海带多糖L01对人脐静脉内皮细胞表达和分泌t-PA的影响

Effect of Polysaccharide of Laminaria L01 on adrenaline-stimulated t-PA expression and secretion in cultured human umbilical vein endothelial cells

目的:利用人脐静脉内皮细胞体外培养模型,探讨肾上腺素刺激状态下海带多糖L01对内皮细胞的保护作用,以及其对其分泌的组织型纤溶酶原激活物(t-PA)的影响,从内皮细胞抗血栓功能方面探讨海带多糖L01对心血管系统的保护作用。方法:体外培养人脐静脉内皮细胞系ECV-304,用肾上腺素刺激24h、48h、72h采样,ELISA法测定HUVECs培养上清液中的t-PA含量,逆转录多聚酶链反应(RT-PCR)检测HUVEC细胞内t-PA mRNA的表达,其扩增产物经电泳后密度扫描及半定量分析。结果:肾上腺素模型组HUVECs培养液中t-PA(24h、48h)含量明显增加,;单给海带多糖对HUVECs t-PA分泌无影响。海带多糖L01在肾上腺素刺激下给药24h,48h后t-PA抗原分泌明显降低(P<0.01),且具有一定的量效关系,但HUVECst-PAmRNA表达各组均无明显差异。结论:肾上腺素可刺激内皮细胞分泌t-PA抗原,海带多糖L01对血管内皮具有一定的保护作用,减少肾上腺素刺激下HUVECs分泌t-PA,而对t-PAmRNA表达无明显影响。

Obeject： To study the effects of Polysaccharide of Laminaria L01 on adrenaline - stimulated tissue plasminogen activator（t-PA） expression and secretion in cultured human umbilical vein endothelial cells （HUVECs）. Methods： Human umbilical vein endothelial cell line ECV-304 exposed to different concentration of Polysaccharide of Laminaria （L01） joined with adrenaline cultured for 24 hours, 48 hours, 72 hourso The levels of t-PA in culture supernatans were measured by enzyme-linked immunosorbent assay （ELISA）; Reverse transcriptase chain reaction（RT-PCR） was used to detect the expression of t-PA messenger RNA（mRNA） in HUVECs. Its expression products were subjected to density scanning after electrophoresis and half-quantiative analysis to reflect the variety of the transcripition level. Results： The model group levels of t-PA in cultured supemants increased significantly for 24h, 48h （P 〈 0.01）. The effect were reduced by different concentration of L01 for 24 and 48h （P 〈 0.01）, and showed some dose-denpendence manner, but the levels of expression of t-PA mRNA was not siginificantly different all the time （P 〉 0.05）. Conclusion： L01 could reduce the effects of adrenaline - stimulated tissue plasminogen activator（t-PA） secretion on cultured human umbilical vein endothelial cells （HUVECs）. L01 could not only have the determinate protective function to the vascular endothelial cells, but also increase fibrinolytic activity and facilitate spontaneous thrombolysis finally.