Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Effect of Human Hydrosalpingeal Fluid on the Development of Mouse Embryo

난관수종액이 생쥐 배아발달에 미치는 영향

  • Park, Joon-Cheol (Department of Obstetrics and Gynecology, Keimyung University School of Medicine) ;
  • Kim, Jeong-A (Department of Obstetrics and Gynecology, Keimyung University School of Medicine) ;
  • Kim, Dong-Ja (Department of Pathology, Rosalind Franklin University of Chicago Medical School) ;
  • Bae, Jin-Gon (Department of Obstetrics and Gynecology, Keimyung University School of Medicine) ;
  • Kim, Jong-In (Department of Obstetrics and Gynecology, Keimyung University School of Medicine) ;
  • Rhee, Jeong-Ho (Department of Obstetrics and Gynecology, Keimyung University School of Medicine)
  • 박준철 (계명대학교 의과대학 산부인과교실) ;
  • 김정아 (계명대학교 의과대학 산부인과교실) ;
  • 김동자 (시카고의과대학 병리학교실) ;
  • 배진곤 (계명대학교 의과대학 산부인과교실) ;
  • 김종인 (계명대학교 의과대학 산부인과교실) ;
  • 이정호 (계명대학교 의과대학 산부인과교실)
  • Received : 2010.01.27
  • Accepted : 2010.05.14
  • Published : 2010.06.30
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Abstract

Objective: The aim of this study was to measure the concentrations of cytokines contained in the hydrosalpingeal fluid and to evaluate the effect on the mouse embryo development with the different cytokine concentration. Methods: The hydrosalpingeal fluids (HSF) were collected during the surgery for hydrosalpinx which was confirmed by hysterosalphingogram. The cytokines in HSF including interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-$\alpha$, Interferon-$\gamma$ (IFN-$\gamma$), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), monocyte chemotactic protein-1 (MCP-1) were measured by ELISA method. HSF were added up to culture media with 5%, 10%, and 30% concentrations. The blastulation rates were compared. Results: IL-$\alpha$, IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-$\alpha$, IFN-$\bamma$, VEGF, EGF, and MCP-1 were detected, but the concentrations were different from each sample. IL-6 and IL-10 were increased in HSF-1 group, and IFN-$\gamma$, MCP-1, VEGF were increased in HSF-2 compared with normal serum range. The Th1/Th2 ratio of HSF-2 (IFN-$\gamma$:IL10) was highly elevated to 61.64, compared with that of HSF-1 (3.69). The blastulation rate was significantly decreased in HSF-2 group (27.7%) compared those of the HSF-1 group (74%) and control group (76.7%). It showed the trend that the blastulation rate was decreased depending on the concentration HSF of culture media in HSF-2 group, but it was not statistically significant. Conclusion: The composition and concentration of cytokines in each HSF were different, and increased proinflammatory cytokines in HSF might be associated with poor embryonic development. Further study will be needed about the effect of each cytokines in HSF.

목 적: 난관수종액내의 사이토카인 농도를 측정하고, 사이토카인 농도가 다른 난관수종액을 이용하여 생쥐 배아 발생에 미치는 영향을 비교하고자 하였다. 연구방법: 난관수종액은 자궁난관 조영술에서 난관수종이 진단되어 복강경을 통한 난관 절제술을 시행하는 경우 난관 절제술 전에 난관으로부터 채취한 다음 3,000 rpm에서 10분간 원심분리시킨 후 상층액만을 $-20^{\circ}C$에서 보관하였다. 난관수종액의 사이토카인의 조성 및 농도를 확인하기 위하여 interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor (TNF)-$\alpha$, interferon (IFN)-$\gamma$, vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), monocyte chemotactic protein (MCP)-1 등을 ELISA 방법으로 측정하였다. 기본 배양액에 난관수종액을 5%, 10%, 30%의 비율로 첨가하여 각 군별로 배반포로의 발달을 관찰하였다. 결 과: 난관수종액내에서 IL-$1{\alpha}$, IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-$\alpha$, IFN-$\gamma$, VEGF, EGF, MCP-1가 검출되었으며, 그 농도에 있어서는 큰 차이를 보였다. 정상 혈청 농도에 비하여 난관수종액-1은 IL-6, IL-10이 증가되어 있었고, 난관 수종액-2는 IFN-$\gamma$, MCP-1 및 VEGF가 증가되어 있었다. 각 난관수종액의 Th1/Th2 비는 HSF-1의 경우 IFN-$\gamma$:IL-10이 3.69로 정상인 데 비하여 HSF-2의 경우 IFN-$\gamma$:IL-10이 61.14로 크게 증가되어 있었다. 난관수종액을 포함하지 않는 배양액에서는 배반포기 발달률은 76.7%이었고, 난관수종액-1군은 74%로 대조군과 차이를 보이지 않았지만, 난관수종액-2군의 경우 27.7%로서 대조군 및 난관수종액-1군과도 유의한 차이를 보였다. 난관수종액-1의 경우 난관수종액 농도에 따른 차이는 없었으며, 난관수종액-2군의 경우 농도에 증가함에 따라 배반포로의 발달이 감소하기는 하였지만 통계적으로 유의하지는 않았다. 결 론: 난관수종액마다 사이토카인의 조성이 다르며 이에 따라 생쥐 배아발달에 미치는 영향이 다를 수 있다. 염증성 사이토카인이 증가된 난관수종액이 배아발달에 악영향을 미칠 것으로 추정된다. 특정 사이토카인에 의한 작용을 규명하기는 위해서는 향후 지속적인 연구가 필요할 것으로 생각된다.

Keywords

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