血管生成素1对大鼠血-脊髓屏障功能的增强作用

摘要

目的:探讨血管生成素1 (Ang1)对大鼠血-脊髓屏障功能的增强作用,及表皮生长因子受体通路底物8(Eps8)在此增强过程的作用.方法:分离、培养大鼠脊髓微血管内皮细胞(SCMEC),建立体外血-脊髓屏障模型.以Ang 1处理的SCMEC为Ang1处理组,未处理的为对照组.于Ang1处理后于不同时间点(0h、4h、8h、12h)测定各组跨内皮电阻(TEER)值,比较2组Eps8蛋白的表达水平.再将SCMEC分为对照siR-NA组、对照siRNA+Ang1组、Eps8 siRNA组及Eps8 siRNA+Angl组,分别给予siRNA干扰Eps8表达及Ang 1处理,比较不同组间Eps8蛋白的表达水平与TEER值.结果:Ang1处理组4h、8h、12 h SCMEC的TEER值均明显高于对照组(P＜0.05),Eps8蛋白的表达水平均高于对照组(P＜0.05).Eps8 siRNA组SCMEC中Eps8蛋白的表达水平明显低于对照siRNA组(P＜0.05),而与Eps8 siRNA+Ang1组间差异无统计学意义(P＞0.05);Eps8 siRNA组不同时间点TEER值均明显低于对照siRNA组(P＜ 0.05);Eps8 siRNA+Ang1组与Eps8 siRNA组间TEER值差异无统计学意义(P＞0.05).结论:Ang1可增强大鼠血-脊髓屏障的功能,其机制可能与Eps8有关.%Objective:To investigate the effect ofAngiopoietin 1 (Ang1) on the function of blood spinal cord barrier (BSB) in rats.Methods:Spinal cord microvessel endothelial cells (SCMECs) of rats were separated and cultured to build the three-dimensional external blood spinal cord barrier model.SCMECs in Ang 1 group were treated with Ang 1 and those in control group were not treated with Ang 1.And then,SCMECs were divided into control siRNA group,control siRNA+Ang group 1,Eps8 siRNA group and Eps8 siRNA+Angl group,in which siRNA was added to interfere Eps8 expression and/or Ang 1 treatment.The transepithelial electrical resistance (TEER) values and the expression of EGF receptor pathway substrate 8 (Eps8) were detected at 0,4,8 and 12 h after treatment.Results:The TEER values and the Eps8 expression of Angl group at 4 h,8 h and 12 h were significantly higher than those in the control group (P＜0.05).The Eps8 expression in the Eps8 siRNA group was significantly lower than that in the control siRNA group (P＜0.05).The TEER values of siRNA group at 4 h,8 h and 12 h were significantly lower than those in the control siRNA group (P＜0.05).Conclusion:Ang1 has an effect on the permeability and barrier function ofBSB.Eps8 maybe plays an important role in it.