Regulation of Matrix Metalloproteinase in Müller Cells in Response to Lipopolysaccharide In Vitro

Abstract

Abstract: : Purpose: To investigate the secretion of matrix metalloproteinase (MMP) from Müller cells in response to lipopolysaccharide. Methods: Cultures of Müller cells were established from dissociated neonatal Wister rat retina as previously described (Hicks and Courtois, 1990). Cells were maintained in culture for three passages. They were treated with 1.0 µg/ml lipopolysaccharide (LPS) for 16 hours, after which cell supernatants were obtained and analyzed by zymography. Zymography was performed by acrylamide gel containing 0.1 mg/ml gelatin. Cell supernatants and cell lysates were examined by Western blotting to identify the type of protease. Results: Zymography showed that Müller cells secrete a protease without LPS stimulation and gelatin degradation activity was decreased by LPS stimulation. This protease was supposed to be matrix metalloproteinase 2 (MMP-2) from its molecular weight. Western blotting confirmed this protease was MMP-2. Conclusion: These data suggest that Müller cells secrete MMP in vitro. There might be other factors to regulate MMP production in response to LPS.

Keywords: 479 Muller cells • 560 retinal culture • 403 extracellular matrix