TNF-α促进内皮源性微体的数量与ICAM-1表达
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国家自然科学基金项目(11172178,11222223,11232010)


TNF-α promotes the quantity and ICAM-1 expression of endothelial microparticles
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    摘要:

    目的 探讨高张应变调控的肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)对内皮源性微体(endothelial microparticles,EMPs)数量与表面细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)表达的作用。方法 采用Flexercell细胞张应变加载系统对大鼠胸主动脉内皮细胞(endothelial cells,ECs)分别施加5%(模拟正常生理状态)和18%(模拟高血压状态)幅度的周期性张应变,加载频率均为1.25 Hz,加载持续时间为24 h,实时PCR检测不同幅度张应变条件下ECs的TNF-α mRNA表达水平。之后应用TNF-α刺激大鼠胸主动脉ECs,收集上清液,超速离心提取得到内皮源性微体(endothelial microparticles,EMPs);用亲脂性苯乙烯基(lipophilic styryl)以及透射电镜对EMPs进行形态鉴定;流式细胞术对TNF-α刺激产生的Annexin V 阳性EMPs进行计数,并检测EMPs表面ICAM-1的表达。结果 与5%正常张应变组相比,18%高张应变条件下ECs的TNF-α表达水平显著上升。TNF-α能够显著上调ECs产生Annexin V阳性的EMPs数量,且TNF-α刺激ECs产生的EMPs表面ICAM-1表达量显著增加。结论 高张应变条件下ECs高表达TNF-α可能介导了EMPs产生和表面ICAM-1高表达。研究结果为后续探讨EMPs在血管重建力学生物学机制中的作用提供新的实验证据。

    Abstract:

    Objective To study the role of cyclic strain-modulated tumor necrosis factor-α (TNF-α) played in the quantity and intercellular cell adhesion molecule-1(ICAM-1) expression of endothelial microparticles (EMPs). Methods The endothelial cells (ECs) primarily cultured from rat aorta were applied with 5% cyclic strain (to simulate normal physiological condition) and 18% cyclic strain (to simulate hyper-tension condition), respectively, by using FX-4000T cyclic stain loading system for 24 hours at the loading frequency of 1.25 Hz. The mRNA expression of TNF-α under different amplitudes of cyclic strain was determined by real time-PCR. The TNF-α was then used to stimulate the ECs from rat aorta, and the supernatants were collected and ultracentrifuged to get endothelial microparticles (EMPs), which were then identified by lipophilic styryl membrane staining and transmission electron microscope for morphological identification. The quantities of Annexin V positive EMPs under TNF-α stimulation were counted by flow cytometer and ICAM-1 expression on EMPs was detected as well. Results Compared with the 5% normal cyclic strain, under 18% high cyclic strain condition,the mRNA expression of TNF-α in ECs increased significantly. TNF-α could then significantly up-regulate the production of Annexin V positive EMPs and promote the expression of ICAM-1 on EMPs. Conclusions The over-expression of TNF-α in ECs under high cyclic strain might mediate the high production of EMPs and over-expression of ICAM-1 on EMPs. The research findings will provide new experiment evidence for further studying the role of EPCs in the mechanobiological mechanism of vascular remodeling.

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吴垒磊,庄飞,韩悦,刘妍妍,齐颖新,姜宗来. TNF-α促进内皮源性微体的数量与ICAM-1表达[J].医用生物力学,2015,30(5):385-391

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  • 收稿日期:2015-02-04
  • 最后修改日期:2015-03-18
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  • 在线发布日期: 2015-10-28
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