Simvastatin and benznidazole block NF-κB activation in Trypanosoma cruzi infection of EA.hy926 endothelial-like cells.

Endothelial-like (EA.hy926) and HUVEC cells were incubated with 5 μM simvastatin or 20 μM benznidazole 0.025% v/v final concentration. After 24 hours, the medium was replaced with drug-free medium, and the cells were at a MOI of 10 for 1 hour. Then, the cells were harvested at the time points indicated in the figure, and total protein expression was determined by Western blot using rabbit monoclonal antibodies against human IKK, p-IKK, IκB, and p-IκB. Representative blots are shown for p-IKK and IKK in untreated (A) and treated (C) cells and for p-IkB and IkB in untreated (B) and treated (D) cells. Bottom panels in A-D correspond to relative blot quantification using α-tubulin as a control. All controls were incubated with DMSO vehicle alone. The data are expressed as the mean ± SD from three independent experiments. One-way ANOVA and Tukey’s post-test analysis were used to find significant differences. ‡ p<0.001 vs. control; * p<0.05; *** p< 0.001 vs. T. cruzi. E. Representative photographs of the p65 localization by confocal microscopy after contact of EA.hy926 cells with T. cruzi for 1 hour with or without previous 5 μM simvastatin and 20 μM benznidazole treatment.