Neuroprotective Effect Of Oxygen-glucose Deprivation Preconditioning On Neuron Cell

Objective:Ischemic Tolerance (IT) means when temporally dealed with the sublethal ischemic or hypoxia , tissue will be tolerant to another long time ischemic attack during a specified period. The temporally sublethal ischemic or hypoxia is called Ischemic Preconditioning (IPC). The IT and IPC may effectively motive auto protection as the damaging stressors, and may prolong tissue's ischemic tolerance ability, which appears death of tissue and cells, the diminution of infarction , obviously lighten of organ dysfunctions. IT usually begins several hours or one day after IPC, and keep on a week or longer. To collate with other culture methods of neonate rat, and improved in drawing the materials, digestion and culture environment, ect, we have got a more ideal result. And at the same time, we have set up a model of oxygen and sugar depriving, so as to further supply some loboratory data for understanding molecular mechanisms of cerebral ischemia and ischemic preconditioning.Method:culture rats cortical neuron with frontal lobe and temporal lobe in vitro.Using Neuronspecific enolase to identification neuron at 3 day to.this oxygen-glucose deprivation model neurons or grouping by cell culture to 12-14 days. Neurons were randomly divided into the following four groups:the control group(group I );the simple oxygen-glucose deprivation group(group II );the oxygen-glucose deprivation preconditioning group(group III);the fatal oxygen-glucose deprivation group(groupIV).and HSP70 c-fos bcl-2 caspase-3 expression in neurons was detected by immunohistochemical technique. Detection apoptosis by TUNEL and AO staining, preconditioning caused almost complete protection of neurons against apoptosis. Result:Culture 3 days, by way of neuronspecific enolase staining to raise neurons. Consepuence, all are the nerve cells. By typan blue staining, It is clear that the cell death of the fatal oxygen-glucose deprivation group(groupIV) is more severe than other groups(group I II III) (P<0.01) .The expression of HSP70/c-fos in both group II and groupIIIwere significantly higher than those of in group I and groupIV (P<0.01) ; The expression of caspase-3/bcl-2 in both group II and group I were no significantly difference (P>0.01) .nevertheless,the expression of bcl-2 in group III were significantly higher than those of in groupIV (P<0.01) .using TUNEL and AO to identification apoptosis,the expression of apoptosis neurons in group I and group II were no significantly difference (P >0.01) ; the expression of apoptosis neurons in groupIIIwere significantly higher than those of in groupIV (P>0.01) .Conclusion:These experiment to set up cortical neuron in vitro culture model, to confirm preconditioning was can stimulate barin tolerance mechanisms,which can cause a higher expression of HSP70 c-fos bcl-2,thus,Caspase-3 expresses to inhibition. The protect mechanism of neuroprotective effect of brain ischemic preconditioning is to raise the synthesis of new protein and to antagonistic neurons apoptosis.