Regulation of MMP-9 production by human corneal epithelial cells

De Quan Li, Balakrishna L. Lokeshwar, Abraham Solomon, Dagoberto Monroy, Zhonghua Ji, Stephen C. Pflugfelder

Research output: Contribution to journalArticle

188 Citations (Scopus)

Abstract

The matrix metalloproteinases. MMP-2 and MMP-9. are known to be critical extracellular-remodeling enzymes in wound healing and other diseases of the ocular surface. This study investigated the regulation of MMP-2 and MMP-9 in human corneal epithelial cells by growth factors and pro-inflammatory cytokines (IL-1β and TNF-α) they are exposed to, and by doxycycline, a medication used to treat ocular surface disease. Primary human corneal epithelial cell cultures were treated with one of the following cytokines (IL-1α, IL-1β, IL-6, IL-8, TNF-α) or growth factors (EGF, HGF, KGF, PDGF-BB, TGF-α, TGF-β), with or without their corresponding inhibitors. The conditioned media were collected after 24 hr for gelatin zymography and MMP-9 activity assay. Total RNA was extracted from the cells treated for 6 hr and was subjected to RT-PCR and Northern hybridization. Between the two gelatinases. MMP-2 and MMP-9, detected by zymography, the 92 kDa MMP-9 in the conditioned medium was markedly up-regulated by the pro-inflammatory cytokines. IL-1β and TNF-α. The MMP-9 protein and activity were dose-dependently stimulated by IL-1β or TNF-α at 0.1, 1.0 and 10 ng ml-1. This upregulation was nearly abolished by neutralizing antibodies (IL-1β and TNF-α) and by IL-1 receptor antagonist. Semi-quantitative RT-PCR and Northern hybridization disclosed that the MMP-9 transcript was also markedly up-regulated in a dose-dependent manner by IL-1β and TNF-α. Doxycycline (10 μg ml-1) suppressed MMP-9 protein level and activity, but not its mRNA, that was stimulated by IL-1β and TNF-α (1 ng ml-1). In contrast, the 72 kDa MMP-2 was not significantly modulated by any of these cytokines. In conclusion, production of MMP-9 is stimulated by the pro-inflammatory cytokines, IL-1β and TNF-α. These factors may play a role in the pathogenesis of MMP-9 mediated corneal matrix degradation. The efficacy of doxycycline in treating ocular surface diseases may be related to its ability to suppress MMP-9 production in the corneal epithelium.

Original languageEnglish (US)
Pages (from-to)449-459
Number of pages11
JournalExperimental eye research
Volume73
Issue number4
DOIs
StatePublished - Jan 1 2001

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Matrix Metalloproteinases
Epithelial Cells
Interleukin-1
Cytokines
Doxycycline
Eye Diseases
Conditioned Culture Medium
Intercellular Signaling Peptides and Proteins
Gelatinases
Polymerase Chain Reaction
Corneal Epithelium
Interleukin-1 Receptors
Gelatin
Neutralizing Antibodies
Interleukin-8
Epidermal Growth Factor
Wound Healing
Interleukin-6
Proteins

Keywords

  • Corneal epithelial cells
  • Gelatinase
  • IL-1β
  • MMP-9
  • Matrix metalloproteinase
  • TNF-α

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Regulation of MMP-9 production by human corneal epithelial cells. / Li, De Quan; Lokeshwar, Balakrishna L.; Solomon, Abraham; Monroy, Dagoberto; Ji, Zhonghua; Pflugfelder, Stephen C.

In: Experimental eye research, Vol. 73, No. 4, 01.01.2001, p. 449-459.

Research output: Contribution to journalArticle

Li, DQ, Lokeshwar, BL, Solomon, A, Monroy, D, Ji, Z & Pflugfelder, SC 2001, 'Regulation of MMP-9 production by human corneal epithelial cells', Experimental eye research, vol. 73, no. 4, pp. 449-459. https://doi.org/10.1006/exer.2001.1054
Li, De Quan ; Lokeshwar, Balakrishna L. ; Solomon, Abraham ; Monroy, Dagoberto ; Ji, Zhonghua ; Pflugfelder, Stephen C. / Regulation of MMP-9 production by human corneal epithelial cells. In: Experimental eye research. 2001 ; Vol. 73, No. 4. pp. 449-459.
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