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Effects of Oyster Shell Extract on Papain-induced Osteoarthritis in C57BL/6J Mice

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

  • Lee, Se-Young (Dept. of Food Science and Nutrition, Pusan National University) ;
  • Kim, Hak-Ju (Seojin Biotech Co., Ltd.) ;
  • Han, Ji-Sook (Dept. of Food Science and Nutrition, Pusan National University)
  • Received : 2013.02.15
  • Accepted : 2013.07.17
  • Published : 2013.08.31

Abstract

We investigated the effects of oyster shell extract (OSE) on papain-induced osteoarthritis in C57BL/6J mice. Osteoarthritis was induced in mice by a papain injection into the knee joint. The mice were divided into a total of five groups (n=8). The normal group was untreated, whereas the papain group (negative control) was induced with osteoarthritis and treated with water daily. The papain+DS group (positive control) was treated with diclofenac sodium. Papain+OSE groups were treated with OSE concentrations of 100 and 200 mg/kg/bw for 20 days. Proteoglycan content in articular cartilage was analyzed through safranine-O fast green staining and H&E staining. The histopathological changes in cartilage were measured by the Rudolphi score approach. The contents of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, and IL-6 in plasma were analyzed by the ELISA method. After experiments, body weights of the treated groups were not significantly different compared with the normal group. Cartilage loss and joint instability significantly improved in a dose-dependent manner in the OSE-treated group compared with the papain group (P<0.05). Proteoglycan content was significantly higher in the OSE-treated group than the papain group (P<0.05). Osteoarthritis scores of the OSE-treated group were significantly decreased compared with the papain group (P<0.05). TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 content in the plasma of the papain+OSE treated groups significantly decreased in a dose-dependent manner compared with the papain group (P<0.05). These results suggest that OSE treatment might have anti-arthritic effects on papain-induced osteoarthritis in C57BL/6J mice.

연구에서는 굴 패각 추출물이 papain으로 유도된 골관절염 동물 모델에 미치는 영향을 살펴보았다. 8주령의 C57BL/6J 마우스 우측 슬관절강에 papain $6{\mu}L$를 투여하여 골관절염을 유발하였다. 실험군은 각 군당 체중이 고르게 8마리씩 배정하여 총 다섯군으로 분류하였다. 정상군은 우측 슬관절강에 생리식염수 $6{\mu}L$ 주사하고 20일간 1일 1회 생리식염수 0.2 mL씩 경구투여 하였다. 대조군, 약물투여군, 굴 패각 추출물 투여군은 골관절염을 유발 후 각각 생리식염수, diclofenac sodium 2 mg/kg/bw, OSE 100, 200 mg/kg/bw를 20일 동안 매일 1회 정해진 시간에 경구투여 하였다. 실험기간 동안 체중을 측정하였으며, 실험 20일에 부검하여 슬관절의 병리조직학적 관찰, 관절연골 내 proteoglycan 함유율, 골관절염 지수의 변화, 혈액내 TNF-${\alpha}$, IL-$1{\beta}$, IL-6 함량을 측정하였다. 체중은 실험기간 동안 모든 개체 간의 유의적 차이가 없었다. 관절연골 내 proteoglycan 함유율은 DS 및 OSE 투여군이 papain 투여 대조군보다 유의성(P<0.05)있게 높았으며, 특히 OSE-200 투여군은 약물 투여군과 유사한 proteoglycan 함유율을 나타냈다. H&E 염색을 통하여 관찰한 결과, 골관절염이 유발된 마우스에 굴 패각추출물을 경구 투여하였을 때 관절연골의 파괴와 골 침식 등 연골의 변성이 약물 처리군과 유사한 것을 보아 굴 패각 추출물이 골관절염 치료에 효과가 있는 것을 확인할 수 있었다. 병리조직학적으로 골관절염 지수는 DS 및 OSE 투여군이 papain 투여 대조군보다 유의성(P<0.05) 있게 낮았으며, OSE-200 투여군이 OSE-100 투여군보다 낮았다. 혈액 내 TNF-${\alpha}$, IL-$1{\beta}$, IL-6 함량은 papain 투여 대조군이 정상군에 비해 유의적(P<0.05)으로 증가하였으며, DS 및 OSE 투여군은 대조군보다 유의성(P<0.05) 있게 낮았다. 특히 TNF-${\alpha}$ 및 IL-6 함량은 OSE-200 투여군이 약물 투여군과 유사한 감소 효과를 나타내었다. 이와 같은 결과는 굴 패각 추출물이 papain으로 유도된 골관절염에서 연골의 변성을 줄이고 염증을 억제함으로써 골관절염 치료에 효과가 있는 것으로 사료된다. 굴 패각은 탄산칼슘이 풍부하기 때문에 골관절염 개선에 효과가 있는 것으로 사료되나, 굴 패각 추출물 중 어떠한 기능성 물질이 골관절염 개선 효과를 나타내었는지에 대해 향후 심도 있는 연구가 필요하다.

Keywords

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