Figure 4
Disulfiram does not act via dopamine-metabolism on ADAM10 gene expression but has additional beneficial side effects in regard to Alzheimer’s disease. (a) SH-SY5Y cells were transiently transfected with a reporter vector for human ADAM10 (hADAM10) and incubated with 0.22 µM disulfiram or dopamine or norepinephrine in the indicated concentration. DMSO served as a solvent-control (values set to 100%). The experiment was conducted three-times independently in duplicates, values were normalized to protein content of the cell lysate and are represented as mean + SD. Statistical analysis: One Way ANOVA with Dunnett’s multiple comparisons test; ***p < 0.001; **p < 0.01. (b) To investigate a potential effect of disulfiram on A-beta aggregation, the fluorescent Thioflavin T test and human A-beta42 peptides were used. Peptides were supplemented with disulfiram as indicated or with the solvent at 37 °C. Three independent experiments were conducted (n ≥ 5) and slopes of fluorescence increase used for quantitation. The slopes obtained for the control were set to 100%. Statistical analysis was performed by One Way ANOVA with Bonferroni’s multiple comparisons test (***p < 0.001). (c) GSK3beta activity was measured by a commercial in vitro kit in two independent experiments performed in duplicates. Measurement values for control-treated reactions were set to 100% and values are presented as mean + SD. Statistical analysis was performed by One Way ANOVA with Dunnett’s multiple comparisons test (***p < 0.001; **p < 0.01).
