Brevilin A Inhibits VEGF-Induced Angiogenesis through ROS-Dependent Mitochondrial Dysfunction

Figure 6

Mito-TEMPOL and N-acetyl-cysteine relieve the consequences of Brevilin A on apoptosis. HUVECs were preincubated with or without Mito-TEMPOL (2 μM) and NAC (3 mM) for 30 min in the presence of VEGF (10 ng/mL), and further administered 12 h treatment with 10 μM BA, and finally subjected to the following analysis. (a) Morphological changes of confluent HUVEC cultures observed under a phase-contrast microscope (). (b) Flow cytometry evaluated different apoptotic phases via using AV-FITC/PI staining reagents (). (c) Bar graphs quantifying the relative distribution of HUVECs in early stage (AV⁺) or late apoptotic stages (AV⁺/PI⁺) (). (d) caspase-3 activation determination (). Western blotting was used to examine (e, f) the protein expression of Bax, Bcl-2, and cleaved caspase-3 (). The graphs summarize the data from at least three times independent experiments. Data are shown as the . One-way ANOVA with Bonferroni post-hoc test and , , and versus solvent (Sol).

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