凝血酶诱导血管内皮细胞生长因子的表达及水蛭素的抑制作用

Influence of hirudin on thrombin-induced expression of VEGF in human umbilical vein endothelial cel1Is

目的:研究凝血酶对血管内皮生长因子(VEGF)表达的影响及重组水蛭素的作用。方法:将原代培养人脐静脉内皮细胞(HUVECs)的第2-5代与凝血酶(4.0u·mL-1)共同孵育,按不同分组加入水蛭素(6.0u·mL-1)和肝素(6.0u·mL-1)。在不同时间,通过MTT比色检测细胞生长活性,以检测细胞损伤程度;用逆转录聚合酶链反应(RT-PCR)方法评价VEGF mRNA的表达。结果:凝血酶对血管内皮细胞损伤的作用从2h即可出现,24h开 始减轻。凝血酶对VEGF的表达呈现双峰状:诱导后1h即有VEGF表达增高,2h减低后再次升高,24h达最高。 重组水蛭素可以有效地阻断凝血酶的上述作用。结论:凝血酶诱导HUVECs损伤并显著上调VEGF mRNA的表 达,该过程可被水蛭素有效抑制。

Objective:To study the effects of thrombin on VEGF (vascular endothelial growth factor) mRNA expression and the reverse effect of hirudin on it. Methods:The 2-5 generations of human umbilical vein endothelial cells (HUVEC) were co-incubated with thrombin(4. 0u·mL-1), then hirudin (6.0u·mL-1) and heparin (6.0u·mL-1) were added respectively to the related active groups. The cells growth activity was determined at different time by MTT colorimetric method, and reverse transcriptase polymerase chain reaction (RT-PCR) technique was adopted to evaluate VEGF mRNA expression. Results:The injury caused by thrombin on HUVEC may be occurred at 2h incubation and relieved after 24h incubation. The VEGF expression was increased after 1h of inducement, increased once again after 2h relief, and increased to maxium at 24h. The effect of thrombin on HUVEC stated above may be effectively inhibited by hirudin. Conclusion:That the injury caused by thrombin on HUVEC and the significant increase in VEGF mRNA expression caused by it may be effectively inhibited by hirudin.