Acetic Acid Infusion Research Articles

MP8-18 THE ROLE OF THE BRAINSTEM IN TIBIAL INHIBITION OF THE MICTURITION REFLEX IN CATS

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Basic Research I1 Apr 2015MP8-18 THE ROLE OF THE BRAINSTEM IN TIBIAL INHIBITION OF THE MICTURITION REFLEX IN CATS Matthew Ferroni, Richard Slater, Bing Shen, Zhiying Xiao, Jicheng Wang, Andy Lee, James Roppolo, William de Groat, and Changfeng Tai Matthew FerroniMatthew Ferroni More articles by this author , Richard SlaterRichard Slater More articles by this author , Bing ShenBing Shen More articles by this author , Zhiying XiaoZhiying Xiao More articles by this author , Jicheng WangJicheng Wang More articles by this author , Andy LeeAndy Lee More articles by this author , James RoppoloJames Roppolo More articles by this author , William de GroatWilliam de Groat More articles by this author , and Changfeng TaiChangfeng Tai More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.285AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Tibial Nerve Stimulation (TNS) has been shown to significantly inhibit the micturition reflex when the bladder is distended by saline infusion or irritated by 0.25% acetic acid (AA) in cats. However, TNS inhibition of AA-induced bladder overactivity is lost after acute spinal cord transection at the level of T9/T10 indicating a supraspinal site of action. This study determines the role of the brainstem in TNS inhibition, which is important for understanding the mechanism of action of tibial neuromodulation therapy for overactive bladder. METHODS 16 cats under isoflurane anesthesia underwent decerebration by surgical transection rostral to the superior colliculi. Then, multiple saline cystometrograms (CMGs) were performed under á-chloralose anesthesia to elicit micturition reflex contractions (>30 cmH2O). 30 minutes of continuous TNS at three times threshold intensity for inducing toe movement was then applied during isovolumetric bladder contractions at frequencies of either 5Hz or 30Hz. After each stimulation, five consecutive CMGs were performed to assess post-stimulation changes of bladder capacity. CMGs were then performed during TNS at either 5Hz or 30Hz. Next, the same stimulation protocol was repeated during AA irritation. At the end of each experiment, naloxone (an opioid receptor antagonist) was administered either intravenously (1 mg/kg) or directly into the brainstem (2.0-4.0 mg in 0.1-0.2 cc) followed by a series of CMGs with/without TNS. ANOVA tests were used to determine statistical significance (p<0.05). RESULTS During saline infusion, 30 minutes of TNS significantly increased post-stimulation bladder capacity above controls with a mean increase of 27.2% (p<0.01) at a frequency of 5 Hz and 44.2% (p<0.01) at a frequency of 30 Hz. However, post-stimulation TNS inhibition was not observed during AA infusion. TNS applied during CMG significantly increased bladder capacity by 27.3% (saline, p<0.01) and 87.6% (AA, p<0.01) respectively over controls at 5 Hz, but failed to achieve significant inhibition at 30 Hz. Naloxone treatments significantly reduced control bladder capacity whether given intravenously (68.1% reduction, p<0.01) or directly into brainstem (58.8% reduction, p<0.01) and completely eliminated the acute TNS inhibition. CONCLUSIONS Our results suggest: (1) the brainstem plays an important role in TNS inhibition of both non-nociceptive (saline distention) and nociceptive (AA irritation) bladder reflexes while the cerebrum is not necessary; (2) opioid receptors in the brainstem are involved in TNS inhibition of the nociceptive bladder reflex. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e79-e80 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Matthew Ferroni More articles by this author Richard Slater More articles by this author Bing Shen More articles by this author Zhiying Xiao More articles by this author Jicheng Wang More articles by this author Andy Lee More articles by this author James Roppolo More articles by this author William de Groat More articles by this author Changfeng Tai More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

A spinal GABAergic mechanism is necessary for bladder inhibition by pudendal afferent stimulation.

Electrical stimulation of pudendal afferents can inhibit bladder contractions and increase bladder capacity. Recent results suggest that stimulation-evoked bladder inhibition is mediated by a mechanism other than activation of sympathetic bladder efferents in the hypogastric nerve, generating α-adrenergic receptor-mediated inhibition at the vesical ganglia and/or β-adrenergic receptor-mediated direct inhibition of the detrusor muscle. We investigated several inhibitory neurotransmitters that may instead be necessary for stimulation-evoked inhibition and found that intravenous picrotoxin, a noncompetitive GABAA antagonist, significantly and reversibly blocked pudendal afferent stimulation-evoked inhibition of bladder contractions in a dose-dependent manner. Similarly, intravenous picrotoxin also blocked pudendal afferent stimulation-evoked inhibition of nociceptive bladder contractions evoked by acetic acid infusion. Furthermore, intrathecal administration of picrotoxin at the lumbosacral spinal cord also blocked bladder inhibition by pudendal afferent stimulation. On the other hand, glycinergic, adrenergic, or opioidergic mechanisms were not necessary for bladder inhibition evoked by pudendal afferent stimulation. These results identify a lumbosacral spinal GABAergic mechanism of bladder inhibition evoked by pudendal afferent stimulation.

Su2073 Neonatal Colonic Irritation Produces Persistent Visceral Hypersensitivity and Altered Affective Behavior in a Mouse Model of Irritable Bowel Syndrome (IBS)

Introduction: A major theme in functional bowel disorders (FBD) such as irritable bowel syndrome (IBS) is that psychological/psychiatric problems have a pathogenic role, based both on the observation that patients with FBD are more anxious and depressed than healthy control, as well as research linking stress and depression to altered gastrointestinal sensory and motor function. However, recent experimental and epidemiological studies suggest that primary visceral disturbances can lead secondarily to persistent behavioral and emotional abnormalities. The aims of the present study were to test the hypothesis that mild chemical irritation of the colon of neonatal mice can produce changes in visceral hypersensitivity and psychological behaviors that persist into adulthood. Methods: Ten-day old C57BL/6 mouse pups received an intracolonic infusion of 20 μl of 0.5% acetic acid (IBS) or saline (control) to mimic a previously validated model of IBS in rats based on neonatal visceral irritation. Psychological behaviors and pain sensitivity of the colon were assessed when these mice were 8-14 weeks of age. Anxiety-like behaviors were tested using elevated plus maze and open field tests. Depression-like behavior was tested by the forced swim test. Colonic hypersensitivity to graded colorectal balloon distension (15, 30, 50 and 70mmHg) was determined by visceromotor responses (VMR) measured by electromyography of external oblique muscle. Resutls: Neonatal colorectal infusion of acetic acid but not saline significantly increased electromyographic activity in response to all of pressure colorectal distension in adults, suggesting the persistence of colonic hypersensitivity. In the elevated plus maze and open field tests, IBS mice showed a significant reduction in time spent in the open arms of elevated plus maze (control: 73.04±5.26s vs IBS: 53.54±6.53s, p<0.05) and in the center of the open field (control: 73.99±6.42s vs IBS: 37.32±4.62s, p<0.05), suggesting an anxietylike phenotype. IBS mice also showed a significant increase in immobility in the forced swim test (control: 90.05±12.08s vs IBS: 140.43±20.06s, p<0.05), suggesting an increase in depression-like behavior. Conclusion: The present results demonstrate that transient colonic irritation in the neonatal period can induce long-lasting depression-like and anxietylike behaviors. These findings provide a new paradigm for the psychological co-morbidity in functional bowel disorders.

Su2074 The Effects of Adenosine A3 Receptor Agonist on Visceral Hypersensitivity of Neonatal Maternally Separated Rat Models

Introduction: A major theme in functional bowel disorders (FBD) such as irritable bowel syndrome (IBS) is that psychological/psychiatric problems have a pathogenic role, based both on the observation that patients with FBD are more anxious and depressed than healthy control, as well as research linking stress and depression to altered gastrointestinal sensory and motor function. However, recent experimental and epidemiological studies suggest that primary visceral disturbances can lead secondarily to persistent behavioral and emotional abnormalities. The aims of the present study were to test the hypothesis that mild chemical irritation of the colon of neonatal mice can produce changes in visceral hypersensitivity and psychological behaviors that persist into adulthood. Methods: Ten-day old C57BL/6 mouse pups received an intracolonic infusion of 20 μl of 0.5% acetic acid (IBS) or saline (control) to mimic a previously validated model of IBS in rats based on neonatal visceral irritation. Psychological behaviors and pain sensitivity of the colon were assessed when these mice were 8-14 weeks of age. Anxiety-like behaviors were tested using elevated plus maze and open field tests. Depression-like behavior was tested by the forced swim test. Colonic hypersensitivity to graded colorectal balloon distension (15, 30, 50 and 70mmHg) was determined by visceromotor responses (VMR) measured by electromyography of external oblique muscle. Resutls: Neonatal colorectal infusion of acetic acid but not saline significantly increased electromyographic activity in response to all of pressure colorectal distension in adults, suggesting the persistence of colonic hypersensitivity. In the elevated plus maze and open field tests, IBS mice showed a significant reduction in time spent in the open arms of elevated plus maze (control: 73.04±5.26s vs IBS: 53.54±6.53s, p<0.05) and in the center of the open field (control: 73.99±6.42s vs IBS: 37.32±4.62s, p<0.05), suggesting an anxietylike phenotype. IBS mice also showed a significant increase in immobility in the forced swim test (control: 90.05±12.08s vs IBS: 140.43±20.06s, p<0.05), suggesting an increase in depression-like behavior. Conclusion: The present results demonstrate that transient colonic irritation in the neonatal period can induce long-lasting depression-like and anxietylike behaviors. These findings provide a new paradigm for the psychological co-morbidity in functional bowel disorders.

ADX71441, a novel, potent and selective positive allosteric modulator of the GABA(B) receptor, shows efficacy in rodent models of overactive bladder.

The GABAB receptor agonist baclofen reduces urethral resistance and detrusor overactivity in patients with spasticity. However, baclofen's side effects limit its use for the treatment of overactive bladder (OAB). Here, we tested a novel GABAB positive allosteric modulator (PAM) ADX71441 in models of OAB in mice and guinea pigs. Mice were left untreated or given (p.o.) vehicle (1% CMC), ADX71441 (1, 3, 10 mg kg(-1) ) or oxybutynin (100 mg kg(-1) ; Experiment 1) or vehicle (1% CMC), baclofen (1, 3, 6 mg kg(-1) ) or oxybutynin (Experiment 2). Treated mice were then overhydrated with water, challenged with furosemide, before being placed into micturition chambers and monitored for urinary parameters. In anaesthetized guinea pigs, intravesical infusion of acetic acid was used to induce OAB and the effects of ADX71441 (1, 3 mg kg(-1) ) or baclofen (1 mg kg(-1) ), administered i.v., on cystometric parameters were monitored. In mice, 10 mg kg(-1) ADX71441 increased urinary latencies, reduced the number of urinary events and the total and average urinary volumes. In guinea pigs, ADX71441 (1 and 3 mg kg(-1) ) increased the intercontraction interval (ICI) and bladder capacity (BC), and reduced micturition frequency (MF) compared to vehicle. At 3 mg kg(-1) ADX71441 completely inhibited the micturition reflex and induced overflow incontinence in five out of 10 animals. Baclofen slightly increased ICI and BC and reduced MF. Our findings demonstrate, for the first time, that a GABAB PAM has potential as a novel approach for the treatment of OAB.

Neuromodulation attenuates bladder hyperactivity in a rat cystitis model

BackgroundWe investigated the regulation of urinary bladder function by electrical stimulation of the L6 spinal nerve (SN) using cystometry in normal rats and in rats with cystitis induced by intravesical infusion of dilute acetic acid.MethodsIn anesthetized rats, a cannula was placed into the bladder dome for saline/acetic acid infusion and intravesical pressure monitoring. Threshold pressure (TP), basal pressure (BP) and inter-contraction interval (ICI) were measured from the bladder pressure recording and void volume (VV) was measured by weighing the voided fluid.ResultsComparison of cystometrograms obtained with infusion of saline or acetic acid showed that acetic acid decreases TP, ICI and VV. These excitatory effects, characteristic of acetic acid induced bladder hyperactivity, were significantly reversed by bilateral SN stimulation (P <0.05, vs pre-stimulation, Student t-test). In saline perfused rats, one hour of bilateral SN stimulation at 10 Hz and at motor threshold (0.19 ± 0.01 milliamps) increased ICI (265%) and VV (217%). In rats perfused with acetic acid, the corresponding increases produced by SN stimulation were 350% for ICI and 383% for VV. The percentage increases in the acetic acid-treated group were not significantly higher than those in saline-treated group.ConclusionsUsing continuous flow cystyometry, we find that SN stimulation can produce effects on micturition consistent with its effects on isovolumetric model, and consistent with the therapeutic effect observed with InterStim® therapy in overactive bladder patients. Although the effect of SN stimulation was slightly greater in bladder irritated over normal rats, the difference was not statistically significant.

Roles of adenosine A1 and A2A receptors in the control of micturition in rats

Adenosine is a neurotransmitter that exerts numerous physiological effects in many organs. However, few studies have focused on the role of adenosine receptors in the control of micturition. Therefore, we examined the role of adenosine A1 and A2A receptors in the control of bladder activity in rats with normal or acetic acid (AA) irritated bladders. Cystometrograms during saline or 0.2% AA infusion were recorded under urethane anesthesia in female Sprague-Dawley rats. After a stabilization period, CCPA (A1 receptor agonist) and/or ZM24138 (A2A receptor antagonist) were administered intravenously (i.v.), intrathecally (i.t.), intracerebroventricularly (i.c.v.), or intravesically. Micturition parameters were recorded and compared before and after drug administration. I.v., i.t., or i.c.v. administration of CCPA or ZM24138 significantly increased intercontraction intervals (ICIs) in both saline and AA infusion groups. During AA infusion, the inhibitory effects induced by i.c.v. CCPA or i.t. ZM24138 were significantly greater than those by i.t. or i.c.v. administration, respectively. Intravesical administration of CCPA, but not ZM24138, significantly increased ICI. These results indicated that: (1) when nociceptive signals from the bladder increase, adenosine A1 receptor-mediated inhibition of micturition is enhanced in the brain, compared to the normal condition, (2) A1 receptor activation also exerts a peripheral inhibitory effect on micturition, and (3) adenosine A2A receptor-mediated excitatory mechanisms are enhanced in the spinal cord following C-fiber bladder afferent stimulation. Thus adenosine A1 receptor agonists and A2A receptor antagonists might be effective for the treatment of overactive bladder and/or bladder hypersensitive disorders, in which C-fiber afferent function is enhanced.

Stability of the acetic acid-induced bladder irritation model in alpha chloralose-anesthetized female cats.

Time- and vehicle-related variability of bladder and urethral rhabdosphincter (URS) activity as well as cardiorespiratory and blood chemistry values were examined in the acetic acid-induced bladder irritation model in α-chloralose-anesthetized female cats. Additionally, bladder and urethra were evaluated histologically using Mason trichrome and toluidine blue staining. Urodynamic, cardiovascular and respiratory parameters were collected during intravesical saline infusion followed by acetic acid (0.5%) to irritate the bladder. One hour after starting acetic acid infusion, a protocol consisting of a cystometrogram, continuous infusion-induced rhythmic voiding contractions, and a 5 min “quiet period” (bladder emptied without infusion) was precisely repeated every 30 minutes. Administration of vehicle (saline i.v.) occurred 15 minutes after starting each of the first 7 cystometrograms and duloxetine (1mg/kg i.v.) after the 8th. Acetic acid infusion into the bladder increased URS-EMG activity, bladder contraction frequency, and decreased contraction amplitude and capacity, compared to saline. Bladder activity and URS activity stabilized within 1 and 2 hours, respectively. Duloxetine administration significantly decreased bladder contraction frequency and increased URS-EMG activity to levels similar to previous reports. Cardiorespiratory parameters and blood gas levels remained consistent throughout the experiment. The epithelium of the bladder and urethra were greatly damaged and edema and infiltration of neutrophils in the lamina propria of urethra were observed. These data provide an ample evaluation of the health of the animals, stability of voiding function and appropriateness of the model for testing drugs designed to evaluate lower urinary tract as well as cardiovascular and respiratory systems function.

Effect of methysergide on pudendal inhibition of micturition reflex in cats

The role of 5-HT2 and opioid receptors in pudendal inhibition of bladder activity induced by intravesical infusion of saline or 0.25% acetic acid (AA) was investigated in anesthetized cats using methysergide (a 5-HT2 receptor antagonist) and naloxone (an opioid receptor antagonist). AA irritated the bladder and significantly (P<0.0001) reduced bladder capacity to 27.0±7.4% of saline control capacity. Pudendal nerve stimulation (PNS) at multiples of the threshold (T) intensity for inducing anal sphincter twitching restored bladder capacity to 60.1±8.0% at 1–2T (P<0.0001) and 92.2±14.1% at 3–4T (P=0.001) of the saline control capacity. Methysergide (0.03–1mg/kg, i.v.) suppressed low intensity (1–2T) PNS inhibition but not high intensity (3–4T) inhibition, and also significantly (P<0.05) increased control bladder capacity at the dosage of 0.3–1mg/kg. During saline infusion without AA irritation, PNS significantly increased bladder capacity to 150.8±9.9% at 1–2T (P<0.01) and 180.4±16.6% at 3–4T (P<0.01) of the saline control capacity. Methysergide (0.1–1mg/kg) significantly (P<0.05) increased saline control bladder capacity and suppressed PNS inhibition at the dosage of 0.03–1mg/kg. After methysergide treatment (1mg/kg), naloxone significantly (P<0.05) reduced control bladder capacity during AA infusion but had no effect during saline infusion. Naloxone also had no influence on PNS inhibition. These results suggest that 5-HT2 receptors play a role in PNS inhibition of reflex bladder activity and interact with opioid mechanisms in micturition reflex pathway. Understanding neurotransmitter mechanisms underlying pudendal neuromodulation is important for the development of new treatments for bladder disorders.

110 NGF SIGNALING MEDIATES INFLAMMATORY CELL RESPONSES OF BLADDER TO THE INTRAVESICAL INSULTS

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Basic Research (II)1 Apr 2013110 NGF SIGNALING MEDIATES INFLAMMATORY CELL RESPONSES OF BLADDER TO THE INTRAVESICAL INSULTS Mahendra Kashyap, Naoki Kawamorita, Michael Chancellor, Naoki Yoshimura, and Pradeep Tyagi Mahendra KashyapMahendra Kashyap Pittsburgh, PA More articles by this author , Naoki KawamoritaNaoki Kawamorita Pittsburgh, PA More articles by this author , Michael ChancellorMichael Chancellor Royal Oak, MI More articles by this author , Naoki YoshimuraNaoki Yoshimura Pittsburgh, PA More articles by this author , and Pradeep TyagiPradeep Tyagi Pittsburgh, PA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1489AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Animals with bladder overactivity induced by intravesical insults such as acetic acid (AA) are an established model for OAB. Evidence for the role of NGF in OAB is mounting owing to rapid expression of NGF in response to bladder insults and the elevation of NGF and chemokines such as MCP-1, CXCL-1, and CXCL-10 in the urine of OAB patients reported by our group. The present work tested the hypothesis that increased bladder tissue levels of NGF drive the overexpression of inflammatory mediators following bladder exposure to AA. METHODS Adult female Sprague-Dawley rats were either exposed to intravesical infusion of saline or saline containing 0.25% acetic acid at the rate of 0.04ml/min for 3h under urethane anesthesia. Rats exposed to acetic acid were instilled 24h earlier for 30 min with vehicle (sham), NGF antisense or scrambled sequence oligonucleotides (ODN) complexed with liposomes. Rat bladders harvested at the end of AA infusion were analyzed for expression of NGF, chemokines, cytokines and adhesion molecules by Luminex xMAP analysis. RESULTS Intravesical AA infusion upregulated the expression of NGF, VEGF, MCP-1, leptin, CXCL-1, CXCL-10, ICAM-1 and E-selectin in the urothelium obtained from sham rats relative to the control group without acetic acid treatment ( p<0.05; ANOVA). NGF and other effectors were elevated in group pretreated with scrambled ODN (p<0.01). However, Sequence specific downregulation with NGF-antisense ODN-liposome complex blunted the expression of NGF and the downstream mediators such as. NGF levels irrespective of group +vely associated with VEGF (r=0.75,p<0.001) and -ively associated with ICAM-1 levels (rs = -0.5, p<0.01, whereas in scrambled group, levels of MCP-1 +vely correlated with NGF (r=0.88, p<0.01) (fig attached). CONCLUSIONS Reduced expression of chemokines accompanying NGF antisense supports the role of NGF as a key mediator responsible for bladder response to chemical stimuli. Expression of adhesion molecules ICAM-1 and E-selectin is prerequisite for the rolling and stable arrest of monocytes, neutrophils, lymphocytes chemoattracted by MCP-1, CXCL-1 and CXCL-10, respectively following AA exposure. Thus, local suppression of NGF in the bladder could be an attractive approach for the treatment OAB, which is often associated with chemokine/cytokine production in the bladder. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e44-e45 Peer Review Report Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Mahendra Kashyap Pittsburgh, PA More articles by this author Naoki Kawamorita Pittsburgh, PA More articles by this author Michael Chancellor Royal Oak, MI More articles by this author Naoki Yoshimura Pittsburgh, PA More articles by this author Pradeep Tyagi Pittsburgh, PA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Effects of Vitamin D Analog on Bladder Function and Sensory Signaling in Animal Models of Cystitis

To measure the effects of nonhypercalcemic vitamin D receptor agonist elocalcitol on bladder function in rats with cyclophosphamide-induced cystitis and on bladder function and sensory nerve activity in a mouse with acetic acid-evoked bladder irritation. Female Wistar rats and male Balb/C mice were gavaged once daily with elocalcitol diluted in miglyol 812 (treatment group) or miglyol alone (control group). On experimental day 12, polyethylene tubing was implanted into the urinary bladder in all the animals. In the mice, a bipolar electrode was positioned under a single postganglionic bladder nerve. At 48 hours after surgery, bladder function was measured in awake, freely moving rats during bladder filling with 0.9% NaCl and both bladder function and sensory nerve activity was measured in awake, restrained mice during continuous intravesical infusion of 0.9% NaCl followed by 0.25% acetic acid. In rats, the treatment group showed a significant increase in bladder capacity and decrease in number of nonvoiding bladder contractions. In mice, the filling pressure during saline infusion was similar in both groups; however, during acetic acid infusion, the average filling pressure was significantly increased (47%) in the control group but not in the elocalcitol treatment group. The firing rate at filling pressure for the treatment group was 3.6-fold and 2.7-fold lower than that in the control group during the saline and acetic acid infusion, respectively. Oral treatment with elocalcitol suppressed signs of detrusor overactivity in both animal models and exerted strong suppressive effect on urinary bladder sensory signaling during filling in mice.

Phytochemical and pharmacological studies on Scorzonera alexandrina Boiss

The present study was designed to investigate secondary metabolite contents of Scorzonera alexandrina growing in the North–Western coast of Egypt. In addition hepatoprotective activity, reduction in blood sugar and ulcerative colitis were also evaluated . Chromatographic methods and spectroscopic analysis were used for isolation and identification of the compounds. In the hepatoprotective activity; twenty-four adult male Wistar albino rats were divided into four groups of six animals each. Rats of the 1st (normal control) and 2nd (CCl4-intoxicated control) groups received the vehicle in a dose of 5mL/kg. The 3rd and 4th groups were treated with the ethanol extract of S. alexandrina in doses of 200 and 400mg/kg, respectively. In the ulcerative colitis study, the same extract was administered orally (200 and 400mg/kg) to rats once a day for 5 consecutive days and the last dose was administered 2h before induction of colitis by intra-rectal infusion of acetic acid. The obtained results revealed that S. alexandrina contains scopoletin, xanthotoxin, apigenin, luteolin, quercetin-7-O-rhaminoside and luteolin-7-O-glucoside, also it showed a significant decrease in blood glucose level, liver functions (ALT, AST, TP), also a significant anti-ulcer effect.

Anti-inflammatory activity of low and high methoxylated citrus pectins

The anti-inflammatory activity of the citrus pectins, CU701 and CU201, with low and high degrees of methylesterification (LM and HM), respectively, was studied after oral administration in mice using three models of inflammation: acetic acid-induced colitis, cytokine production by blood leukocytes in response to lipopolysaccharide (LPS) and endotoxin shock. Oral administration of either CU701 or CU201 24h before the infusion of acetic acid into the colon was found to prevent the progression of colitis. However, CU701 was found to decrease the adhesion of peritoneal granulocytes and their production of reactive oxygen species in colitic mice, whereas CU201 had no effect on granulocyte activity. Only CU701 was found to decrease tumour necrosis factor-α (TNF-α) release and to increase production of the anti-inflammatory cytokine interleukin-10 (IL-10) 24h after oral administration in mice. CU201 had no effect on cytokine production. CU701, but not CU201, was found to improve the survival rate in mice that were subjected to a lethal dose of LPS. The present study demonstrates that LM pectin would be a more efficient anti-inflammatory agent than HM pectin upon oral administration. LM pectin was found to inhibit both local and systemic inflammation, whereas HM pectin was shown to prevent intestinal inflammation.

Male lower urinary tract symptoms and α1D‐adrenoceptors

Historically, α(1)-adrenoceptors have been classified into three subtypes (α(1)(A), α(1)(B) and α(1)(D)) that are widely distributed in various organs. Research on the α(1)(D)-adrenoceptors in the bladder, urethra and prostate has focused on the relationship between expression levels and symptoms of bladder outlet obstruction, and the implications and functional roles of α(1)(D)-adrenoceptors subtypes in these organs. The α(1)(D)-adrenoceptor messenger ribonucleic acid and protein seem to be increased in obstructed bladders or small capacity bladders. In contrast, α(1)(D)-adrenoceptor subtype knock-out mice have been found to have a prolonged voiding interval. Interestingly, an α(1)(D)-adrenoceptor antagonist was found to inhibit the facilitation of afferent nerve activity for the micturition reflex induced by intravesical infusion of acetic acid. Clinically, patients who felt urgency at low filling volumes and had a small bladder capacity were found to have more α(1)(D)-adrenoceptor messenger ribonucleic acid in their bladder mucosa than patients who felt urgency at high filling volumes and had a large bladder capacity. An α(1)(D)-adrenoceptor antagonist was found to increase the first desired volume and the maximum desired volume while decreasing detrusor overactivity in pressure flow studies. Thus, α(1)(D)-adrenoceptors in the lower urinary tract might play an important role in the pathophysiology of lower urinary tract disorders.

JTS-653 Blocks Afferent Nerve Firing and Attenuates Bladder Overactivity Without Affecting Normal Voiding Function

We evaluated the role of TRPV1 in bladder overactivity based on afferent nerve firing and urodynamic parameters using the selective TRPV1 antagonist JTS-653. We evaluated the effects of JTS-653 on the increased pelvic nerve discharge and intravesical pressure induced by intravesical infusion of 100 μM capsaicin in anesthetized rats. The effects of JTS-653 on the urodynamic parameters of bladder overactivity induced by intravesical infusion of 30 nM resiniferatoxin or 0.2% acetic acid, or on normal bladder activity were evaluated by cystometry in conscious rats. The effects of JTS-653 on carbachol induced contraction were investigated using bladder muscle strips. JTS-653 significantly suppressed the capsaicin induced increase in nerve discharge and intravesical pressure. Intravesical infusion of resiniferatoxin or acetic acid decreased the intercontraction interval and voided volume. JTS-653 significantly increased the intercontraction interval and voided volume in rats with resiniferatoxin or acetic acid induced bladder overactivity without affecting maximal voiding pressure. The antimuscarinic agent propiverine significantly decreased maximal voiding pressure but did not affect the intercontraction interval or voided volume in rats with acetic acid induced bladder overactivity. In normal rats JTS-653 showed no significant effects on the intercontraction interval, voided volume or maximal voiding pressure. JTS-653 did not affect carbachol induced contraction of the bladder muscle. Our findings suggest that TRPV1 is involved in bladder overactivity via afferent nerve activation but it is not associated with normal voiding function. A TRPV1 antagonist would be a useful drug for bladder overactivity with a different pharmacological profile than antimuscarinic agents.

A mouse model for interstitial cystitis/painful bladder syndrome based on APF inhibition of bladder epithelial repair: a pilot study

BackgroundInterstitial 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.MethodsThe 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.ResultsBladder 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).ConclusionsThis 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.

253 INVOLVEMENT OF METABOTROPIC GLUTAMATE RECEPTOR MGLU5 IN PUDENDAL INHIBITION OF BLADDER OVERACTIVITY IN CATS

You have accessJournal of UrologyBladder and Urethra: Anatomy, Physiology and Pharmacology I1 Apr 2012253 INVOLVEMENT OF METABOTROPIC GLUTAMATE RECEPTOR MGLU5 IN PUDENDAL INHIBITION OF BLADDER OVERACTIVITY IN CATS Jeffrey Larson, P. Dafe Ogagan, Jicheng Wang, Bing Shen, James Roppolo, Changfeng Tai, and William C. de Groat Jeffrey LarsonJeffrey Larson Pittsburgh, PA More articles by this author , P. Dafe OgaganP. Dafe Ogagan Pittsburgh, PA More articles by this author , Jicheng WangJicheng Wang Pittsburgh, PA More articles by this author , Bing ShenBing Shen Pittsburgh, PA More articles by this author , James RoppoloJames Roppolo Pittsburgh, PA More articles by this author , Changfeng TaiChangfeng Tai Pittsburgh, PA More articles by this author , and William C. de GroatWilliam C. de Groat Pittsburgh, PA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.310AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Neuromodulation is a useful treatment modality in patients with refractory lower urinary tract symptoms, but the mechanisms of action are poorly understood. Metabotropic glutamate receptors are implicated in neuropathic pain syndromes but their involvement in micturition is poorly understood. We hypothesized that pudendal inhibition of bladder overactivity could be due to activation of the mGlu5 receptor. We used a selective mGluR5 antagonist (MTEP) to assess this receptor's function during neuromodulation. METHODS MTEP was administered to 11 cats in increasing cumulative doses. Saline (n=5) or 0.25% acetic acid (n=6) were utilized to irritate the bladder and induce overactivity. Bladder capacity and the degree of inhibition of bladder activity induced by pudendal nerve stimulation were assessed by cystometrogram. RESULTS In the absence of pudendal nerve stimulation, MTEP significantly increased saline bladder capacity. Infusion of acetic acid (AA) significantly reduced bladder capacity compared to saline. During saline CMGs, pudendal nerve stimulation significantly inhibited bladder contraction and increased bladder capacity by 196 ±37.4%; a smaller increase in bladder capacity was observed with AA infusion (98.4 ± 55.2%). Administration of MTEP doses of ≥ 1mg/kg significantly blocked the pudendal nerve inhibition of bladder contractions during AA infusion. At all doses tested during saline infusion, MTEP significantly increased baseline bladder capacity without neuromodulation. CONCLUSIONS MTEP can suppress reflex micturition as well as pudendal nerve stimulation induced inhibition of bladder activity. These results indicate that glutamic acid is an excitatory transmitter in the micturition pathway essential to the pudendal nerve stimulation evoked inhibitory mechanism. The mGlu5 receptor could serve as a novel and important potential new target for the treatment of OAB. Potential mGluR5 locations include spinal cord inhibitory interneurons, spinal tract neurons, or upper tract CNS. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e102-e103 Peer Review Report Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jeffrey Larson Pittsburgh, PA More articles by this author P. Dafe Ogagan Pittsburgh, PA More articles by this author Jicheng Wang Pittsburgh, PA More articles by this author Bing Shen Pittsburgh, PA More articles by this author James Roppolo Pittsburgh, PA More articles by this author Changfeng Tai Pittsburgh, PA More articles by this author William C. de Groat Pittsburgh, PA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

497 DIFFERENTIAL ROLE OF OPIOID RECEPTORS IN TIBIAL NERVE INHIBITION OF NOCICEPTIVE AND NON-NOCICEPTIVE BLADDER REFLEXES IN CATS

You have accessJournal of UrologyBladder and Urethra: Anatomy, Physiology and Pharmacology II1 Apr 2012497 DIFFERENTIAL ROLE OF OPIOID RECEPTORS IN TIBIAL NERVE INHIBITION OF NOCICEPTIVE AND NON-NOCICEPTIVE BLADDER REFLEXES IN CATS Abhijith Mally, Jeffrey Larson, P. Dafe Ogagan, Changfeng Tai, Bing Shen, Jicheng Wang, James Roppolo, and William C. de Groat Abhijith MallyAbhijith Mally Pittsburgh, PA More articles by this author , Jeffrey LarsonJeffrey Larson Pittsburgh, PA More articles by this author , P. Dafe OgaganP. Dafe Ogagan Pittsburgh, PA More articles by this author , Changfeng TaiChangfeng Tai Pittsburgh, PA More articles by this author , Bing ShenBing Shen Pittsburgh, PA More articles by this author , Jicheng WangJicheng Wang Pittsburgh, PA More articles by this author , James RoppoloJames Roppolo Pittsburgh, PA More articles by this author , and William C. de GroatWilliam C. de Groat Pittsburgh, PA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.567AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Electrical stimulation of somatic afferent pathways in the tibial nerve can inhibit bladder activity and therefore aid in the management of overactive bladder symptoms. Opioid peptides have a well known role in controlling pain mechanisms and are involved in somatic afferent inhibition of non-nociceptive bladder reflexes. We hypothesized that inhibition of irritation-induced bladder overactivity by tibial nerve stimulation could be due to activation of opioid receptors. We used naloxone to block the opioid receptors and assess the receptor function during tibial neuromodulation. METHODS Naloxone was administered to 20 cats in increasing cumulative doses. Saline (n=10) or .25% acetic acid (n=10) were utilized to irritate the bladder and induce overactivity. Bladder capacity and the degree of inhibition of bladder activity induced by tibial nerve stimulation were assessed by cystometrogram. RESULTS Infusion of acetic acid (AA) significantly reduced bladder capacity compared to saline. During CMG, tibial nerve stimulation (TNS) significantly inhibited bladder contraction and increased bladder capacity by 41.5±9.9% and 46.1±7.9% of the saline control capacity. Naloxone completely eliminated TNS inhibition of bladder overactivity. Naloxone did not change bladder capacity during AA irritation. However, during saline infusion, naloxone significantly reduced bladder capacity to 66.5±8.1% of the control capacity. During saline infusion TNS induced an acute increase in bladder capacity and an increase that persisted following stimulation. Naloxone administered during the post-stimulation inhibition still decreased bladder capacity. CONCLUSIONS These results indicate that opioid receptors have different roles in modulation of bladder reflexes induced by nociceptive and non-nociceptive stimuli and in somatic afferent inhibition of these reflexes, raising the possibility that opioid receptors may be a target for pharmacological treatment of lower urinary tract disorders. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e204 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Abhijith Mally Pittsburgh, PA More articles by this author Jeffrey Larson Pittsburgh, PA More articles by this author P. Dafe Ogagan Pittsburgh, PA More articles by this author Changfeng Tai Pittsburgh, PA More articles by this author Bing Shen Pittsburgh, PA More articles by this author Jicheng Wang Pittsburgh, PA More articles by this author James Roppolo Pittsburgh, PA More articles by this author William C. de Groat Pittsburgh, PA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

In VivoandIn VitroAntinociceptive Effect ofFagopyrum cymosum(Trev.) Meisn Extracts: A Possible Action by Recovering Intestinal Barrier Dysfunction

Fagopyrum cymosum (Trev.) Meisn (Fag) is a herb rhizome which has been widely used to treat diseases. To investigate the effects and mechanisms of the Fag on irritable bowel syndrome (IBS), in vivo neonatal pups maternal separation (NMS) combined with intracolonic infusion of acetic acid (AA) was employed to establish IBS rat models. Fag reduced their visceral hyperalgesia and the whole gut permeability, ameliorated colonic mucosa inflammation and injury, and upregulated the expression of decreased tight junction proteins (TJs) of claudin-1, occludin, and ZO-1 (except ZO-2) in colonic epithelium. Caco-2 monolayer cells were incubated with TNF-α and IFN-γ in vitro to establish an epithelial barrier dysfunction model whose transepithelial electrical resistance (TER) depended more on dose of Fag than that of the controls, and whose TJs levels were lower than those of the controls. Fag upregulated the NP-40 insoluble and soluble components of the four TJs markedly in a dose-dependent manner. These data suggest that Fag alleviated the hyperalgesia of IBS rats by reducing intestinal inflammation and enhancing mucosal epithelial function after regulating the structure and function of TJs.

Inhibition of bladder overactivity by stimulation of feline pudendal nerve using transdermal amplitude‐modulated signal (TAMS)

• To develop a non-invasive neuromodulation method targeting the pudendal nerve. • Bladder overactivity induced by acetic acid (AA) irritation was partially suppressed by electrical stimulation of the pudendal nerve in α-chloralose anaesthetized cats using a transdermal amplitude-modulated signal (TAMS). • During cystometrography (CMG), intravesical infusion of 0.25% AA significantly decreased the mean (se) bladder capacity to 28.8 (5.9)% of the capacity measured during saline infusion. • The TAMS stimulation inhibited AA-induced bladder overactivity at 5, 7 and 10 Hz, and significantly increased the mean (se) bladder capacity to 61.8 (9.9)%, 51.3 (14.5)%, 53.6 (14.9)%, respectively, of the control capacity during saline infusion, whereas stimulation at 20-40 Hz had no effect. • Under isovolumetric conditions at a bladder volume ranging between 130 to 160% of the bladder capacity measured during AA infusion, TAMS stimulation at all frequencies (5-40 Hz) significantly suppressed the irritation-induced rhythmic bladder contractions, reduced the area under the bladder pressure curve, and decreased the frequency of bladder contractions. However, the amplitude of rhythmic bladder contractions was only significantly decreased at stimulation frequencies of 5-20 Hz. • At bladder volumes above the AA control capacity, TAMS stimulation with frequencies of 20-30 Hz had an excitatory effect, resulting in large amplitude (>25 cmH(2) O) bladder contractions. • TAMS stimulation targeting the cat pudendal nerve can inhibit C-fibre afferent-mediated bladder overactivity. • Thus, clinical research seems warranted to explore the usefulness of this technology for patients with overactive bladder symptoms.