Cargando…

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...

Descripción completa

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
_version_ 1783386636995788800
author Lu, Yan-Ting
Ma, Xiu-Li
Xu, Yu-Hui
Hu, Jing
Wang, Fang
Qin, Wan-Ying
Xiong, Wen-Yong
author_facet Lu, Yan-Ting
Ma, Xiu-Li
Xu, Yu-Hui
Hu, Jing
Wang, Fang
Qin, Wan-Ying
Xiong, Wen-Yong
author_sort Lu, Yan-Ting
collection PubMed
description 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.
format Online
Article
Text
id pubmed-6328422
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Springer Singapore
record_format MEDLINE/PubMed
spelling pubmed-63284222019-01-25 A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells Lu, Yan-Ting Ma, Xiu-Li Xu, Yu-Hui Hu, Jing Wang, Fang Qin, Wan-Ying Xiong, Wen-Yong Nat Prod Bioprospect Original Article 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. Springer Singapore 2018-11-01 /pmc/articles/PMC6328422/ /pubmed/30387082 http://dx.doi.org/10.1007/s13659-018-0188-4 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Lu, Yan-Ting
Ma, Xiu-Li
Xu, Yu-Hui
Hu, Jing
Wang, Fang
Qin, Wan-Ying
Xiong, Wen-Yong
A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells
title A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells
title_full A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells
title_fullStr A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells
title_full_unstemmed A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells
title_short A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells
title_sort fluorescent glucose transport assay for screening sglt2 inhibitors in endogenous sglt2-expressing hk-2 cells
topic Original Article
url 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
work_keys_str_mv AT luyanting afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT maxiuli afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT xuyuhui afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT hujing afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT wangfang afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT qinwanying afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT xiongwenyong afluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT luyanting fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT maxiuli fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT xuyuhui fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT hujing fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT wangfang fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT qinwanying fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells
AT xiongwenyong fluorescentglucosetransportassayforscreeningsglt2inhibitorsinendogenoussglt2expressinghk2cells