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Purpose: To determine whether (1) the in vitro expression of epithelial basement membrane components nidogen-1, nidogen-2, and perlecan by keratocytes, corneal fibroblasts, and myofibroblasts is modulated by cytokines/growth factors, and (2) perlecan protein is produced by stromal cells after photorefractive keratectomy.

Methods: Marker-verified rabbit keratocytes, corneal fibroblasts, myofibroblasts were stimulated with TGF-β1, IL-1α, IL-1β, TGF-β3, platelet-derived growth factor (PDGF)-AA, or PDGF-AB. Real-time quantitative RT-PCR was used to detect expression of nidogen-1, nidogen-2, and perlecan mRNAs. Western blotting evaluated changes in protein expression. Immunohistochemistry was performed on rabbit corneas for perlecan, alpha-smooth muscle actin, keratocan, vimentin, and CD45 at time points from 1 day to 1 month after photorefractive keratectomy (PRK).

Results: IL-1α or-1β significantly upregulated perlecan mRNA expression in keratocytes. TGF-β1 or-β3 markedly downregulated nidogen-1 or-2 mRNA expression in keratocytes. None of these cytokines had significant effects on nidogen-1,-2, or perlecan mRNA expression in corneal fibroblasts or myofibroblasts. IL-1α significantly upregulated, while TGF-β1 significantly downregulated, perlecan protein expression in keratocytes. Perlecan protein expression was upregulated in anterior stromal cells at 1 and 2 days after− 4.5 or− 9 diopters (D) PRK, but the subepithelial localization of perlecan became disrupted at 7 days and later time points in− 9-D PRK corneas when myofibroblasts populated the anterior stroma.

Conclusions: IL-1 and TGF-β1 have opposing effects on …