December 1997
Volume 38, Issue 13
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Articles  |   December 1997
Prostaglandins increase matrix metalloproteinase release from human ciliary smooth muscle cells.
Author Affiliations
  • R N Weinreb
    Glaucoma Center, University of California San Diego, La Jolla 92093, USA.
  • K Kashiwagi
    Glaucoma Center, University of California San Diego, La Jolla 92093, USA.
  • F Kashiwagi
    Glaucoma Center, University of California San Diego, La Jolla 92093, USA.
  • S Tsukahara
    Glaucoma Center, University of California San Diego, La Jolla 92093, USA.
  • J D Lindsey
    Glaucoma Center, University of California San Diego, La Jolla 92093, USA.
Investigative Ophthalmology & Visual Science December 1997, Vol.38, 2772-2780. doi:
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    • Get Citation

      R N Weinreb, K Kashiwagi, F Kashiwagi, S Tsukahara, J D Lindsey; Prostaglandins increase matrix metalloproteinase release from human ciliary smooth muscle cells.. Invest. Ophthalmol. Vis. Sci. 1997;38(13):2772-2780.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: To identify matrix metalloproteinases (MMPs) released by ciliary smooth muscle cells in vitro and to determine whether MMP release is altered by exposure to prostaglandins (PGs). METHODS: Human ciliary smooth muscle cells were grown to confluence in monolayer cultures and treated with PGF2 alpha, 11-deoxy-PGE1, or PhXA85 (the nonesterified analogue of PhXA41) for 12 to 72 hours. The activity of MMP in the medium was assayed using gelatin and casein zymography. Identification of the specific MMP associated with each band was made by Western blot analysis. Band intensity, which reflects activity, was measured with a scanning laser densitometer. RESULTS: Three major bands appeared in the gelatin zymographs at positions corresponding to molecular weights of 62 kDa, 68 kDa, and 97 kDa. A single band at 50 kDa predominated in the casein zymograms. Substitution of EDTA for calcium and zinc in the development solution eliminated the appearance of these bands, indicating that they reflect MMP activity. Immunoblotting, using MMP-specific antibodies, confirmed that the three bands in the gelatin zymographs were MMP-1, MMP-2, and MMP-9, respectively; the single band in the casein zymographs was MMP-3. Treatment with 200 nM PGF2 alpha, 11-deoxy-PGE1, or PhXA85 for 72 hours increased the combined density scores for MMP-1 and MMP-2 by 37%, 64%, and 27%; the density scores for MMP-9 by 268%, 253%, and 125%; and the density scores for MMP-3 by 35%, 71%, and 22%, respectively. CONCLUSIONS: These results indicate that ciliary smooth muscle cells can secrete MMP-1, MMP-2, MMP-3, and MMP-9. In addition, exposure to PGF2 alpha, 11-deoxy-PGE1, or PhXA85 increases production of all four MMPs. These observations support the hypothesis that increased MMP production by ciliary muscle cells has a role in increasing uveoscleral outflow facility after topical PG administration.

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