人肾小球系膜细胞葡萄糖转运蛋白的鉴定及功能研究

摘要

Objective　To evaluate the role of glucose transporter 1 (GLUT1) in the glucose uptake of glomerular mesangial cells.rnMethods　Cultured human glomerular mesangial cells were used. The expressions of glucose transporter 1 mRNA and protein were detected with reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry and immunofluorescence staining. Glucose uptake was determined with 2-deoxy-［3H］-D-glucose uptake assay.rnResults　The expressions of GLUT1 mRNA and proteins were detected in human mesangial cells. Glucose uptake and its kinetics assay showed that GLUT1 is a functional glucose transporter in cultured human mesangial cells, and that its function could be inhibited by the specific inhibitor, Phloretin. rnConclusion　GLUT1 is the predominant glucose transporter in human mesangial cells, which has the kinetic characteristics of high affinity and low capacity for D-glucose. This suggests that in order for mesangial cells to take up excessive quantities of glucose, as in diabetes, changes in glucose transporter expression, translocation or activity may be required.%目的研究葡萄糖转运蛋白在肾小球系膜细胞糖摄取中的作用。rn方法体外培养人肾小球系膜细胞, 采用RT-PCR,细胞免疫荧光染色及流式细胞仪分析技术检测葡萄糖转运蛋白1(GLUT1)mRNA和蛋白质的表达。用2-脱氧-葡萄糖摄入法与根皮素抑制实验对GLUT1的功能进行测定。rn结果 RT-PCR,细胞免疫荧光染色及流式细胞仪分析证实系膜细胞存在GLUT1mRNA和蛋白质的表达糖摄取实验及其动力学分析表明系膜细胞GLUT1是一种高亲和性糖转运蛋白, 其功能能被根皮素所抑制。rn结论 GLUT1是人肾小球系膜细胞的主要葡萄糖转运蛋白,这为进一步研究GLUT1在糖尿病肾病发病机理中的作用奠定了基础。