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Tobias Meyer-ter-Vehn, Susanne Gebhardt, Walter Sebald, Mathias Buttmann, Franz Grehn, Günther Schlunck, Petra Knaus; p38 Inhibitors Prevent TGF-β–Induced Myofibroblast Transdifferentiation in Human Tenon Fibroblasts. Invest. Ophthalmol. Vis. Sci. 2006;47(4):1500-1509. doi: 10.1167/iovs.05-0361.
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purpose. The role of mitogen-activated protein kinase (MAPK) pathways in TGF-β–induced myofibroblast transdifferentiation of human tenon fibroblasts (HTFs) was investigated to identify potential pharmacologic targets for the inhibition of scarring after glaucoma surgery.
methods. TGF-β–dependent activation of Smad2, p38, and Erk-1/2 was examined by Western blot analysis. TGF-β–induced mRNA expression of collagen Iα1, fibronectin, and the myofibroblast transdifferentiation marker alpha smooth muscle actin (α-SMA) was analyzed by real-time RT-PCR. α-SMA protein expression and subcellular distribution were determined by Western blot analysis and immunofluorescence cytochemistry. Fibroblast contractility was assessed in three-dimensional collagen gel contraction assays, stress fiber assembly with rhodamine-phalloidin stains, and confocal microscopy. Cell proliferation was measured with an MTT assay. Specific pharmacologic kinase inhibitors were used to characterize the involvement of MAPK-dependent pathways.
results. TGF-β stimulation of HTF induced a rapid and transient activation of Smad2 and Erk, whereas p38 activation was biphasic and sustained. After 24 hours of TGF-β stimulation, increased levels of collagen Iα1, fibronectin, and α-SMA transcripts were detected. After 3 days of stimulation, HTF displayed increased α-SMA protein levels, enhanced contractility, and assembly of actin stress fibers. TGF-β also induced HTF proliferation. Specific p38 inhibitors prevented all these aspects of TGF-β–induced myofibroblastic transdifferentiation.
conclusions. Pharmacologic inhibition of p38 abrogates TGF-β–induced myofibroblast transdifferentiation, reduces extracellular matrix protein expression and HTF proliferation, and may therefore serve to inhibit scarring after glaucoma surgery.
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