The Effect Of Qma In PC12 Under Oxidative Stress Injury

Objective: To investigate the protective effect of polyamine analogues on the hydrogen peroxide-induced injury in rat pheochromocytoma cells(PC12 cells),and its possible mechanism.Methods: Different concentrations of hydrogen peroxide have cytotoxicity on PC12 cells.By detecting the damage rate with MTT,we determine the best concentrations of damage of hydrogen peroxide。By observing cell morphology by means of inverted microscope,we investigate the protective function of polyamine analogues、NAC、Nimodipine on PC12 cells against the cytotoxicity induced by hydrogen peroxide in the experiment.By Acridine Orange and Ethidium Bromide(AO/EB) double staining combined with living cells high intension imaging system to observe apoptosis morphology and characteristic. With rhodamine 123(Rh123) staining combined with living cells high intension imaging system and flow cytometry instrument to test cells mitochondrial membrane potential change and morphological characteristics. Through the DCFH- DA combined with flow cytometry instrument detection of intracellular ROS levels, as the polyamine compounds added antioxidant effect. By living cells high intension imaging system with immune histochemical method determined the expression of activated caspase 3, we study the possible mechanisms of antioxidant effect of polyamine compounds.Results:1. After being cultured with final concentration of 500μmol/L H2O2 after 24 h,the cell survival rate decreased significantly, and the final concentration of 1μmol/L 、 5μmol/L 、25μmol/L polyamine compounds as QMA can obviously improve the damaged cells form, the survival rate of PC12 cells increased significantly, and presents the concentration dependence.2. PC12 cells was damaged by 500μmol/L H2O2 for 24 hours, which triggered the intracellular oxidative stress response, causing cell oxidative stress injury, induced mitochondrial membrane potential decreased, and increased number of intracellular ROS.3. Polyamine compounds can significantly reduce the level of ROS in cells, inhibition of the mitochondrial membrane potential induced by H2O2 is reduced, it can significantly improve the H2O2 induction of apoptosis, inhibiting the expression of activated caspase- 3.Conclusions: By in vitro experiments show that polyamine compounds as QMA have protective effect on PC12 cell injury induced by H2O2, Which provide experiment basis for the furtherstudy on polyamine analogues in alzheimer’s disease.