Objective: To investigate the regulation of PTEN on apoptosis and the active roles in blocking Ca2+ influx and neuroprotection.Methods: In the OGD model with anaerobic gas mixture and deoxygenated

glucose-free extracellular solution

shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively.Intracellular free calcium concentration were measured with Fura-2 AM fluorescent indicator

the expression of PTEN were analyzed by immunoblotting(blot) and neuron apoptosis were detected by AO/EB and Flow Cytometry.Results: Apoptosis rate and calcium concentration after OGD was significantly higher than that of normal group(P<0.05).But intracellular free calcium concentration and apoptosis with shRNAspten-GFP treatment after OGD was significantly lower than that in OGD group and PshGFP group(P<0.05).Conclusion: shRNAspten-GFP plasmid can been expressed successfully in cultured neurons.PTEN down-regulation could effectively suppress apoptosis occurrence induced by OGD and the increase of intracellular free calcium concentration

and then protect neuron from anoxia injury.【