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Shahid Husain, Farahdiba Jafri, Craig E. Crosson; Acute Effects of PGF2α on MMP-2 Secretion from Human Ciliary Muscle Cells: A PKC- and ERK-Dependent Process. Invest. Ophthalmol. Vis. Sci. 2005;46(5):1706-1713. doi: 10.1167/iovs.04-0993.
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purpose. Studies were designed to evaluate the cellular mechanisms associated with prostaglandin (PG)F2α-induced matrix metalloproteinase (MMP)-2 secretion from human ciliary muscle (HCM) cells.
methods. The secretion and activity of MMP-2 was determined by Western blot analysis and zymography, using conditioned medium and HCM cells. ERK1/2 activity was measured by in-gel kinase assay and Western blot analysis with anti-phospho-ERK1/2 antibodies.
results. PGF2α increased the secretion of MMP-2 in a dose-dependent manner with an EC50 of 2.7 × 10−8 M. The addition of 1 μM PGF2α also increased MMP-2 secretion in a time-dependent manner with maximum secretion occurring at 4 hours after administration. At 4 hours, the maximum increase in MMP-2 secretion and activity were 112% ± 32% and 88% ± 18%, respectively. The secretory action of PGF2α was inhibited by pretreatment with a protein kinase C (PKC) inhibitor, chelerythrine chloride; the FP receptor antagonist, AL-8810; and the MEK inhibitor, PD-98059. The addition of PGF2α and latanoprost acid increased ERK1/2 activity by 117% ± 12% and 75% ± 9%, respectively. The PGF2α- and latanoprost-acid–induced ERK1/2 activation was blocked by the presence of PKC inhibitors and downregulation of PKC by prolonged incubation with a phorbol ester.
conclusions. These data provide evidence that FP receptor activation leads to an increase in the secretion and activation of MMP-2 through PKC- and ERK1/2-dependent pathways. FP-agonist–induced activation of ERK1/2 was blocked by PKC inhibitors, indicating that PKC activation is required for ERK1/2 activation and MMP-2 secretion from HCM cells. In the ciliary muscle, the functional responses to ERK1/2 activation include secretion of MMP-2, supporting the hypothesis that increases in uveoscleral outflow facility induced by PG administration involves the secretion and activation of MMP-2.
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