Preliminary Investigation on the Mechanism of C_2C_(12) Cell Apoptosis induced by Hydrogen Peroxide

ObjectiveTo investigate the molecular mechanism that apoptosis of mouse embryonic myogenic cell line C 2C 12 cells was induced by oxidative stress.Apoptosis of mouse C 2C 12 cells was induced by exposure to 0.5 mmol/L hydrogen peroxide (H 2O 2) for different durations, and their apoptotic changes were detected by Hoechst 33258 fluorescence staining. The activities of caspase -3, -8, -9 were examined by caspase colorimetric assay kit and Western-blotting. The intracellular distribution of cytochrome C and its release from mitochondria were observed by indirect immunofluorescence and Western-blotting.Exposure to 0.5 mmol/L H 2O 2 for 24 hours markedly induced C 2C 12 cell apoptosis as shown by Hoechst 33258 fluorescence staining. The activities of caspase -3, -8, -9 significantly increased after 4 hours of H 2O 2 treatment, and reached their peaks at 8～12 hour. The release of cytochrome C from mitochondria to cytoplasm was detected after exposure to H 2O 2 for 1～2 hours.[Conclusions]Oxidative stress can induce apoptosis of C 2C 12 cells through simultaneous activation of mitochondria and death receptor signal pathways. This result provides new information for clinical prophylaxis and treatment of apoptosis related cardiovascular diseases.