流体剪切应力对人脐静脉内皮细胞Pim-1基因表达的影响

发布时间：2018-02-11 04:59

本文关键词： 内皮细胞 Pim- 剪切应力基因表达　出处：《中国临床解剖学杂志》2017年06期 　论文类型：期刊论文

【摘要】：目的探讨流体剪切应力对人脐静脉内皮细胞(HUVECs)中Pim-1基因表达的影响及可能的信号机制。方法酶消化法分离健康产妇新鲜脐静脉获取原代HUVECs,应用平行平板流动腔系统给HUVECs加载不同时间(1、4、6 h)和不同大小(0.5、1.5、3.0 Pa)的层流剪切应力,用实时定量RT-PCR检测Pim-1基因的表达水平,用蛋白质印迹法检测p-Akt和p-STAT3的表达水平,利用Wortmannin(PI3K抑制剂)、Deguelin(Akt抑制剂)和AG490(JAK抑制剂)信号阻断剂探讨信号转导途径。结果 HUVECs在1.5 Pa剪切应力作用4 h后Pim-1基因表达明显增加。不同强度的切应力均会刺激Pim-1基因的表达,其中3.0 Pa表达最强。切应力能显著激活p-Akt和p-STAT3的表达。AG490可以明显抑制Pim-1的表达,而Wortmannin和Deguelin增强Pim-1的表达。结论流体剪切应力可诱导HUVECs中Pim-1基因表达,其表达量与刺激时间和剪切应力的强度密切相关,这种作用可能通过JAK/STAT3和PI3K/Akt信号通路调节。
[Abstract]:Objective to investigate the effect of fluid shear stress on the expression of Pim-1 gene in human umbilical vein endothelial cells (HUVECs) and its signal mechanism. The laminar shear stress of HUVECs is loaded at different times (1 / 4 / 6 h) and 0. 5 / 1. 5 / 3. 0 Pa). The expression levels of Pim-1 gene and p-Akt and p-STAT3 were detected by real-time quantitative RT-PCR and Western blot respectively. The signal transduction pathway was studied by using Wortmannin(PI3K inhibitor Deguelinon Akt inhibitor and AG490(JAK inhibitor). Results the expression of Pim-1 gene increased significantly after HUVECs was exposed to 1.5 Pa shear stress for 4 h. The expression of Pim-1 gene was stimulated by shear stress of different intensities. Shear stress could significantly activate the expression of p-Akt and p-STAT3. AG490 could significantly inhibit the expression of Pim-1, while Wortmannin and Deguelin could enhance the expression of Pim-1. Conclusion fluid shear stress can induce the expression of Pim-1 gene in HUVECs. The expression level is closely related to the stimulation time and shear stress intensity, which may be regulated by JAK/STAT3 and PI3K/Akt signaling pathways.
【作者单位】：湖北医药学院附属随州医院转化医学研究中心;
【基金】：国家自然科学基金项目(31670961) 湖北省科技厅自然基金项目(2015CFB186)
【分类号】：R318.01

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