Catalpol protects rat pheochromocytoma cells against oxygen and glucose deprivation-induced injury.
OBJECTIVE
Catalpol has been identified to have neuroprotective effect on gerbils subjected to transient global cerebral ischemia. However, the mechanism that catalpol prevents ischemia is still unclear. In the present study, PC12 cells, exposed to oxygen and glucose deprivation (OGD) followed by reperfusion, were used as an in vitro model of ischemia. The protective effects of catalpol were investigated in ischemic-induced apoptosis in PC12 cells.


METHODS
After OGD for 3 hours and reoxygenation for 18 hours, cell survival was quantified by the reduction of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were determined using commercially available kits. Caspase-3 assay was performed using caspase-3 assay kit. Microplate reader was used to detect the intensities of rhodamine123 (Rh123) and reactive oxygen species (ROS). The level of Bcl-2 protein was measured by flow cytometry.


RESULTS
Catalpol attenuated ischemia-induced apoptotic death via preventing the decrease in the level of Bcl-2 protein and the activities of SOD and GSH-PX, inhibiting the reduction of mitochondrial membrane potential and suppressing activation of caspase-3.


CONCLUSIONS
The results suggest that the catalpol has the potential to prevent ischemic-induced apoptosis.
Wang
Zhuo
Z
Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116023, China.
An
Li-Jia
LJ
Duan
Yan-Long
YL
Li
Ya-Chen
YC
Jiang
Bo
B
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002452:Cell Count; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000627:therapeutic use*; D005947:Glucose / Q000172:deficiency*; D005960:Glucosides / Q000627:therapeutic use*; D005979:Glutathione Peroxidase / Q000378:metabolism; D000860:Hypoxia / Q000517:prevention & control*; D057888:Iridoid Glucosides; D039823:Iridoids / Q000627:therapeutic use*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D016716:PC12 Cells; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism
D004365:Drugs, Chinese Herbal; D005960:Glucosides; D057888:Iridoid Glucosides; D039823:Iridoids; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; C078040:catalpol; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D053148:Caspase 3; D005947:Glucose