Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Conditioned Medium of Soybean Extract Treated Osteoblasts Inhibits RANKL Induced Differentiation of Osteoclasts

대두추출물을 처리한 조골세포 조건배양액은 RANKL에 의해 유도된 파골세포 분화를 억제

  • Park, Kyung-Ho (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Ju, Won-Chul (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Yeo, Joo-Hong (Dept. of Agricultural Biology, National Academy of Agricultural Science) ;
  • Lee, Kwang-Gill (Dept. of Agricultural Biology, National Academy of Agricultural Science) ;
  • Cho, Yun-Hi (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University)
  • 박경호 (경희대학교 동서의학대학원 의학영양학과) ;
  • 주원철 (경희대학교 동서의학대학원 의학영양학과) ;
  • 여주홍 (농촌진흥청 농업생물부) ;
  • 이광길 (농촌진흥청 농업생물부) ;
  • 조윤희 (경희대학교 동서의학대학원 의학영양학과)
  • Published : 2010.01.30
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Abstract

Soybean is of particular interest as a food supplement of isoflavones for inhibiting bone resorption in postmenopausal woman. These beneficial effects of isoflavones are caused by functioning as partial agonists or antagonists of estrogen, of which anti-resorptive effect is mediated indirectly through paracrine factors produced by osteoblasts that act on osteoclasts. In this study, the indirect effect of soybean on osteoclastic differentiation of RAW264.7 cells were investigated. The conditioned medium was collected from MC3T3-E1 osbeoblasts treated with 0.001 mg/mL~0.1 mg/mL soybean extracts for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of paracrine factors in the conditioned medium, the protein expression of osteoprotegerin (OPG) with soybean extract was specifically higher in a dose dependent manner than with $10^{-9}$ M~$10^{-6}$ M of estrogen, genistein or daidzein standards. In RAW264.7 cells, the conditioned medium with soybean inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrateresistant alkaline phosphatase (TRAP)-positive osteoclasts and protein expression of MMP-9 were significantly decreased. Coupled with the low expression of estrogen receptor $\alpha$ and $\beta$ proteins in RANKL treated RAW264.7 cells, we demonstrate that the conditioned medium of soybean treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with the selective expression of OPG in osteoblasts.

본 연구는 MC3T3-E1 조골 전구세포에 대두추출물을 처리 후 그 조건배양액에서 파골세포분화 관련 국소인자 중파골세포분화 촉진인자(IL-$\beta$, IL-6, RANKL, TNF-$\alpha$, M-CSF) 및 파골세포로의 분화억제에 관여하는 국소인자인 OPG의 발현변화를 조골세포 조건배양액에서 살펴보았으며, 이 조골세포 조건배양액을 RAW264.7 파골전구세포에 처리 시 파골세포로의 분화를 억제정도를 TRAP 염색 및 관련 분화 지표의 발현을 통해 알아보았다. 대두추출물 처리한 조골세포 조건배양액에서 OPG의 발현이 농도 의존적으로 현저히 증가하였다. 그러나 강력한 파골세포 분화촉진인자로 알려진 IL-1$\beta$의 발현 역시 고농도의 대두추출물 처리 시 현저히 증가하여 고농도의 대두추출물을 처리한 CM 처리 시 TRAP 염색된 일부 파골세포를 확인하였고, 파골세포분화 관련지표인 MMP-9의 발현 또한 저농도 대두 추출물을 처리한 CM 처리에 비해 증가하였다. 이는 저농도의 대두추출물이 고농도의 대두추출물에 비해 파골세포분화억제에 효과적임을 의미하나, 추후 파골세포수준에서 분화억제 관련 기전연구가 필요할 것으로 생각되어진다.

Keywords

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