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.
N1 - Funding Information:
Supported in part by Public Health Service Research Grants EY11915 (SCP), Department of Health and Human Service, National Eye Institute, CA61038 (BLL), National Cancer Institute, Bethesda, MD, U.S.A., an unrestricted Grant from Research to Prevent Blindness and the Drs David and Maureen Smith Ocular Surface and Tear Research Fund.
PY - 2001
Y1 - 2001
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
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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 -