Partial Bladder Outlet Obstruction Group Research Articles

HucMSC exosomes attenuate partial bladder outlet obstruction-induced renal injury and cell proliferation via the Wnt/β-catenin pathway

Bladder outlet obstruction (BOO) can cause serious complications including kidney damage; nevertheless, there are currently no animal models for studying BOO-induced kidney damage. Mesenchymal stem cells (MSCs) are widely used in therapeutic studies of renal fibrosis. However, MSC-derived exosomes show improved safety profile and more controllable characteristics compared with those of MSCs. Herein, we established a kidney injury mouse model of partial bladder outlet obstruction (PBOO) and evaluated the effects of human umbilical cord MSC-derived exosomes (hucMSC-Exos) on PBOO-induced reflux kidney injury in this model. Exosomes were isolated from a hucMSC-conditioned medium, purified by ultracentrifugation, and examined. Living image was performed to indicate the distribution of hucMSC-Exos. The PBOO-treated mice interacted with PBS (phosphate-buffered saline) or hucMSC-Exos. Morphologic changes and expression of interstitial-fibrosis—related, cell proliferation and Wnt/β-catenin signaling-pathway indices were evaluated. At 7 days after induction of PBOO, structural destruction of renal tubules was observed. Expression of the interstitial markers and the cellular-proliferation index increased significantly in the PBOO group compared with the control group. The isolated exosomes were 30–150 nm in diameter, showing a round shape and bilayer membrane structure with CD63, TSG101, Alix expressed, enriched in the kidney of the PBOO group. Administering hucMSC-Exos to post-PBOO mice reversed renal injury and suppressed expression of Wnt/β-catenin signaling pathway-related proteins. hucMSC-Exos inhibited PBOO-induced kidney injury and cellular proliferation and suppressed the Wnt/β-catenin signaling pathway. Our findings will spur the development of novel hucMSC-Exo-mediated therapies for treating patients with renal fibrosis.

Sodium butyrate ameliorates erectile dysfunction through fibrosis in a rat model of partial bladder outlet obstruction.

In different animal models, a histone deacetylase (HDAC) inhibitor, sodium butyrate (NaBu) reduced inflammation, oxidative stress and fibrosis which were involved in the pathogenesis of erectile dysfunction (ED), but whether NaBu could improve ED in an experimental animal model of benign prostate hyperplasia (BPH) was not known. To investigate the preventive effect of NaBu on ED in a partial bladder outlet obstruction (PBOO) rat model. PBOO was induced by partial urethral obstruction. NaBu (20mg/kg/day) was administered orally to rats for 6 weeks after creation of PBOO. In vivo erectile responses, in vitro relaxation and contraction responses in cavernosal tissue were measured. Real-time polymerase chain reaction (RT-PCR) and Western blot were performed to determine the gene and protein expression. Inflammation, fibrosis, and localization of proteins were evaluated using histological techniques. HDAC activity and tumor necrosis factor (TNF)-α levels were measured in penile tissues. NaBu improved decreased intracavernosal pressure/mean arterial pressure, nitrergic and endothelium-dependent relaxation responses, and contractile responses to phenylephrine and electrical field stimulation in the PBOO group without affecting increased bladder weight. Increased endothelial nitric oxide synthase (eNOS), transforming growth factor (TGF)-β1, and nuclear factor kappa B (NF-κB) gene levels in PBOO group were ameliorated by NaBu treatment. The administration of NaBu to PBOO rats significantly increased neuronal NOS (nNOS) and decreased TGF-β1 protein expression. The nuclear/cytosolic ratio of NF-κB demonstrated a decrease in PBOO and all treatment groups compared to control. A significant increase in the nuclear-to-cytoplasmic ratio of nuclear factor erythroid 2-related factor 2 (Nrf2) after PBOO was reduced by the treatment. Both eNOS and inducible NOS (iNOS) protein expression, together with TNF-α levels did not differ in the penile tissue of all groups. In histological analysis, increased TGF-β1 protein expression and fibrosis, as well as decreased nNOS protein in PBOO, were reversed by the treatment. NaBu did not normalize moderate inflammation in obstructed rats. An increase in the HDAC activity in PBOO was significantly suppressed by NaBu. Inhibition of the HDAC activity by NaBu in penile tissue could ameliorate fibrosis-associated changes induced by PBOO. NaBu promotes recovery of erectile function, and also significantly prevents penile fibrosis and normalizes TGF-β1 and nNOS protein expression in a rat model of PBOO. The HDAC pathway may present a promising target to prevent ED in patients with BPH.

Establishment of an Animal Model of Vesicoureteral Reflux Renal Injury through Partial Bladder Outlet Obstruction

To establish an animal model of reflux renal damage through bladder outlet obstruction. Sixty male C57BL/6 mice aged 6-8 weeks were randomly assigned to a control group, a sham operation group, and a partial bladder outlet obstruction (PBOO) group, with 20 mice in each group. Laparotomy were performed on the PBOO mice under anesthesia in order to separate the bladder necks and to perform guided partial ligation of the bladder neck with a metal rod of 0.3 mm diameter. Mice in the sham operation group had laparotomy and had their bladder necks separated without ligation. The control group did not receive any treatment. 7 days after the surgery, 12 surviving mice were randomly selected from each group to observe the general changes of the bladder, ureter, renal pelvis and kidney. Retrograde urography was performed through the bladder. Kidney tissues were extracted for histopathological analysis. The expression levels of Vimentin, proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) were examined with Western blot, immunohistochemistry and immunofluorescence staining tests, respectively. Compared with the control and sham operation group, the bladder, ureter, and renal pelvis of the mice in the PBOO group were significantly enlarged, vesicoureteral reflux was more obvious, the kidney volume and mass increased ( P<0.001), and renal parenchyma became thinner ( P<0.000 1). Histopathological staining showed glomerular atrophy, renal tubule expansion, tubulointerstitial inflammatory cell infiltration, glomerular basement membrane hyperplasia and obvious interstitial fibrosis. Western blot, immunofluorescence and immunohistochemistry staining showed that the expression levels of Vimentin, PCNA and α-SMA in kidney tissue were elevated ( P<0.000 1). After PBOO, the bladder, ureter, and kidney of the mice showed obvious morphological alteration and presented reflux renal fibrosis-like damage. This can be used as an animal model to study the pathological alteration mechanism and therapeutic measures of renal fibrosis caused by bladder outlet obstruction.

Long-term administration of alpha-1 blocker can reverse the micturition pattern in a bladder outlet obstruction murine model.

To investigate the effect of chronic administration of an alpha-1 blocker on micturition patterns in long-term partial bladder outlet obstruction. Mice were divided into three groups: a normal group, in which animals were fed a standard diet; a partial bladder outlet obstruction group, in which the proximal urethra was tied and animals were fed a standard diet; and a partial bladder outlet obstruction+naftopidil group, in which the proximal urethra was tied and animals were fed a standard diet containing naftopidil. Micturition behavior was evaluated in all groups for 6months after partial bladder outlet obstruction surgery. The parameters evaluated included voided volume, time per void, urination frequency and total urine volume. Quantitative assessment of gene expression was also carried out. Total urine volume, as well as total and average voided volume during night, was significantly decreased in partial bladder outlet obstruction+naftopidil mice compared with partial bladder outlet obstruction animals. The levels of transcripts encoding 5-hydroxytryptamine2A and tissue inhibitor of metalloproteinase2 were significantly decreased in the partial bladder outlet obstruction+naftopidil group compared with the partial bladder outlet obstruction group. Long-term administration of an alpha-1 blocker seems to reverse the disturbance of the micturition pattern caused by partial bladder outlet obstruction. Mechanistically, this effect might be mediated by changes in the expression of a serotonin receptor and/or in the activity of the fibrogenesis pathway.

Changes in Apoptosis-Related Proteins in The Urothelium of Rat Bladder Following Partial Bladder Outlet Obstruction and Subsequent Relief.

Partial bladder outlet obstruction (PBOO) induces sustained bladder over-distension, leading to ischemia/reperfusion (I/R)-related oxidative damage of the urothelium via apoptosis. The present study aimed to investigate the sequential course of apoptosis in the urothelium of rat bladder and identify the changes in apoptosis-related proteins during PBOO and subsequent relief. Materials and Methods: The study was conducted using 60 female Sprague-Dawley rats divided into three groups: sham-operated, PBOO only, and PBOO plus subsequent relief. PBOO was induced for 2 weeks, and then the obstruction was relieved by removal of the ligature. The urothelium was assessed by a histological analysis, and expression levels of apoptosis-related proteins were detected by quantitative PCR and immunoblotting. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL)-positive cells were significantly increased in the PBOO only group when compared with the sham-operated group, and decreased in the PBOO relief group when compared with the PBOO group (P < 0.001). From the quantitative PCR and the western blot analyses, expression of Bax, caspase-3, P38, and Jnk was significantly increased in the PBOO group (P < 0.001). However, expression of Erk, Bcl-2 significantly decreased in the PBOO group (P < 0.001). The expression of Erk and Bcl-2 significantly increased in the PBOO relief group when compared with the PBOO group (P < 0.001). In comparison to the sham-operated group, expression levels of survivin significantly increased in both the PBOO and PBOO plus relief groups (P < 0.001). In addition, the expression levels were significantly different between the PBOO and PBOO plus relief groups (P < 0.001). PBOO induced apoptosis of urothelium is related to alterations in the MAPK signaling pathways and apoptosis-related protein change. These results may also suggest that the pro-survival Erk signaling cascade and the expression survivin are activated in response to ischemic bladder injury and associated with initiation of bladder restoration in PBOO and subsequent relief. However, the mechanism of survivin as anti-apoptotic protein in ischemic bladder injuries remains unclear.

The effects of oral administration of the novel muscarinic receptor antagonist DA-8010 on overactive bladder in rat with bladder outlet obstruction

BackgroundDA-8010 is a novel compound developed for the treatment of overactive bladder (OAB) and urinary incontinence. The aims of this study were to investigate the effects of DA-8010 on OAB in a rat model.MethodsStudy animals were divided into the following five groups of seven animals each: a sham-operated control group, a control group with partial bladder outlet obstruction (BOO) (OAB group), and three DA-8010 (doses of 0.3 mg/kg/day, 1 mg/kg/day, and 3 mg/kg/day, respectively) with partial BOO groups. Oral administration of the drugs was continued for 14 days after 2 weeks of partial BOO. After 4 weeks of partial BOO, cystometrography was performed in all groups. Additionally, pro-inflammatory cytokines, Rho-kinases, and histology of the bladder were analyzed.ResultsThere was a significant increase in the contraction interval and a decrease in contraction pressure in the 3 mg/kg/day DA-8010 group versus those in the OAB group. Rho kinase was also significantly decreased in the DA-8010 3 mg/kg/day dosage treatment group. The increased ratio of collagen to smooth muscle after partial BOO was significantly attenuated in the DA-8010 3 mg/kg/day dosage group.ConclusionsOral administration of DA-8010 at 3 mg/kg/day improved findings in an OAB rat model induced by partial BOO. Our results suggest that the novel muscarinic receptor antagonist DA-8010 may be a promising drug for treating patients with OAB.

The Antimuscarinic Agent Tolterodine Regulates Bladder Extracellular Matrix in Partial Bladder Outlet Obstruction in Rats

Background/Aims: Antimuscarinic agents can delay the progression of bladder dysfunction caused by bladder outlet obstruction (BOO). To date, the relationship between muscarinic receptor activity and the bladder extracellular matrix (ECM) remains unclear. Thus, an animal model of partial BOO (PBOO) in female rats was established to explore the variation in bladder wall ECM proteins under PBOO conditions with antimuscarinic agent administration. Methods: Rats were randomly divided into three groups: sham, PBOO, and PBOO plus tolterodine. Picrosirius red staining was used to examine the smooth muscle and collagen content of bladder samples. Gene microarray and RT-PCR were performed to survey the expression of ECM proteins, receptors, and metabolism regulators in the rat bladder. Positive results were further evaluated by immunohistochemistry. Results: Picrosirius red staining showed that smooth muscle volume significantly increased in the PBOO and PBOO plus tolterodine groups (p < 0.05), while collagen significantly increased in the PBOO group (p < 0.05) but not in the PBOO plus tolterodine group. Gene microarray and RT-PCR revealed that none of the collagen subtypes exhibited significant changes after PBOO establishment and tolterodine administration. However, matrix metalloproteinases (MMPs) increased significantly in the PBOO plus tolterodine group (p < 0.05). Additionally, PBOO inhibited the expression of non-collagen ECM proteins in the rat bladder wall, while tolterodine induced the expression of non-collagen ECM proteins and ECM receptors. Conclusions: Tolterodine decreased the volume of collagen in PBOO rat bladder wall, possibly via MMPs, and regulated the expression of ECM proteins and receptors.

Effects of varying degrees of partial bladder outlet obstruction on urinary bladder function of rats: A novel link to inflammation, oxidative stress and hypoxia.

The aim of the present study was to investigate the effects of different degrees of obstruction, and the roles of inflammation, oxidative stress, and hypoxia parameters on bladder function. Thirty male Sprague-Dawley rats were divided into 3 groups (n = 10 in each group): (i) sham-operated control; (ii) severe partial bladder outlet obstruction (PBOO); and (iii) moderate PBOO. Severe and moderate PBOO were induced by urethral ligation using 3-Fr and 4-Fr catheters, respectively, for 6 weeks. After 6 weeks, the in vitro contractile responses to carbachol, electrical field stimulation, ATP and KCl were measured in bladder strips. In addition, mRNA and protein expression of nuclear factor kappa B (NF-κB) hypoxia-inducible factor (HIF) and nuclear factor, erythroid 2-like 2 (Nrf2) in bladder were determined by real-time polymerase chain reaction and western blotting. Malondialdehyde (MDA) levels in bladder tissues were also determined. Rats in the severe PBOO group had the highest bladder weight. Detrusor strips from rats in the severe PBOO group exhibited 61%-82% smaller contractile responses to all four stimuli than those from the sham-operated group. Activity of NF-κB as an inflammatory marker was increased in the severe PBOO group, whereas HIF-1α and HIF-2β protein levels were increased significantly in the moderate PBOO group. A master regulator of oxidative stress, Nrf2 expression was increased in all obstructed rats. MDA levels were higher in the severe PBOO group than in sham-operated group. The results of the present study reveal the importance of oxidative stress-induced NF-κB signaling in bladder dysfunction with severe obstruction. Altered HIF signaling may contribute to the functional impairment after PBOO. Novel and evolving therapies targeting oxidative and/or inflammatory pathways may be a reasonable strategy for the management of lower urinary tract symptoms or benign prostatic hyperplasia.

Increased expression of neuregulin 1 in the urothelium of rat bladder with partial bladder outlet obstruction

BackgroundThis study determined whether changes in the expression of neuregulin (NRG) 1, erbB2 tyrosine kinase (ErbB2) and the M2 muscarinic receptor in the urothelium and detrusor muscle of the rat bladder were associated with partial bladder outlet obstruction (PBOO).MethodsMale Sprague-Dawley rats (body weight 250–300 g) were used and subdivided into control (n = 10) and PBOO groups (n = 20). PBOO was induced for 21 days, and the expression of NRG1, ErbB2 and M2 muscarinic receptor mRNA and protein was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting, respectively.ResultsIn the urothelium of rat bladder samples, protein expression and mRNA expression of NRG1, ErbB2 and M2 muscarinic receptor were significantly increased in the PBOO group compared to the control group (p < 0.05). Only mRNA expression levels of NRG1/ ErbB2 were higher in the detrusor muscle of the PBOO group compared to the control group (p < 0.05).ConclusionsOur study demonstrated remarkable changes in the expression of NRG1/ErbB2 receptor mRNA and protein in the urothelium and muscle layer. These results suggest that NRG1 overexpression plays some kind of role against the PBOO-induced upregulated muscarinic receptors in detrusor overactivity.

Evaluation of the effects of omega-3 & interferon alpha-2b administration on partial bladder outlet obstruction in a rat model.

Background & objectives:In bladder outlet obstruction-induced rat models, the transforming growth factor-beta (TGF-β) and collagen ratios have been shown to be increased. Increased TGF-β leads to fibrosis. In this study, the effect of omega-3 and interferon alpha-2b (IFN α-2b) was investigated on oxidative stress, inflammation and fibrosis in bladder structure in a partial bladder outlet obstruction (PBOO) rat model.Methods:A total of 35 male Wistar albino rats, weighing 300-350 g, were used in the study. The rats were randomly divided into five groups. At the end of the experimental period, bladders were harvested from all the rats, and pathological analysis of the rat bladder tissues was performed. In addition, investigations were carried out with enzymatic and non-enzymatic antioxidant systems to study the antioxidant properties of omega-3 fatty acid and IFN alpha-2b.Results:Increased bladder weight in the PBOO group, in comparison to the control group, was decreased by the administration of omega-3 and IFN α-2b (P=0.002). Significantly higher superoxide dismutase (SOD) levels were detected in group 2 in comparison to the control group. It was also detected that serum SOD, glutathione peroxidase and nitric oxide (NO) levels were significantly higher in group 2 when compared to the control group (P<0.05). In the pathologic evaluation, group 2 showed significantly increased inflammation and fibrosis compared to the control group. Omega-3 treatment significantly decreased inflammation. It was shown that IFN α-2b application partially decreased inflammation.Interpretation & conclusions:The results of the present study showed that in addition to the standard primary approaches to prevent the damage to the upper urinary tract secondary to PBOO, omega-3 fatty acid and IFN α-2b could be beneficial as adjunct treatment in clinical practice. However, this needs to be further investigated with prospective, randomized clinical trials with larger sample sizes.

MP52-13 THE EFFECT OF PARTIAL BLADDER OUTLET OBSTRUCTION (PBOO) SEVERITY AND HYPERCHOLESTEROLEMIA ON ERECTILE FUNCTION IN RATS

You have accessJournal of UrologySexual Function/Dysfunction/Andrology: Basic Research I1 Apr 2015MP52-13 THE EFFECT OF PARTIAL BLADDER OUTLET OBSTRUCTION (PBOO) SEVERITY AND HYPERCHOLESTEROLEMIA ON ERECTILE FUNCTION IN RATS Serap Gur, Nur Bayatli, and Didem Yilmaz Serap GurSerap Gur More articles by this author , Nur BayatliNur Bayatli More articles by this author , and Didem YilmazDidem Yilmaz More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.1730AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Erectile dysfunction (ED) and lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) are commonly coexist and very prevalent in men aged over 50 years. One of the leading theories of this interrelation is atherosclerosis-induced pelvic injury. We investigated the degree of partial bladder outlet obstruction (PBOO) induced by a ligation (with 5F or 4F catheter [c]) of the bladder neck and the role of atherosclerosis induced by hypercholesterolemic diet (HCD) on erectile function in rats. METHODS A total of 96 Sprague-Dawley male rats were divided into 6 groups: 1) Sham-operated; 2) PBOO (5Fc); 3) PBOO (4Fc); 4) HCD 5)HCD-PBOO (5Fc) and 6) HCD-PBOO (4Fc). HCD groups were fed 4% cholesterol and PBOO groups were subjected by ligation of the urethra for 6 wk. Erectile response was measured using cavernosal nerve stimulation by evaluating ratios of intracavernosal pressure (ICP)/mean arterial pressure. The relaxant and contractile properties of corpus cavernosum smooth muscle (CCSM) strips were evaluated in tissue baths. Immunohistochemical (IHC) staining for neuronal nitric oxide synthase (nNOS), endothelial NOS (eNOS), Rho-activated kinase (ROCK) and collagen deposition was performed. RESULTS The weight of bladder enhanced gradually in obstructed and combination groups and peaked in HCD-PBOO (4F)(Table). Relaxant and contractile responses of CCSM decreased depending on the severity of PBOO with HCD (Table). Rho-kinase inhibitor Y-27632 (10 μM) inhibited peak and sustained contractile responses to phenylephrine in groups (Table). We found lower nNOS, eNOS, higher Rho-kinase and collagen expressions in PBOO and combined groups. CONCLUSIONS The PBOO caused the more marked ED manifesting by considerable lowering of erectile response and the shift HCD in rats promoted. Our results suggest that the PBOO causes the functional and molecular changes, penile remodeling, accumulation of extracellular collagen in penile tissue, which may become more stressed with the HCD. The management of atherosclerosis may be beneficial in the treatment and continued research are needed for PBOO with HCD and will undoubtedly contribute to the future development of novel therapies for LUTS and ED. Sham-control PBOO(5F) PBOO(4F) HCD HCD-PBOO(5F) HCD- PBOO(4F) Bladder weight (g) 0.12 ± 0.01 0.25 ± 0.008*** 0.34 ± 0.004*** 0.53 ± 0.02*** 0.67 ± 0.004***### 0.91 ± 0.01 ***††† ICP/MAP (7.5V) 1.15 ± 0.06 0.66 ± 0.007*** 0.50 ± 0.020*** 0.52 ± 0.010*** 0.32 ± 0.01***### 0.10 ± 0.002***††† ACh (10-3M) 91.5 ± 1.5 78.9 ± 0.7*** 63.7 ± 2.5*** 46.7 ± 1.3*** 18.9 ± 1.1*** ### 6.2 ± 0.2*** ††† EFS relaxation (20Hz) 96.7 ±1.4 86.9 ± 1.6* 70.5 ± 2.1*** 62.6 ± 0.5*** 55.3 ± 1.9***### 16.6 ± 1.6***††† SNP (10-3M) 100 ± 0 100 ± 0 97.4 ± 0.9 91.4 ± 0.3 81.4 ± 0.3*** ### 66.1 ± 0.3*** ††† EFS contraction (40Hz) 69.5 ± 0.2 51.4 ± 0.8*** 43.8 ± 0.1*** 39.6 ± 0.02*** 29.0 ± 0.3***### 18.4 ± 0.9***††† Phe (10-3M) 90.9 ± 4.4 60.3 ± 2.6*** 43.5 ± 1.3*** 36.7 ± 0.8*** 28.1 ± 1.1***### 13.0 ± 1.1***††† Y-27 632(1μM) 100 ± 0 93.8 ± 0.3 43.5 ± 0.2*** 36.7 ± 0.5*** 28.13 ± 1.1***### 13.0 ± 0.8***††† Data are mean ± SEM (n=12-15). *p<0.05, **p<0.01, ***p<0.001 vs sham-control; ###p<0.001 vs PBOO (5F) and †††p<0.001 vs PBOO (4F). © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e632 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Serap Gur More articles by this author Nur Bayatli More articles by this author Didem Yilmaz More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...

MP31-14 CHANGES IN APOPTOSIS RELATED PROTEINS IN THE BLADDER AFTER PARTIAL BLADDER OUTLET OBSTRUCTION RELIEF

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 2015MP31-14 CHANGES IN APOPTOSIS RELATED PROTEINS IN THE BLADDER AFTER PARTIAL BLADDER OUTLET OBSTRUCTION RELIEF Ki Hak Song, Chong koo Sul, Yong gil Na, Jae sung Lim, Ju Hyun Shin, and Jong mok Park Ki Hak SongKi Hak Song More articles by this author , Chong koo SulChong koo Sul More articles by this author , Yong gil NaYong gil Na More articles by this author , Jae sung LimJae sung Lim More articles by this author , Ju Hyun ShinJu Hyun Shin More articles by this author , and Jong mok ParkJong mok Park More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.1369AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Partial bladder outlet obstruction (PBOO) induces prolonged bladder over-distension, leading to bladder dysfunction and structural alterations via induction of apoptosis. The aim of present study was to investigate sequential course of the apoptosis in bladder tissue, and changes of apoptosis related proteins in PBOO and subsequent relief. METHODS The study was conducted using 60 female Sprague-Dawley rats (body weight 250–300 g), divided into sham operated group (n=20), PBOO only (n=20), and PBOO group plus subsequent relief (n=20). PBOO rats were induced for 2 weeks, the obstruction has relived by removal of ligation after 2 weeks. After PBOO and relief surgery, urodynamic studies measuring contraction interval and contraction pressure were done. The tissues of rats were assessed by histologic anaylsis (H&E and TUNNEL stain) and expression of apoptosis related receptors (such as caspase-3, Erk, Jnk, P38, Bax, Bcl-2 and Survivin) have detected by RT-PCR and Western blotting. RESULTS Filling cystometry studies showed a significant decrease in contraction interval and increase in contraction pressure in the PBOO group. TUNNEL positive cells were significantly increased in PBOO group compared to sham operated group and decreased in PBOO relief group compared to PBOO group. On RT-PCR and Western blotting, expression of Bax, caspase-3, P38 and Jnk was significantly increased in PBOO group. But, expression of Erk, Bcl-2 was significantly decreased in PBOO group. The expression of Erk and Bcl-2 was significantly increased in PBOO relief group compared to PBOO group. CONCLUSIONS PBOO induced apoptosis of bladder tissue is associated with imbalance of MAPK pathways and apoptosis related protein. These results may also imply that pro-survival Erk kinase cascade is activated in response ischemic bladder injury and associated with initiation of bladder protection after PBOO. However, the mechanism of Survivin as anti-apoptotic factor in ischemic bladder tissue remains unclear. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e361 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ki Hak Song More articles by this author Chong koo Sul More articles by this author Yong gil Na More articles by this author Jae sung Lim More articles by this author Ju Hyun Shin More articles by this author Jong mok Park More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Effect of short- and long-term sildenafil treatment on erectile dysfunction in rats with partial bladder outlet obstruction.

Lower urinary tract symptoms (LUTS) and erectile dysfunction (ED) are frequent problems in older men worldwide. We evaluated the effect of short- and long-term sildenafil treatment on erectile function in rats with surgically induced partial bladder outlet obstruction (PBOO). A total of 60 male Sprague-Dawley rats were randomized in five groups: (1) control (sham-operated); (2) PBOO for 3 weeks; (3) PBOO for 6 weeks; (4) sildenafil (1.5 mg/rat/day) treated PBOO for 3 weeks; and (5) sildenafil treated PBOO for 6 weeks. We assessed erectile function by measuring intracavernous pressures (ICP), mean arterial pressure (MAP) and total ICP after cavernous nerve stimulation. Corpus cavernous smooth muscle (CCSM) strips were isolated and evaluated for relaxation responses using organ-bath preparation. Neuronal nitric oxide synthase (nNOS) expression was determined immunohistochemically. Experimental PBOO at 3 and 6 weeks showed decreased erectile response based on ICP/MAP ratio, total ICP and decreased expression of nNOS, which returned to normal after prolonged daily treatment with sildenafil. CCSM strips from PBOO rats displayed reduced relaxation responses to both electrical field stimulation (EFS) and acetylcholine (ACh) as well as nNOS enzyme intensity when compared to untreated PBOO group, which was reversed by treatment with sildenafil for 6 weeks. Daily sildenafil treatment prevents development of ED in PBOO rats in a time dependent manner. Further studies are needed to explore the effectiveness of sildenafil in patients with BPH/LUTS in association with ED.

Contribution of bone marrow-derived mesenchymal stem cells to the morphological changes in the bladder after partial outlet obstruction: A preliminary study

To investigate the contribution of bone marrow-derived mesenchymal stem cells to the changes in bladder morphology in response to partial bladder outlet obstruction. Allogenic bone marrow cells were transplanted from transgenic rats expressing green fluorescent protein into female Sprague-Dawley rats 1 day after their bone marrow-derived mesenchymal stem cells had been destroyed by irradiation. This generated chimeric rats in which green fluorescent protein labeled bone marrow-derived mesenchymal stem cells replaced host bone marrow-derived mesenchymal stem cells. The animals received partial bladder outlet obstruction or sham surgery 6 weeks later. The animals were killed 6 weeks after the surgery, and bladder tissue was prepared for immunofluorescence with antibodies against a urothelium marker (AE1/AE3), a myofibroblast marker (vimentin), a smooth muscle marker and green fluorescent protein. More labeled bone marrow-derived mesenchymal stem cells were found in the partial bladder outlet obstruction group than in the in the sham group. Most bone marrow-derived mesenchymal stem cells were present around the basement membrane (laminin) and lamina propria below the urothelium. Bone marrow-derived mesenchymal stem cells were also found in the urothelium layer, and some of them were double-stained with green fluorescent protein and AE1/AE3. Some bone marrow-derived mesenchymal stem cells, which were located in the interstitial tissue, were double-stained with green fluorescent protein and vimentin. Bone marrow-derived mesenchymal stem cells, which migrated into the smooth muscle layer, showed fusiform morphology, and some were double-stained with green fluorescent protein and smooth muscle marker. Bone marrow-derived mesenchymal stem cells home to the partial bladder outlet obstruction bladder, and these cells have the potential to differentiate into the several components of bladder tissue including the urothelium, myofibroblasts and smooth muscle cells. Thus, bone marrow-derived mesenchymal stem cells contribute to the morphological changes of the bladder in response to partial bladder outlet obstruction.

Expression and electrophysiological characteristics of P2X3 receptors in interstitial cells of Cajal in rats with partial bladder outlet obstruction

WHAT'S KNOWN ON THE SUBJECT? AND WHAT DOES THE STUDY ADD?: In the urinary bladder, histological studies suggest a network of functionally connected interstitial cells of Cajal (ICCs) are located between the urothelium and sensory nerve endings, which might transfer signals directly between them. Purinergic P2X3 receptors may play roles in the micturition reflex pathway, and its expression profiles in ICCs could be altered in urinary bladder dysfunction. The present experiments showed a novel time-dependent P2X3 receptor up-regulation in ICCs in an experimental rat model of partial bladder outlet obstruction. To investigate the expression and electrophysiological characteristics of purinergic P2X3 receptors in interstitial cells of Cajal (ICCs) at different time points after partial bladder outlet obstruction (PBOO) in rats. In all, 48 female Sprague-Dawley rats were randomly divided into four groups: 4, 6 and 8 week PBOO groups and sham-operated controls. At 4 weeks after surgery, cystometry was performed to investigate bladder function in vivo. Subsequently, the rats were humanely killed at 4, 6 or 8 weeks and the bladders were harvested for measurements. P2X3 expression in ICCs of bladder was investigated by immunofluorescent staining. The level of P2X3 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). Inward currents in corresponding ICCs after PBOO were investigated electrophysiologically. Cystometrography showed a valid increase in maximum detrusor pressure in rats subjected to PBOO. The bladder weight in the PBOO group was significantly higher than that in the control group. In contrast to sham rats, there was a significant increase in the intensity of P2X3 staining in the ICCs in all PBOO groups. C-kit labelled isolated ICCs were strongly stained with the P2X3 antibody. RT-PCR showed that the expression of P2X3 mRNA was significantly up-regulated at 4, 6 and 8 weeks in the ICCs from the PBOO rats. In the ICCs, the mean peak amplitude of inward currents was significantly increased in the PBOO groups compared with the control group. The expression of P2X3 receptors showed a time dependent up-regulation in the ICCs of the bladder in rats with PBOO. PBOO induced the potentiation of P2X3 receptors function, as evidenced by α, β-methylene ATP-enhanced inward currents in the ICCs of the rat bladder.

Changes in Angiotensin II Type 1 Receptor Expression in the Rat Bladder by Bladder Outlet Obstruction

Purpose: To demonstrate the change in the expression of angiotensin II type 1 receptor (AT1) in the rat bladder with partial bladder outlet obstruction (P-BOO). Materials and Methods: Bladder specimens were obtained from 12-week-old Wistar female rats that were divided into two groups, a P-BOO group and a control group. The rats of the P-BOO group were divided into six groups: a sham-operated control group, 1 day postoperatively, 2 days postoperatively, 4 days postoperatively, 7 days postoperatively and 14 days postoperatively. The cystometric findings and immunohistochemical staining of the detrusor muscle with the AT1 antibody were compared in each group. Results: AT1 localized on the cell membrane of the detrusor smooth muscle and in cytoplasm of suburothelial myofibroblasts in the control rats. The expression of AT1 disappeared in the detrusor muscle and suburothelial myofibroblasts in P-BOO, but AT1 was highly expressed in urothelial cells 1 day after surgery. The expression of AT1 in urothelial cells gradually decreased with time after surgery. AT1 completely disappeared in urothelial cells 14 days after surgery. Conclusions: The present study demonstrated that the site of AT1 expression changes in response to the mechanical stress caused by P-BOO, and finally there was no expression of AT1 in rat bladder tissue following P-BOO. These data suggest the change in AT1 expression may play a role in bladder function.

The Relaxant Effect of Ginseng Saponin on the Bladder and Prostatic Urethra: An in vitro and in vivo Study

Aim: To assess the effects of ginseng saponin on relaxation of the bladder and prostatic urethra and to determine its mechanism of action. Materials and Methods: For the in vitro study, prostatic urethra muscle strips were harvested from 18 male New Zealand rabbits. The strips were mounted in organ baths and connected to force displacement transducers. After stabilization, maximal tissue contractions were obtained by the application of phenylepinephrine to the urethra strips, and a dose-response curve for ginseng saponin was constructed (10^–6–10^–2M). After pretreatment of urethra strips with N-nitro-L-arginine methyl ester (L-NAME), another dose-response curve for ginseng saponin was constructed. For the in vivo study, we used adult male Sprague-Dawley rats divided into three groups [control, partial bladder outlet obstruction (PBOO) and saponin-fed groups], and we monitored the vesical pressure (P_ves) and urethral perfusion pressure (UPP). Results: The ginseng saponin induced a significant dose-dependent relaxant effect on the prostatic urethra strips. A significant relaxant effect of ginseng saponin was observed from 10^–3M, and ginseng saponin significantly relaxed urethra strips by 50.2 ± 20.26% at 10^–2M. The relaxant effect was partially inhibited with L-NAME pretreatment. In the in vivo study, the change in UPP between baseline and relaxation was significantly higher in the saponin group than in the control or PBOO group (p < 0.001). The saponin group showed a significantly lower baseline P_ves than the PBOO group. Conclusions: We observed a significant relaxation effect of ginseng saponin on the bladder and prostatic urethra in both in vitro and in vivo studies. The mechanism by which ginseng saponin induces relaxation appears to involve the nitric oxide/nitric oxide synthase pathway.

1006 IMPAIRMENT OF SENSORY INNERVATION IN THE BLADDER NECK BY OUTLET OBSTRUCTION CONTRIBUTES TO THE DEVELOPMENT OF ERECTILE DYSFUNCTION

You have accessJournal of UrologySexual Function/Dysfunction/Andrology: Basic Research II1 Apr 20121006 IMPAIRMENT OF SENSORY INNERVATION IN THE BLADDER NECK BY OUTLET OBSTRUCTION CONTRIBUTES TO THE DEVELOPMENT OF ERECTILE DYSFUNCTION Qi Lei, Joseph Hypolite, Shaohua Chang, Robert Seftel, Samuel Chacko, Alan Wein, Allen Seftel, and Anna Malykhina Qi LeiQi Lei Glenolden, PA More articles by this author , Joseph HypoliteJoseph Hypolite Glenolden, PA More articles by this author , Shaohua ChangShaohua Chang Camden, NJ More articles by this author , Robert SeftelRobert Seftel Glenolden, PA More articles by this author , Samuel ChackoSamuel Chacko Glenolden, PA More articles by this author , Alan WeinAlan Wein Philadelphia, PA More articles by this author , Allen SeftelAllen Seftel Camden, NJ More articles by this author , and Anna MalykhinaAnna Malykhina Glenolden, PA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1108AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Benign prostatic hyperplasia (BPH) and erectile dysfunction (ED) are common problems in aging males worldwide. Clinical evidence of impaired sensation in ED suggests a neurogenic component of ED development, especially when ED is co-morbid with lower urinary tract symptoms. The objective of this work was to evaluate the role of the damage to bladder neck nerves during partial bladder outlet obstruction (PBOO) on sensory innervation of the corpus cavernosum (CC) and CC smooth muscle (CCSM) function using an animal model of PBOO. METHODS Four groups of adult male Sprague Dawley rats (N=32) were included in the study: sham operated (control), PBOO surgery, sham and PBOO groups with injected retrograde tracers. PBOO was surgically induced by a partial ligation of the bladder neck during laparotomy. Retrograde labeling technique was used to label L6-S2 dorsal root ganglion (DRG) neurons which innervate the urinary bladder and CC. The contractility and relaxation of the CC smooth muscle strips was studied in vitro in response to different stimuli. RESULTS Urinary bladders from animals with PBOO weighted 308.75±61.83 mg (N=13, 2 weeks PBOO) when compared to sham group (126.60±12.80 mg, N=13, p≤0.05). L6-S1 DRG ganglia were removed bilaterally and sectioned to count bladder and CC labeled sensory neurons. The obtained results showed that L6-S2 DRG ganglia receive input from both the urinary bladder and CC and 2-3% of all labeled neurons were double labeled from both organs suggesting some overlap in sensory innervation. The contractile response of CC muscle strips to electric stimulation was significantly reduced in PBOO group (2.223±0.3262 g/mg, p≤0.05) when compared to control (3.6367±0.3577 g/mg). Force generation of CCSM to 125 mM KCl was also significantly reduced in the PBOO group (2.61±0.35 g/mg vs 4.94±0.91 g/mg in control, p≤0.05). After CCSM was pre-contracted with phenylephrine, relaxation induced by nitroprusside was significantly diminished in PBOO group (2.76±0.29 g/mg vs 3.66±0.27 g/mg in the control, p≤0.05). There was also a significant decrease in n-NOS expression in PBOO animals when compared to the sham group (47%, N=4, p≤0.05). CONCLUSIONS PBOO leads to the impairment of bladder neck innervation followed by an increase in CCSM tone and a down-regulation of nNOS expression in the CC smooth muscle. These results provide evidence that nerve damage in PBOO may contribute to the development of ED causing secondary changes in the contraction/relaxation mechanisms of CCSM. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e409 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Qi Lei Glenolden, PA More articles by this author Joseph Hypolite Glenolden, PA More articles by this author Shaohua Chang Camden, NJ More articles by this author Robert Seftel Glenolden, PA More articles by this author Samuel Chacko Glenolden, PA More articles by this author Alan Wein Philadelphia, PA More articles by this author Allen Seftel Camden, NJ More articles by this author Anna Malykhina Glenolden, PA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Effect of doxazosin on expression of eNOS in rat kidney in the presence of partial bladder outlet obstruction

The role of nitric oxide in the pathogenesis of renal injury has begun to be appreciated. We therefore designed this study to demonstrate the relationship between endothelial nitric oxide synthase (eNOS) expression and doxazosin in the kidneys of rats with surgically created partial bladder outlet obstruction (BOO), to further understand the role of doxazosin in the prevention of renal parenchymal damage by partial BOO. A total of 35 adult female Wistar rats, mean weight 250 g, were randomly allocated to 3 experimental groups: group1, sham-operated (n=10); group 2, partial BOO group (n=14) and group 3, partial BOO group treated with doxazosin (n=11). Partial BOO in rats was surgically induced. Results were assessed by eNOS immunohistochemistry. eNOS staining in kidneys in group 1 (16.45 ± 1.63) was significantly higher than in group 2 (5.09 ± 0.61) (p<0.05). After 15 days of doxazosin treatment in addition to partial BOO (group 3), eNOS staining in the kidney (11.80 ± 1.63) was significantly higher than in group 2 (5.09 ± 0.61) (p<0.05). In samples taken after 15 days of doxazosin treatment in addition to partial BOO, eNOS staining in kidneys (11.80 ± 1.63) was lower than in the sham-operated group (16.45 ± 1.63), but the difference was not significant (p>0.05). These findings may provide insight into the beneficial and restorative effects of α(1)-adrenoceptor antagonists on eNOS expression in the kidney, when used to treat symptoms of benign prostate hyperplasia and hypertension.

Increased penile expression of transforming growth factor and elevated systemic oxidative stress in rabbits with chronic partial bladder outlet obstruction

There is a growing body of evidence to support the direct link between obstructive bladder dysfunction and erectile dysfunction (ED). However, there have been few pathophysiological studies to determine the relationship between lower urinary tract syndrome (LUTS) and ED. As the transforming growth factor-β1 (TGF-β1) that induces the synthesis of collagen in the penile tissues is critical for the development of ED, the first aim of this study was to investigate the expression of TGF-β1 in the penis from male rabbits with chronic partial bladder outlet obstruction (PBOO). Besides, it has been suggested that oxidative stress plays a significant role in the pathophysiological mechanism of ED. Thus, the second aim of this study was to further investigate whether the urinary or serum oxidative stress markers are involved in chronic PBOO-induced penile dysfunction. A total of 16 male New Zealand White rabbits were separated equally into four groups: a control group and PBOO groups obstructed for 2, 4 and 8 weeks respectively. Using the RT-PCR and Western blot analysis, a progressive increase of TGF-β1 in penis was found at 2, 4 and 8 weeks after obstruction. Moreover, the biomarkers for oxidative stress or oxidative damage were significantly detected in the penis of rabbits after PBOO, which include the enhancement of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in urine and plasma, plasma malondialdehyde (MDA) and total antioxidant capacity (TAC), as well as reduction of glutathione (GSH). On the basis of our results, the increase of TGF-β1 and elevated systemic oxidative stress may play key roles to contribute to penile dysfunction after chronic PBOO.