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Nonlethal Oxidant Injury to Human Retinal Pigment Epithelium Cells Causes Cell Membrane Blebbing but Decreased MMP-2 Activity

By Maria E. Marin-castaño, Karl G. Csaky and Scott W. Cousins


PURPOSE. This study was undertaken to determine whether transient or sustained nonlethal oxidant injury can induce RPE cell membrane blebbing and alter RPE expression of matrix metalloproteinase (MMP)-2 and type IV collagen, two molecules that are necessary for regulation of the turnover of the RPE basal lamina. METHODS. The ARPE-19 cell line stably expressing green fluorescent protein (GFP) targeted to the cell membrane was bleb injured by exposure to myeloperoxidase (MPO; 10 microunits) and H 2O 2 (100 �M). Sustained (�6 hours) or transient (up to 6 hours) exposure to MPO/H 2O 2 was evaluated. An MTS assay conversion and cell counts were used to detect cell viability. Supernatants and the cell homogenates were collected from cultured ARPE-19 to assess fluorescent GFP-derived blebs, MMP-2 protein by Western blot, MMP-2 activity by zymography

Year: 2013
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