Protection Of Erythropoietin On Oxygen-glucose Deprivation Induced Injury In Primary Cultured Rat Cortical Neurons

Background: Erythropoietin has significant neuroprotective effect on hypoxic-ischemic brain damage.The mechanisms include anti-apoptosis effect,anti-oxidant effect,anti-inflammatory effect and stimulate angiogenesis and neurogenesis,etc.The results of the animal experiments or clinical researches are different.The differences may be related to duration of hypoxia,timing,dose and frequency of erythropoietin administration.Erythropoietin has shown significant neuroprotective potential in hypoxic-ischemic brain injury.Therefore,in order to realize the translation from laboratory to the bedside of erythropoietin,each of these factors mentioned above must be studied and evaluated.Objective: To explore the protective effects of rhEPO against hypoxic-ischemic brain injury and the effective dose and duration of action of rhEPO.Method: Isolate cortical neurons of neonatal Wistar rats and the cells were primarily cultured.Use the treptavidin-perosidase to evaluate the purification rate of neurons.Wistar rats after 7-11 days’ primary culture were randomly divided into control group,OGD group and rhEPO treatment group.The OGD model of cortical neurons was established in the rhEPO treatment group.After OGD,cortical neurons were treated with different doses of rhEPO(0.1、1、10、100U/ml)and were given with 10U/ml rhEPO on each time point after OGD 0h,24 h,48h.The changes of morphology of the cells were observed under microscope.Then detected cells’ survival rate by MTT assay.Results: 1.The positive rate of primary cultured cerebra cortex neurons was more than 90%,which met the experiment requirements.2.Compared with OGD group,the survival rate of neurons was higher in the rhEPO treatment group(0.1、1、10U/ml)(P<0.05).The protection was the most significant when the concentration of rhEPO was 10U/ml(P<0.05).And the cortical neurons’ survival rate was the highest when given with 10U/ml rhEPO after OGD immediately(P<0.05).Whereas,the survival rate of cells treated with the same dosage was not increased after OGD 24 h,48h.Conclusion: Erythropoietin protected neonatal rat cortical neurons against injury induced by oxygen-glucose deprivation.And the effective concentration was 0.1-10U/ml,of which the dose of 10U/ml was the most effective.Moreover,the protection was the most obvious when cortical neurons were given with 10U/ml rhEPO after OGD immediately.