Netrin-1促进氧糖剥夺后脑微血管内皮细胞存活

发布时间：2018-10-15 08:50

【摘要】：目的通过建立体外氧糖剥夺模型模拟缺血缺氧环境,观察轴突导向因子netrin-1对脑微血管内皮细胞氧糖剥夺损伤后的作用。方法体外培养脑微血管内皮细胞,免疫荧光法检测其netrin-1受体结直肠癌缺失基因(deleted in colorectal cancer,DCC)及uncoordinated(UNC)5H2的表达;并使用不同浓度netrin-1作用于氧糖剥夺损伤的脑微血管内皮细胞,Cell Counting Kit-8(CCK-8)试剂盒及Annexin V-FITC/PI细胞凋亡双染试剂盒检测其细胞活性。结果脑微血管内皮细胞表达UNC5H2受体,无DCC受体表达;与对照组相比,浓度为10、50、100 ng/m L的netrin-1作用于氧糖剥夺损伤的脑微血管内皮细胞后细胞活性增强(10 ng/m L:1.25±0.20,存活率63.33%,P0.05;50 ng/m L:1.41±0.17,存活率79.40%,P0.05;100 ng/m L:1.33±0.27,存活率75.98%,P0.05),且在其浓度为50 ng/m L时细胞活性最强。结论 Netrin-1对氧糖剥夺损伤的脑微血管内皮细胞具有剂量相关的保护作用,这种保护作用可能由其受体UNC5H2介导。
[Abstract]:Objective to investigate the effects of axon guidance factor (netrin-1) on cerebral microvascular endothelial cells (MECs) after oxygen-glucose deprivation injury by establishing an in vitro oxygen-glucose deprivation model. Methods brain microvascular endothelial cells were cultured in vitro. The expression of (deleted in colorectal cancer,DCC and uncoordinated (UNC) 5H2 was detected by immunofluorescence. The cell activity of, Cell Counting Kit-8 (CCK-8) kit and Annexin V-FITC/PI cell apoptosis double staining kit were detected by different concentrations of netrin-1 on brain microvascular endothelial cells (CCK-8) after oxygen glucose deprivation. Results brain microvascular endothelial cells expressed UNC5H2 receptor, but no DCC receptor expression. The cell activity of 10 ng/m / 50 100 ng/m / L netrin-1 was enhanced (10 ng/m L: 1.25 卤0.20, 63.33% P 0.05 ng/m L: 1.41 卤0.17, 79.40% P0.05 ng/m L 1 33 卤0.27, 75.98% P0.05), and the cell activity was the highest when the concentration of 10 ng/m L was 1.25 卤0.20, the survival rate was 63.33 ng/m L: 1.17, and the survival rate was 79.40% P0.05% 100 ng/m L 1. 33 卤0.27, and the survival rate was 75.98% (P0.05). Conclusion Netrin-1 has a dose-related protective effect on cerebral microvascular endothelial cells injured by oxygen glucose deprivation, which may be mediated by its receptor UNC5H2.
【作者单位】：中山大学附属第一医院神经科;
【基金】：国家自然科学基金(编号:81371276) 广东省自然科学基金自由申请项目(编号:2014A030313065) 中山大学青年教师培育计划基金(编号:13ykpy16)
【分类号】：R965

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