脉络宁对原代大鼠海马神经元在氧糖剥夺/复氧中的保护作用

目的:观察脉络宁对氧糖剥夺/复氧(oxygen glucose deprivation/reoxygenation,OGD/R)诱导的SD大鼠原代培养海马神经元损伤的保护作用及可能机制。方法:原代培养从新生24 h内SD乳鼠取材的海马神经元,建立氧糖剥夺30 min/复氧8 h海马神经元损伤模型。实验分为正常对照组、OGD/R模型组、脉络宁低、中、高剂量组(5,10和20μg.mL-1)。采用Hoechest33258染色测定细胞凋亡,免疫细胞化学检测各组细胞中脑红蛋白(neuroglobin,NGB)及Bcl-2的表达,PCR检测其mRNA的表达情况。结果:与OGD/R模型组比较,各剂量脉络宁组神经元凋亡率均不同程度降低(P<0.01),且随药物浓度的升高而降低;NGB和Bcl-2蛋白表达不同程度地升高(P<0.01),且随药物浓度升高而升高;NGB及Bcl-2蛋白的mRNA亦升高(P<0.01)。结论:脉络宁发挥神经保护作用可能的机制之一是抑制OGD/R诱导的海马神经元细胞凋亡,促进NGB和Bcl-2蛋白的表达。

Neuroprotective effect of Mailuoning on injury after oxygen glucose deprivation/reoxygenation in primary cultures of rat hippocampal neurons

Objective: To determine the neuroprotective effect of Mailuoning against oxygen glucose deprivation/reoxygenation(OGD/R)-induced injury and the possible mechanisms.Methods: Primarily cultured hippocampal neurons from newborn SD rats(<24 h) were subjected to 30-min OGD/8-h R.The neurons were treated with Mailuoning 5,10,20 μg·mL-1.Apoptotic cells were analyzed by Hoechest33258 staining.Neuroglobin(NGB)and Bcl-2 expression were analyzed by immunocytochemistry.The mRNA expression of NGB and Bcl-2 was analyzed by RT-PCR.Results: Compared with OGD/R control,Mailuoning at 3 concentrations significantly decreased neuronal apoptosis(P<0.01) concentration-dependently,and increased the expression of NGB protein with the increasing of drug concentration(P<0.01).It also significantly increased the mRNA expression of NGB and Bcl-2(P<0.01).Conclusion: Mailuoning has a neuroprotective effect on OGD/R injury in hippocampal neurons,which is related to inhibiting apoptosis,and increasing NGB and Bcl-2 expression.