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 language | English (US) |
|---|---|
| Pages (from-to) | 449-459 |
| Number of pages | 11 |
| Journal | Experimental eye research |
| Volume | 73 |
| Issue number | 4 |
| DOIs | |
| State | Published - Jan 1 2001 |
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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 journal › Article
}
TY - JOUR
T1 - Regulation of MMP-9 production by human corneal epithelial cells
AU - Li, De Quan
AU - Lokeshwar, Balakrishna L.
AU - Solomon, Abraham
AU - Monroy, Dagoberto
AU - Ji, Zhonghua
AU - Pflugfelder, Stephen C.
PY - 2001/1/1
Y1 - 2001/1/1
N2 - 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.
AB - 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.
KW - Corneal epithelial cells
KW - Gelatinase
KW - IL-1β
KW - MMP-9
KW - Matrix metalloproteinase
KW - TNF-α
UR - http://www.scopus.com/inward/record.url?scp=0035172290&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035172290&partnerID=8YFLogxK
U2 - 10.1006/exer.2001.1054
DO - 10.1006/exer.2001.1054
M3 - Article
C2 - 11825017
AN - SCOPUS:0035172290
VL - 73
SP - 449
EP - 459
JO - Experimental Eye Research
JF - Experimental Eye Research
SN - 0014-4835
IS - 4
ER -