螺旋藻激酶对人脐静脉内皮细胞t-PAmRNA、PAI-1mRNA表达的影响

Effects of Spirulina Kinase on the Expression of t-PAmRNA、PAI-1mRNA in human Umbilical Vein Endothelial Cell

目的研究螺旋藻激酶(Spirulina kinase,SPK)对人脐静脉内皮细胞t-PAmRNA、PAI-1mRNA表达的影响,探讨其抗血栓作用机制,为开发新型溶栓药物提供理论依据。方法体外培养人脐静脉内皮细胞(HUVECs),建立肾上腺素(Adr)损伤模型加入不同浓度的SPK,并以肝素(Hep)为阳性对照,与空白组对照,经过12,24,48 h后,逆转录多聚酶链式反应(RT-PCR),扩增产物经凝胶电泳分离后,凝胶图像分析仪照相和扫描,测定各组t-PAmRNA、PAI-1mRNA与内参基因三磷酸甘油醛脱氢酶(GAPDH)mRNA的相对表达量。结果在Adr损伤模型中,2 mg/ml SPK可以上调t-PAmRNA表达,超大剂量SPK降低t-PAmRNA表达。SPK还可以随着剂量的增加明显的抑制PAI-1mRNA的表达,效果强于大剂量的肝素。结论 SPK可能通过小部分活化t-PAmRNA的表达,抑制PAI-1mRNA的表达,从而达到抗血栓的作用。

Objective To study the effects of spirulina kinase from spirulina on the expression of t - PAmRNA, PAI- 1 mRNA in human umbilical vein endothelial cells （HUVECs） and to explore the mechanism of its anti - thrombotic and provide theoretical basis for the development of new thrombolytic drugs. Methods The third to fifth generation of HUVECs were incubated respective- ly with adrenaline （Adr） damage model - different concentrations of SPK and heparin （Hep） as a positive control, for 12, 24, 48hours. Meanwhile , cells were also accordingly exposed to 2mg/mlSPK alone. After reverse transcription - polymerase chain reac- tion （ RT - PCR ） , PCR products were separated by gel electrophoresis, and gel images were analyzed and scanned to measure the relative expression of t- PAmRNA, PAl -lmRNA with the reference gene GAPDHmRNA. Results In Adr injury models, 2rag/ mlSPK could increase the expression of t - PAmRNA, concentration of SPK to reduce the expression of t - PAmRNA. SPK also sig- nificantly inhibited the expression of PAI - 1 mRNA with increasing concentration. The effect was stronger than high concentrationof heparin. Conclusion SPK possibly activate the expression of t - PAmRNA, inhibit the expression of PAI- 1 mRNA to achieve the antithrombotic effect.