A mouse model for interstitial cystitis/painful bladder syndrome based on APF inhibition of bladder epithelial repair: a pilot study.
BACKGROUND
Interstitial cystitis/painful bladder syndrome (IC/PBS) is a chronic bladder disorder with bladder epithelial thinning or ulceration, pain, urinary frequency and urgency. There is no reliably effective therapy for IC/PBS, and no generally accepted animal model for the disorder in which potential therapies can be tested. Bladder epithelial cells from IC/PBS patients make a small glycopeptide antiproliferative factor or "APF" that inhibits proliferation, decreases tight junction protein expression, increases paracellular permeability, and induces changes in gene expression of bladder epithelial cells in vitro that mimic abnormalities in IC/PBS patient biopsy specimens in vivo. We therefore determined the ability of a synthetic APF derivative to inhibit bladder epithelial repair in mice.


METHODS
The bladder epithelium of female CBA/J mice was stripped by transurethral infusion of 3% acetic acid, and mice were subsequently treated daily with one of three intravesical treatments [synthetic as-APF, inactive unglycosylated control peptide, or phosphate buffered saline carrier (PBS)] for 1-21 days. Fixed bladder sections were either stained with haematoxylin and eosin for determination of epithelial area by image analysis, or incubated with anti-uroplakin III (UPIII) or anti-zonula occludens type 1 (ZO-1) antibodies for immunofluorescence microscopy. Epithelial measurement data were analyzed by a two-way analysis of variance (ANOVA); post hoc comparisons of multiple groups were carried out using the Tukey-Kramer method.


RESULTS
Bladder epithelial repair was significantly attenuated in as-APF-treated mice as compared to control mice on days 3-21 (p < 0.05); the mean epithelial/total area over all measured days was also significantly lower in as-APF-treated mice vs. mice in either control group by post hoc analysis (p < 0.0001 for both comparisons). UPIII and ZO-1 expression was also decreased in as-APF-treated mice as compared to mice in either control group by day 7 (UPIII) or day 14 (ZO-1).


CONCLUSIONS
This model demonstrates in vivo effects of as-APF which abrogates bladder epithelial repair and expression of UPIII and ZO-1 in CBA/J mice following transurethral acetic acid infusion. As bladder epithelial thinning, decreased UPIII expression, and decreased ZO-1 expression are histopathologic features of IC/PBS patient biopsies, this model may be useful for studying the pathophysiology of IC/PBS and the effect of potential therapies.
Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA. skeay@medicine.umaryland.edu
Susan
S
Keay
Samantha
S
Leitzell
Ashley
A
Ochrzcin
George
G
Clements
Min
M
Zhan
David
D
Johnson
D000818:Animals; D018856:Cystitis, Interstitial / Q000378:metabolism / Q000473:pathology*; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology*; D005260:Female; D006023:Glycoproteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D010865:Pilot Projects; D001743:Urinary Bladder / Q000378:metabolism / Q000473:pathology*
D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; C490708:antiproliferative factor APF, human