内皮祖细胞对氧糖剥夺/复氧损伤神经元的保护作用

Protective effect of endothelial progenitor cells on oxygen and glucose deprivation / reoxygenation injured neurons

目的:研究内皮祖细胞(EPCs)共培养对氧糖剥夺/复氧(OGD/R)损伤神经元是否有保护作用。方法:分离培养新生小鼠的海马神经元,利用缺氧培养室建立神经元OGD/R模型。分离培养小鼠骨髓源性EPCs,采用Transwell小室建立EPCs与OGD/R损伤神经元的共培养系统。将神经元随机分为4组,即对照组、OGD/R组、EPCs共培养组及EPCs共培养并加入一氧化氮合酶(e NOS)抑制剂(L-NAME)组。神经元凋亡率用TUNEL荧光法检测。用Western blot法检测神经元caspase 3及内皮型e NOS活性。结果:OGD/R组神经元的细胞凋亡率和caspase 3活性较对照组显著增加。与OGD/R组相比,EPCs共培养组神经元凋亡情况显著降低(P<0.05),且e NOS磷酸化水平明显增高(P<0.05)。而与EPCs共培养组相比,EPCs+LNAME组神经元凋亡情况明显增加(P<0.05),且e NOS磷酸化水平也明显下降(P<0.05)。结论:与EPCs共培养可以通过提高神经元e NOS的活性来保护OGD/R损伤的神经元。

Objective： To investigate whether endothelial progenitor cells（EPCs） co-culture has protective effect on oxygen and glucose deprivation / reoxygenation（ OGD / R） injured neurons. Methods： Hippocampal neurons were isolated and cultured from newborn mice,and OGD / R model of neurons was performed using a hypoxia chamber.EPCs were isolated from the bone marrow of mice,and Transwell was used to build the co-culture system of EPCs and the OGD / R injured neurons. Neurons were randomly divided into four groups：the control group,the OGD / R group,the EPCs co-culture group and the EPCs ＋ nitric oxide synthase inhibitor（ L-NAME） group. Neuronal apoptosis rate was detected by TUNEL assay. Western blot was used to detect caspase 3 and endothelial nitric oxide synthase（ e NOS） activity. Results： Neuronal caspase 3 activity and apoptosis rate were significantly increased in the OGD / R group compared with the control group. Neuronal apoptosis was significantly decreased and e NOS phosphorylation was increased in the EPCs co-culture group compared with the OGD / R group. However,the EPCs ＋ L-NAME group showed increased neuronal apoptosis and decreased e NOS phosphorylation compared with the EPCs co-culture group. Conclusion： EPCs co-culture can protect OGD / R injured neurons by promoting the activation of e NOS.