Activation of extracellular signal-regulated kinase in trabecular meshwork cells.
A number of different agents, such as growth factors, cytokines and phorbol esters have been shown to modulate trabecular meshwork cell function. These studies were designed to evaluate the role extracellular signal-regulated kinase (ERK) pathway plays in mediating the responses to platelet-derived growth factor-BB (PDGF-BB) and phorbol 12-myristate 13-acetate (PMA) in trabecular meshwork cells. The human trabecular meshwork cell line, HTM-3, and the bovine trabecular meshwork (BTM) cells were treated with either PDGF-BB or PMA and the activation of ERK 1/2 evaluated. The effects of the MAP kinase kinase (MEK) inhibitor U0126, and the PKC inhibitor chelerythrine on ERK 1/2 were also determined. In a separate group of experiments, cells were treated with PDGF-BB or PMA and the secretion of matrix metalloproteinase-2 (MMP-2) evaluated. The addition of PDGF-BB or PMA produced time- and dose-dependent activation of ERK 1/2. Pretreatment with U0126 or chelerythrine significantly reduced ERK 1/2 activation induced by PDGF-BB or PMA. The addition of PDGF-BB or PMA stimulated the secretion of MMP-2. This secretory response was inhibited by pretreatment with the MEK inhibitor U0126. In trabecular meshwork cells, PDGF-BB and PMA activate ERK 1/2 by a PKC-dependent mechanism. Activation of ERK 1/2 by these agents in trabecular meshwork cells leads to the secretion of MMP-2. These studies provide evidence that ERK pathway is an important mechanism for integrating various signals that regulate trabecular function.
Shearer
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Ola B. Williams Glaucoma Therapeutic Development Center, Department of Ophthalmology, Medical University of South Carolina, 167 Ashley Avenue, Charleston, SC 29425, U.S.A.
Crosson
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D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D002070:Butadienes / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D020935:MAP Kinase Signaling System / Q000502:physiology*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors; D009570:Nitriles / Q000494:pharmacology; D010617:Phenanthridines / Q000494:pharmacology; D011233:Precipitin Tests; D020574:Proto-Oncogene Proteins c-sis / Q000502:physiology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014129:Trabecular Meshwork / Q000502:physiology*
D000470:Alkaloids; D053119:Benzophenanthridines; D002070:Butadienes; D004791:Enzyme Inhibitors; D009570:Nitriles; D010617:Phenanthridines; D020574:Proto-Oncogene Proteins c-sis; C113580:U 0126; C016299:chelerythrine; D020929:Mitogen-Activated Protein Kinase Kinases; D020778:Matrix Metalloproteinase 2; D013755:Tetradecanoylphorbol Acetate