有机阴离子转运蛋白1在人血管平滑肌细胞表达的意义

Detection of Organic Anion Transporter 1 in Human Vascular Smooth Muscle Cells and Its Clinical Significance

目的检测有机阴离子转运蛋白1(OAT1) mRNA和蛋白质在人血管平滑肌细胞的表达以及尿酸刺激对其表达的影响。方法分离培养人脐动脉血管平滑肌细胞,提取细胞总RNA,RT-PCR方法扩增特异性OAT1cDNA片断,制备探针,用Northern blot方法检测OAT1 mRNA的表达,提取细胞膜蛋白和总蛋白,用Western blot方法检测OAT1的蛋白表达;用800μmol/L的尿酸刺激细胞,观察OAT1的表达变化。结果从人血管平滑肌细胞检测到OAT1 mRNA和蛋白的表达。尿酸可以明显增强OAT1的表达。结论人血管平滑肌细胞有明确的OAT1表达,OAT1可能部分介导了血管平滑肌细胞对尿酸的摄取过程。

[Objective]To detect expression of organic anion transporter 1 （OATI） mRNA and protein in human vascular smooth muscle cells and find the effects of uric acid to its expression. [Methods]Total RNA was isolated from cultured human vascular smooth muscle cells which was obtained from arteria umbilicalis. The specific human OATI cDNA fragment was amplified by RT-PCR and a probe was produced using this am- plified product. The OAT1 mRNA was detected by Northern blot and its protein expression level was measured by Western blot in human vascular smooth muscle cells, At the same time, we also detected OAT1 mRNA and protein expression in human vascular smooth muscle cells with or without uric acid stimulation. [Results]The OAT1 mRNA and protein were detectible in human vascular smooth muscle cells, and uric acid could significantly induce OAT1 mRNA and protein upregulation. [Conclusion]The OAT1 is exactly expressed in human vascular smooth muscle cells. This transporter may be involved in uric acid uptake.