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

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Effects of Calcium Citrate Isolated from Oyster Shell on Papain-induced Osteoarthritis in C57BL/6J Mice

굴 패각 구연산칼슘이 Papain으로 유도된 골관절염 C57BL/6J Mice에 미치는 영향

  • Choi, Eun-Young (Department of Food Science and Nutrition, Pusan National University) ;
  • Kim, Hak-Ju (Seojin Biotech Co., Ltd.) ;
  • Han, Ji-Sook (Department of Food Science and Nutrition, Pusan National University)
  • Received : 2014.06.16
  • Accepted : 2014.07.17
  • Published : 2014.10.31
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Abstract

This study investigated the effects of calcium citrate on papain-induced osteoarthritis in C57BL/6J mice. Osteoarthritis was induced by injecting $6{\mu}L$ of papain into the knee joints of mice. Calcium citrate was made by crushing the centrifuged precipitate after reacting 0.5 M citric acid with 1 kg of oyster shell extract. The mice were divided into five groups (n=8). The normal group was untreated, whereas the papain group was induced to have osteoarthritis and treated with $200{\mu}L$ of water per day. The papain+DS group was treated with diclofenac sodium. The papain+calcium citrate groups were treated with calcium citrate at 150 and 300 mg/kg/bw for 28 days. Proteoglycan contents in articular cartilages were measured by safranin O/fast green staining and hematoxylin & eosin staining. Histopathological changes in cartilages were analyzed by the Rudolphi score approach. Contents of pro-inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in plasma, were measured by the ELISA method. Body weights among the treated groups were not significantly different compared with that of the normal group. Cartilage loss and joint instability in the calcium citrate group improved significantly (P<0.05) in a dose-dependent manner compared with the papain group. Further, proteoglycan content of the calcium citrate group was considerably (P<0.05) higher than that of the papain group. Osteoarthritis scores in the calcium citrate group were considerably (P<0.05) reduced compared with the papain group. In the group treated with calcium citrate, contents of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in plasma were significantly (P<0.05) reduced in a dose-dependent manner in comparison with the normal group. Based on these results, we suggest that calcium citrate is effective for treatment of osteoarthritis.

본 연구에서는 구연산칼슘 투여가 papain으로 유도된 골관절염 동물모델에 미치는 영향을 살펴보았다. 8주령의 C57 BL/6J 마우스의 우측 슬관절강에 papain $6{\mu}L$를 투여하여 골관절염을 유발하였다. 실험군은 각 군당 8마리씩 체중이 고르게 배정하여 총 5군으로 나누었다. 정상군은 우측 슬관절강에 생리식염수 $6{\mu}L$를 주사하였고, 대조군, 약물투여군, 구연산칼슘 투여군은 papain $6{\mu}L$로 골관절염을 유발하였다. 정상군과 대조군은 0.2 mL의 생리식염수를, 약물투여군은 diclofenac sodium 2 mg/kg/bw를, 구연산칼슘 투여군은 150, 300 mg/kg/bw를 28일간 1일 매일 정해진 시간에 1회 경구 투여하였다. 실험기간 동안 체중을 측정하였으며, 실험 종료일에는 부검하여 슬관절의 병리조직학적 관찰을 하였고 관절연골 내 proteoglycan 함유율, 골관절염 지수의 변화, 혈액 내 TNF-${\alpha}$, IL-$1{\beta}$, IL-6 함량을 측정하였다. 체중은 실험기간 동안 모든 개체 간의 유의적인 차이는 없었다. 혈액 내 TNF-${\alpha}$, IL-$1{\beta}$, IL-6 함량은 papain 투여 대조군이 정상군에 비해 유의적으로 증가하였으며 약물 및 구연산칼슘 투여군은 대조군보다 유의성 있게 낮았다. 관절연골 내 proteoglycan의 함유율은 약물 및 구연산칼슘 투여군이 papain 투여 대조군보다 유의성 있게 높았다. Hematoxylin과 eosin 염색 관찰 결과, 골관절염이 유발된 마우스에 구연산칼슘을 경구 투여하였더니 관절연골의 파괴와 골 침식 등 연골의 변성이 약물 투여군과 유사하게 나타났다. 이와 같은 결과는 구연산칼슘이 papain으로 유도된 골관절염에서 연골의 변성을 줄이고 염증을 억제함으로써 골관절염 치료에 효과가 있은 것으로 사료되며, 추후 더 자세한 연구가 필요하다.

Keywords

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