目的用RT-PCR和Northern杂交技术,评价层流低切应力刺激对人脐静脉血管内皮细胞Toll样受体(TLR)表达的影响。方法将0.42Pa切应力为低切应力组处理内皮细胞1h,未作任何刺激的内皮细胞为对照组,从两组血管内皮细胞提取总RNA,采用RT-PCR和Northern杂交技术明确对照和切应力刺激1h人脐静脉血管内皮细胞TLR-2和TLR-4 mRNA的表达情况。结果RT-PCR和Northern杂交均显示,层流低切应力刺激1h后血管内皮细胞TLR-4表达增强、TLR-2几乎无变化。结论层流低切应力可引起人脐静脉内皮细胞TLR-4 mRNA表达增加,提示炎性信号受体TLR-4可能介导层流切应力诱导血管内皮细胞某些效应基因的表达。
Objective To assess the expression of Toll-like receptor-2(TLR-2) and TLR-4 in human umbilical vein endothelial cells (HUVECs) after low laminar shear stress stimulation. Methods Total RNA was isolated from HUVECs untreated and treated with 0.42 Pa for 1 hour. RT-PCR and Northern hybridization were used to determine the change of TLR-2 and TLR-4 mRNAs expression after 1-hour stimulation of laminar shear stress. Results The results of RT-PCR and Northem hybridization showed that the expression of TLR-4 in HUVECs was increased after 1-hour shear stress stimulation, while TLR-2 had no change after stimulation. Conclusion low laminar shear stress can induce TLR-4 mRNA expression, suggesting that the inflammatory signal receptor TLR-4 may be involved in laminar shear stress-induced effective gene expression in human vascular endothelial cells.