December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Degradation Of Timp-1 And Increased Gelatinase Activity In Human Stromal Cell Cultures Treated With Nitric Oxide By-products
Author Affiliations & Notes
  • MC Kenney
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • B Lin
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • M Chwa
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • M Saghizadeh
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • S Rosenberg
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • AV Ljubimov
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • DJ Brown
    Ophthalmology Research Cedars-Sinai Medical Center Los Angeles CA
  • Footnotes
    Commercial Relationships   M.C. Kenney, None; B. Lin, None; M. Chwa, None; M. Saghizadeh, None; S. Rosenberg, None; A.V. Ljubimov, None; D.J. Brown, None. Grant Identification: NIH EY06807, the Schoellerman Charitable Foundation, the Discovery Fund for Eye Research
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 3208. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      MC Kenney, B Lin, M Chwa, M Saghizadeh, S Rosenberg, AV Ljubimov, DJ Brown; Degradation Of Timp-1 And Increased Gelatinase Activity In Human Stromal Cell Cultures Treated With Nitric Oxide By-products . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3208.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose: To determine whether peroxynitrite and/or nitric oxide donors (SIN-1 and SNAP, respectively) altered the tissue inhibitor of metalloproteinase-1 (TIMP-1) and/or matrix metalloproteinase-2 (MMP-2) produced by normal and keratoconus stromal cells in vitro. Methods: A total of 22 normal and 24 keratoconus corneas were analyzed in this study. The RNA levels of TIMPs and MMPs from intact keratoconus and age-matched normal corneas were evaluated by semi-quantitative RT-PCR and Southern blotting. Normal and keratoconus stromal cells were incubated in vitro with 1 mM of SIN-1 or SNAP. RNA levels for TIMP-1, MMP-2, and beta-actin were measured by Northern blot analyses. Peroxynitrite and TIMP-1 levels were measured by Western blots and dot blot analyses. Culture media were analyzed by gelatin zymography and a collagenolytic fibril assay. Results: Keratoconus corneas had decreased TIMP-1 mRNA (p<0.005) and protein levels compared to normal intact corneas. Within 4 hours in SIN-1 treated cultures, there was an accumulation of peroxynitrite (NT positive proteins). After 18 hours, SIN-1 treated cultures showed degradation of the TIMP-1 protein and increased TIMP-1 RNA levels. The MMP-2 RNA levels increased with SIN-1 but remained the same with SNAP treatment. Media from the SIN-1 and SNAP treated samples had increased gelatinase activities. Conclusion: Peroxynitrite was associated with altered RNA levels for MMP-2/TIMP-1 and fragmentation of TIMP-1 protein. In light of the multiple cellular functions of TIMP-1 (i.e., inhibition of MMP activity, suppression of apoptosis) this relationship may play an important role in the pathogenesis of keratoconus.

Keywords: 450 keratoconus • 374 cornea: stroma and keratocytes 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×