Abstract Objective To explore the influence of oxidative stress induced by hydrogen peroxide (H2O2) on age-related maculopathy susceptibility 2 (ARMS2) in the transcription and protein levels of human retinal pigment epithelial (ARPE-19) cells, and to investigate the effects of oxidative stress on the ARMS2 gene in retinal pigment epithelial cells. Methods Experimental study. Hydrogen peroxide in concentrations of 0, 100, 300, 500, 700 μmol/L was used for ARPE-19 cells for specified time periods, and then methyl thiazolyhetrazolium (MTF) was used to detect the effects at 2 and 4 hours. Changes of ARMS2 in transcription and protein levels were detected by Realtime-PCR and immunofluorescence. The comparisons which between groups were analyzed by One-way ANOVA. The least significant difference was used to compare the means. The correlation between H2O2 concentration and cell activity were measured with estimates of curve. Results With concentrations of 0-700 μmol/L of hydrogen peroxide for 4 hours, the optical density were 0.531±0.037, 0.370+0.017, 0.371±0.016, 0.33±0.006, and 0.297±0.012, respectively, the activity of ARPE-19 decreased with increasing concentrations (F=6.782, P<0.01). The result of estimates of curve revealed that activity of ARPE-19 was negative correlated with the concentrations of hydrogen peroxide (r=-0.99, P=0.036). With concentrations of 100-700 μmol/L of hydrogen peroxide for 4 hours, Reahime-PCR results were 1.154±0.007, 1.324±0.022, 1.350±0.011, and 1.280±0.031, respectively, showed that hydrogen peroxide caused ARMS2 gene mRNA expression to increase (F=33.409, P=0.000). When the concentration of hydrogen peroxide increased to 300-500 μmol/L, ARMS2 gene mRNA expression reached a higher level, and then decreased with the increase of hydrogen peroxide. Immunofluorescence results were 7320±493, 14 300±848, 22 400±1596, 23 400±2405, and 19 200±561, respectively, showed that hydrogen peroxide caused ARMS2 protein expression to increase (F=22.843, P<0.01). When the concentration of hydrogen peroxide increased to 300-500 μmol/L, ARMS2 protein expression reached a higher level, and then decreased with an increase in hydrogen peroxide. The change tendency for transcription and protein levels were the same. Conclusion ARMS2 in ARPE-19 cells can increase in transcription and protein levels with an increase in a low concentration of hydrogen peroxide.The expression of ARMS2 decreases when the concentration of hydrogen peroxide higher than the tolerance concentration of ARPE-19.
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