난소관련병변에서 복강체액내의 cytokine 농도

전상식(Sang Sik Chun),조영래(Young Lae Cho),구태본(Tae Bon Koo),서장수(Jang Soo Suh) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.2

목적 : 최근에 난소암을 비롯한 종양성 질환과 자궁내막증등의 부인과 내분비 질환에서 면역반응의 활성에 관여하는 cytokine에 대한 연구가 많이 되고 있어서 저자들은 난소관련병변들의 병인분석과 감별진단의 가능성 및 치료적 응용에 있어서 복강체액내에 cytokine의 이용가능성을 조사하여 보았다. 연구방법 : 조직학적으로 진단이된 양성난소낭선종 11예, 성숙낭성기형종 10예, 자궁내막증 23예, 그리고 난소암 11예를 대상으로 하여 복강내 체액을 채취하여 IFN-γ, IL-1β, IL-5, IL-6, IL-10 및 TNF-α의 농도를 측정하였으며 통계처리는 일원분산분석법과 Scheffe 법 그리고 Mann-Whitney 법으로 하였다. 결과 : 난소암 환자에서 IL-6와 IL-10의 농도는 다른 양성병변들에 비해서 의미있게 증가하였으며(P<0.05) 자궁내막증 환자에서 대조군에 비하여 IL-6와 IL-10의 농도가 높았으며 특히 자궁내막증 병기Ⅲ과 IV인 경우에서 병기Ⅰ과 Ⅱ에 비교하여 IL-6와 IL-10의 농도가 의미있게 증가되었다. 결론 : 복수내의 cytokine 환경은 난소관련병변의 병인과 질병의 진행과정과 관계가 있을 것으로 사료되며 복강내체액의 cytokine 농도 측정은 난소암과 양성병변과의 감별진단 및 자궁내막증 환자의 중증도의 진단과 병인규명에 도움을 줄 수 있을것으로 생각된다. Objective : Recent studies have shown that multifunctional cytokines have been implicated in the pathogenesis and pathophysiology of various ovarian lesions including ovarian cancer and endometriosis. This study was performed to determine the possibility of the clinical usefulness of peritoneal cytokines in patients with various ovarian lesions. Methods : Peritoneal fluid was obtained from patients with benign cystic adenoma(n=11), benign cystic teratoma(n=10), endometriosis(n=23), malignant ovarian tumor(n=11), and women without evidence of any pathology(n=7) at the time of laparotomy or operative laparoscopic surgery and were examined for the levels of IFN-γ, IL-1β, IL-5, IL-6, IL-10 and TNF-α using enzyme-linked immunosorbent assay(ELISA). Statistical analysis was performed with the one way ANOVA, Scheffe test or Mann-Whitney test. Results : The mean concentration of IL-6 and IL-10 was significantly higher in peritoneal fluid specimens with ovarian cancer than other benign pathology(p<0.05). Both IL-6 and IL-10 levels in peritoneal fluid specimens with endometriosis tended to be higher than normal, and the levels of IL-6 and IL-10 were significantly higher in peritoneal fluid of women with severe endometriosis compaired to women with mild endometriosis (p<0.05). Conclusion : This study supports the concept that regional immunologic dysfunction might be involved in the disease process of various ovarian pathology such as ovarian cancer and endometriosis. A larger study would help in evaluating the potential use of local peritoneal cytokines in differentiating ovarian cancer from other benign pathology, and demonstrating any association between concentrations of cytokines and severity of endometriosis.

불임환자에서 sonohysterography의 이용

이춘희(Chun Hee Lee),김광수(Gwang Soo Kim),이동영(Dong Young Lee),박철민(Chul Min Park),오영은(Young Eun Oh),이택후(Taek Hoo Lee),전상식(Sang Sik Chun) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.9

목적: 이 연구는 불임환자의 자궁강내의 병변을 조사하는 데 있어서 기존에 이용되던 질식초음파, 자궁난관조영술 및 자궁경의 경우와 비교하여 새로이 시도한 sonohysterography의 유용성을 알아보고자 하였다. 연구방법: 불임을 주소로 본원 산부인과를 방문한 33명의 환자를 대상으로 병력조사 및 질식초음파를 시행하여 불임의 원인으로 예측되어지는 자궁강내의 병변, 즉 자궁내막용종, 점막하근종 또는 자궁내유착증이 의심되는 환자에서 sonohysterography, 자궁난관조영술 및 자궁경을 시행하였다. 자궁경의 진단을 최종의 근거로 각 검사의 각 진단에 대한 민감도, 특이도, 양성예측치 및 음성예측치를 구하여 비교하였다. 결과: 질식초음파와 자궁난관조영술의 경우 각각의 민감도는 41.7%와 83.3%였으며 sonohysterography의 경우는 100%였다. 각 병변별로 비교하여도 sonohysterography의 민감도가 높게 나타났다. 결론: Sonohysterography가 불임환자 뿐만 아니라 일반 부인과 환자의 자궁강내의 병변을 찾아내는 데 있어서 기존의 다른 진단방법을 대체할 수 있는 우수한 진단방법이라고 사료된다. Objective: The purpose of this study was to assess the usefulness of sonohysterography in the detection of abnormalities of the uterine cavity in infertile patients, compared with other diagnostic methods, such as transvaginal sonography, hysterosalpingography and hysteroscopy. Methods: Three intrauterine pathologies including intrauterine synechiae, endometrial polyp and submucosal myoma, that could be possible causes of infertility were diagnosed. Results: Transvaginal ultrasonography and hysterosalpingography were able to detect 41.7% and 83.3% of uterine pathologies respectively comparing with sonohysterography to detect all of the uterine pathologies. Conclusion: Our results show that sonohysterography is easy, inexpensive, well-tolerated and non-invasive technique with high sensitivity in the detection of uterine pathologies. Therefore the use of sonohysterography for the diagnosis of intrauterine pathologies in infertile patients is highly recommended.

자궁내유착의 자궁경 치료와 그 결과

주응식(Eung Sik Ju),김일규(Il Gyu Kim),오영은(Young Eun Oh),이현정(Hyun Jung Lee),이택후(Taek Hu Lee),전상식(Sang Sik Chun) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.2

본원 산부인과에서 최근 10년간 각종 질환으로 수술적 자궁경술을 시행받은 163명을 분석하여 이들 중에서 불임을 주소로하는 자궁내 유착 환자 98명을 대상으로 수술 후의 reproductive outcome을 조사 분석하였다. 자궁내 유착의 정도는 미국불임학회의 기준을 따라 분류하였고 분류에 의하면 자궁내 유착 환자 98명은 각각 mild 29.6%, moderate 43.9%, severe 26.5%의 빈도를 보였다. 자궁내 유착 경우 선행 원인으로는 인공 혹은 자연 임신중절이 대부분을 차지하고 그외에 제왕절개술, 포도상 기태, 골반결핵 등이 있었으나 원인을 정확히 밝힐 수 없는 경우도 일부 있었다. 유착제거술 후 임신율은 전체로는 53.3%, mild, moderate, severe의 경우 각각 84.6%, 45.0%, 33.3%의 임신율을 보였다. 환자에 따라서는 자궁내막증이나 난관폐쇄 등 다른 불임인자를 포함하는 경우도 있어 그 결과의 분석이 어렵고 또한 많은 환자가 추적에서 이탈되어 45.9%(45/98명)만이 추적 조사가 가능하였다. 추적기간은 1.0년 이상을 대상으로 하였고 최장 8.8년간 추적이 가능하였다. 합병증으로는 자궁천공 5례, 호흡곤란 내지 폐부종 2례, 출혈과 방광손상 각각 1례 등으로 나타났다. One hundred sixty three patients were hysteroscoped for various reasons at our department over 10 year period. Ninety eight patients out of 163 who were diagnosed and treated for their intrauterine adhesions(IUAs) by hysteroscopy were analysed retrospectively. Patients were classified as having mild, moderate or severe stage of IUAs according to American Fertility Society classification. Twenty nine patients(29.6%) had mild IUAs, 43 patients (43.9%) had moderate IUAs, and 26 patients(26.5%) were classified as having severe stage IUAs. The predisposing causal factors of IUAs were abortion, full-term delivery, H. mole, pelvic tuberculosis and myomectomy in order of frequency but in eight cases no cause was identified. After hysteroscopic surgery, although high rate(83.9%) of restoration of normal menstruation was attained but term pregnancy rate was only 53.3% and rather high rate (32%) of preterm or abortion were identified and those pregnancy outcomes were correlated with the stage of IUAs. Complications of surgery consisted of uterine or bladder perforation, pulmonary edema and hemorrhage.

마우스에서 배반포 형성, 세포 수 및 ICM의 비율에 미치는 배양액의 효과

박기상,이택후,전상식,송해범,Park, Kee-Sang,Lee, Taek-Hoo,Chun, Sang-Sik,Song, Hai-Bum 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.4

Objective : The aim of this study was to evaluate the influence of different media on blastulation, mean cell number, percentage of inner cell mass (ICM) of total cells and ICM : trophectoderm (TE) ratio in mice. Materials and methods: A total 552 two cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection (mated just after hCG injection) and cultured in MEM (n=276) or TCM (n=276) supplemented with 20% hFF. The grading of blastocysts from zona-intact (ZiB) to -escape (hatching and hatched, ZeB) was performed at 72 hours after culture. Total, TE and ICM cell numbers were analyzed by differential staining of blastocyst. Statistical analysis was performed using t-test with SigmaPlot-2001. P-values < 0.05 were accepted as statistically significant. Results: The blastulation rate in MEM ($64.9{\pm}4.95%$) was significantly higher (p=0.0031) than that in TCM ($57.2{\pm}5.22%$). No differences were found in the number of ZiB and ZeB between MEM ($31.9{\pm}2.62$, $33.0{\pm}4.58%$), and TCM ($27.2{\pm}4.28$, $30.1{\pm}4.58%$). A total 314 blastocysts (MEM=166; TCM=148) were stained differentially. Mean cell number of blastocysts was significantly higher (p=0.0002) in TCM ($73.1{\pm}3.3$) than in MEM ($61.7{\pm}2.5$). Differential staining was successfully performed in 155 blastocysts (MEM=77; TCM=78). The percentage of ICM was significantly higher in MEM than in TCM ($20.9{\pm}1.3$ vs. $17.1{\pm}1.2%$, p=0.0281). The ICM : TE ratio was higher in TCM than in MEM (1 : $4.85{\pm}0.68$ vs. 1 : $3.78{\pm}0.78$, NS). Conclusion: These results show that MEM increase the blastocyst formation and percentage of ICM of total cells comparing with TCM in mice.

The Clinical Outcomes after Embryo Transfer (ET) on Day 2 and Day 5 or Subsequent ET on Day 2-5, 2-6, 2-7, 3-5 and 4-7 in In Vitro Fertilization-ET Cycles

김대원,박기상,송해범,이택후,전상식,Kim, Dae-Won,Park, Kee-Sang,Song, Hai-Bum,Lee, Taek-Hoo,Chun, Sang-Sik The Korean Society for Reproductive Medicine 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.1

연구목적: 일반적으로 IVF-ET에서 가장 높은 임신율을 얻는 방법은 5 day ET (배반포기 배아 이식)이지만 장기간 배양이 적절하지 못한 경우에는 $2{\sim}4$일째에 ET를 실시하고 나서 $5{\sim}7$일째에 배반포기에 도달한 배아를 재이식 (SET)하여, SET의 효용성에 대하여 조사하고자 실시하였다. 연구재료 및 방법: 48주기의 환자에서 회수한 난자와 수정란은 10%와 20% hFF가 첨가한 DMEM에서 이식 직전까지 각각 공배양하였다. 채란 2일 (group I, day 2 ET), 5일째 이식 (group II, day 5 ET) 또는 재이식 (group III, SET; 2-5, 2-6, 2-7, 3-5, 4-7일)을 실시하면서 수정률, 할구분할률 및 임신율을 각각 비교하였다. 결과에 대한 통계 분석을 SAS (version 6.2)를 이용한 Duncan's Multiple Range Test를 이용하여 p값이 0.05 보다 작을 때 통계적으로 유의차가 있는 것으로 하였다. 결 과: 수정률은 group II (90.5%)가 다른 군에 비하여 높게 (p<0.05) 나타났다 (group I: 80.6%; group III: 82.9%). 할구분할률은 군간에 차이가 없었다 (수정란 당 $93.3{\sim}99.1%$). 임상적 임신율은 group II와 III (각각 58.3%)가 group I (33.3%) 보다 높게 나타났다. 그러나 처리군이 적어서 통계적인 차이는 없었다. 결 론: 배반포기 배아를 단독 이식하는 것이 임신율을 높일 수 있는 최선의 방법으로 나타났지만, 채란수가 적거나 수정률이 저조한 경우에는 $2{\sim}4$일째에 ET를 실시한 후 여분의 배아를 배반포기까지 배양한 다음 $5{\sim}7$일에 재이식 (SET)하면 blastocyst ET에서 나타날 수 있는 이식 자체의 실패를 방지할 수 있으면서 임신율을 높일 수 있는 이식 기법이 될 것이다.

생쥐 난소에서 Preantral Follice의 단순 분리법

김주환,박기상,송해범,전상식,Kim, Ju-Hwan,Park, Kee-Sang,Song, Hai-Bum,Chun, Sang-Sik 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3

Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1) enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at $37^{\circ}C$ and 5% $CO_2$ incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 lU/ml and DNAse 20 lU/ml. After 20 min., follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when ${\rho}$ value was less than 0.05. Results: In handling time, mincing or scraping method ($28{\pm}3.42$ min or $16{\pm}1.58$ min) were significantly (p<0.00001) shorter than enzymatical method ($72{\pm}1.69$ min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method ($49.8{\pm}3.91$) than in mincing or scraping method ($25.3{\pm}2.33$ or $20.5{\pm}1.75$). Isolated follicles in ${\leq}$90${\mu}m$ were significantly (p<0.005) higher in enzymatical method ($15{\pm}1.71$) than in mincing or scraping method ($7.8{\pm}0.98$ or $8.1{\pm}1.31$). In 91~130 ${\mu}m$, isolated follicles were significantly (p<0.0005) higher in enzymatical method ($33{\pm}3.27$) than in mincing or scraping method ($16.3{\pm}1.82$ or $10.7{\pm}1.38$). In ${\geq}$ 131 ${\mu}m$, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ${\leq}$ 90 ${\mu}m$ was highest in scraping method (39.6% vs. enzymatical method: 30.1%, p<0.05; mincing method: 30.9%, p=0.11719, NS). Rate of follicles in $91{\sim}130$ ${\mu}m$ was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ${\geq}$131 ${\mu}m$ was highest in scraping method (8.3% vs. enzymatical or scraping method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handling time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91 ~ 130 ${\mu}m$ was highest in all methods.

대구.경북 지역 한국인 다낭성 난소 증후군 환자의 임상 양상

배진영,김미연,성수경,오정은,전상식,이택후,Bae, Jin-Young,Kim, Mi-Yeon,Sung, Su-Kyung,Oh, Jeong-Eun,Chun, Sang-Sik,Lee, Taek-Hoo 대한생식의학회 2009 Clinical and Experimental Reproductive Medicine Vol.36 No.1

목 적: 다낭성 난소 증후군은 생식기 여성에서 나타나는 가장 흔한 내분비 질환 중 하나이며 다양한 임상 양상을 보이고 있다. 본 연구에서는 한국인 다낭성 난소 증후군 환자 특유의 임상 양상 및 혈액학적 소견을 밝히고자 하였으며 그 결과를 터키와 미국의 다낭성 난소 증후군 환자와 비교 분석하였다. 연구방법: 2000년 1월부터 2008년 4월까지 경북대학교 병원 불임치료실을 찾은 88명의 다낭성 난소 증후군 환자를 대상으로 하여 후향적으로 임상 양상 및 혈액학적 소견을 연구하였다. 결 과: 한국인 다낭성 난소 증후군 환자군 내에서는 통계학적으로 유의한 연령에 따른 임상 양상의 차이가 없었으나 터키와 미국의 다낭성 난소 증후군 환자와 비교했을 때 고안드로겐혈증의 빈도는 유의하게 낮았다. 결 론: 다낭성 난소 증후군 환자 내에서도 인종간의 임상 양상 및 혈액학 소견의 차이가 있다고 생각되며 유전 및 생활 습관의 차이 등이 그 원인을 설명할 수 있을 것이다. 그러나 연구 대상인 환자의 수가 적고 연령군의 분포가 좁아 향후 좀 더 큰 규모의 연구가 필요하다. Objective: PCOS is one of the most common endocrine disorder affecting reproductive age women. It shows wide range of reproductive and metabolic manifestations. This study was to determine the differences of clinical and laboratory characteristics between adolescent and adult Korean Daegu Kyungpook PCOS patients. We also compared clinical and laboratory values of Korean PCOS patients to Turkish and American PCOS patients. Methods: 88 cases were diagnosed and enrolled as PCOS patients, based on 2003 Rotterdam criteria, who visited KNUH Reproductive Endocrinology division between Jan. 2000 and Apr. 2008. We retrospectively reviewed clinical characteristics, serum hormone levels, ultrasono-graphic findings. And we divided into 2 groups, adolescent and adult. We analyzed them with Chi-square test and Student's t-test statistically. Results: There were no statistically significant differences between adult and adolescent Korean PCOS patients. The prevalence of hyperandrogenism such as facial acne or hirsutism, obesity, and insulin resistance of Korean PCOS women was significantly lower than Turkish and American PCOS women. Conclusion: From the results of this study, we can suggest that it is sure that there are differences of clinical and laboratory characteristics of PCOS patients between races. Genetic background, dietary habits and life style affect the expression of clinical symptoms of PCOS. But we could not find out the difference between adolescent and adult PCOS patients of same race. It might be due to the limitation of small patient number and narrow range of age distribution. So, there must be large scale and multi-center and multi-regional study.

인간양수에 의한 생쥐 난자 투명대의 정자수용능력 억제의 관찰

박기상,이택후,송해범,전상식,Park, Kee-Sang,Lee, Taek-Hoo,Song, Hai-Bum,Chun, Sang-Sik 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.1

Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

Glutamine 함유 배양액에 첨가한 에너지원이 마우스의 배 발달에 미치는 영향

김주환,박기상,이택후,전상식,송해범,Kim, Ju-Hwan,Park, Kee-Sang,Lee, Taek-Hoo,Chun, Sang-Sik,Song, Hai-Bum 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.1

Objective: Mammalian embryos undergo changes of energy environment for transfer from oviduct to uterus. Also, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration (oviduct - glucose: 0.5 mM, pyruvate: 0.32 mM, lactate: 10.5 mM; uterus - goucose: 3.15 mM, pyruvate: 0.1mM, lactate: 5.87 mM, respectively). This study was conducted to examine the effect of these energy sources added in DMEM with glutamine on the mouse embryo development. Methods: There was used ICR female mouse. Two cell embryos of mouse are collected by method of 'flushing'. Flushing fluid was used Ham's F-10 added to 20% FBS. The collected 2 cell embryos were cultured in media such as Control (only DMEM), group A and B (DMEM supplemented with 0.5 mM and 3.15 mM glucose), and group C and D (DMEM supplemented with 0.1 mM and 0.32 mM pyruvate), and group E and F (DMEM supplemented with 5.87 mM and 10.5 mM lactate). All experimental media supplemented with 20% hFF, respectively. Pattern of embryo development was observed to interval at 24hr during 96hr. Results : The media with glutamine added glucose (group A: 51.0%; group B: 48.4%) was significantly (p<0.05) higher than other experimental group in development into the morula stage after 24 hr in culture, but not significantly different compared with control and the rate of development into the blastocyst was significantly (p<0.05) low in the both of pyruvate (group C: 7.9% group D: 6.8%) and lactate (group E: 7.1%, group F: 7.1%) treatment group after 48 hr in culture. Development into the blastocyst and hatched balstocyst after 72 hr in culture revealed similarly in control (81.9%) and glucose treatment group (group A: 83.3%, group B: 82.8%). However, development into the hatched and attached blastocyst after 96hr in culture revealed significantly (p<0.05) development in the glucose treatment group (group A: 82.3%, group B: 78.5%) than control (63.2%), and its of pyruvate (group C: 34.1%, group D: 34.1%) and lactate (group E: 25.9%, group F: 33.3%) treatment group were significantly (p<0.05) lower than control similar to previous observations. Conclusion : The glucose added to the DMEM with only glutamine, as energy source, was highly to the rate of development compared with control, but the other energy sources were not, synthetically. Above refer to, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration. Thus, further studies are will examine continuously to effects by interaction of different energy sources in the mouse embryo development, and these results will provide to foundation on the human embryo culture.

자궁내막증 환자와 대조군에서의 자궁내막 유전자 발현의 차이: Microarray를 이용한 연구

정민지,정은정,이신제,김문규,전상식,이택후,Chung, Min-Ji,Chung, Eun-Jung,Lee, Shin-Je,Kim, Moon-Kyu,Chun, Sang-Sik,Lee, Taek-Hoo 대한생식의학회 2007 Clinical and Experimental Reproductive Medicine Vol.34 No.1

목 적: 자궁내막증은 자궁내부에 존재하여야 할 자금내막조직이 자궁 외에 존재하는 질환으로 그 발생기전은 아직 명확하게 밝혀져 있지 않다. 이에 저자들은 자궁내막증 환자와 정상 대조군의 자궁내막조직 간의 유전자 발현의 차이가 자궁내막증의 발병과 관련이 있을 것이라는 가정 하에 DNA microarray 기술을 도입하여 연구를 시행하였다. 연구방법: 2002년 1월부터 2002년 12월까지의 기간 동안 본원 산부인과에서 자궁내막증 환자와 자궁내막증 이외의 다른 부인과적 질환으로 수술을 시행한 환자들을 대상으로 채취한 자궁내막 조직으로 KNU 4.8K cDNA chip을 이용하여 유전자 발현을 비교 연구하였다. 유전자칩으로 자궁내막증 조직에서 발현의 증감을 보였던 유전자 중에서 8종의 유전자를 대상으르 RT-PCR이나 real time RT-PCR 법을 통하여 그 발현 양상을 검증하였다. 결 과: 자궁내막증에 이환된 여성의 자궁내막조직에서 대조군에 비하여 높게 발현되고 있는 것으로 나타난 유전자들은 ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ trarsporting (ATP5C1), LPS induced TNF-$\alpha$ factor 등으로 세포의 에너지 생성과 대사과정 및 신호전달에 관여하는 유전자들이었다. 한편 자궁내막중 환자의 자궁내막조직에서 대조군에 비하여 낮게 발현된 유전자들은 insulin like growth factor II associated protein, EGF-containing fibulin-like EMP1, matrix Gla protein, TGF beta-induced, TGF beta receptor 1(activin A receptor type II-like kinase), cystallin alpha B, fibulin 5, tissue inhibitor of metalloproteinase 3, collage type XII, alpha 1, tissue inhibitor of metalloproteinase 1, decorin 등으로 세포외기질의 구성 및 기능에 관련이 있었다. 결 론: 이상의 DNA mirroarry 및 RT-PCR을 통해 얻어진 결과에서 자궁내막증의 자궁내막조직에서 대조군에 비하여 유전자들의 발현에 차이가 있음을 확인하였다. Objective: Pathogenesis of the endometriosis is very complex and the etiology is still unclear. Our hypothesis is that there may be some difference in gene expression patterns between eutopic endometriums with or without endometriosis. In this study, we analyzed the difference of gene expression profile with cDNA microarray. Methods: Endometrial tissues were gathered from patients with endometriosis or other benign gynecologic diseases. cDNA microarray technique was applied to screen the different gene expression profiles from early and late secretory phase endometria of those two groups. Each three mRNA samples isolated from early and late secretory phase of endometrial tissues of control were pooled and used as master controls and labeled with Cy3-dUTP. Then the differences of gene expression pattern were screened by comparing eutopic endometria with endometriosis, which were labeled with Cy5-dUTP. Fluorescent labeled probes were hybridized on a microarray of 4,800 human genes. Results: Twelve genes were consistently over-expressed in the endometrium of endometriosis such as ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ transporting (ATP5C1) and TNF alpha factor. Eleven genes were consistently down-regulated in the endometriosis samples. Many extracellular matrix protein genes (decorin, lumican, EGF-containing fibulin-like extracellular matrix protein 1, fibulin 5, and matrix Gla protein) and protease/protease inhibitors (serine proteinase inhibitor, matrix metalloproteinase 2, tissue inhibitor of metalloproteinase 1), and insulin like growth factor II associated protein were included. Expression patterns of selected eight genes from the cDNA microarray were confirmed by quantitative RT-PCR or real time RT-PCR. Conclusion: The result of this analysis supports the hypothesis that the endometrium from patients with endometriosis has distinct gene expression profile from control endometrium without endometriosis.