This research focused on the induction of cytotoxic effects by
danthron, a natural
anthraquinone derivative on C6 rat
glioma cells through exploring the means of cell death and the effects on mitochondrial function. We found that
danthron decreased the percentage of viable C6 cells and induced cell morphological changes in a dose-and time-dependent manner. The morphological and nuclei changes (
DAPI staining) in C6 cells were observed using a contrast-microscope and fluorescence microscopy, respectively. The results suggest that cell death of C6 cells which are induced by
danthron is closely related to apoptotic death.
Danthron decreased the level of mitochondrial membrane potential (ΔΨ( m )), stimulated the release of
cytochrome c from mitochondria to cytosol and promoted the levels of
caspase-9 and
caspase-3, or induced the release of AIF and Endo G from mitochondria. Based on both observations, we suggest that the
danthron-provoked apoptotic death of C6 cells is mediated through the mitochondria-dependent pathway. Furthermore, our results also indicated that
danthron triggered apoptosis through
reactive oxygen species (ROS) production which were increased after 1 h exposure of
danthron, which was reversed by the ROS scavenger
N-acetyl-L: -cysteine (NAC). As a consequence,
danthron-mediated cell death of C6 cells via ROS production, mitochondrial transmembrane potential collapse and releases of
cytochrome c, AIF and Endo G. Taken together,
danthron was demonstrated to be effective in killing C6 rat
glioma cells via the ROS-promoted and mitochondria-dependent apoptotic pathways.