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A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells

ABSTRACT: The sodium-dependent glucose transporters 2 (SGLT2) plays important role in renal reabsorption of urinal glucose back to plasma for maintaining glucose homeostasis. The approval of SGLT2 inhibitors for treatment of type 2 diabetes highlights the SGLT2 as a feasible and promising drug targe...

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Detalles Bibliográficos
Autores principales: Lu, Yan-Ting, Ma, Xiu-Li, Xu, Yu-Hui, Hu, Jing, Wang, Fang, Qin, Wan-Ying, Xiong, Wen-Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Singapore 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6328422/
https://www.ncbi.nlm.nih.gov/pubmed/30387082
http://dx.doi.org/10.1007/s13659-018-0188-4
Descripción
Sumario:ABSTRACT: The sodium-dependent glucose transporters 2 (SGLT2) plays important role in renal reabsorption of urinal glucose back to plasma for maintaining glucose homeostasis. The approval of SGLT2 inhibitors for treatment of type 2 diabetes highlights the SGLT2 as a feasible and promising drug target in recent years. Current methods for screening SGLT2 inhibitors are complex, expensive and labor intensive. Particularly, these methods cannot directly measure nonradioactive glucose uptake in endogenous SGLT2-expressing kidney cells. In present work, human kidney cells, HK-2, was incubated with a fluorescent d-glucose derivant 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose (2-NBDG) and the fluorescent intensity of 2-NBDG was employed to measure the amount of glucose uptake into the cells. By optimizing the passages of HK-2 cells, 2-NBDG concentration and incubation time, and by measuring glucose uptake treated by Dapagliflozin, a clinical drug of SGLT2 inhibitors, we successfully developed a new assay for measuring glucose uptake through SGLT2. The nonradioactive microplate and microscope-based high-throughput screening assay for measuring glucose can be a new method for screening of SGLT2 inhibitors and implied for other cell assays for glucose measurement extensively. GRAPHICAL ABSTRACT: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13659-018-0188-4) contains supplementary material, which is available to authorized users.