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血管内皮细胞位于血流和血管壁之间,除受化学因素的调节外还受力学因素的影响。切应力可通过刺激相应的力学感受系统来调节内皮细胞某些基因的表达,其中包括诱导内皮细胞表达IL- 8。为了阐明MAPK信号途径中的ERK1/ 2信号通路是否参与调控低切应力上调人脐静脉内皮细胞IL- 8基因表达,采用Western blot分析了低切应力(4.2 0 dyne/ cm2 )处理不同时间内皮细胞ERK1/ 2的磷酸化水平及TPK抑制剂Genistein,MEK抑制剂PD980 5 9对其磷酸化的影响;采用定量RT- PCR检测内皮细胞经低切应力刺激或给予阻断剂后再行切应力刺激等处理后IL- 8基因的表达。结果显示:(1)低切应力处理可引起内皮细胞ERK1/ 2蛋白磷酸化水平上调,其磷酸化水平与切应力作用时间有关,具有快速、双向性的特点(在刺激10 min时达到高峰,2 h左右降至未刺激水平) ,阻断剂Genistein和PD980 5 9处理后,ERK1/ 2磷酸化水平与低切应力刺激10 min比较明显降低;(2 )阻断剂Genistein,PD980 5 9可显著抑制低切应力所致的内皮细胞IL- 8m RNA上调。结果表明低切应力可通过ERK1/ 2信号途径上调人脐静脉内皮细胞IL- 8基因的表达。
Vascular endothelial cells located between the bloodstream and the vessel wall, in addition to being regulated by chemical factors are also subject to mechanical factors. Shear stress regulates the expression of certain genes in endothelial cells by stimulating the corresponding mechanosensory system, including the induction of endothelial cells to express IL-8. To investigate whether ERK1 / 2 signaling pathway in MAPK signaling pathway is involved in the regulation of IL-8 gene expression in human umbilical vein endothelial cells by low-shear stress, Western blot was used to analyze the effect of low shear stress (4.2 0 dyne / cm2) The phosphorylation of ERK1 / 2 and the effect of TPK inhibitor Genistein and MEK inhibitor PD98059 on its phosphorylation were detected by quantitative RT-PCR. The endothelial cells were stimulated by low-shear stress or blocked with shear stress After treatment of IL-8 gene expression. The results showed that: (1) Low shear stress induced the phosphorylation of ERK1 / 2 in endothelial cells. The phosphorylation level of ERK1 / 2 was related to the shear stress time, which was rapid and bidirectional (peaked at 10 min, 2 h), the phosphorylation level of ERK1 / 2 was significantly decreased after 10 min treatment with Genistein and PD98059 (2) blockers Genistein and PD98059 Significant inhibition of low shear stress induced endothelial cells IL-8m RNA up-regulation. The results show that low shear stress can up-regulate the expression of IL-8 gene in human umbilical vein endothelial cells by ERK1 / 2 signaling pathway.