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D.-N. Hu, M. Chen, J. E. Roberts, I. Aass, M. P. Patel, S. A. McCormick, R. Rosen; Protective Effect of HGF Against Hydrogen Peroxide Induced Damage in Human RPE Cells in vitro. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1845.
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To test the protective effects of hepatocyte growth factor (HGF) against hydrogen peroxide (H2O2) induced damage in human RPE cells in vitro.
Confluent cultured human retinal pigment epithelial (RPE) cells were subjected to oxidative stress induced by H2O2 ( 0.4 mM ). Quantity of viable cells was measured using the MTT assay. To determine the protective antioxidant effect of HGF, some wells of RPE cells were pretreated with 10-100 ng/ml HGF. Cells cultured without HGF or H2O2 were used as controls. All tests were performed intriplicate.
. HGF at concentrations of 10-100 ng/ml did not influence the amount of viable cells in confluent cultures of human RPE. H2O2 at 0.4 mM decreased viable cells to 22% of the negative control. Viable cells cultured with 10, 30 and 100 ng/ml HGF and H2O2 were 108%, 126% and 169% of cells cultured with H2O2 but without HGF. The difference between cells cultured with and without HGF was statistically significant at 30-100 ng/ml (0.05>P>0.01 at 30 ng/ml and P < 0.01 at 100 ng/ml). As compared to the controls (without H2O2), viable cells cultured with 10, 30 and 100 ng/ml HGF and H2O2 were 23.8%, 27.7% and 37.2% of the controls. All of these differences were statistically significant.
HGF at concentrations of 30-100 ng/ml has a partial protective effect against oxidative damage of human RPE cells induced by H2O2. Human RPE are constantly exposed to oxidative stress, such as sunlight, smoking and phagocytosis that results in the formation of reactive oxygen species which may damage these cells. HGF is a growth factor normally present in the eye (0.5-2.5 ng/ml in the aqueous humor and vitreous). High levels of exogenous HGF (30-100 ng/ml) have a protective effect on the RPE against oxidative stress.
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