The relevance of immune responses to partial bladder outlet obstruction and reversal.

AIMS: Partial bladder outlet obstruction (PBOO) causes tissue inflammation, a significant increase in markers of systemic oxidative stress, and proliferation of circulating myeloid-derived suppressor cells. Here, we investigated the regulatory mechanisms underlying inflammation and helper T cell involvement in PBOO. METHODS: Surgical PBOO was performed in four groups of rats: control (C), obstruction at 2 (O2) and 4 (O4) weeks, and 4 weeks after the relief of PBOO (R4) (n = 6 each). The urinary levels of prostaglandin E metabolite (PGEM), expression of inflammatory cytokines (IL-6 and IL-17) in the bladder, numbers of peripheral blood regulatory T cells (Treg cells), and levels of TGF-ß1 were assessed via immunohistochemistry, flow cytometry, or ELISA. RESULTS: The levels of urinary PGEM, bladder IL-17, and TGF-ß1 and the numbers of peripheral Treg cells (Foxp3) were all significantly increased at 2 and 4 weeks after PBOO. PGEM, IL-17, and Treg cells (Foxp3) were decreased after the relief of PBOO, while the levels of TGF-ß1 continued to increase. CONCLUSIONS: Transient PBOO triggers an acute, reversible increase in inflammatory cytokines and Treg cells. The distinct dynamics of individual inflammatory markers support their potential use as markers for monitoring bladder inflammation.

Inflammasomes in the urinary tract: a disease-based review.

Inflammasomes are supramolecular structures that sense molecular patterns from pathogenic organisms or damaged cells and trigger an innate immune response, most commonly through production of the proinflammatory cytokines IL-1ß and IL-18, but also through less understood mechanisms independent of these cytokines. Great strides have been made in understanding these structures and their dysfunction in various inflammatory diseases, lending new insights into urological and renal problems. From a clinical perspective, benign urinary pathology almost universally involves the inflammatory process, and understanding how inflammasomes translate etiological conditions (diabetes, obstruction, stones, urinary tract infections, etc.) into acute and chronic inflammatory responses is critical to understanding these diseases at a molecular level. To date, inflammasome components have been found in the bladder, prostate, and kidney and have been shown to be activated in response to several infectious and noninfectious insults. In this review, we summarize what is known regarding inflammasomes in both the upper and lower urinary tract and describe several common disease states where they potentially play critical roles.

The NLRP3 Inflammasome Mediates Inflammation Produced by Bladder Outlet Obstruction.

PURPOSE: While bladder outlet obstruction is well established to elicit an inflammatory reaction in the bladder that leads to overactive bladder and fibrosis, little is known about the mechanism by which this is initiated. NLRs (NOD-like receptors) and the structures that they form (inflammasomes) have been identified as sensors of cellular damage, including pressure induced damage, and triggers of inflammation. Recently we identified these structures in the urothelium. In this study we assessed the role of the NLRP3 (NACHT, LRR and PYD domains-containing protein 3) inflammasome in bladder dysfunction resulting from bladder outlet obstruction. MATERIALS AND METHODS: Bladder outlet obstruction was created in female rats by inserting a 1 mm outer diameter transurethral catheter, tying a silk ligature around the urethra and removing the catheter. Untreated and sham operated rats served as controls. Rats with bladder outlet obstruction were given vehicle (10% ethanol) or 10 mg/kg glyburide (a NLRP3 inhibitor) orally daily for 12 days. Inflammasome activity, bladder hypertrophy, inflammation and bladder function (urodynamics) were assessed. RESULTS: Bladder outlet obstruction increased urothelial inflammasome activity, bladder hypertrophy and inflammation, and decreased voided volume. Glyburide blocked inflammasome activation, reduced hypertrophy and prevented inflammation. The decrease in voided volume was also attenuated by glyburide mechanistically as an increase in detrusor contraction duration and voiding period. CONCLUSION: Results suggest the importance of the NLRP3 inflammasome in the induction of inflammation and bladder dysfunction secondary to bladder outlet obstruction. Arresting these processes with NLRP3 inhibitors may prove useful to treat the symptoms that they produce.

M3 muscarinic receptor-like immunoreactivity in sham operated and obstructed guinea pig bladders.

PURPOSE: Type 3 muscarinic receptors, which are present in the bladder wall, are important for bladder function. However, their role in the context of the urothelium is not well defined. Understanding the role of type 3 muscarinic receptors has been limited by the lack of specific type 3 muscarinic receptor antibodies. Thus, we identified a specific type 3 muscarinic receptor antibody and investigated the site of type 3 muscarinic receptors in sham operated and obstructed guinea pig bladders. MATERIALS AND METHODS: The specificity of 4 commercially available type 3 muscarinic receptor antibodies was determined. Immunohistochemistry was then done in bladder tissue from sham operated and obstructed guinea pig bladders. RESULTS: One of the 4 antibodies examined had the needed specificity in terms of blocking peptide and Western blot characterization. Using this antibody type 3 muscarinic receptor immunoreactivity was associated with muscle cells, nerves and interstitial cells. Four types of interstitial cells were identified, including suburothelial, lamina propria, surface muscle and intramuscular interstitial cells. In the obstructed model the bladder wall was hypertrophied and there was nerve fiber loss. The number of lamina propria, surface muscle and intramuscular interstitial cells was increased but not the number of suburothelial interstitial cells. Also, surface muscle interstitial cells appeared to form clusters or nodes with type 3 muscarinic receptor immunoreactivity. CONCLUSIONS: Nerve loss and the up-regulation of interstitial cells with type 3 muscarinic receptor immunoreactivity may underlie major functional changes in the pathological bladder. This indicates that type 3 muscarinic receptor specific anticholinergic drugs may affect not only the detrusor muscle, as previously thought, but also interstitial cells and nerve fibers.

Effect of Serenoa repens, lycopene, and selenium on proinflammatory phenotype activation: an in vitro and in vivo comparison study.

OBJECTIVES: To investigate the antiinflammatory activity of Serenoa repens (SeR), LY, and) on proinflammatory phenotype in rat peritoneal macrophages (Ms) stimulated with Salmonella enteritidis lipopolysaccharide (LPS) and in the prostate of rats with partial bladder outlet obstruction. SeR, combined with other compounds, such as LY and Se is used to relieve symptoms associated with benign prostatic hyperplasia (BPH). Inflammation plays a pivotal role in the pathogenesis of BPH and represents a target for anti-BPH drugs. METHODS: After stimulation with 1 µg/mL of LPS, peritoneal rat MΦs were coincubated with LY (2 µg/mL), Se (0.03 µg/mL), and SeR (10 µg/mL), alone or in association (LY-Se-SeR) and with RPMI. Inducible cyclooxygenase (COX-2), 5-lypoxygenase (5-LOX), inducible nitric oxide synthase (iNOS), and inhibitor κBα (IκB-α) protein were evaluated by Western blot. Nuclear factor-kappa B (NF-κB) binding activity was measured by electrophoretic mobility shift assay. Tumor necrosis factor-α (TNF-α) gene expression was investigated by real-time polymerase chain reaction. We also evaluated malondialdehyde (MDA) and nitrite levels. RESULTS: LPS stimulation produced a proinflammatory phenotype in rat peritoneal MΦs. LY, Se, and SeR inhibited the inflammatory cascade, but the Ly-Se-SeR association caused a greater inhibitory effect on the expression of COX-2, 5-LOX, and iNOS. The Ly-Se-SeR association showed a higher efficacy in reducing the loss of IκB-α, the increased NF-κB binding activity, the enhanced mRNA levels of TNF-α, the elevated MDA, and nitrite content. The LY-Se-SeR association in vivo caused a greater inhibitory effect on prostate inflammation induced in rats by partial bladder outlet obstruction. CONCLUSIONS: The LY-Se-SeR association might be useful in the treatment of BPH.

Immunoexpression of adrenergic receptors in detrusor from patients with prune belly syndrome: a digital quantification.

INTRODUCTION: Prune belly syndrome (PBS) presents with large-capacity bladders, high compliance and post-void residual volumes. Operative and conservative treatments are controversial. When histologically compared to normal bladder, bladder outlet obstruction results in an up- or down-regulation of adrenoceptors. Our goal was to study the immunoexpression of adrenoceptors in detrusor from patients with PBS. MATERIALS AND METHODS: Bladder domes from PBS patients (n=14) were studied (PBG). For normal controls, bladder specimens were obtained at adult surgery (n=13) (CG1) and at child autopsy (n=5) (CG2). Staining was performed using antibodies to alpha1a, alpha1b, alpha1d and beta3 adrenoceptors. Five to 10 images were captured on an optic microscope with a digital camera and analysed with Photoshop. The immunocyhistochemical index with arbitrary units was calculated and compared. RESULTS: Mean age was 1.28, 64 and 1.41 years for PBG, CG1 and CG2, respectively. The immunohistochemical index with arbitrary units of alpha1a receptors was 0.06 in PBG, 0.16 in CG1 and 0.14 in CG2 (p=0.008); of alpha1b 0.06, 0.06 and 0.07 (p=0.781); and of alpha1d 0.04, 0.04 and 0.05 (p=0.618). Regarding beta3 the respective values were 0.07, 0.14 and 0.10 (p=0.378). CONCLUSION: Our results show a decrease in alpha1a-adrenoceptor immunostaining intensity in detrusor from children with PBS. Further in vitro studies are needed to determine whether these observations are physiologically significant.

Phenotypic transition of the collecting duct epithelium in congenital urinary tract obstruction.

Epithelial-mesenchymal transition (EMT) has emerged in recent years as an important process in the development of organ fibrosis in many human diseases. Our previous experience in a nonhuman primate model of obstructive nephropathy suggested that EMT of collecting duct epithelium contributes to the development of interstitial fibrosis. In this study we demonstrate for the first time in humans that obstructed fetal collecting duct epithelium undergoes transition to mesenchymal phenotype, characterized by decreased expression of epithelial markers, de novo expression of mesenchymal markers with subsequent loss of cell-cell interaction, disruption of the basement membrane, and increased deposition of extracellular matrix into the expanded interstitium of the obstructed kidney. The results of this study therefore support the previous findings from animal studies and suggest that EMT of the collecting duct epithelium might contribute to the development of interstitial fibrosis in human fetal obstructive nephropathy.

[The staged medical rehabilitation of patients with secondary chronic pyelonephritis in infravesical obstructions]. / Etapnaia meditsinskaia reabilitatsiia bol'nykh vtorichnym khronicheskim pielonefritom pri infravezikal'nykh obstruktsiiakh.

The authors believe that chronic pyelonephritis patients operated for infravesical obstructions need staged postoperative rehabilitation. In antibacterial postoperative treatment, a significant response was seen in 7.9%, a partial response in 18.4% and no response in 73.7% of patients. After combined physiotherapy--in 38%, 41.7% and 20.3% of patients, respectively.

Immunoglobulin levels in fetal anatomic compartments.

Immunoglobulins G, M, and A were found in varying concentrations in four different human fetal anatomic compartments of 17 fetuses in the absence of apparent infection. The highest levels were noted in the thoracic cavity.