天津医药

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血小板微颗粒对人脐静脉内皮细胞VEGF表达的影响

解畅1,李曦铭2,贾红丹3,张迎怡4,毛用敏1,崔让庄1,丛洪良2   

  1. 1. 天津市胸科医院
    2. 天津胸科医院
    3. 河北省秦皇岛市第一医院
    4. 胸科医院心内科
  • 收稿日期:2011-09-30 修回日期:2011-10-28 出版日期:2012-02-15 发布日期:2012-02-15
  • 通讯作者: 丛洪良

Platelet-derived Microparticles Affects Vascular Endothelial Growth Factor Expression in Endothelial Cells

  • Received:2011-09-30 Revised:2011-10-28 Published:2012-02-15 Online:2012-02-15

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摘要： 摘要 目的：探讨血小板微颗粒(PMPs)影响内皮细胞血管内皮生长因子（VEGF）释放的可能机制及其临床意义。方法：常规培养人脐静脉内皮细胞（HUVECs）CRL-1730，配制不同干预浓度的PMPs与HUVECs共培养不同时间，应用半定量逆转录聚合酶链反应技术测定内皮细胞VEGF、VEGFⅡ型受体Flt/KDR、磷脂酰肌醇-3激酶(phosphatidylinositol-3 kinase，PI3K)以及胞外信号调节激酶(extracellular signal-regulated kinase，ERK) mRNA的表达水平。结果：（1）VEGF mRNA的表达水平在0 mg/L组显著高于PMPs10、30、50、100 mg/L干预组(均P<0.05)，VEGFⅡ型受体Flt/KDR mRNA表达水平在0 mg/L组显著低于PMPs10、30、50、100 mg/L (均P<0.05)。ERK mRNA表达水平在PMPs 50 mg/L组(1.141±0.093)显著高于0 mg/L组（0.749±0.205），P=0.004，PI3K mRNA表达水平在PMPs100 mg/L组(1.344±0.133)显著高于0 mg/L组(0.999±0.246)，P=0.004。（2）50 mg/L PMPs干预细胞，VEGF mRNA的表达水平在0 h组(0.746±0.179)显著高于24 h组(0.318 ±0.091)，P＜0.001，KDR mRNA的表达水平在0 h组显著低于PMPs干预4 h、24 h组（均P<0.05），PI3K mRNA的表达水平在0 h组(0.999±0.246)显著低于24 h组(2.622±1.213)，P=0.004。结论：PMPs可能通过VEGF受体，激活其下游信号转导通路，发挥促进血管和血管发生为基础的生物学效应。

关键词: 血小板活化, 内皮细胞, 血管内皮生长因子类, 1-磷脂酰肌醇3-激酶, 细胞外信号调节MAP激酶类, 信号传导

Abstract: Abstract Objective: To investigate the possible mechanism of platelet-derived microparticles (PMPs) effecting on vascular endothelial growth factor（VEGF）releasing and its clinical significance in endothelial cells.Methods: Human umbilical vein endothelial cells (HUVECs) CRL-1730 were commonly cultured. Different concentrations of PMPs intervene HUVECs and incubate for different time. We used semi-quantitative reverse transcription polymerase chain reaction techniques to measure VEGF、phosphatidylinositol-3 kinase(PI3K)、extracellular signal-regulated kinase(ERK)and VEGF type II receptor (KDR) mRNA levels. Results: (1)VEGF mRNA level in 0 mg/L group was significantly higher than the 10、30、50、100 PMPs mg/L groups(P<0.05,respectively).In contrast, Flt/KDR mRNA mRNA level in 0 mg/L group was significantly lower than the 10、30、50、100 mg/L PMPs groups(P<0.05,respectively).ERK mRNA level(1.141±0.093) in 50 mg/L PMPs group was significantly higher than that of the 0 mg/L group(0.749±0.205）,P=0.004.PI3K mRNA level(1.344±0.133) in 100mg/L PMPs group was significantly higher than that of the 0 mg/L group(0.999±0.246),P=0.004.(2)Just as the effect of different concentration PMPs on mRNA expression, VEGF mRNA level(0.746±0.179) in 0 hours group was significantly higher than that of the 24 hours intervention group(0.318 ±0.091),P＜0.001.KDR mRNA level in 0 hours group was significantly lower than the 4hours and 24hours groups (P<0.05, respectively).PI3K mRNA level(0.999±0.246) in 0 hours group was significantly lower than that of the 24hours group (2.622±1.213),P=0.004)..Conclusions: PMPs may induce the biological processes of blood vessel and angiogenesis via VEGF receptor and its downstream signal transduction pathways.

Key words: Platelet Activation, Endothelial Cells, Vascular Endothelial Growth Factors 1-Phosphatidylinositol 3-Kinase, Extracellular Signal-Regulated MAP Kinases, Signal Transduction