【摘要】：目的通過建立體外氧糖剝奪模型模擬缺血缺氧環(huán)境,觀察軸突導向因子netrin-1對腦微血管內(nèi)皮細胞氧糖剝奪損傷后的作用。方法體外培養(yǎng)腦微血管內(nèi)皮細胞,免疫熒光法檢測其netrin-1受體結(jié)直腸癌缺失基因(deleted in colorectal cancer,DCC)及uncoordinated(UNC)5H2的表達;并使用不同濃度netrin-1作用于氧糖剝奪損傷的腦微血管內(nèi)皮細胞,Cell Counting Kit-8(CCK-8)試劑盒及Annexin V-FITC/PI細胞凋亡雙染試劑盒檢測其細胞活性。結(jié)果腦微血管內(nèi)皮細胞表達UNC5H2受體,無DCC受體表達;與對照組相比,濃度為10、50、100 ng/m L的netrin-1作用于氧糖剝奪損傷的腦微血管內(nèi)皮細胞后細胞活性增強(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),且在其濃度為50 ng/m L時細胞活性最強。結(jié)論 Netrin-1對氧糖剝奪損傷的腦微血管內(nèi)皮細胞具有劑量相關的保護作用,這種保護作用可能由其受體UNC5H2介導。
[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 鹵0.20, 63.33% P 0.05 ng/m L: 1.41 鹵0.17, 79.40% P0.05 ng/m L 1 33 鹵0.27, 75.98% P0.05), and the cell activity was the highest when the concentration of 10 ng/m L was 1.25 鹵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 鹵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.
【作者單位】： 中山大學附屬第一醫(yī)院神經(jīng)科;
【基金】：國家自然科學基金(編號:81371276) 廣東省自然科學基金自由申請項目(編號:2014A030313065) 中山大學青年教師培育計劃基金(編號:13ykpy16)
【分類號】：R965

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本文編號：2272007