血管内皮细胞损伤时TLR9激活与ICAM-1、E-选择素表达的研究

TLR9 Activation and Expression of ICAM-1,E-selectin in Vascular Enthelial Cell Damage

目的:通过CpG ODN诱导血管内皮细胞炎症损伤,观察其TLR9 mRNA以及细胞间粘附分子-1(ICAM-1)、E-选择素表达的变化,探讨炎症反应时血管内皮损伤的可能机制。方法 :使用10μM CpG ODN(实验组)和生理盐水(对照组)分别处理正常人脐静脉血管内皮细胞(HUVEC),分别于0 h、6 h、12 h、24 h、48 h、72 h时间点,以RT-qPCR法检测TLR9 mRNA的表达,免疫细胞化学法检测ICAM-1、E-选择素表达情况。结果:TLR9 mRNA表达在实验组6 h、12 h、48 h、72 h时水平明显较对照组高(P<0.05),在6h、48h表达水平明显高于其他时间组;实验组ICAM-1和E-选择素表达水平除0h时外,均明显高于对照组,其中ICAM-1在24h组时最高,E-选择素12h表达水平最高。结论:Cp G ODN损伤血管内皮细胞,激活TLR9,可呈双峰样高水平表达,且ICAM-1和E-选择素均显著升高。

Objective ： To investigate TLR9 mRNA, intercellular adhesion molecule-1 and E-selectin expression in the vascular endothelial cells inflammatory injury induced by Cp G ODN, explore possible mechanisms of vascular endothelial cell damage during inflammation. Methods ： Normal human umbilical vein endothelial cells（HUVEC）were treated with 10μM CpG ODN（experimental group） and normal saline（control group） respectively. TLR9 mRNA expression are tested by RT-q PCR, ICAM-1, E-selectin expression are tested by immunocytochemistry at 0 h, 6 h,12 h, 24 h, 48 h, 72 h. Results ： TLR9 mRNA expressions in the experiment group were significantly higher than that of control group at 6 h, 12 h, 48 h, 72 h. TLR9 mRNA expression of 6h and 48 h were significantly higher than 0 h, 12 h, 24 h, 72 h in the experiment group. ICAM-1 and E-selection expression were significantly higher than those of control group at all times expect 0 h, and the former expressed highest level at 24 h. The latter expressed highest level at 12 h. Conclusion ： TLR9 of HUVECs can be activated by Cp G ODN, which express a ＂bimodal ＂ high level, and ICAM-1 and E-selection expressed high.