Rahmoune Hassan, Thompson Paul W, Ward Joanna M, Smith Chari D, Hong Guizhu, Brown John
Clinical Pharmacology Unit, GlaxoSmithKline, Translational Medicine and Technology, Human Biomarkers Centre, Addenbrooke's Hospital, Cambridge, CB2 2GG, UK.
Diabetes. 2005 Dec;54(12):3427-34. doi: 10.2337/diabetes.54.12.3427.
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.
经肾小球滤过的大部分葡萄糖被近端曲管上皮细胞的葡萄糖转运体重吸收。然而，由于无法在不进行肾活检的情况下从患者中分离出原代肾细胞，在2型糖尿病等疾病中对此进行研究一直很困难。我们在此报告一种从新鲜尿液中免疫磁珠分离和新型原代培养人脱落近端肾小管上皮细胞（HEPTECs）的方法。原代分离物高度富集且分化，并表达特征性近端肾小管表型标志物。它们在后续多达六次传代培养中持续表达近端肾小管标志物CD13/氨肽酶-N、钠葡萄糖共转运蛋白（SGLT）2和碱性磷酸酶，其方式与从肾活检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HEPTECs比健康个体的细胞表达显著更多的SGLT2和易化性葡萄糖转运体GLUT2。我们还证明，与健康对照受试者相比，从2型糖尿病患者分离的HEPTECs中肾葡萄糖摄取明显增加。我们的研究结果首次在人类细胞模型中表明，肾葡萄糖转运体表达和活性增加与2型糖尿病有关。
