Anthocyanin-rich Aronox extract from Aronia melanocarpa E protects against 7beta-hydroxycholesterol-induced apoptosis of endothelial cells.
OBJECTIVE
It has been previously reported that dietary polyphenolic compounds have a positive effect in cardiovascular diseases, although the underlying mechanisms remain unknown. In this study, the effects of Aronox, an anthocyanin-rich extract from Aronia melanocarpa E, on human umbilical vein endothelial cell (HUVEC) apoptosis was investigated.


METHODS
Apoptosis was measured in HUVEC pretreated with Aronox and than treated with 7beta-hydroxycholesterol (20 microg/ml). For the evaluation of apoptosis we used the cell binding of annexin V, the propidium iodide uptake method and the TUNEL assay. We also measured the generation of reactive oxygen species (ROS) in the cells, using fluorescent probe 2',7'-dichlorofluorescein acetate. Moreover, alteration of the mitochondrial membrane potential with subsequent cytochrome c release, activation of caspase-3, and the level of anti-apoptotic Bcl-2 were investigated.


RESULTS
Aronox significantly decreased 7beta-hydroxycholesterol-induced apoptosis and ROS generation, and ameliorated the collapse of the mitochondrial transmembrane potential with subsequent inhibition of cytochrome c release. Furthermore, down-regulation of Bcl-2 and up-regulation of caspase-3 activation were reversed by Aronox.


CONCLUSIONS
The protective action of the extract against apoptosis in endothelial cells may contribute to the potential benefits drawn from consumption of plant polyphenols.
Zapolska-Downar
Danuta
D
National Institute of Food and Nutrition, Warsaw, Poland. dzapolska@izz.waw.pl
Nowicka
Grazyna
G
Sygitowicz
Grazyna
G
Jarosz
Mirosław
M
D017304:Annexin A5 / Q000378:metabolism; D000872:Anthocyanins / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D045304:Cytochromes c / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D006888:Hydroxycholesterols / Q000633:toxicity; D020287:In Situ Nick-End Labeling; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D046936:Photinia / Q000737:chemistry*; D010936:Plant Extracts / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D017304:Annexin A5; D000872:Anthocyanins; D006888:Hydroxycholesterols; D010936:Plant Extracts; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; C011724:cholest-5-en-3 beta,7 alpha-diol; D045304:Cytochromes c; D053148:Caspase 3