Expression of degradative enzymes and protease inhibitors in corneas with keratoconus

Lili Zhou, Shoichi Sawaguchi, Sally S. Twining, Joel Sugar, Robert S Feder, Beatrice Y.J.T. Yue*

*Corresponding author for this work

Research output: Contribution to journalArticle

177 Scopus citations

Abstract

PURPOSE. Keratoconus is characterized by thinning and scarring of the central region of the cornea. Previous research showed that, in corneas obtained from patients with keratoconus, lysosomal enzyme activities are elevated, whereas levels of protease inhibitors such as α1-proteinase inhibitor are reduced. This study was undertaken to examine further the expression of a spectrum of proteolytic enzymes and protease inhibitors. METHODS. Corneal buttons were collected from patients with keratoconus, healthy subjects, and patients with other corneal diseases. Immunohistochemical staining was performed on paraffin sections. Enzymatic assays and western blot analysis were carried out for cathepsins B and G. In addition, an in situ zymography procedure was used to examine the gelatin- and casein-digesting activities in corneas with keratoconus. RESULTS. An enhanced staining was found with antibodies to cathepsins B and G. Enzymatic assays and western blotting confirmed that the levels of these two enzymes were elevated in corneas with keratoconus. No alteration was noted with any of the matrix metalloproteinase (MMP) family members and other enzymes and inhibitors examined, although in situ zymography did indicate an increase in net gelatin- and casein-digesting activities in corneas with keratoconus. These activities were mostly abolished by inhibitors for serine and cysteine proteinases, but not by those for MMPs and aspartic proteinases. CONCLUSIONS. Levels of cathepsins B and G are increased in corneas with keratoconus. These enzymes may contribute to the heightened in situ gelatin- and casein- digesting activities, leading to abnormalities in keratoconus.

Original languageEnglish (US)
Pages (from-to)1117-1124
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume39
Issue number7
StatePublished - Jan 1 1998

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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    Zhou, L., Sawaguchi, S., Twining, S. S., Sugar, J., Feder, R. S., & Yue, B. Y. J. T. (1998). Expression of degradative enzymes and protease inhibitors in corneas with keratoconus. Investigative Ophthalmology and Visual Science, 39(7), 1117-1124.