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Daniel Meller, De–Quan Li, Scheffer C. G. Tseng; Regulation of Collagenase, Stromelysin, and Gelatinase B in Human Conjunctival and Conjunctivochalasis Fibroblasts by Interleukin-1β and Tumor Necrosis Factor-α. Invest. Ophthalmol. Vis. Sci. 2000;41(10):2922-2929.
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purpose. Overexpression and increased activities of matrix metalloproteinases
(MMPs) have recently been reported in cultured conjunctival fibroblasts
from patients with conjunctivochalasis. The role of inflammatory
cytokines in modulating expression of MMPs, their tissue inhibitors
(TIMPs), and urokinase plasminogen activator (uPA) as potential
contributors to the pathogenesis of conjunctivochalasis was
methods. Interleukin-1β (IL-1β) or tumor necrosis factor-α (TNF-α) was
added at 10 ng/ml to a serum-free medium. Expression of transcripts and
proteins of MMPs, TIMPs, and uPA by cultured normal human conjunctival
and conjunctivochalasis fibroblasts was determined by Northern
hybridization, enzyme-linked immunosorbent assay (ELISA) and Western
blot analysis, respectively. Gelatin and casein zymographies were
performed in serum-free conditioned media with and without the
respective enzyme inhibitors.
results. Without challenging the cells, conjunctivochalasis fibroblasts showed
mRNA and protein overexpression of MMP-1 and MMP-3 compared with normal
conjunctival fibroblasts, which showed minor or no expression of these
enzymes. IL-1β markedly and TNF-α to lesser extent increased mRNA
and protein expression of MMP-1 and MMP-3 in conjunctivochalasis
fibroblasts from 2 subjects when compared with normal conjunctival
fibroblasts from 2 subjects and with their nonstimulated counterparts.
In conjunctivochalasis fibroblasts and normal conjunctival fibroblasts,
TNF-α, but not IL-1β, induced a gelatinolytic activity of MMP-9,
which was further confirmed by Western blot analysis and ELISA.
Expression of MMP-2, TIMP-1, and TIMP-2 mRNA and protein was not
influenced by IL-1β or TNF-α, and no difference was found in the
gelatinolytic activity of MMP-2 between both cell types.
conclusions. Inflammatory cytokines such as IL-1β and TNF-α, which can
potentially be derived from the ocular surface and tears, may be
responsible for increased expression of MMPs in cultured
conjunctivochalasis fibroblasts. Ocular inflammation might be one
important denominator in the pathogenesis of
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