Glucose transporters in human renal proximal tubular cells isolated from the urine of patients with non-insulin-dependent diabetes
Glucose transporters in human renal proximal tubular cells isolated from the urine of patients with non-insulin-dependent diabetes
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DOI：
10.2337/diabetes.54.12.3427
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发表时间：
2005-12-01
期刊：
影响因子：
7.7
通讯作者：
Brown, J
中科院分区：
文献类型：
--
作者：
Rahmoune, H;Thompson, PW;Brown, J
被肾小球过滤的大部分葡萄糖被近端曲管上皮的葡萄糖转运体重新吸收。然而，在2型糖尿病等疾病中研究这一点一直很困难，因为无法从没有肾脏活检的患者身上分离出原始肾脏细胞。我们报道了一种从新鲜尿液中分离和原代培养人近端肾小管上皮细胞(HEPTECs)的方法。初级分离株高度丰富和分化，表达特有的近端小管表型标记。经过6次传代培养，它们继续表达近端小管标志物CD13/氨基肽酶-N、钠葡萄糖共转运体(SGLT)2和碱性磷酸酶，其表达方式与从肾活检组织中分离的人近端细胞相似。在高血糖环境中，从2型糖尿病患者分离的HEPTEC比来自健康个体的细胞显著表达更多的SGLT2和促进葡萄糖转运蛋白GLUT2。我们还发现，与健康对照组相比，从2型糖尿病患者分离的HEPTECs对肾脏葡萄糖的摄取显著增加。我们的发现首次表明，在人类细胞模型中，肾脏葡萄糖转运体表达和活性增加与2型糖尿病有关。
The bulk of glucose that is filtered by the renal glomerulus is reabsorbed by the glucose transporters of the proximal convoluted tubular epithelium. However, it has been difficult to investigate this in diseases such as type 2 diabetes because of the inability to isolate primary renal cells from patients without a renal biopsy. We report here a method for the immunomagnetic isolation and novel primary culture of human exfoliated proximal tubular epithelial cells (HEPTECs) from fresh urine. The primary isolates are highly enriched and differentiated and express characteristic proximal tubular phenotypic markers. They continue to express the proximal tubular markers CD13/aminopeptidase-N, sodium glucose cotransporter (SGLT) 2, and alkaline phosphatase through up to six subsequent subcultures in a similar way to human proximal cells isolated from renal biopsies. In a hyperglycemic environment, HEPTECs isolated from patients with type 2 diabetes expressed significantly more SGLT2 and the facilitative glucose transporter GLUT2 than cells from healthy individuals. We also demonstrated a markedly increased renal glucose uptake in HEPTECs isolated from patients with type 2 diabetes compared with healthy control subjects. Our findings indicate for the first time in a human cellular model that increased renal glucose transporter expression and activity is associated with type 2 diabetes.