基于EphA4/ephrinA3信号通路研究安寐丹对星形胶质细胞和神经元氧糖剥夺/复氧损伤的保护作用及机制OA

Objective:To investigate the protective effects and underlying mechanisms of Anmeidan(AMD)against oxygen-glucose deprivation/reoxygenation(OGD/R)-induced injury in astrocytes and neurons by regulating the EphA4/ephrinA3 signaling pathway.Methods:An invitro OGD/R model was established using mouse astrocytes(C8-D1A)and hippocampal neurons(HT22 cells).Control serum and AMD-containing serum were prepared.Cells were divided into control,OGD/R,OGD/R+AMD,and OGD/R+melatonin groups.Cell proliferation and viability were assessed via CCK-8 assay to determine the optimal AMD serum concentration.Apoptosis rates were measured by flow cytometry.Immunofluorescence staining was performed to evaluate fluorescence intensity of neuronal marker NeuN,astrocytic marker GFAP,and neurogenesis marker BrdU.Protein expression of ephrinA3,EphA4,EAAT1,EAAT2,and SYN was analyzed by Western blot.Cellular ultrastructure was observed via transmission electron microscopy(TEM).Results:Compared with the control group,the OGD/R group showed reduced fluorescence intensity of NeuN and GFAP(P<0.01),abnormal neuronal morphology,upregulated EphA4/ephrinA3 protein expression(P<0.01),and decreased EAAT1/EAAT2 levels(P<0.05,P<0.01).AMD treatment significantly enhanced GFAP and NeuN fluorescence intensity,improved neuronal morphology,downregulated EphA4/ephrinA3 expression(P<0.01),upregulated EAAT1,EAAT2,SYN(P<0.05),and reduced apoptosis rates(P<0.01).Conclusion:Anmeidan alleviates OGD/R-induced astrocyte and neuronal injury by downregulating the EphA4/ephrinA3 signaling pathway and modulating glutamate transport,which may underlie its therapeutic potential in sleep deprivation-related neural damage.