Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe

Abstract Intestinal bacterial β-glucuronidase (βG) hydrolyzes glucuronidated metabolites to their toxic form in intestines, resulting in intestinal damage. The development of a method to inhibit βG is thus important but has been limited by the difficulty of directly assessing enzyme activity in live...

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Autores principales:	Michael Chen, Kai-Wen Cheng, Yi-Jou Chen, Chang-Hung Wang, Ta-Chun Cheng, Kuo-Chien Chang, An-Pei Kao, Kuo-Hsiang Chuang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2d0f1bd83aa0414da3acd81e41ce2509
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spelling	oai:doaj.org-article:2d0f1bd83aa0414da3acd81e41ce25092021-12-02T12:32:33ZReal-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe10.1038/s41598-017-03252-42045-2322https://doaj.org/article/2d0f1bd83aa0414da3acd81e41ce25092017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03252-4https://doaj.org/toc/2045-2322Abstract Intestinal bacterial β-glucuronidase (βG) hydrolyzes glucuronidated metabolites to their toxic form in intestines, resulting in intestinal damage. The development of a method to inhibit βG is thus important but has been limited by the difficulty of directly assessing enzyme activity in live animals. Here, we utilized a fluorescent probe, fluorescein di-β-D-glucuronide (FDGlcU), to non-invasively image the intestinal bacterial βG activity in nude mice. In vitro cell-based assays showed that the detection limit is 104 colony-forming units/well of βG-expressing bacteria, and that 7.81 ng/mL of FDGlcU is enough to generate significant fluorescent signal. In whole-body optical images of nude mice, the maximum fluorescence signal for βG activity in intestines was detected 3 hours after gavage with FDGlcU. Following pretreatment with a bacterial βG inhibitor, the fluorescence signal was significantly reduced in abdomens and excised intestines images. For a 4-day antibiotic treatment to deplete intestinal bacteria, the FDGlcU-based images showed that the βG activity was decreased by 8.5-fold on day 4 and then gradually increased after treatment stopped. The results suggested that FDGlcU-based imaging revealed the in vitro and in vivo activity of intestinal bacterial βG, which would facilitate pharmacodynamic studies of specific bacterial βG inhibitors in animal studies.Michael ChenKai-Wen ChengYi-Jou ChenChang-Hung WangTa-Chun ChengKuo-Chien ChangAn-Pei KaoKuo-Hsiang ChuangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Michael Chen Kai-Wen Cheng Yi-Jou Chen Chang-Hung Wang Ta-Chun Cheng Kuo-Chien Chang An-Pei Kao Kuo-Hsiang Chuang Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
description	Abstract Intestinal bacterial β-glucuronidase (βG) hydrolyzes glucuronidated metabolites to their toxic form in intestines, resulting in intestinal damage. The development of a method to inhibit βG is thus important but has been limited by the difficulty of directly assessing enzyme activity in live animals. Here, we utilized a fluorescent probe, fluorescein di-β-D-glucuronide (FDGlcU), to non-invasively image the intestinal bacterial βG activity in nude mice. In vitro cell-based assays showed that the detection limit is 104 colony-forming units/well of βG-expressing bacteria, and that 7.81 ng/mL of FDGlcU is enough to generate significant fluorescent signal. In whole-body optical images of nude mice, the maximum fluorescence signal for βG activity in intestines was detected 3 hours after gavage with FDGlcU. Following pretreatment with a bacterial βG inhibitor, the fluorescence signal was significantly reduced in abdomens and excised intestines images. For a 4-day antibiotic treatment to deplete intestinal bacteria, the FDGlcU-based images showed that the βG activity was decreased by 8.5-fold on day 4 and then gradually increased after treatment stopped. The results suggested that FDGlcU-based imaging revealed the in vitro and in vivo activity of intestinal bacterial βG, which would facilitate pharmacodynamic studies of specific bacterial βG inhibitors in animal studies.
format	article
author	Michael Chen Kai-Wen Cheng Yi-Jou Chen Chang-Hung Wang Ta-Chun Cheng Kuo-Chien Chang An-Pei Kao Kuo-Hsiang Chuang
author_facet	Michael Chen Kai-Wen Cheng Yi-Jou Chen Chang-Hung Wang Ta-Chun Cheng Kuo-Chien Chang An-Pei Kao Kuo-Hsiang Chuang
author_sort	Michael Chen
title	Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
title_short	Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
title_full	Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
title_fullStr	Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
title_full_unstemmed	Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
title_sort	real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/2d0f1bd83aa0414da3acd81e41ce2509
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Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe

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