Protective effects of edaravone against cobalt chloride-induced apoptosis in PC12 cells.
OBJECTIVE
To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms.


METHODS
PC12 cells impaired by CoCl2 were used as the cell model of hypoxia. MTT (methyl thiazolyl tetrazolium) was used to assay the viability of the PC12 cells exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer.


RESULTS
CoCl2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. CoCl2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 micromol/L.


CONCLUSIONS
These results indicated that Eda could protect PC12 cells from CoCl2-induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities.
Chen
Ji-Xiang
JX
Department of Neurology, Union hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. chen_jix@126.com
Zhao
Ting
T
Huang
Dan-Xia
DX
D000818:Animals; D016587:Antimutagenic Agents / Q000633:toxicity; D000983:Antipyrine / Q000031:analogs & derivatives* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002452:Cell Count / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D003035:Cobalt / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D000077553:Edaravone; D016166:Free Radical Scavengers / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D016716:PC12 Cells / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D016587:Antimutagenic Agents; D016166:Free Radical Scavengers; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D003035:Cobalt; D053148:Caspase 3; C018021:cobaltous chloride; D000077553:Edaravone; D000983:Antipyrine