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.

Figures
Products
嗨！很高兴为您提供帮助！

尊敬的 MCE 客户您好，
请您选择所在区域，我们将转接对应客服为您服务！

热门区域

A

B

C

F

G

H

J

L

N

Q

S

T

X

Y

Z

热 A B C F G H J L N Q S T X Y Z