자궁내막증의 병인: 초기 병변 형성을 중심으로

나용진,Na, Yong-Jin 대한생식의학회 2007 Clinical and Experimental Reproductive Medicine Vol.34 No.2

폐경여성의 호르몬 치료가 사망률에 미치는 영향

나용진 ( Yong Jin Na ),송용중 ( Yong Jung Song ),구윤희 ( Yun Hee Koo ) 대한폐경학회 2012 대한폐경학회지 Vol.18 No.3

호르몬 치료의 궁극적인 결과로서 사망률의 감소에 관하여 현재까지 많은 논란이 있다. 대규모 전향적 코호트 연구에서는 교란 변수들을 보정한 이후에도 호르몬 치료를 시행한 경우가 시행하지 않은 경우에 비해 전체 사망률을 의미 있게 감소시켰다. WHI 연구결과의 두 번째 분석 (second analysis)에서는 호르몬 치료군에서 30%의 사망률 감소를 보고하였고, 과거의 관찰연구들도 이와 유사한 결과를 보여주었다. WHI 연구결과의 두 번째 분석을 포함한 메타분석에서는 젊은 폐경여성에서의 호르몬 치료는 사망률을 감소시키는 것으로 나타났다. 비록 득과 실에 대한 이해에 있어 엄청난 동요가 있어 왔지만 적어도 젊은 폐경여성에서의 호르몬 치료는 심혈관질환의 이환율 및 사망률을 줄이고 나아가 전체 사망률을 줄일 수 있을 것으로 생각된다. There was controversy about decrease in mortality as the eventual result of hormone replacement therapy (HRT). The reduction of mortality was known in large observational-cohort studies in women with HRT compared with non-HRT after correction of confounding factors. The second analysis of Women`s Health Initiative (WHI) results revealed that the 30% decrease in mortality was observed in HRT users. These findings resemble with the results of past observational studies. The meta-analysis including the second analysis of WHI results showed that HRT use in young menopausal women decrease the mortality. Although there was a chaos about the risk and benefit of HRT since the first announcement of WHI study in 2002, HRT in young menopausal women could decrease the morbidity and mortality related to coronary heart disease and stroke. (J Korean Soc Menopause 2012;18: 133-138)

중증 자궁내막증 환자의 복강액이 단핵구의 HLA-DR 발현에 미치는 영향

나용진 ( Yong Jin Na ),박종훈 ( Jong Hoon Park ),곽종영 ( Jong Young Kwak ),이규섭 ( Kyu Sup Lee ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.12

목적: 자궁내막증은 자궁강 밖에 자궁내막 조직이 존재하는 질환으로 가임기 여성에서 흔한 질환이다. 발생기전으로는 월경혈의 역류, 자궁내막 세포의 이소성 착상, 유전학적 인자, 그리고 호르몬 이상 등이 제시되고 있으나 아직 명확하지 않다. 최근에는 자궁내막 세포의 이소성 착상 기전을 면역학적 내성과 관련하여 설명하고 있는 등, 자궁내막증의 발생에서 면역학적 기전의 중요성이 점차 강조되고 있다. 본 연구는 자궁내막증의 발생과 관련된 면역학적 인자에 관한 분석을 위하여 자궁내막증 환자의 복강액이 단핵구에서의 HLA-DR 발현에 미치는 영향을 알아보고자 하였다. 연구 방법: 가임기 여성으로 복강경 검사를 받은 환자들 중 자궁내막증 환자의 복강액 (n=20)과 대조군의 복강액 (n=10)을 본 수술 시작 전에 캐뉼러를 이용하여 채취하였다. 건강한 공여자로부터 얻은 말초혈액 단핵구는 분리배지 (Histopaque- 1077)를 이용하여 분리, 배양하였고, THP-1 세포는 RPMI 1640 배지를 이용하여 배양하였다. 단핵구에서의 HLA-DR의 발현은 동일초점 현미경으로 관찰하였고, IL-10, TGF-β와 VEGF의 농도는 효소결합면역흡착검사를 이용하여 측정하였다. 결과: 자궁내막증 환자의 복강액은 단핵구에서의 HLA-DR 발현을 감소시켰으며 (P<0.01), 첨가된 자궁내막증 환자의 복강액 농도가 증가할수록 THP-1 세포의 HLA-DR 발현이 용량 의존성으로 감소하였다. 또한 자궁내막증 환자의 복강액 내 IL-10 농도와 단핵구의 HLA-DR 발현은 역상관관계를 보였다 (r=-0.518, P=0.019). 결론: 단핵구의 HLA-DR 발현은 자궁내막증 환자의 복강액에 의해 IL-10과 연관되어 억제되며, 이는 자궁내막증 환자의 복강액 내 IL-10이 단핵구의 HLA-DR의 발현을 조절하는 중요한 요소(들) 중 하나라는 것을 의미한다. Objective: Endometriosis is a common gynecologic disorder characterized by the presence of endometrial tissue outside the uterine cavity. Many theories have been suggested to explain the mechanism for the development of this disease. Although no single theory can explain all cases of endometriosis, retrograde menstruation theory has gained the widest acceptance. But the discrepancy between the incidence of retrograde menstruation and prevalence of endometriosis suggests that additional factor(s) may determine susceptibility to endometriosis in the peritoneal cavity and the survival and implantation of endometrial cells seem to be related with immunologic factors. For evaluation of the immunologic factors, we investigated the effects of peritoneal fluid from patients with advanced endometriosis (EPF) on the HLA- DR expression of monocytes. Methods: EPF (n=20) and peritoneal fluid from the control group (CPF) (n=10) were collected prior to main laparoscopic operation. CPF were obtained from fertile women with various gynecologic problem other than endometriosis, such as dermoid cyst and carcinoma in situ of the uterine cervix. Peripheral blood mononuclear cells were collected by density-gradient centrifugation of whole blood over a peripheral blood mononuclear cell separation medium (Histopaque-1077), and THP-1 cells derived from a monocyte/macrophage cell line (Korean Cell Line Bank, Seoul, Korea) were cultured in RPMI-1640 medium. The expression of HLA-DR was evaluated by confocal microscopy, and the levels of IL-10, TGF-β and VEGF were measured by standard enzyme-linked immunosorbent assay kits (R and D system). Results: Compared to CPF, the addition of 10% EPF (n=10) to culture medium significantly reduced the percentage of HLA-DR positive cells in cultures of a THP-1, monocytic cell line at 48 hours. The effect of EPF was dose-dependent, and similar effect was observed in the peripheral blood monocytes. An inverse correlation was found between the HLA-DR expression level and IL-10 concentration in EPF (r=-0.518; P=0.019) and in the supernatant of peripheral blood monocyte cultured in EPF (n=20) (r=-0.459; P=0.042). IL-neutralizing antibody significantly abrogated the effect of EPF on HLA-DR expression level. Conclusion: These results suggest that HLA-DR expression level on monocytes is down-regulated by EPF, and that IL-10 may be one of the factors in EPF to modulate HLA-DR expression level.

Effects of peritoneal fluid from endometriosis patients on the release of vascular endothelial growth factor by neutrophils and monocytes

나용진 ( Yong Jin Na ),( Seung Hong Yang ),( Dae Won Baek ),( Dong Hyung Lee ),( Ki Hyung Kim ),( Young Min Choi ),( Sung Tack Oh ),( Young Seoub Hong ),( Jong Young Kwak ),( Kyn Sup Lee ) 대한산부인과학회 2007 대한산부인과학회 학술대회 Vol.93 No.-

cDNA 미세배열법과 조직배열법을 이용한 자궁경부암의 유전자 분석: MTA1을 중심으로

최진국 ( Jin Kook Choi ),장성규 ( Sung Kyu Jang ),이동형 ( Dong Hyung Lee ),김기형 ( Ki Hyung Kim ),나용진 ( Yong Jin Na ),최경운 ( Kyung Un Choi ),이창훈 ( Chang Hoon Lee ),윤만수 ( Man Soo Yoon ) 대한산부인과학회 2005 Obstetrics & Gynecology Science Vol.48 No.11

체외수정시술의 결과를 예측할 수 있는 인자로서 난구세포의 활용에 관한 연구

김광대,김기형,나용진,이규섭,Kim, Kwang-Dae,Kim, Ki-Hyung,Na, Yong-Jin,Lee, Kyu-Sup 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.3

Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.

최신임상강좌 : 다낭성난소증후군 환자의 불임치료

구윤희 ( Yun Hee Koo ),나용진 ( Yong Jin Na ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.10

다낭성난소증후군 환자에서 불임에 대한 적절한 치료방법은 아직까지 명확히 정립되어 있지 않다. 가장 중요하게 고려할 사항은 치료를 시작하기 전 충분한 상담을 통해 다른 건강상의 문제점을 찾고, 비만한 여성은 운동 및 식이요법을 통해 체중을 감량해야 한다는 점이다. 다낭성난소증후군 환자에서 배란 유도를 위한 일차 치료제는 클로미펜이며 클로미펜에 반응이 없거나 임신에 실패한 경우 생식샘자극호르몬 투여를 고려할 수 있다. 생식샘자극호르몬을 투여할 경우 다태임신의 위험이 있으므로 난소의 반응을 초음파 검사를 통해 철저히 관찰하도록 하여야 한다. 복강경하 난소수술은 임신율 및 생존아 출생율에 있어서 생식샘자극호르몬과 비슷한 효과를 보이므로 이차적 치료로 고려할 수 있으나 수술적 위험 및 추가적인 배란유도제 사용의 필요성 때문에 배란유도의 목적만으로 시행하기에는 적절하지 않다. 체외수정은 불임 치료의 마지막 단계로 고려되며 단 한 개의 배아만 이식을 하는 방법으로 다태아 임신의 위험을 줄일 수 있다는 점에서 다낭성난소증후군 여성에서 효과적인 치료방법이 될 수 있다. Metformin과 같은 인슐린반응 개선제는 배란유도를 위한 일차 치료제 또는 클로미펜과 병합치료 시 역할에 있어 논란의 여지가 있다. 방향화효소 억제제는 배란 유도에 있어 클로미펜만큼 효과적이나 아직까지 불임치료제로서 승인이 되지 않은 상태로 향후 연구를 통해 효과 및 장기적 안정성이 입증되어야 할 것이다. The treatment of infertile women with polycystic ovary syndrome (PCOS) has not yet been established. Before any intervention is initiated, lifestyle modification with diet and exercise leading to weight loss should be emphasized in overweight women. The recommended first-line treatment for ovulation induction remains clomiphene citrate. If clomiphene citrate use fails to result in ovulation or pregnancy, the recommended second-line intervention is exogenous gonadotropins. The use of exogenous gonadotropins is related to an increased risk for multiple pregnancies, and, therefore, intense ovarian response monitoring is required. Laparoscopic ovarian surgery is an alternative to gonadotropin therapy because this surgery is as effective as gonadotropin in pregnancy rate or live birth rate. However, the surgery may require additional ovulation induction drug and should be performed by well-trained surgeon, its use solely to induce ovulation is unwarranted. Recommended third-line treatment is in vitro fertilization (IVF). IVF reduces the risk of multiple pregnancies by using single-embryo transfer in women with PCOS. The use of metformin alone as first-line infertility therapy or in combination with clomiphene is surrounded by controversies. Aromatase inhibitors appear to be as effective as clomiphene citrate for induction of ovulation, but the drug is currently not approved for treatment of infertility. Further studies should demonstrate the efficacy and long term safety.

Nitric oxide 의 자궁내막 세포 증식 억제효과에 관한 연구

김기형(Ki Hyung Kim),나용진(Yong Jin Na),이규섭(Kyu Sup Lee),김원희(Won Whe Kim) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.8

N/A Objective : Development of endometriosis is associated with endometrial stromal cell proliferation. The effects of nitric oxide (NO) on the proliferation of endometrial cells and the effects of peritoneal fluid from women with and without endometriosis on the production of NO and NO-induced changes were investigated. Methods : Endometrial stromal cells were prepared from women with endometriosis (n=10) and healthy control (n=10). Peritoneal fluid from 10 patients were collected at the beginning of laparoscopic procedures. The proliferation of cells was assessed by [3H]thymidine. Nitrite, a stable NO product, was measured with Griess reagents. Expression level of endothelial nitric oxide synthase (eNOS) was analyzed by quantitative RT-PCR. Results : Sodium nitroprusside, an NO donor, reduced proliferation of the endometrial and endometriosis cells from primary culture in a dose-dependent manner. The increased production of NO and inhibitory effect of NO on the endometrial stromal cell proliferation were reduced by peritoneal fluid from patients with endometriosis but not by interleukin-8, 17-β estradiol or TGF-β1. Transcripts of the endothelial NOS (eNOS) gene were detected in unstimulated endometrial cells and the expression patterns were not changed by exposure of cells to SNP or peritoneal fluid for 24 hours. Conclusion : It is postulated that these findings show NO inhibits proliferation of endometrial cells and the inhibitory effect of NO on endometrial cells was abrogated by certain factor (s) of peritoneal fluid from patients with endometriosis. In conclusion, NO appears to be involved in the pathogenesis of endometriosis.

생쥐 내세포괴의 분리방법과 지지세포의 종류와 Mitomycin C 처리 시간이 내세포괴 Colony 형성률에 미치는 영향

장호진,고경래,김미경,나용진,이규섭,Jang, Ho-Jin,Ko, Kyung-Rae,Kim, Mi-Kyung,Na, Yong-Jin,Lee, Kyu-Sup 대한생식의학회 2006 Clinical and Experimental Reproductive Medicine Vol.33 No.4

목 적: 본 연구는 생쥐 포배기 배아로부터 내세포괴를 분리하는 방법과 지지세포의 종류와 mitomycin C 처리 시간이 내세포괴 colony 형성률에 미치는 영향을 관찰하기 위해 시행되었다. 연구방법: 일반적인 면역절제술, 주사바늘을 이용한 부분 영양막세포 절개법, 포배기 배아 공배양법으로 내세포괴를 분리한 후, 상업적으로 구입이 가능한 STO 또는 직접 제조한 생쥐 배아섬유아세포 (pMEF)를 지지세포로 이용하여 배양하였다. 또한, mitomycin C를 1, 2, 3시간 동안 처리한 각각의 지지세포에서 7일 동안 배양한 후, 내세포괴 colony 형성률을 살펴보았다. 결 과: STO 지지세포에서는 부분 영양막세포 절개법을 사용한 경우 (52%)가 면역절제술 (12%)이나 포배기 배아 공배양법 (16%)을 사용한 경우보다 내세포괴 colony 형성률이 유의하게 높았다 (p<0.05). pMEF 지지세포에서의 형성률은 부분 영양막세포 절개법을 사용한 경우 (88%)와 포배기 배아 공배양법 (82%)을 사용한 경우가 면역절제술 (16%)을 사용한 경우보다 높았다 (p<0.05). STO와 pMEF 모두에서, 2시간 mitomycin C 처리군 (52%, 88%)이 1시간 처리군 (9%, 42%)과 3시간 처리군 (18%, 76%)보다 높은 내세포괴 colony 형성률을 보여주었다 (p<0.05). 결 론: 이상의 결과는 부분 영양막세포 절개법이 생쥐 포배기 배아로부터 내세포괴를 분리하는 가장 효과적인 방법이며, 가장 적절한 mitomycin C 처리 시간은 2시간이라는 것을 보여준다. 그러나 이와 같은 부분 영양막세포 절개법의 효용성을 보다 명확하게 확인하기 위해서는 분리한 내세포괴를 계대배양하여 줄기세포주로서의 특성을 확인하는 실험이 추가적으로 필요할 것으로 생각된다. Objective: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony. Methods: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days. Result: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions. Conclusion: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.

유전자결핍 생쥐를 이용한 aquaporin-3의 태반 내 분포 및 기능 연구

김하정 ( Ha Jeong Kim ),나용진 ( Yong Jin Na ),김기형 ( Ki Hyung Kim ),이동형 ( Dong Hyung Lee ),김휘곤 ( Hwi Gon Kim ),조무성 ( Mu Seong Jo ),정준양 ( Jun Yang Jung ),전미련 ( Mei Lian Quan ),배혜란 ( Hae Rahn Bae ),최욱환 ( Ook 대한산부인과학회 2009 Obstetrics & Gynecology Science Vol.52 No.8

목적: Aquaporin (AQP) 3은 세포막을 통한 선택적인 수분과 글리세롤 이동을 촉진시키는 수분통로 단백질이다. 본 연구는 AQP3 유전자결핍 생쥐를 이용하여 태반에서 AQP3을 통한 수분 및 글리세롤 이동이 양수 및 태아 성장에 미치는 영향을 조사하고자 하였다. 연구 방법: 재태 14.5일된 야생형 및 AQP3 유전결핍생쥐 각각 30마리에서 임신낭을 적출하여 양수량을 비교 분석하였다. AQP3 발현 및 분포는 면역조직화학법과 western blot을 이용하여 조사하였다. 결과: 재태 14.5일된 생쥐 태반에서 AQP3는 태아막의 난황막세포의 기저측면막과 미로 태반의 융합영양막세포에 가장 높게 발현되었다. 반면 AQP1은 태아막의 난황막세포의 관강막과 미로 태반의 혈관조직에 고농도로 발현되었다. AQP3 유전자결핍 생쥐의 재태 14.5일이 태반은 야생형 CD1 생쥐와 비교할 때 태반 형성 및 태반 세포의 분화에 있어서 유의한 차이는 관찰되지 않았다. 그러나 AQP3 유전자결핍 생쥐는 야생형 CD1 생쥐에 비해 태아 단위 무게 당 양수량은 증가되었으며, 양수의 삼투질 농도는 감소되었고, 이로 인해 양수 내 이온과 다른 용질의 농도가 낮았다. 또한 AQP3 유전자결핍 생쥐 태아 평균 체중은 야생형 생쥐의 70% 수준으로 감소되어 있었다. 결론: AQP3는 모체 혈액 및 난황과 접하고 있는 태반과 태아막에 풍부하게 발현되어 이를 통한 수분과 글리세롤의 이동을 촉진시킴으로써 양수 조절 및 태아 성장에 중요한 역할을 담당하고 있을 것으로 추정된다. Objective: Aquaporin (AQP) 3 is a small integral membrane protein that functions as a facilitated transporter of water and glycerol. To elucidate a role of AQP3 in placenta, changes in amniotic fluid composition and fetal growth were investigated using AQP3 null mice. Methods: Embryonic day 14,5 gestational sacs of wild-type and AQP3 kncok-out pregnant mice, thirty each, were used for this study. AQP3 localization and expression were assessed by immunohistochemistry and western blot. Results: AQP3 was highly expressed in basolateral membrane of visceral yolk sac cells of fetal membrane and syncytiotrophoblast cells of labyrinthine placenta. In contrast, AQP1 was expressed in apical membrane of visceral yolk sac cells and endothelial cells lining vasculature. There was no significant difference in normal placentation and differentiation from trophoblast stem cells between wild type and AQP3 null mice. However, AQP3 null mice had increased amount of amniotic fluid per gram of body weight and decreased osmorality of amniotic fluid with low concentrations of ions and solutes in amniotic fluid. In addition, AQP3 null mice pups were smaller than CD1 wild type mice. Conclusion: AQP3 plays an important role in amniotic water balance and nutrient supply to developing fetus by facilitating transplacental transport of water and glycerol.