Esculetin (dihydroxycoumarin) inhibits the production of matrix metalloproteinases in cartilage explants, and oral administration of its prodrug, CPA-926, suppresses cartilage destruction in rabbit experimental osteoarthritis

Harumoto Yamada, Koju Watanabe, Tsuyoshi Saito, Haruhisa Hayashi, Yoshiaki Niitani, Toshiyuki Kikuchi, Akira Ito, Kyosuke Fujikawa, L. Stefan Lohmander

Research output: Contribution to journalArticle

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Abstract

Objective. To investigate the in vitro effects of 6,7-dihydroxycoumarin (esculetin) on the production of matrix metalloproteinases (MMP) in rabbit articular cartilage, and the in vivo effects of orally administered CPA-926, a prodrug of esculetin, on cartilage destruction in rabbit experimental osteoarthritis (OA). Methods. In vitro studies were performed using rabbit articular cartilage explants. Esculetin 10-100 μM was added to cartilage explants in the presence or absence of interleukin 1α (IL-1α). Effects of esculetin on cartilage metabolism were assessed. Proteoglycan release into medium was determined by dye precipitation with 1,9-dimethylmethylene blue, synthesis of proMMP-1 (interstitial procollagenase) and proMMP-3 (prostromelysin 1) by Western blotting, and collagen degradation activity using FITC labeled collagen. In vivo experimental OA was induced in the knee joints of 15 Japanese adult white rabbits by partial lateral meniscectomy. Ten rabbits were orally administered 200 or 400 mg/kg/day of CPA-926 from the day of surgery for 14 days. The size of the macroscopic erosive area on the femoral condyle and tibial plateau was measured, and cartilage destruction was histologically evaluated. Collagenolytic activities in synovial fluid were measured using FITC labeled collagen as a substrate. Results. In vitro, esculetin inhibited the IL-1α induced release of proteoglycan into the medium in a dose dependent manner. The collagenolytic activities in cartilage explant medium induced by IL-1α were also suppressed with the addition of 33-100 μM esculetin (p = 0.0209 at 33 and 100 μM, p = 0.0202 at 66 μM). Western blotting of cartilage explant medium showed a decrease in the levels of proMMP-1 and proMMP-3 in the medium by treatment with esculetin. In vivo: At 14 days after surgery, the femoral condyle and tibial plateau in the control group showed macroscopic erosions of cartilage. Compared with the control group, the rabbits treated with CPA-926 at the dose of 400 mg/kg exhibited reduction of the size of the erosive area on the tibial plateau (p = 0.009). Histological evaluation indicated protection against the development of destructive changes in the tibial plateau cartilage at a dose of 200 mg/kg (p = 0.0442) and 400 mg/kg (p = 0.0446) of CPA-926. Conclusion. These results indicate that esculetin inhibits matrix degradation in rabbit joint cartilage explants through the suppression of MMP synthesis, secretion, or activity. Prophylactic administration of its prodrug, CPA-926, appears to provide some protection against cartilage destruction in a short term rabbit experimental OA model.

Original languageEnglish
Pages (from-to)654-662
Number of pages9
JournalJournal of Rheumatology
Volume26
Issue number3
Publication statusPublished - 22-03-1999

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Prodrugs
Matrix Metalloproteinases
Osteoarthritis
Cartilage
Oral Administration
Rabbits
Interleukin-1
Matrix Metalloproteinase 1
Collagen
Fluorescein-5-isothiocyanate
Articular Cartilage
Proteoglycans
Thigh
Ambulatory Surgical Procedures
Western Blotting
6((2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl)oxy)-7-hydroxycoumarin
esculetin
Bone and Bones
Control Groups
Synovial Fluid

All Science Journal Classification (ASJC) codes

  • Rheumatology
  • Immunology and Allergy
  • Immunology

Cite this

Yamada, Harumoto ; Watanabe, Koju ; Saito, Tsuyoshi ; Hayashi, Haruhisa ; Niitani, Yoshiaki ; Kikuchi, Toshiyuki ; Ito, Akira ; Fujikawa, Kyosuke ; Stefan Lohmander, L. / Esculetin (dihydroxycoumarin) inhibits the production of matrix metalloproteinases in cartilage explants, and oral administration of its prodrug, CPA-926, suppresses cartilage destruction in rabbit experimental osteoarthritis. In: Journal of Rheumatology. 1999 ; Vol. 26, No. 3. pp. 654-662.
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title = "Esculetin (dihydroxycoumarin) inhibits the production of matrix metalloproteinases in cartilage explants, and oral administration of its prodrug, CPA-926, suppresses cartilage destruction in rabbit experimental osteoarthritis",
abstract = "Objective. To investigate the in vitro effects of 6,7-dihydroxycoumarin (esculetin) on the production of matrix metalloproteinases (MMP) in rabbit articular cartilage, and the in vivo effects of orally administered CPA-926, a prodrug of esculetin, on cartilage destruction in rabbit experimental osteoarthritis (OA). Methods. In vitro studies were performed using rabbit articular cartilage explants. Esculetin 10-100 μM was added to cartilage explants in the presence or absence of interleukin 1α (IL-1α). Effects of esculetin on cartilage metabolism were assessed. Proteoglycan release into medium was determined by dye precipitation with 1,9-dimethylmethylene blue, synthesis of proMMP-1 (interstitial procollagenase) and proMMP-3 (prostromelysin 1) by Western blotting, and collagen degradation activity using FITC labeled collagen. In vivo experimental OA was induced in the knee joints of 15 Japanese adult white rabbits by partial lateral meniscectomy. Ten rabbits were orally administered 200 or 400 mg/kg/day of CPA-926 from the day of surgery for 14 days. The size of the macroscopic erosive area on the femoral condyle and tibial plateau was measured, and cartilage destruction was histologically evaluated. Collagenolytic activities in synovial fluid were measured using FITC labeled collagen as a substrate. Results. In vitro, esculetin inhibited the IL-1α induced release of proteoglycan into the medium in a dose dependent manner. The collagenolytic activities in cartilage explant medium induced by IL-1α were also suppressed with the addition of 33-100 μM esculetin (p = 0.0209 at 33 and 100 μM, p = 0.0202 at 66 μM). Western blotting of cartilage explant medium showed a decrease in the levels of proMMP-1 and proMMP-3 in the medium by treatment with esculetin. In vivo: At 14 days after surgery, the femoral condyle and tibial plateau in the control group showed macroscopic erosions of cartilage. Compared with the control group, the rabbits treated with CPA-926 at the dose of 400 mg/kg exhibited reduction of the size of the erosive area on the tibial plateau (p = 0.009). Histological evaluation indicated protection against the development of destructive changes in the tibial plateau cartilage at a dose of 200 mg/kg (p = 0.0442) and 400 mg/kg (p = 0.0446) of CPA-926. Conclusion. These results indicate that esculetin inhibits matrix degradation in rabbit joint cartilage explants through the suppression of MMP synthesis, secretion, or activity. Prophylactic administration of its prodrug, CPA-926, appears to provide some protection against cartilage destruction in a short term rabbit experimental OA model.",
author = "Harumoto Yamada and Koju Watanabe and Tsuyoshi Saito and Haruhisa Hayashi and Yoshiaki Niitani and Toshiyuki Kikuchi and Akira Ito and Kyosuke Fujikawa and {Stefan Lohmander}, L.",
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Esculetin (dihydroxycoumarin) inhibits the production of matrix metalloproteinases in cartilage explants, and oral administration of its prodrug, CPA-926, suppresses cartilage destruction in rabbit experimental osteoarthritis. / Yamada, Harumoto; Watanabe, Koju; Saito, Tsuyoshi; Hayashi, Haruhisa; Niitani, Yoshiaki; Kikuchi, Toshiyuki; Ito, Akira; Fujikawa, Kyosuke; Stefan Lohmander, L.

In: Journal of Rheumatology, Vol. 26, No. 3, 22.03.1999, p. 654-662.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Esculetin (dihydroxycoumarin) inhibits the production of matrix metalloproteinases in cartilage explants, and oral administration of its prodrug, CPA-926, suppresses cartilage destruction in rabbit experimental osteoarthritis

AU - Yamada, Harumoto

AU - Watanabe, Koju

AU - Saito, Tsuyoshi

AU - Hayashi, Haruhisa

AU - Niitani, Yoshiaki

AU - Kikuchi, Toshiyuki

AU - Ito, Akira

AU - Fujikawa, Kyosuke

AU - Stefan Lohmander, L.

PY - 1999/3/22

Y1 - 1999/3/22

N2 - Objective. To investigate the in vitro effects of 6,7-dihydroxycoumarin (esculetin) on the production of matrix metalloproteinases (MMP) in rabbit articular cartilage, and the in vivo effects of orally administered CPA-926, a prodrug of esculetin, on cartilage destruction in rabbit experimental osteoarthritis (OA). Methods. In vitro studies were performed using rabbit articular cartilage explants. Esculetin 10-100 μM was added to cartilage explants in the presence or absence of interleukin 1α (IL-1α). Effects of esculetin on cartilage metabolism were assessed. Proteoglycan release into medium was determined by dye precipitation with 1,9-dimethylmethylene blue, synthesis of proMMP-1 (interstitial procollagenase) and proMMP-3 (prostromelysin 1) by Western blotting, and collagen degradation activity using FITC labeled collagen. In vivo experimental OA was induced in the knee joints of 15 Japanese adult white rabbits by partial lateral meniscectomy. Ten rabbits were orally administered 200 or 400 mg/kg/day of CPA-926 from the day of surgery for 14 days. The size of the macroscopic erosive area on the femoral condyle and tibial plateau was measured, and cartilage destruction was histologically evaluated. Collagenolytic activities in synovial fluid were measured using FITC labeled collagen as a substrate. Results. In vitro, esculetin inhibited the IL-1α induced release of proteoglycan into the medium in a dose dependent manner. The collagenolytic activities in cartilage explant medium induced by IL-1α were also suppressed with the addition of 33-100 μM esculetin (p = 0.0209 at 33 and 100 μM, p = 0.0202 at 66 μM). Western blotting of cartilage explant medium showed a decrease in the levels of proMMP-1 and proMMP-3 in the medium by treatment with esculetin. In vivo: At 14 days after surgery, the femoral condyle and tibial plateau in the control group showed macroscopic erosions of cartilage. Compared with the control group, the rabbits treated with CPA-926 at the dose of 400 mg/kg exhibited reduction of the size of the erosive area on the tibial plateau (p = 0.009). Histological evaluation indicated protection against the development of destructive changes in the tibial plateau cartilage at a dose of 200 mg/kg (p = 0.0442) and 400 mg/kg (p = 0.0446) of CPA-926. Conclusion. These results indicate that esculetin inhibits matrix degradation in rabbit joint cartilage explants through the suppression of MMP synthesis, secretion, or activity. Prophylactic administration of its prodrug, CPA-926, appears to provide some protection against cartilage destruction in a short term rabbit experimental OA model.

AB - Objective. To investigate the in vitro effects of 6,7-dihydroxycoumarin (esculetin) on the production of matrix metalloproteinases (MMP) in rabbit articular cartilage, and the in vivo effects of orally administered CPA-926, a prodrug of esculetin, on cartilage destruction in rabbit experimental osteoarthritis (OA). Methods. In vitro studies were performed using rabbit articular cartilage explants. Esculetin 10-100 μM was added to cartilage explants in the presence or absence of interleukin 1α (IL-1α). Effects of esculetin on cartilage metabolism were assessed. Proteoglycan release into medium was determined by dye precipitation with 1,9-dimethylmethylene blue, synthesis of proMMP-1 (interstitial procollagenase) and proMMP-3 (prostromelysin 1) by Western blotting, and collagen degradation activity using FITC labeled collagen. In vivo experimental OA was induced in the knee joints of 15 Japanese adult white rabbits by partial lateral meniscectomy. Ten rabbits were orally administered 200 or 400 mg/kg/day of CPA-926 from the day of surgery for 14 days. The size of the macroscopic erosive area on the femoral condyle and tibial plateau was measured, and cartilage destruction was histologically evaluated. Collagenolytic activities in synovial fluid were measured using FITC labeled collagen as a substrate. Results. In vitro, esculetin inhibited the IL-1α induced release of proteoglycan into the medium in a dose dependent manner. The collagenolytic activities in cartilage explant medium induced by IL-1α were also suppressed with the addition of 33-100 μM esculetin (p = 0.0209 at 33 and 100 μM, p = 0.0202 at 66 μM). Western blotting of cartilage explant medium showed a decrease in the levels of proMMP-1 and proMMP-3 in the medium by treatment with esculetin. In vivo: At 14 days after surgery, the femoral condyle and tibial plateau in the control group showed macroscopic erosions of cartilage. Compared with the control group, the rabbits treated with CPA-926 at the dose of 400 mg/kg exhibited reduction of the size of the erosive area on the tibial plateau (p = 0.009). Histological evaluation indicated protection against the development of destructive changes in the tibial plateau cartilage at a dose of 200 mg/kg (p = 0.0442) and 400 mg/kg (p = 0.0446) of CPA-926. Conclusion. These results indicate that esculetin inhibits matrix degradation in rabbit joint cartilage explants through the suppression of MMP synthesis, secretion, or activity. Prophylactic administration of its prodrug, CPA-926, appears to provide some protection against cartilage destruction in a short term rabbit experimental OA model.

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