitric Oxide on Embryo Development

박세희,주보선,문화숙,김윤연,조재동,강호성,김한도,Park, Sea-Hee,Joo, Bo-Sun,Moon, Hwa-Sook,Kim, Yoon-Yeon,Cho, Jae-Dong,Kang, Ho-Sung,Kim, Han-Do The Korean Society for Reproductive Medicine 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.2

목적: 일산화질소 (nitric oxide; NO)는 생식계를 비롯한 여러 생체내 기관에서 다양하고도 중요한 작용을 하는 것으로 알려져 있으며, 복강액은 난관내강과 연결되어 복강액 내의 세포 성분의 변화는 난관의 미세환경을 변화시켜 수정과 초기 배아 발생에 영향을 줄 수 있다. 본 연구는 배아 발생에 있어서 일산화질소의 역할을 이해하고 복강액 내의 NO농도 변화가 배아 발생에 미치는 역할을 조사하기 위해 수행되었다. 방법: 과배란시킨 1세대 잡종 암컷 생쥐 (C57BL${\times}$CBA/Ca)로부터 1세포기 배아를 얻어 10% synthetic serum substitute가 첨가된 modified human tubal fluid 배양액에서 4일 동안 체외배양하였다(대조군). 실험을 위해 이러한 배양조건에 sodium nitroprusside (SNP)를 $0{\sim}1mM$의 다양한 농도로 배양초기부터 첨가하거나, $200{\mu}M$ SNP를 2-, 4-, 8-세포기의 각기 다른 배아시기에 첨가하였으며, 복강경수술을 받는 42명의 여성으로부터 채취한 복강액을 SSS대신 단백질원으로 사용하여 포배아까지의 배아 발달율을 관찰하였으며, 복강액 내의 NO농도를 Griess방법에 의해 측정하였다. 배아의 apoptotic body는 H33342 염색법으로 조사하였으며 배아 발달율은 3회 이상 반복 실험한 결과의 mean${\pm}$SEM으로 나타내었다. 결과: SNP는 농도에 의존적으로 배발생을 억제하였으나 배아 단계에 대한 특이성은 관찰할 수 없었으며, 특히 $100{\mu}M$ 이상의 고농도의 SNP는 2-세포기 단계에서 배아 발생을 정지시켰다. 또한 단백질원으로 복강액 이용시 배 발생율은 복강액 내의 NO 농도에 따라 현저한 차이가 발견되었으며, $2.5{\mu}M$이상의 NO를 함유한 복강액에서 배양한 배아의 발생율은 현저하게 감소하였다. cGMP analogue인 8-bromo-cGMP를 배양액에 첨가시 배아 발생에는 변화가 없었으며, SNP에 의해 배발생이 정지된 2-세포기 배아에서 apoptotic body를 발견할 수 없었다. 결론: 이상의 결과로 보아 NO는 고농도에서 배아 발생을 저해하며, 복강액 내의 NO와 같은 성분의 변화는 배아 발생에 유해한 효과를 유발할 것으로 사료된다. 이러한 NO의 배아 발생 억제효과는 cGMP로 중재되는 경로나 apoptosis유발과는 관계가 없는 것 같다.

Alendronate에 의한 MC3T3-E1 골모세포주의 분화와 Id-1과 Id-2의 발현 증가

박민정 ( Min Jung Park ),주보선 ( Bo Sun Joo ),이지은 ( Ji Eun Lee ),김현정 ( Hyun Jung Kim ),문성은 ( Sung Eun Moon ),문화숙 ( Hwa Sook Moon ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.3

목적: Alendronate는 대표적인 비스포스트네이트로서 파골세포의 활성을 떨어뜨려서 과도한 골흡수를 억제하는 약제로 알려져 왔으나, 최근에는 골세포 증식 및 분화를 증가시킨다고 보고되었다. 그러나 alendronate가 조골세포 분화에 미치는 기전에 대한 연구는 매우 부족하다. Id (Inhibitors of DNA binding/differentiation)은 helix-loop-helix (HLH) 계열에 속하는 전사인자이며, BMP에 의해 유도되는 골세포 분화과정동안 주요한 역할을 하는 것으로 알려져 있다. 그러므로 본 연구는 alendronate가 골세포 분화와 Id-1 및 Id-2 유전자의 발현에 미치는 효과를 조사하고자 한다. 연구 방법: 조골모세포인 MC3T3-E1 세포주에 alendronate를 다양한 농도 (10-9 M-10-4 M)와 시간 (24, 48 및 72시간)으로 처리한 후 조골세포로의 분화 정도를 ALP 활성도와 조골세포 분화표지자인 ALP, Col 1, OCN에 대한 발현을 역전사-중합효소반응 (RT-PCR) 방법으로 조사하였다. 또한, alendronate을 처리 후 Id-1 및 Id-2의 유전자 발현을 RT-PCR 방법으로 조사하였다. 결과: Alendronate는 ALP 활성도뿐만 아니라 ALP, Col 1 및 OCN의 mRNA 발현을 증가시키며, Id-1 및 Id-2의 발현도 증가시켰다. 이러한 alendronate에 의한 골세포 분화는 고농도보다 저농도에서 골세포 분화에 효과적이었다. 결론: 본 연구는 골형성과 관련된 전사인자 Id-1과 Id-2의 발현이 alendronate에 의한 조골세포 분화 과정동안 촉진됨을 보여주고 있으며, 이러한 alendronate 효과는 농도 의존적으로 골세포 분화에 영향을 미치는 것으로 사료된다. Objective: Alendronate, a widely used bisphosphonates, acts to inhibit bone resorption by interfering with the activity of osteoclasts. Recently, it has been reported that alendronate also may increase bone proliferation and osteoblastic differentiation. However, little is known about mechanism of the action of alendronate on osteoblast differentiation, especially in transcription level. Inhibitors of DNA binding/differentiation (Ids) are helix-loop-helix (HLH) transcription factors and play an important role in BMP-induced osteoblast lineage-specific differentiation. Therefore, this study was aimed to investigate the effect of alendronate on osteoblast differentiation and expression of Id-1 and Id-2. Methods: MC3T3-E1, pre-osteoblast cell line, were treated with alendronate of various concentrations (10-9 M-10-4 M) and time periods (24, 48 and 72 hours). And then, the effect of alendronate on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and RT-PCR for osteoblast differentiation markers such as ALP, type 1 collagen (Col 1), and osteocalcin (OCN). The expressions of Id-1 and Id-2 were measured by RT-PCR. Results: Alendronate treatment increased not only ALP activity, but also expressions of ALP, Col 1, and OCN. Also, alendronate treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This alendronate-induced osteoblastic differentiation is more effective in lower doses rather than high doses. Conclusion: This study shows that the expression of transcription factor Id-1 and Id-2 was increased in a dose-dependent manner during alendronate-induced osteoblast differentiation.

조기난소부전 여성에서 자연 임신 2예

이수경 ( Soo Kyung Lee ),주보선 ( Bo Sun Joo ),문성은 ( Sung Eun Moon ),김상갑 ( Sang Kap Kim ),구자성 ( Ja Sung Koo ),문화숙 ( Hwa Sook Moon ) 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.11

Premature ovarian failure (POF) is a syndrome defined as hypergonadotropic hypogonadism associated with amenorrhea, oligomenorrhea or other forms of menstrual irregularity for at least 3 consecutive months before the age of 40. The management of POF is approached by HRT, emotional support and infertility treatment. Women with premature ovarian failure who desire to become pregnant are best treated by assisted reproductive technology with donor oocyte. However, POF has the possibility of a 5-10% spontaneous pregnancy. The physician should recommend the patient to consult with their physician if they have any symptoms of pregnancy or no withdrawal bleeding after HRT. Therefore we report two cases of spontaneous pregnancies in women with premature ovarian failure.

혈중 E2 농도가 체외수정술의 임신율에 미치는 영향

박세희 ( Sea Hee Park ),주보선 ( Bo Sun Joo ),송수진 ( Soo Jin Song ),김경서 ( Kyung Sue Kim ),문성은 ( Sung Eun Moon ),문화숙 ( Hwa Sook Moon ) 대한산부인과학회 2008 Obstetrics & Gynecology Science Vol.51 No.11

목적: hCG 투여 당일 혈중 E2 농도가 체외수정술의 임신결과에 미치는 효과를 조사하고자 한다. 방법: 체외수정시술을 시행받은 455 주기를 대상으로 hCG 주사 당일 측정한 E2 농도에 따라 A군 (<1,000 pg/mL), B군 (1,000~2,000 pg/mL), C군 (2,000~3,000 pg/mL), D군 (3,000~4,000 pg/mL), E군 (≥4,000 pg/mL)으로 나눈 다음 체외수정술 결과를 비교하였다. 결과: hCG 투여 당시 혈중 E2 농도는 평균 3,745.3 pg/mL였으며, 채취된 평균 난자수는 10.1개였다. 148주기 (32.5%)에서 임상적 임신이 확인되었으며, 착상률 12.2%, 분만율 18.7%였다. E2 농도가 증가할수록 채취된 난자수는 증가된 경향을 나타냈다. 임신율 또한 E2 농도가 증가함에 따라 점차 증가하다가 D군 (50.0%)에서 가장 높은 다음, E군에서 다시 감소하였다. 38세 미만 여성에서는 전체 환자 군에서와 유사한 결과를 보인 반면, 38세 이상 여성에서는 임신율과 분만율 모두 C군에서 유의하게 가장 높았다. 결론: 이러한 결과는 혈중 E2 농도가 농도 의존적으로 임신결과에 영향을 미칠 수 있으며, 임신결과와 양성적 상관관계를 가지는 최적의 E2 농도 범위가 있음을 시사하고 있다. 과도한 E2 농도에 의한 임신결과의 저해는 여성 나이가 38세 이상일 경우 더욱 민감하였다. Objective: To evaluate the effects of serum E2 levels on the day of hCG administration on the pregnancy outcome of IVF-ET after controlled ovarian hyperstimulation (COH). Methods: Data of 455 cycles of fresh IVF-ET with COH were retrospectively investigated. Serum E2 levels on the day of hCG administration were categorized into 5 groups; A (<1,000 pg/mL), group B (1,000~2,000 pg/mL), group C (2,000~3,000 pg/mL), group D (3,000~4,000 pg/mL), and group E (≥ 4,000 pg/mL). Results: Mean E2 levels on the day of hCG administration were 3,745.3 pg/mL and mean number of retrieved oocytes were 10.1. Of 455 cycles, 148 (32.5%) cycles were clinically pregnant. Implantation rate was 12.2% and delivery rate was 18.7%. The number of obtained oocytes increased with increasing levels of serum E2. Pregnancy rate gradually increased as E2 levels increased up to the group D, but began to fall in the group E. In younger women (<38 yrs), the IVF-ET outcomes were similar to those of total patients but in older women (≥38 yrs), pregnancy rate and delivery rate were significantly higher in the group C than other groups. Conclusions: This result shows that serum E2 levels have a concentration-dependent effect on the pregnancy outcome and there is an optimal range of E2 levels to achieve for a successful pregnancy. Excessive E2 levels seem more deleterious to the pregnancy outcome in older women aged ≥38 years.

비스포스포네이트가 인간 양수유래 중간엽 줄기세포로부터 조골세포의 분화에 미치는 영향

안준협 ( Joon Hyop An ),안병민 ( Byeong Min An ),최태영 ( Tae Young Choi ),김상국 ( Sang Gook Kim ),문화숙 ( Hwa Sook Moon ),주보선 ( Bo Sun Joo ) 대한산부인과학회 2009 Obstetrics & Gynecology Science Vol.52 No.3