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内皮祖细胞对氧糖剥夺/复氧损伤神经元的保护作用
作者：李子惠1  柏盈盈1  居胜红2 
单位：1. 东南大学医学院 分子与功能影像实验室, 江苏 南京 210009;
2. 东南大学附属中大医院 放射科, 江苏 南京 210009
关键词：内皮祖细胞 神经元 氧糖剥夺/复氧 共培养 内皮型一氧化氮合酶 
分类号：R743.3
出版年·卷·期（页码）：2015·34·第三期（347-352）
摘要：

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(eNOS)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eNOS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eNOS磷酸化水平明显增高(P< 0.05).而与EPCs共培养组相比,EPCs+L-NAME组神经元凋亡情况明显增加(P< 0.05),且eNOS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eNOS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(eNOS) 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 eNOS 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 eNOS phosphorylation compared with the EPCs co-culture group. Conclusion: EPCs co-culture can protect OGD/R injured neurons by promoting the activation of eNOS.
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