Downregulation of WNT11 is associated with bladder tissue fibrosis in patients with interstitial cystitis/bladder pain syndrome without Hunner lesion

Abstract

This study assessed the functional role of WNT genes and the association between WNT signalling cascades and fibrosis in interstitial cystitis/bladder pain syndrome (IC/BPS) patients. Twenty-five patients (3 males, 22 females; mean age 59.7 ± 10.9 years), included 7 non-Hunner-type IC (NHIC), 18 Hunner-type IC (HIC), and 5 non-IC (control) groups. The expression of sonic hedgehog, WNT gene family, and genes previously reported as biomarkers for IC/BPS were examined using RT-PCR in biopsy specimens from the mucosa and submucosa layer of the bladder. WNT2B, WNT5A, WNT10A, and WNT11 functions in the urothelium were evaluated by silencing in an HBlEpC cell line. Pelvic Pain and Urgency/Frequency Patient Symptom Scale scores, O’Leary-Sant Symptom and Problem Index scores, and Visual Analogue Scores did not differ between the NHIC and HIC groups. However, HIC patients had significantly shorter symptom duration (30.9 vs 70.8 months, p = 0.046), higher daily urinary frequency (16.1 versus 8.5 times, p = 0.006), and smaller bladder capacity (208.6 versus 361.4 ml, p = 0.006) than NHIC patients. Overall WNT gene expression was lower in NHIC than HIC patients. Bladder epithelial tissues from HIC patients were characterised by the downregulation of WNT11. Silencing of WNT11, WNT2B, WNT5A, and WNT10A in HBlEpCs resulted in fibrotic changes, indicated by fibrotic morphology, increased fibrosis-related gene expression, and nuclear localisation of phosphorylated SMAD2, and increased vimentin and fibronectin levels. Downregulation of WNT11 results in fibrotic changes of bladder epithelial cells and is associated with the pathogenesis and differential diagnosis of NHIC. Decreased expression of WNT11 is a potential biomarker for predicting NHIC.

N-acetylcysteine prevents bladder tissue fibrosis in a lipopolysaccharide-induced cystitis rat model

“…Histological analysis of the expression in the bladder tissues rats was performed two weeks after the final instillation of PS/LPS to evaluate epithelial denudation, mast cell infiltration, tissue fibrosis, and apoptosis with cytokeratin immunostaining (Keratin, Pan Ab-1; Thermo Scientific, Foster City, CA, USA), Masson’s trichrome staining (Junsei Chemical, Tokyo, Japan), toluidine blue staining (Toluidine blue-O; Daejung Chemicals & Metals, Seoul, Korea), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining (Roche, Mannheim, Germany), respectively, as previously described 38,39 . For lymphoplasmacytic infiltration analysis, we used the antibodies CD3 (sc-80668; Santa Cruz Biotechnology, Dalla, TX, USA), CD20 (sc-393894; Santa Cruz Biotechnology, Dallas, TX, USA) and CD138 (ab34164; Abcam, Cambridge, MA, USA) to detect T-lymphocytes, B-lymphocytes, plasma cells, respectively.…”

“…Using Taqman reverse transcription reagents (Applied Biosystems), mRNA (400 ng) was reverse transcribed per manufacturer’s instructions. Quantitative assessment of the target genes’ expression levels was performed using real-time quantitative PCR (RQ-PCR) with a PikoReal™ Real-Time PCR System (Thermo Scientific) with iQ™ SYBR Green PCR Master Mix (Bio-Rad, Hercules, CA, USA), as described previously 39,40 . Gene expression data were quantified using duplicated RQ-PCR assays (n = 10) from five independent animals.…”

Induction of detrusor underactivity by extensive vascular endothelial damages of iliac arteries in a rat model and its pathophysiology in the genetic levels

“…For the RQ-PCR analysis, reverse transcription of the isolated total RNA was performed using TaqMan reverse transcription reagents (Applied Biosystems) and the expression level of the indicated transcripts was quantified by RQ-PCR with the PikoReal RT-PCR System (Thermo Fisher Scientific, Inc., Waltham, MA, USA) with iQ SYBR Green PCR Master Mix (Bio-Rad, Hercules, CA), as described previously 44,45 . Two randomly chosen areas from each slide or duplicated RQ-PCR assays (n = 10) using five independent animals were used to quantify the digital image or gene expression data.…”

Functional, histological and molecular characteristics of human exstrophy detrusor

“…However, expression of cyclin-D1, another target of β-catenin transcription was not significantly altered in exstrophy samples. Little is known about the particular Wnt-proteins that regulate normal human bladder development and generation of fibrosis, but down-regulation of Wnt11 is associated with fibrosis in patients with bladder pain syndrome [28]. Overall, downregulation of a Wnt-related pathway in a congenital bladder anomaly is a novel observation [29] and may result in increased differentiation into fibroblasts rather than a smooth muscle lineage.…”