中文摘要：

目的通过研究5-单硝酸异山梨酯对斑马鱼血管发育的影响,探讨硝酸酯类药物促进血管新生的作用机制。方法以血管内皮具有绿色荧光蛋白标记的转基因斑马鱼胚胎为研究对象,将以100、300、500、1 000μg/ml 5-单硝酸异山梨酯处理的胚胎作为实验组,以0.5%二甲基亚砜（DMSO）处理的胚胎作为对照组,从受精卵形成后0.5～1 h开始药物处理,24～26 h后观察药物对转基因斑马鱼血管发育的影响。使用荧光显微镜观察新生血管状态并记录血管长度。分别从药物处理后0、6、12和24 h的斑马鱼胚胎中提取总RNA,采用实时定量RT-PCR方法检测血小板源性生长因子受体β（PDGFRβ）的表达变化。结果 5-单硝酸异山梨酯可促进斑马鱼血管发育（P〈0.01）,且在100、300、500μg/ml 3个浓度梯度范围内其促进作用与药物浓度成正比。实验组较对照组PDGFRβ表达量升高,在药物作用12和24 h时差异有统计学意义（P〈0.05,P〈0.01）。结论 5-单硝酸异山梨酯可上调PDGFRβ的表达,促进斑马鱼血管发育。

英文摘要：

Objective To investigate the pro-angiogenesis mechanism of isosorbide mononitrate by observeing its effect on the angiogenesis in Tg（flk1：EGFP） transgenic zebrafish.Methods The embryos incubated with isosorbide mononitrate（100,300,500,and 1 000 μg/ml） were taken as the experiment groups,and the embryos exposed to 0.5% DMSO were identified as the controls.The embryos were treated one hour before the postfertilization stage,and the status and length of the newly-formed blood vessels were observed under fluorescence microscope 24-26 h after treatments.The total RNAs were extracted from zebrafish embryos at 0,6,12,and 24 h after treatment;real-time quantitative RT-PCR was performed to examine the level of platelet-derived growth factor receptor β（PDGFRβ）.Results Isosorbide mononitrate greatly promoted the growth of the intersegmental vessels in zebrafish（P0.01）.It also dose-dependently promoted the growth of vascular at concentrations 100,300,and 500 μg/ml.The expression levels of PDGFRβ gene were increased in the experiment groups compared with DMSO group,with significant difference found at 12 h and 24 h after treatment（P0.05,P0.01）.Conclusion Our data suggest that isosorbide mononitrate can up-regulate PDGFRβ expression and promote angiogenesis in zebrafish.