• Clinical and Experimental Reproductive Medicine
• /
• 제33권2호
• /
• pp.105-113
• /
• 2006

목 적: 본 연구를 통해 $TGF-{\beta}1$에 의해 유도된 인간자궁내막의 탈락막화 과정에서 ERK와 $PPAR{\gamma}$의 역할을 규명하고자 하였다. 연구방법: 자궁내막 기질세포는 DMEM/F12 (10% FBS, 1 nM E2 and 100 nM P4) 조건에서 배양하였다. 연구 목적에 따라 $TGF-{\beta}1$ (5 ng/ml), Rosiglitazone (50 nM)와 PD98059 ($20{\mu}M$)를 배양액에 첨가하였다. Trypan-Blue와 hematocytometer를 이용하여 현미경하에서 세포의 개수를 측정하였다. Enzyme-linked immunosorbent assay (ELISA)와 western blotting 방법을 사용하여 단백질의 발현 정도를 관찰하였다. 결과 및 결론: 배양액에 $TGF-{\beta}1$을 첨가하여 세포의 증식 정도를 측정한 결과 $TGF-{\beta}1$이 세포의 증식을 억제하는 것을 알 수 있었다. 또한 배양된 세포로부터 PGE2 및 prolactin의 발현을 유도하는 것을 알 수 있었다. 이러한 $TGF-{\beta}1$의 작용은 Smad 및 ERK의 활성화를 통하여 일어남을 알 수 있었다. $PPAR{\gamma}$의 기질인 rosiglitazone을 배양액에 첨가한 결과 $TGF-{\beta}1$에 의한 세포 증식의 억제가 역전되는 것을 알 수 있었다. 뿐만 아니라, 세포 내 ERK의 활성 역시 억제 시켰으며 이 결과 PGE2와 prolactin의 발현이 억제 되는 것을 관찰할 수 있었다. 따라서 본 연구를 통해 $TGF-{\beta}1$에 의한 자궁내막 기질세포의 탈락막화는 Smad와 ERK의 활성화를 통하여 이루어지며 이러한 과정은 $PPAR{\gamma}$에 의해 억제됨을 알 수 있었다.

• 대한한방부인과학회지
• /
• 제34권4호
• /
• pp.30-45
• /
• 2021

Objectives: This study was performed to investigate the effects of pregnancy-related four herbal medicines (Samul-tang, Onkyung-tang, Chokyungjongok-tang and Moutan Radicis Cortex) on the endometrial and placental cells. Methods: In this study, we examined viability and decidualization of telomerase immortalized human endometrial stromal cell lines (T-HESCs) and viability and invasion ability of human first trimester trophoblast cell lines Sw.71 by four herbal medicines (Samul-tang, Onkyung-tang, Chokyungjongok-tang and Moutan Radicis Cortex) Results: In the study, we showed that Samul-tang, Onkyung-tang, Chokyungjongok-tang increased decidualization marker prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1) in T-HESCs. Moutan Radicis Cortex decreased the mRNA level of PRL and IGFBP1, and the protein level of PRL and IGFBP1 had no significant effect. Moreover, four herbal medicines reduced invasion ability of Sw.71 cells. Conclusions: These results suggest that Samul-tang, Onkyung-tang, and Chokyungjongok-tang have beneficial effects on successful embryo implantation and pregnancy maintenance by increasing decidualization markers such as PRL and IGFBP1. Moutan Radicis Cortex reduces the mRNA levels of PRL and IGFBP1, which may adversely affect pregnancy. Further investigations are needed to elucidate the significance of the decreased invasive ability of Sw.71 cells induced by four herbal medications.

• Clinical and Experimental Reproductive Medicine
• /
• 제30권1호
• /
• pp.65-75
• /
• 2003

Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

• Clinical and Experimental Reproductive Medicine
• /
• 제37권3호
• /
• pp.207-216
• /
• 2010

목 적: Small interfering RNA (siRNA)를 이용하여 homeobox (HOXA) 10 유전자의 발현이 억제된 일차배양 자궁내막 세포를 이용하여 자궁내막 탈락막화 (decidualization)에 HOXA유전자를 포함한 세포 내 신호전달기전을 분석하고자 하였다. 연구방법: 본원 산부인과에서 자궁내막 질환 이외의 이유로 전자궁 적출술을 받은 환자의 자궁내막 조직을 채취한다. $37^{\circ}C$에서 20분간 Trypsin-EDTA를 처리하여 단일세포로 분리한 후 10% fetal bovine serum이 첨가된 DMEM/F12 배지를 이용하여 24시간 동안 $37^{\circ}C$ 5% $CO_2$ 배양기 안에서 배양한다. 배양된 자궁내막 세포를 HOXA10 siRNA로 첨가한 후 TGF-${\beta}1$을 10 ng/mL 농도로 48시간 첨가하여 탈락막화를 유도한다. 배양된 자궁내막 세포에서 reverse transcription polymerase chain reaction을 이용하여 HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferator-activated receptor (PPAR)-$\gamma$ 및 wingless-type MMTV integration site family (Wnt)의 발현을 관찰하였다. 결 과: HOXA10의 경우 transforming growth factor (TGF)-${\bata}1$과 HOXA10 siRNA를 처리하지 않은 대조군에 비하여 TGF-${\beta}1$을 처리한 군에서 약 1.8배 가량 발현양의 증가를 보였다. 자궁내막 탈락막 표지인자로 알려져 있는 prolactin의 경우 TGF-${\beta}1$을 처리한 경우 대조군에 비하여 유의한 발현의 증가를 보였으며 HOXA10 siRNA를 처리한 군에 있어서는 TGF-${\beta}1$을 첨가하더라도 prolactin mRNA의 발현양의 증가를 관찰할 수 없었다. 또한 자궁내막 세포의 분화인자로 알려져 있는 COX-2의 발현 역시 HOXA10 siRNA를 처리한 군에 있어서 mRNA 발현양이 유의하게 감소하였으며 TGF-${\beta}1$을 처리하여도 발현의 증가를 관찰할 수 없었다. Wnt4의 경우 HOXA10 siRNA를 이용하여 HOXA10의 발현을 억제한 경우 대조군에 비하여 유의하게 mRNA의 발현양이 감소하였으며 이러한 발현양의 감소는 TGF-${\beta}1$을 처리하여도 증가됨을 관찰할 수 없었다. PPAR$\gamma$의 발현은 HOXA10 siRNA의 처리와 관계없이 TGF-${\beta}1$에 의하여 감소하는 것을 관찰할 수 있었다. 결 론: Progesterone에 의하여 자궁내막 상피세포에서 분비되는 것으로 알려져 있는 TGF-${\beta}1$에 의한 자궁내막 기질세포의 분화 (탈락막화)는 HOXA10 및 Wnt에 의하여 조절되는 것으로 생각된다.

• 한국발생생물학회:학술대회논문집
• /
• 한국발생생물학회 2003년도 전기 한국발생생물학회 제16차 학술대회논문집
• /
• pp.29-29
• /
• 2003

No Abstract, See Full Text

• Clinical and Experimental Reproductive Medicine
• /
• 제22권2호
• /
• pp.155-161
• /
• 1995

The present investigation has been undertaken to elucidate the differentiation mechanism the uterus which is the environment of the embryo development, by demonstrating the role of ovarian steroids hormone in the decidualization of the pseudopregnant rat uterus. To determine the effect of ovarine steroids and artificial stimulation (trauma) on the differenciation of the uterine endometrium and decidualization for implantation, attempt was made to measure concentrations of serum estradiol($E_2$), progesterone($P_4$) and nuclear $P_4$ receptor in the traumatized and non-traumatized uterine tissue of the pseudopregnant rat. The results obtained are as followings : The concentration of serum $E_2$ on day 9(implantation stage) was similar in both of intact pseudopregnant rat(47.63pg/ml) and normal pregnant rat(40.71pg/ml). And among the treated groups, $E_2$ concentration was highest in the $E_2$ treated group in comparision with intact control group(relative value; 73.27%). The concentration of serum $P_4$ was also highest in the $P_4$ treated group(23.12pg/ml). Relative value of $P_4$ treated group in comparision with intact group(24.88pg/ml) was 92.93%. The nuclear $P_4$ receptor levels in the artificial traumatized groups were higher compared with the non-traumatized control groups. This study, therefore, clearly demonstrates that the methods for inducing pseudopregnant (vagina tapping;120/min) and inducing decidualization(oil injection; 0.1ml/uterine horn) appear to be effective, $P_4$ appears to be effective in the differenciation of the uterine endometrial tissue for the implantation process. Concentration of serum $P_4$ seems to be well correlated with the level of the nuclear $P_4$ receptor during the early embryo development. These results seem to be well correlated with ALPase activities in the normal and pseudopregnant rat uterus shown in the previous study.

• 대한한방부인과학회지
• /
• 제34권3호
• /
• pp.1-14
• /
• 2021

Methods: We investigated the ability to induce decidualization of human endometrial stromal cells and invasive ability of human trophoblast cells by water extract of three types herbal medicines (Dangguijakyak-san, Siryung-tang, Antae-eum) and ethanol extract on Scutellariae Radix. Results: In the process of decidualization of endometrial stromal cells, three herbal medicines including Dangguijakyak-san, Siryung-tang, and Scutellariae Radix increased the production of decidual markers such as prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). However, Antae-eum increased mRNA levels of PRL and IGFBP and secretion of IGFBP in decidual stromal cells, but not PRL. Four herbal medicines inhibited the invasion of trophoblast cells. Conclusions: Four herbal medicines may play a role in implantation in women with reproductive failures. However, further studies are warranted to elucidate whether these medications are helpful in the maintenance of pregnancy.

• Clinical and Experimental Reproductive Medicine
• /
• 제29권4호
• /
• pp.295-302
• /
• 2002

Uterine cells carry out proliferation and differentiation for preparation the embryonic implantation during pregnancy. Therefore regulation of the cell proliferation is an essential step for uterine preparation, but there is not much information about the proliferation related genes in pregnant uterus. To identify these implantation specific genes, a PCR-select cDNA subtraction method was employed and got a few genes. One of the identified genes is a novel gene encoding oral tumor suppressor doc-1. To detect the doc-1 expression on the pregnant uterus, dot blotting, RT-PCR, and in situ hybridization were employed. Dot blotting revealed that doc-1 mRNA expression increase after implantation. During normal pregnancy, doc-1 mRNA expression was detected as early as day 1 of pregnancy with RT-PCR. Its expression was increased about 15 times after embryonic implantation. doc-1 transcript was localized in luminal epithelial cells but it was very faint during preimplantation. After starting the implantation, it localized in the stromal cells; heightened expression of doc-1 correlates with intense stromal cell proliferation surrounding the implanting blastocyst on day 6 morning. However in the decidualized cells, the intensity of localized doc-1 mRNA was weak. From those results, it is revealed that doc-1 express at pregnant uterus of the mouse. In addition it is suggested that doc-1 is the gene regulating the proliferation of the luminal epithelial cells and stromal cells during early implantation and decidualization.

• Clinical and Experimental Reproductive Medicine
• /
• 제38권3호
• /
• pp.119-125
• /
• 2011

Implantation of an embryo occurs during the mid-secretory phase of the menstrual cycle, known as the "implantation window." During this implantation period, there are significant morphologic and functional changes in the endometrium, which is followed by decidualization. Many immune cells, such as dendritic and natural killer (NK) cells, increase in number in this period and early pregnancy. Recent works have revealed that antigen-presenting cells (APCs) and NK cells are involved in vascular remodeling of spiral arteries in the decidua and lack of APCs leads to failure of pregnancy. Paternal and fetal antigens may play a role in the induction of immune tolerance during pregnancy. A balance between effectors (i.e., innate immunity and helper T [Th] 1 and Th17 immunity) and regulators (Th2 cells, regulatory T cells, etc.) is essential for establishment and maintenance of pregnancy. The highly complicated endocrine-immune network works in decidualization of the endometrium and at the fetomaternal interface. We will discuss the role of immune cells in the implantation period and during early pregnancy.

• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 2005년도 제49차 추계학술대회
• /
• pp.114-114
• /
• 2005