Liu Z, Li Y, Zhang J
Institute of Nephrology, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002.
Zhonghua Yi Xue Za Zhi. 1999 Oct;79(10):780-3.
To identify the glucose transporter-1 (GLUT1) in human glomerular mesangial cell and to evaluate the modulatory role of TGF-beta 1 and Rhein on the function of GLUT1 in mesangial cells.
GLUT1 in human glomerular mesangial cells was identified by RT-PCR for its mRNA expression. Cell surface protein of GLUT1 had been demonstrated by immunofluorescence staining and flow cytometry analysis using specific antibody. The uptake rate of glucose by mesangial cells was detected using the [3H]-2-DG. Glucose uptake specificity was confirmed by the co-incubation with unlabeled 2-DG or GLUT1 inhibitor Phlorhizin.
Human glomerular mesangial cells expressed functional GLUT1. TGF-beta 1 stimulated [3H]-2-DG uptake and GLUT mRNA expression in mesangial cells. The increase in [3H]-2-DG uptake and GLUT1 mRNA expression by TGF-beta 1 in mesangial cells were markedly abolished by the addition of Rhein in a dose dependent manner, while Rhein showed no effect on the glucose uptake in mesangial cells cultured in normal glucose medium.
Functional GLUT1 does present in human mesangial cells. TGF-beta 1 stimulates the glucose uptake by enhancing the GLUT1 mRNA expression in mesangial cells. This effect could be antagonized by Rhein. These results suggest that Rhein might be a hopeful drug for patients with diabetic nephropathy.
鉴定人肾小球系膜细胞中的葡萄糖转运蛋白1(GLUT1),并评估转化生长因子-β1(TGF-β1)和大黄酸对系膜细胞中GLUT1功能的调节作用。
通过逆转录聚合酶链反应(RT-PCR)检测人肾小球系膜细胞中GLUT1的mRNA表达,以鉴定GLUT1。使用特异性抗体通过免疫荧光染色和流式细胞术分析证实GLUT1的细胞表面蛋白。使用[3H]-2-脱氧葡萄糖([3H]-2-DG)检测系膜细胞对葡萄糖的摄取率。通过与未标记的2-DG或GLUT1抑制剂根皮苷共同孵育来确认葡萄糖摄取的特异性。
人肾小球系膜细胞表达功能性GLUT1。TGF-β1刺激系膜细胞中[3H]-2-DG摄取和GLUT mRNA表达。在系膜细胞中,加入大黄酸以剂量依赖性方式显著消除了TGF-β1引起的[3H]-2-DG摄取增加和GLUT1 mRNA表达增加,而大黄酸对在正常葡萄糖培养基中培养的系膜细胞的葡萄糖摄取没有影响。
功能性GLUT1确实存在于人系膜细胞中。TGF-β1通过增强系膜细胞中GLUT1 mRNA表达来刺激葡萄糖摄取。这种作用可被大黄酸拮抗。这些结果表明大黄酸可能是糖尿病肾病患者的一种有希望的药物。