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Young Jeung Park, Yeoun-Hee Kim, Se-Hyung Lee, Jae-Chang Jung, Kyoo Won Lee; Effect of Cyclosporin A (Cys A) to the Matrix Metalloproteinase (MMP)-3 and MMP-13 on the Migration of Cultured Pterygial Fibroblast Cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2117.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effects of matrix metalloproteinase (MMP) 3 and MMP-13 expression in cultured human pterygial fibroblast cells according to the concentration and exposure time of Cyclosporine A (Cys A) after being scratched.
Scratched-human pterygial fibroblast cells were exposed to a Cys A concentrations of 1 µg/ml (0.0001%) and 100 µg/ml (0.01%) for 3 or 10 minutes and then cells were washed with D-PBS and re-incubated or exposed to a MMP-3 specific inhibitor (MMP-3 VII inhibitor) or MMP-9/13 inhibitor with serum depletion DMEM-F-12 (1:1) media for 48 hours. Cells were lysed for Western blotting or extracted mRNA for RT-PCR. Supernatant media were used to perform TCA precipitation for Western blotting. MMP-3 and MMP-13 expression of fibroblast after the stimulation were studied.
The cellular migrations were markedly suppressed by pre-treatment with Cys A in a time- and dose- dependent manner on the cultured pterygial fibroblast cells (P < 0.01). Interestingly, this migration was also blocked by MMP-3 specific inhibitor or MMP-9-13 inhibitor (P < 0.01). Furthermore, pre-treatment of Cys A reduced MMP-3 mRNA (P < 0.01) and MMP-13 protein (P < 0.02) expression level.
Previous studies reported that the expression of several types of MMPs was increased in pterygium, and Cys A can be effective in preventing recurrences after primary pterygium surgery. Our results suggested that Cys A inhibited cell migration after being scratched by reducing expression of MMPs. These findings reveal therapeutic potential for pterygium progression.
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