2. Academic Validation
  3. Real-time imaging of intestinal bacterial β-glucuronidase activity by hydrolysis of a fluorescent probe

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

  • Sci Rep. 2017 Jun 9;7(1):3142. doi: 10.1038/s41598-017-03252-4.
Michael Chen 1,  Kai-Wen Cheng 2,  Yi-Jou Chen 1,  Chang-Hung Wang 1,  Ta-Chun Cheng 3,  Kuo-Chien Chang 4,  An-Pei Kao 5,  Kuo-Hsiang Chuang 6 7 8 9
Affiliations

Affiliations

  • 1 Ph.D. Program for the Clinical Drug Discovery from Botanical Herbs, Taipei Medical University, Taipei, Taiwan.
  • 2 Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan.
  • 3 Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, Kaohsiung, Taiwan.
  • 4 Department of Orthopedics, Tri-service General Hospital, Taipei, Taiwan.
  • 5 Stemforce Biotechnology Co., Ltd, Chiayi City, Taiwan.
  • 6 Ph.D. Program for the Clinical Drug Discovery from Botanical Herbs, Taipei Medical University, Taipei, Taiwan. khchuang@tmu.edu.tw.
  • 7 Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei, Taiwan. khchuang@tmu.edu.tw.
  • 8 Ph.D Program in Biotechnology Research and Development, Taipei Medical University, Taipei, Taiwan. khchuang@tmu.edu.tw.
  • 9 The Ph.D. Program of Translational Medicine, Taipei Medical University, Taipei, Taiwan. khchuang@tmu.edu.tw.
PMID: 28600512 DOI: 10.1038/s41598-017-03252-4
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.

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