Studies Of NO、SHD And Caspase-3 Correlation On Oxygen-glucose Deprivation Model In Neurons

Objective: To observe the changes of nitric oxide(NO) content, the activity of succinate dehydrogenase(SDH) and the expression of caspase-3 on oxygen-glucose deprivation model in rat neurons.Methods:(1)The cortical tissue was digested and the cells were planted in the medium with 10% fetal bovine serum,and the culture medium was replaced by neurobasal medium with 2%B27 after 4 hours.(2)The morphological changes were observed with phase-contrast microscope.Immunofluorescence was applied to identify the neurons.(3)The cortial neurons cultured for 8 days were randomly divided into three groups. There were the control group, the oxygen-glucose deprivation group and the aminoguanidine group.(4)The content of NO in culture medium was detected by nitrate reduction method.The activity of SDH in culture medium was determined by spectrophotometry.The expression of caspase-3 was measured by western blot.Results:(1) The neurons were cultured successfully, and the purity of neurons was above 90%.(2)The content of NO in oxygen-glucose deprivation group was significantly higher than the control group(P<0.05).Compared with the oxygen-glucose deprivation group,the content of NO in aminoguanidine group was significantly decreased.The difference was statistically significant(P<0.05).(3)The activity of SDH in oxygen-glucose deprivation group was significantly lower than the control group(P<0.05).Compared with the oxygen-glucose deprivation group, The activity of SDH in aminoguanidine group was significantly increased,the difference was statistically significant(P<0.05).(4)The expression of caspase-3 in oxygenglucose deprivation group were higher than the control group(P<0.05).Compared with oxygen-glucose deprivation group, the expression of caspase-3 in aminoguanidine group was significantly decreased, the difference was statistically significant(P<0.05).Conclusions: Aminoguanidine can inhibit the formation of NO, the decline of SDH activity and the protein expression of Caspase-3 in rat cortical neurons. So aminoguanidine can inhibit the apoptosis of the cortical neurons, which has a protective effect on the neurons.