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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 the changes of TLR9 mRNA, ICAM-1 and E-selectin in vascular endothelial cells induced by CpG ODN, and to explore the possible mechanism of vascular endothelial injury induced by inflammatory reaction. Methods: Human umbilical vein endothelial cells (HUVECs) were treated with 10μM CpG ODN (experimental group) and normal saline (control group) respectively. The cells were exposed to HUVEC at 0 h, 6 h, 12 h, 24 h, 48 h and 72 h The expression of TLR9 mRNA was detected by RT-qPCR and the expression of ICAM-1 and E-selectin by immunocytochemistry. Results: The expression of TLR9 mRNA in the experimental group was significantly higher than that in the control group at 6 h, 12 h, 48 h and 72 h (P <0.05), and the expression of TLR9 mRNA at 6 h and 48 h was significantly higher than that in the control group And E-selectin expression levels were significantly higher than those of the control group except 0h, ICAM-1 was the highest in 24h group and E-selectin 12h was the highest. CONCLUSION: CpG ODN can damage vascular endothelial cells and activate TLR9, showing a high level of bimodal expression, and both ICAM-1 and E-selectin are significantly increased.