Advanced

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 and Stefan Lohmander, L. LU (1999) In Journal of Rheumatology 26(3). p.654-662
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... (More)

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.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animal model, Cartilage, Coumarin derivative, Esculetin, Matrix metalloproteinases, Osteoarthritis
in
Journal of Rheumatology
volume
26
issue
3
pages
9 pages
publisher
J Rheumatol Publ Co
external identifiers
  • Scopus:0032983177
ISSN
0315-162X
language
English
LU publication?
yes
id
06d0a79a-b3cf-4912-8717-2b7012ffa686
date added to LUP
2016-05-05 00:04:40
date last changed
2016-06-09 14:30:01
@misc{06d0a79a-b3cf-4912-8717-2b7012ffa686,
  abstract     = {<p>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.</p>},
  author       = {Yamada, Harumoto and Watanabe, Koju and Saito, Tsuyoshi and Hayashi, Haruhisa and Niitani, Yoshiaki and Kikuchi, Toshiyuki and Ito, Akira and Fujikawa, Kyosuke and Stefan Lohmander, L.},
  issn         = {0315-162X},
  keyword      = {Animal model,Cartilage,Coumarin derivative,Esculetin,Matrix metalloproteinases,Osteoarthritis},
  language     = {eng},
  number       = {3},
  pages        = {654--662},
  publisher    = {ARRAY(0x7f12608)},
  series       = {Journal of Rheumatology},
  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},
  volume       = {26},
  year         = {1999},
}