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R.C. Tripathi, B.J. Tripathi, J. Chen, J. Li, K.V. Chalam, S. Gotsis, V.A. Pakalnis; Growth Factors in Secondary Aqueous Humor Modulate Trabecular Cell Expression of MMP-3 . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1167.
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Purpose: To determine the mRNA and protein expression of matrix metalloprotease (MMP)-3 (stromelysin-1) by trabecular cells treated with growth factors present in secondary aqueous humor encountered in glaucomatous disease of the eye. Methods: Serum-starved, first passage porcine trabecular cells were incubated from 48 hours to 7 days in medium containing the combination and concentration of growth factors that are present in secondary aqueous humor and, as controls, in serum-free medium. We extracted total RNA, performed RT-PCR using primer pairs specific for porcine MMP-3 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and quantified the products. We utilized Western blotting to detect and quantify MMP-3 protein. Results: Compared to controls, expression of MMP-3 mRNA by trabecular cells was down-regulated 70% after incubation for 48 hours as well as 7 days in medium containing growth factors present in secondary aqueous humor. MMP-3 protein was not detectable by western blotting. Conclusions: Matrix metalloproteases have a crucial role in the regulated turnover of the extracellular matrix (ECM) in the trabecular meshwork. The down-regulation of MMP-3 mRNA by trabecular cells exposed to growth factors present in secondary aqueous humor implies a decreased proteolytic activity that contributes to accumulation of components of the ECM, such as glycoproteins and proteoglycans. Our findings indicate that induction of MMP-3 gene expression in the trabecular meshwork of glaucomatous eyes could effectively reduce buildup of ECM molecules in the aqueous outflow pathway to increase outflow facility.
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