24-Epibrassinolide, a Phytosterol from the Brassinosteroid Family, Protects Dopaminergic Cells against MPP+-Induced Oxidative Stress and Apoptosis
Figure 4
Selective detection of O2•− by MitoSOX Red in neuronal PC12 cells. Once in the cells, this fluorogenic dye is oxidized by superoxide and exhibits red fluorescence. (a) Fluorescence microphotographs at 3-hour or 24-hour treatment. Cells were pretreated with 24-Epi or vehicle (Ctrl) for 3 h. Then, they were treated or not with MPP+ for 3 h or 24 h. A significant signal of MitoSOX Red is apparent in neuronal cells treated with MPP+. This signal is much less intense in cells exposed to vehicle (Ctrl) or 24-Epi alone (24-Epi). A marked reduction of red fluorescence is also clear in cells pretreated with 24-Epi and then treated with MPP+ (24-Epi + MPP+). DDC: cells were treated with 80 M DDC for 3 h or 24 h. 24-Epi + DDC: cells were pretreated with 10−9 M 24-Epi, then with 80 M DDC for 3 h or 24 h. (b) Semiquantitative image analysis of fluorescence at 3 h and 24 h. Nuclei were counterstained in blue with Hoechst 33342. Data are expressed as relative fluorescent units and are means ± S.E.M. Magnification 400x. . versus Ctrl, versus MPP+, and versus DDC.
