Acute hyperglycemia results in dynamic changes in the superoxide anion concentration of HeLa cells.

<p>(A) 5x10⁵ HeLa cells were exposed to glucose concentrations ranging from 4 mmol/L (control) to 20 mmol/L and sequentially imaged. Representative image is 15 minutes post treatment. (B) Quantification of coelenterazine chemiluminescence at each time point for each sample (n = 3). (C) 5x10⁵ RIN-5F cells were exposed to glucose concentrations ranging from 4 mmol/L (control) to 20 mmol/L and sequentially imaged. Representative image is 15 minutes post treatment. (D) Quantification of coelenterazine chemiluminescence at each time point for each sample (n = 3). (E) 5x10⁵ INS-1 cells were exposed to glucose concentrations ranging from 4 mmol/L (control) to 20 mmol/L and sequentially imaged. Representative image is 15 minutes post treatment. (F) Quantification of coelenterazine chemiluminescence at each time point for each sample (n = 3). (G) 5x10⁵ HeLa cells were exposed to glucose concentrations ranging from 13.2 mmol/L (control) to 27.8 mmol/L and sequentially imaged. Representative image is 120 minutes post treatment. (H) Quantification of coelenterazine chemiluminescence for HeLa, RIN-5F and INS-1 cells treated as described in G at 120 minutes post-treatment (n = 3). (Error bars represent the mean ± SEM, * denotes a significant difference from the 13.2 mmol/L HeLa cells of P < 0.015, + denotes a significant difference from the 13.2 mmol/L RIN-5F cells of P < 0.015, and # denotes a significant difference from the 13.2 mmol/L INS-1 cells of P < 0.015.) (I) Dihydroethidium assay of 5x10⁵ HeLa cells exposed to glucose concentrations of 4 mmol/L (control) or 20 mmol/L with the quantification of 2-hydroxyethidium fluorescence as each time point minus the pre-treatment (-5 minutes) fluorescence intensity for each sample (n = 3). (Error bars represent the mean ± SEM, * denotes a significant difference from the 4 mmol/L control of P < 0.04.) (J) Coelenterazine assay of 5x10⁵ HeLa cells exposed to glucose concentrations of 4 mmol/L (control) or 20 mmol/L with the quantification as the cumulative area under the curve at each time point minus the pre-treatment (-5 minutes) chemiluminescence for each sample (n = 3). (Error bars represent the mean ± SEM, * denotes a significant difference from the 4 mmol/L control of P < 0.004.)