Empirical Srudy Of The Polymorphisms Of Apolipoprotein E Gene (APOE) And The Excitatory Amino Acids After Injury

Objective To investigate the correlation between the polymorphisms of apolipoprotein E gene (poprotein:apoE,gene:APOE) and excitatory amino acids(EAAs).Methods Astrocytes were separated from APOE gene-knockout newborn rat cerebral cortex and identified by GFAP-FITC immunofluorescence staining. The recombinant plasmids (pEGFP-N1-APOEε2, pEGFP-N1-APOEε3, pEGFP-N1-APOEε4) were respectively transfected into the isolated astrocytes with Lipofectamine 2000.Subcellular localizatin of APOE and transfection efficiency were observed by fluorescent microscope,Western blot and RT-PCR technique was applied to detected the expression of APOE in astrocytes after transfected. Scratched models of three genotypes astrocytes were produced, Excitatory amino acids were measured by Amino acid analyzer, Apoptosis was detected by flow cytometry (FCM).Results The level of Glu and Asp after astrocytes scratched reach peak at 30 min,APOEε4 transfected astrocytes release higher level of Glu and Asp, compared with those in the other 2 groups at 30 and 60 min after injury (P<0.05).FCM showed the apoptosis ratio reach peak at at 24h,APOEε4 group was higher significantly than those in the other 2 groups at 12h and 24h after injury (P<0.05).Conclusion APOE affect EAAs released from the astrocytes after injury in an isoform-specific manner.APOEε4 carriers may have more severe deterioration and poorer prognosis due to EAAs toxicity.