ERK1/2信号通路在内皮微粒诱导人脐静脉内皮细胞ICAM-1表达中的作用
Endothelial microparticle-induced ICAM-1 expression in human umbilical vein endothelial cells via ERK1/2 signaling pathway
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摘要: 目的:探讨细胞外信号调节激酶1/2（ERK1/2）信号通路在内皮微粒（EMPs）诱导人脐静脉内皮细胞（HUVECs）表达细胞间黏附分子-1（ICAM-1）中的作用。方法：体外培养HUVECs，选取生长良好的第4～5代细胞，分为EMPs不同浓度作用组、EMPs不同时间作用组及ERKl/2特异性抑制剂组。应用蛋白免疫印迹法（Western blot）检测ICAM-1和磷酸化ERK1/2蛋白的表达。用实时荧光定量PCR（qRT-PCR）检测ICAM-1 mRNA的表达。结果：EMPs作用HUVECs后，ICAM-1 mRNA和其蛋白以及磷酸化ERK1/2蛋白的表达显著高于对照组，且呈浓度和时间依赖性的关系（均P<0.01）；而ERKl/2特异性抑制剂PD98059对ICAM-1 mRNA和其蛋白以及磷酸化ERK1/2蛋白的表达，均有明显的抑制作用（均P<0.01）。结论：EMPs可诱导HUVECs中ICAM-1表达，其机制可能与激活ERK1/2信号通路有关。Abstract: AIM:To investigate the role of extracellular signal-regulated kinases 1/2 (ERK1/2) signaling pathway in endothelial microparticle (EMP)-induced expression of intercellular adhesion molecule 1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were cultured in vitro and fourth to fifth generation cells were chosen for the test. HUVECs were incubated with EMPs at different concentrations and at different times or pretreated with inhibitors of ERK1/2 (PD98059) for 60 min before incubation. Protein expression of ERK1/2 and ICAM-1 was measured by Western blot in HUVECs. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was used to analyze the mRNA expression of ICAM-1 in HUVECs. RESULTS: Exposure to EMPs caused a significant increase in mRNA and protein expressions of ICAM-1 and resulted in a rapid activation of ERK1/2 in a dose- and time-dependent manner (all P<0.01). The effect of EMPs was attenuated by PD98059 (all P<0.01). CONCLUSION: ERK1/2 signaling pathways are involved in EMP-induced expression of ICAM-1 in HUVECs.
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