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(Investigative Ophthalmology and Visual Science. 2004;45:2998-3004.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-0051
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Dexamethasone Inhibition of IL-1–Induced Collagen Degradation by Corneal Fibroblasts in Three-Dimensional Culture

Ying Lu,1 Ken Fukuda,2 Yang Liu,1 Naoki Kumagai,1 and Teruo Nishida1

1From the Departments of Biomolecular Recognition and Ophthalmology and 2Ocular Pathophysiology, Yamaguchi University School of Medicine, Yamaguchi, Japan.

PURPOSE. Corticosteroids regulate the functions of inflammatory cells. The purpose of the present study was to investigate the effect of dexamethasone on collagen degradation by corneal fibroblasts, an underlying cause of corneal ulceration.

METHODS. Rabbit corneal fibroblasts were cultured in three-dimensional gels of type I collagen and in the absence or presence of IL-1ß or dexamethasone. The extent of collagen degradation was determined by measurement of the amount of hydroxyproline generated by acid-heat hydrolysis of culture supernatants. The expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) was evaluated by immunoblot analysis, gelatin zymography, and reverse transcription and real-time polymerase chain reaction. The phosphorylation of mitogen-activated protein kinases (MAPKs) in corneal fibroblasts was assessed by immunoblot analysis.

RESULTS. Dexamethasone inhibited IL-1ß–induced collagen degradation by corneal fibroblasts in a dose-dependent manner. Both the synthesis and activation of MMPs and the expression of TIMPs were inhibited by dexamethasone, as was the activity of plasmin in culture supernatants. Dexamethasone also inhibited the IL-1ß–induced phosphorylation of the MAPKs extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), but not that of p38.

CONCLUSIONS. Dexamethasone exerted multiple effects on the MMP-TIMP system in corneal fibroblasts and thereby inhibited IL-1ß–induced collagen degradation by these cells. Inhibition of the IL-1ß–induced activation of ERK and JNK may contribute to these effects of dexamethasone.




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