Precision-cut intestinal slices
as an ex vivo model to study
NSAID-induced intestinal toxicity
Xiaoyu Niu
The research presented in this thesis was carried out mainly in the Division of Pharmacokinetics,
Toxicology and Targeting, University of Groningen, the Netherlands, and partly in the
Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Chiba
University, Japan.
This research project was financially supported by the Ubbo Emmius Scholarship of University
of Groningen. Publication of this thesis was financially supported by the University of
Groningen library and The Faculty of Mathematics and Natural Sciences.
Cover design: Xiaoyu Niu, the original work is attributed to Matt Might, a professor in Computer
Science at the University of Utah, created The Illustrated Guide to a Ph.D. to explain what a
Ph.D. is to new and aspiring graduate students. http://matt.might.net/articles/phd-school-inpictures/
Page layout: Xiaoyu Niu
Printed by: Off Page, Amsterdam
Copyright © 2014 Xiaoyu Niu
ISBN printed version: 978-90-367-7322-5
ISBN electronic version: 978-90-367-7323-2
No part of this book may be reproduced or transmitted in any form or by any means without
written permission of the author and the publisher holding the copyright of the published articles.
Precision-cut intestinal slices as an ex vivo model
to study NSAID-induced intestinal toxicity
Proefschrift
ter verkrijging van de graad van doctor aan de
Rijksuniversiteit Groningen
op gezag van de
rector magnificus prof. dr. E. Sterken
en volgens besluit van het College voor Promoties.
De openbare verdediging zal plaatsvinden op
vrijdag 24 oktober 2014 om 11.00 uur
door
Xiaoyu Niu
geboren op 2 november 1982
te Hebei, China
Promotor:
Prof. dr. G.M.M. Groothuis
Co-promotor:
Dr. I.A.M. de Graaf
Beoordelingscommissie:
Prof. dr. N.P.E. Vermeulen
Prof. dr. H.W. Frijlink
Prof. dr. G. Dijkstra
Paranimfen:
Miriam Langelaar-Makkinje
Andreia F. Batista de Almeida
To all the supportive friends on my way to the PhD
Contents
Abbreviations
Chapter 1
General introduction: Evaluation of the intestinal toxicity and
transport of xenobiotics utilizing precision-cut slices
8
11
(Xenobiotica 2013; 43(1): 73-83)
Chapter 2
Aim and outline of the thesis
35
Chapter 3
Precision cut intestinal slices are an appropriate ex vivo
model to study NSAID-induced intestinal toxicity in rats
39
(Toxicology in vitro 2014; 28(7): 1296–1305)
Chapter 4
Diclofenac toxicity in human intestine ex vivo is not related
to the formation of intestinal metabolites
65
(Arch Toxicol 2014; in press)
Chapter 5
Consequences of Mrp2 deficiency for diclofenac toxicity in
the rat intestine ex vivo
87
(Submitted)
Chapter 6
The influence of ischemia-reperfusion damage on the 107
viability of intestinal precision cut slices ex vivo
(To be submitted)
Chapter 7
Summary, general discussion and future perspectives
127
Appendices
Samenvatting
137
Acknowledgement
142
List of publications
145
Curriculum vitae
146
Abbreviations
Abbreviations
3Rs
4’-OH DCF
5-OH DCF
ACN
ADRs
AMV-RT
AP
Bach1
BiP
BSA
Chop
Ct
CYP
DAG
DCF
DMEs
DMSO
DNPH
DTNB
DTs
EHBR
ER
GI
GSH
GSSG
GST
HE
HO-1
HSF1
HSP-70
I-FABP
IPC
IR
KHB
KRB
LCA
LDH
mPT
Mrp2NRS
8
replacement, refinement and reduction
4’-hydroxyl diclofenac
5-hydroxyl diclofenac
acetonitrile
adverse drug reactions
avian myeloblastosis virus reverse transcriptase
alkaline phosphatase
BTB and CNC homologue 1
binding immunoglobulin protein
bovine serum albumin
CCAAT-enhancer-binding protein homologous protein
comparative threshold cycle
cytochrome P450
diclofenac acyl-glucuronide
diclofenac
drug metabolizing enzymes
dimethyl sulfoxide
2,4-dinitrophenylhydrazine
5,5'-dithiobis-(2-nitrobenzoic acid)
drug transporters
eisai hyperbilirubinemic rat
endoplasmic reticulum
gastro-intestinal
reduced glutathione
oxidized glutathione
glutathione transferases
hematoxylin-eosin
heme oxygenase 1
heat shock transcription factor 1
heat shock protein 70
intestinal fatty acid binding protein
ischemic preconditioning
ischemia-reperfusion
Krebs-Henseleit buffer
Krebs-Ringer buffer
lithocholic acid
lactate dehydrogenase
permeability transition pore
Mrp2 deficient
normal rat serum
Abbreviations
NSAIDs
PBS
PCIS
PPPD
Rnasin
ROI
ROS
RT-PCR
SD
SMA
SOD
SULT
TRTris
UGT
UW
WmE
WT
γ-GCS
non-steroidal anti-inflammatory drugs
phosphate buffered saline
precision cut intestinal slice
pylorus-preserving pancreaticoduodenectomy
recombinant RNasin® ribonuclease inhibitor
regions of interest
reactive oxygen species
quantitative real-time polymerase chain reaction
sprague-dawley rat
superior mesenteric artery
super oxide dismutase
sulfotransferases
groningen-yellow transporter-deficient rats
trishydroxymethyl-aminomethane
UDP-glucuronosyltransferase
University of Wisconsin medium
Williams medium E
wild type
γ-glutamylcysteine synthetase
9
10