RESUMEN
OBJECTIVE: Previous studies have shown that Astragalus polysaccharides (APS) and Astragaloside IV (AS-IV) protect against inflammation-related cell damage and exhibit immune enhancement. Since urothelial injury may result in an overactive bladder (OAB), the aim of this study was to investigate the efficacy of APS and AS-IV on urothelial injury in an experimental animal model. MATERIALS AND METHODS: The effects of APS and AS-IV on the proliferation and migration of primary human urothelial cells (HUCs) or primary human fibroblast cells (HFCs) were assessed using an in vitro wounding model and colorimetric thiazolyl blue assays. Sixty virgin female mice were randomized into five groups: group 1-saline-injected plus treatment with H2O, group 2-cyclophosphamide (CYP) plus treatment with H2O, group 3-CYP plus treatment with solifenacin succinate (SS; 10 mg/kg), group 4-CYP plus treatment with AS-IV (100 mg/kg), and group 5-CYP plus treatment with APS (100 mg/kg). Cystometry assessment was conducted and cell junction-associated protein zonula occludens-2 (ZO-2) expression was measured. Voiding interval values (time between voids) were assessed in mice under anesthesia. Lastly, immunohistochemistry analysis was used to confirm the location and level, respectively, of ZO-2 expression. RESULTS: APS and AS-IV did not influence the cell viability but increased migration in HFCs compared with the controls. The OAB mice showed significantly lower voiding interval values. Voiding interval values were significantly higher in the CYP plus treatment with APS (100 mg/kg) and AS-IV (100 mg/kg) groups than in the CYP-induced OAB group. Additionally, the expression of ZO-2, a tight junction protein, was increased in the CYP plus treatment APS (100 mg/kg) and AS-IV (100 mg/kg) groups compared with the CYP-induced OAB group. CONCLUSION: These findings suggest that APS and AS-IV modulate urothelial wound healing, which ameliorates urinary frequency of mice treated with CYP. APS or AS-IV may have the potential benefit of acting as urothelial wound healing modulators.
Asunto(s)
Planta del Astrágalo/química , Polisacáridos/farmacología , Saponinas/farmacología , Triterpenos/farmacología , Vejiga Urinaria Hiperactiva/tratamiento farmacológico , Urotelio/lesiones , Animales , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ciclofosfamida , Modelos Animales de Enfermedad , Femenino , Fibroblastos/efectos de los fármacos , Humanos , Ratones , Vejiga Urinaria Hiperactiva/inducido químicamente , Micción/efectos de los fármacos , Urotelio/citologíaRESUMEN
The results of recent studies on the mechanisms of kidney damage are presented in the review of literature. The role of the immune system in the occurrence, development and outcome of damage to the epithelium of the renal tubules, as well as molecular, genetic and metabolic changes which determine the extent and consequences of renal trauma are described in details. The mechanisms of restoration of the renal parenchyma and the development of fibrosis following the cessation of injury are given.
Asunto(s)
Enfermedades Renales/fisiopatología , Túbulos Renales/fisiopatología , Riñón/fisiopatología , Urotelio/fisiopatología , Fibrosis/fisiopatología , Humanos , Riñón/lesiones , Enfermedades Renales/etiología , Túbulos Renales/lesiones , Urotelio/lesionesRESUMEN
Based on the hypothesis that bioscaffold permeability is a major factor in determining the outcome of histologically complete and functional bladder regeneration, we evaluated regeneration processes of four-layer porcine small intestinal submucosa (SIS) construct; and compared results between rat bladders augmented with single-layer SIS bioscaffolds. Sprague-Dawley female rats were subjected to hemi-cystectomy followed by anastomosis of a patch of either single- or four-layer porcine SIS. Permeability was analyzed in situ using magnetic resonance imaging (MRI) at post-operative days 7 and 14. Bladder sections excised at days 7, 14, 28, and 56 post-operation Samples were assessed by H&E and Masson's trichrome stains. Urothelial differentiation was analyzed using cytokeratin AE1/AE3, and uroplakin III (UPIII). In addition, quantitative and qualitative evaluations of neutrophils, mast cells, eosinophils, and macrophages were performed using anti-myeloperoxidase, Alcian blue, Giemsa stain, and anti-CD68 staining methods, respectively. Four-layer SIS was consistently impermeable as evidenced by the absence of intravesical administered gadolinium with diethylenetriaminepentacetate (Gd-DTPA) contrast signal in peripheral regions of augmented bladders compared with single-layer SIS bioscaffold. Elevated and sustained eosinophil and neutrophil infiltrations were prominent in four-layered SIS-augmented bladders compared with single-layer SIS with comparable impermeability. Delayed but consistent urothelial regeneration and differentiation were observed in four-layer SIS-augmented bladders; and urothelial differentiation was observed at day 56 post-augmentation. In conclusion, four-layer SIS enacts an elevated inflammatory response along with extended urothelial regeneration. Four-layer SIS may activate a different but yet to be identified mechanism for inflammatory responses. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1960-1969, 2019.
Asunto(s)
Inmunomodulación , Mucosa Intestinal/química , Intestino Delgado/química , Regeneración , Andamios del Tejido/química , Vejiga Urinaria , Urotelio , Animales , Femenino , Ratas , Ratas Sprague-Dawley , Porcinos , Vejiga Urinaria/lesiones , Vejiga Urinaria/fisiología , Urotelio/lesiones , Urotelio/fisiologíaRESUMEN
The urothelium is an epithelia barrier lined by a luminal layer of binucleated, octoploid, superficial cells. Superficial cells are critical for production and transport of uroplakins, a family of proteins that assemble into a waterproof crystalline plaque that helps protect against infection and toxic substances. Adult urothelium is nearly quiescent, but rapidly regenerates in response to injury. Yet the mechanism by which binucleated, polyploid, superficial cells are produced remains unclear. Here, we show that superficial cells are likely to be derived from a population of binucleated intermediate cells, which are produced from mononucleated intermediate cells via incomplete cytokinesis. We show that binucleated intermediate and superficial cells increase DNA content via endoreplication, passing through S phase without entering mitosis. The urothelium can be permanently damaged by repetitive or chronic injury or disease. Identification of the mechanism by which superficial cells are produced may be important for developing strategies for urothelial repair.
Asunto(s)
Citocinesis , Endorreduplicación , Mitosis , Poliploidía , Urotelio/fisiopatología , Animales , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Femenino , Masculino , Ratones , Urotelio/lesionesRESUMEN
BACKGROUND: The kidney is considered to be a structurally stable organ with limited baseline cellular turnover. Nevertheless, single cells must be constantly replaced to conserve the functional integrity of the organ. PDGF chain B (PDGF-BB) signaling through fibroblast PDGF receptor-ß (PDGFRß) contributes to interstitial-epithelial cell communication and facilitates regenerative functions in several organs. However, the potential role of interstitial cells in renal tubular regeneration has not been examined. METHODS: In mice with fluorescent protein expression in renal tubular cells and PDGFRß-positive interstitial cells, we ablated single tubular cells by high laser exposure. We then used serial intravital multiphoton microscopy with subsequent three-dimensional reconstruction and ex vivo histology to evaluate the cellular and molecular processes involved in tubular regeneration. RESULTS: Single-tubular cell ablation caused the migration and division of dedifferentiated tubular epithelial cells that preceded tubular regeneration. Moreover, tubular cell ablation caused immediate calcium responses in adjacent PDGFRß-positive interstitial cells and the rapid migration thereof toward the injury. These PDGFRß-positive cells enclosed the injured epithelium before the onset of tubular cell dedifferentiation, and the later withdrawal of these PDGFRß-positive cells correlated with signs of tubular cell redifferentiation. Intraperitoneal administration of trapidil to block PDGFRß impeded PDGFRß-positive cell migration to the tubular injury site and compromised the recovery of tubular function. CONCLUSIONS: Ablated tubular cells are exclusively replaced by resident tubular cell proliferation in a process dependent on PDGFRß-mediated communication between the renal interstitium and the tubular system.
Asunto(s)
Desdiferenciación Celular , Células Epiteliales/fisiología , Túbulos Renales Proximales/fisiología , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Regeneración , Urotelio/fisiología , Animales , Calcio/metabolismo , Comunicación Celular , Movimiento Celular/efectos de los fármacos , Femenino , Microscopía Intravital , Riñón/citología , Túbulos Renales Proximales/citología , Túbulos Renales Proximales/lesiones , Linfocinas/metabolismo , Masculino , Ratones , Inhibidores de Fosfodiesterasa/farmacología , Factor de Crecimiento Derivado de Plaquetas/metabolismo , Recuperación de la Función , Trapidil/farmacología , Urotelio/lesionesRESUMEN
Augmentation enterocystoplasty remains the gold standard surgical bladder reconstruction procedure to increase the capacity and compliance of dysfunctional bladders. Since the use of the patient's intestine has severe risks of complications, alternative biodegradable matrices have been explored. Porcine small intestinal submucosa (SIS) has gained immense interests in bladder reconstruction due to its favorable properties. However, trials have shown inconsistent regeneration with SIS, attributed to the heterogeneity in microstructures and mechanical properties. We hypothesize that uneven SIS permeability to urine is a factor responsible for the inconsistency. We measured permeability to urine in situ using a contrast enhanced-magnetic resonance imaging (MRI), and evaluated urothelium regeneration using immunohistochemical staining of urothelial cell markers in SIS-augmented rat bladders. Results showed significant differences in permeability among SIS-augmented rat bladders. Commercial SIS scaffolds were then categorized into nonleaky and leaky groups based on MRI results. Hematoxylin and eosin staining showed higher numbers of inflammatory cells in leaky SIS on day 14 relative to nonleaky SIS. In addition, trichrome staining showed major changes in the distribution of collagen on day 28 between SIS-augmented bladder groups. Furthermore, expressions of urothelium-associated markers (cytokeratins AE1/AE3, claudin 4, and uroplakin III) were completed in bladders augmented with nonleaky SIS, whereas limited urothelial differentiation was noticed in leaky SIS-augmented bladders at post-augmentative day 14. These results show that scaffold permeability to urine may be responsible for variations in regenerative capacity of porcine SIS. Applications of MRI technique will be helpful to understand a relationship between biomaterial property and regenerative capacity. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1778-1787, 2018.
Asunto(s)
Materiales Biocompatibles , Mucosa Intestinal/química , Procedimientos de Cirugía Plástica , Regeneración , Vejiga Urinaria , Urotelio , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Diferenciación Celular/efectos de los fármacos , Femenino , Imagen por Resonancia Magnética , Permeabilidad , Ratas , Ratas Sprague-Dawley , Porcinos , Vejiga Urinaria/lesiones , Vejiga Urinaria/fisiología , Vejiga Urinaria/cirugía , Urotelio/lesiones , Urotelio/fisiología , Urotelio/cirugíaRESUMEN
The bladder urothelium functions as a urine-blood barrier and consists of basal, intermediate, and superficial cell populations. Reconstructive procedures such as augmentation cystoplasty and focal mucosal resection involve localized surgical damage to the bladder wall whereby focal segments of the urothelium and underlying submucosa are respectively removed or replaced and regeneration ensues. We demonstrate using lineage-tracing systems that urothelial regeneration following augmentation cystoplasty with acellular grafts exclusively depends on host keratin 5-expressing basal cells to repopulate all lineages of the de novo urothelium at implant sites. Conversely, repair of focal mucosal defects not only employs this mechanism, but in parallel host intermediate cell daughters expressing uroplakin 2 give rise to themselves and are also contributors to superficial cells in neotissues. These results highlight the diversity of urothelial regenerative responses to surgical injury and may lead to advancements in bladder tissue engineering approaches.
Asunto(s)
Queratina-5/genética , Regeneración/genética , Vejiga Urinaria/crecimiento & desarrollo , Uroplaquina II/genética , Urotelio/crecimiento & desarrollo , Animales , Diferenciación Celular/genética , Linaje de la Célula/genética , Rastreo Celular/métodos , Regulación del Desarrollo de la Expresión Génica/genética , Humanos , Complicaciones Intraoperatorias/metabolismo , Complicaciones Intraoperatorias/patología , Ratones , Ingeniería de Tejidos , Vejiga Urinaria/lesiones , Vejiga Urinaria/metabolismo , Orina/fisiología , Urotelio/lesiones , Urotelio/metabolismoRESUMEN
Homeostatic maintenance and repair of the urothelium upon injury are required for a functional bladder in both healthy and disease conditions. Understanding the cellular and molecular mechanisms underlying the urothelial regenerative response is key to designing strategies for tissue repair and ultimately treatments for urologic diseases including urinary tract infections, voiding dysfunction, painful bladder syndrome, and bladder cancer. In this article, we review studies on urothelial ontogeny during development and regeneration following various injury modalities. Signaling pathways involved in urothelial regeneration and in urothelial carcinogenesis are also discussed. Developmental Dynamics 246:336-343, 2017. © 2016 Wiley Periodicals, Inc.
Asunto(s)
Regeneración , Neoplasias Urológicas/patología , Urotelio/crecimiento & desarrollo , Animales , Humanos , Transducción de Señal , Vejiga Urinaria/fisiología , Enfermedades Urológicas/fisiopatología , Urotelio/lesiones , Urotelio/fisiologíaRESUMEN
PURPOSE: To evaluate and compare the acute effect of electromagnetic and electrohydraulic extracorporeal shockwave lithotripsy (SWL) on the urothelial layers of kidney and ureter. MATERIALS AND METHODS: Fifty patients, 29 males (58%) and 21 females (42%), with an average age of 51.68 years (range: 37-70) who underwent SWL application in two different centers were included. Twenty-eight patients (56%) were treated with electrohydraulic and 22 (44%) were treated with electromagnetic lithotripsy. Urinary cytologic examinations were done immediately before and after SWL therapy and 10 days later. The average numbers of epithelial cells, red blood cells (RBC), and myocytes were counted under 40 × magnification. RESULTS: There were significant differences in the number of epithelial cells and RBC before and after immediate application of SWL: 1.66 and 14.9 cells/field, (p = 0.001), 5.44 and 113.45 cells/field, respectively (p = 0.001). The number of RBC was significantly higher in patients treated with electromagnetic lithotripsy than those treated with electrohydraulic: 141.9 and 93.4 cells/field, respectively (p = 0.02). No myocyte or basement membrane elements were detected in any of the cytologic examinations. Cytologic examinations done after 10 days of SWL therapy revealed recovery of all abnormal cytologic findings. CONCLUSIONS: The acute increments in the number of epithelial cells and RBC after SWL were statistically significant but it was not permanent. SWL-induced urinary urothelial lesion is limited to the mucosal layer and there was no evidence of damage to the basal membrane or muscle layer. Electromagnetic lithotripsy caused high numbers of RBC than the electrohydraulic device on the postimmediate urine cytologic examination.
Asunto(s)
Radiación Electromagnética , Litotricia/efectos adversos , Litotricia/métodos , Urotelio/lesiones , Adulto , Anciano , Células Epiteliales/citología , Recuento de Eritrocitos , Femenino , Humanos , Litotricia/instrumentación , Masculino , Persona de Mediana Edad , Células Musculares/citología , Orina/citología , Urotelio/patologíaRESUMEN
No disponible
Asunto(s)
Humanos , Femenino , Anciano , Enfermedades de la Vejiga Urinaria/complicaciones , Enfermedades de la Vejiga Urinaria , Síndrome , Urotelio , Urotelio/lesiones , Cistoscopía/métodos , Cistitis Intersticial/complicaciones , Cistitis Intersticial/fisiopatología , Dolor/complicaciones , Dolor/etiología , Vejiga Urinaria/patología , Cistoscopía/instrumentación , Vejiga Urinaria , Enfermedades de la Vejiga Urinaria/patología , Cistitis Intersticial , Manejo del Dolor , Analgésicos/uso terapéutico , Corticoesteroides/uso terapéutico , Antiinflamatorios/uso terapéutico , Toxinas Botulínicas Tipo A/uso terapéuticoRESUMEN
INTRODUCTION: The study proposed a technique for early detection of the damaging effect of shock waves on the urinary tract tissues, for monitoring the state of urothelial cells in the early post-procedure period and choosing an adequate method for preventing and managing possible complications. MATERIALS AND METHODS: The study analyzed the urine samples of 300 patients aged 20 to 50 years, who for the first time underwent ESWL for kidney stones. The urine sediment smears were fixed in the May-Grunwald stain and stained with azure eosin solution according to Pappenheim. Besides evaluating general cytologic characteristics, a morphometric examination of urine sediment was performed. RESULTS AND DISCUSSION: To define quantitative parameters, the total number of epithelial cells (ep) was determined with further calculation of the mean number of epithelial cells in the field of view (ep= ep/10). Correlation between the number of abnormal epithelial cells (cp) and the total number of epithelial cells (ep) was analyzed. The resulted "destruction index" (DI) was the "pool" of all cytopathological changes in epithelial cells DI = cp/ep. Immediately after ESWL, DI markedly increased with significant difference (<0.05) in numeric values. Two hours after the procedure, the DI reduced compared to the previous value. And only at day 4 DI was close to the norm, although remaining somewhat elevated. CONCLUSION: Extracorporeal shock waves lithotripsy produces the acute urothelial damage resulting in cytopathological changes of varying severity. The above-mentioned technique provides objective and highly significant clinical and diagnostic information on the state of the urothelium after the exposure to shock waves.
Asunto(s)
Litotricia/efectos adversos , Cálculos Urinarios/terapia , Urotelio/lesiones , Adulto , Hematuria/etiología , Hematuria/patología , Hematuria/orina , Humanos , Litotricia/métodos , Persona de Mediana Edad , Cálculos Urinarios/orina , Sistema Urinario/lesiones , Sistema Urinario/patología , Urotelio/patología , Adulto JovenRESUMEN
Epithelial tissue structure is the emergent outcome of the interactions between large numbers of individual cells. Experimental cell biology offers an important tool to unravel these complex interactions, but current methods of analysis tend to be limited to mean field approaches or representation by selected subsets of cells. This may result in bias towards cells that respond in a particular way and/or neglect local, context-specific cell responses. Here, an automated algorithm was applied to examine in detail the individual calcium transients evoked in genetically homogeneous, but asynchronous populations of cultured non-immortalized normal human urothelial cells when subjected to either the global application of an external agonist or a localized scratch wound. The recorded calcium transients were classified automatically according to a set of defined metrics and distinct sub-populations of cells that responded in qualitatively different ways were observed. The nature of this variability in the homogeneous cell population was apportioned to two sources: intrinsic variation in individual cell responses and extrinsic variability due to context-specific factors of the environment, such as spatial heterogeneity. Statistically significant variation in the features of the calcium transients evoked by scratch wounding according to proximity to the wound edge was identified. The manifestation of distinct sub-populations of cells is considered central to the coordination of population-level response resulting in wound closure.
Asunto(s)
Calcio/metabolismo , Citosol/química , Células Epiteliales/metabolismo , Urotelio/citología , Cicatrización de Heridas/fisiología , Señalización del Calcio/fisiología , Células Cultivadas , Técnica del Anticuerpo Fluorescente , Humanos , Urotelio/lesionesRESUMEN
BACKGROUND: Irrigation or washouts of the bladder are usually performed in various clinical settings. In the 1980s Elliot and colleagues argued that urothelial damage could occur after washouts and irrigations of the bladder. The exact mechanism underlying urothelial damage has not yet been discovered. To our knowledge, this is the first report of fatal fluid overload and pulmonary edema, due to urothelium disruption occurring during bladder irrigation, approached performing complete histological and immunohistochemical investigation on bladder specimens. The exposed case deserves attention since it demonstrates that, although very rarely, irrigation or washouts of the bladder may have unexpected serious clinical consequences. CASE PRESENTATION: An 85 year-old Caucasian man, unable to eat independently and whose fluid intake was controlled, underwent continuous bladder irrigation with a 3-way catheter due to a severe episode of macrohematuria. During the third day of hospitalization, while still undergoing bladder irrigation, he suddenly experienced extreme shortness of breath, breathing difficulties, and cough with frothy sputum. His attending nurse immediately noted that there was no return of the fluid (5 liters) introduced through bladder irrigation. He was treated urgently with hemodialysis. At the beginning of the dialysis treatment, the patient had gained 7.4 kg since the previous measurement (24 hours prior) without any clear explanation. Although a significant weight loss (from 81 to 76 kg) due to the dialysis procedure, the patient died shortly after the final treatment. The autopsy revealed that the brain and the lungs were heavily edematous. Microscopic examination of bladder specimens revealed interstitial and mucosal swelling, and loss of the superficial cell layer. Intermediate and basal urothelial cells were preserved. Altogether the above mentioned findings were suggestive of a diffuse disruption of the urothelium. In conclusion the death of the man was attributed to an acute severe pulmonary edema due to massive fluid absorption. CONCLUSION: Our case demonstrates that urothelium disruption may occur during irrigation and washouts of the bladder, also in the absence of other well-known predisposing conditions. Inappropriate use of bladder irrigation should be avoided and a close attention is required of the fluid balance is mandatory when irrigating the bladder.
Asunto(s)
Edema Pulmonar/etiología , Irrigación Terapéutica/efectos adversos , Vejiga Urinaria , Urotelio/lesiones , Anciano de 80 o más Años , Resultado Fatal , Humanos , MasculinoRESUMEN
OBJECTIVE: Analysis of renal excretory system integrity and efficacy of radiofrequency ablation with and without irrigation with saline at 2°C (SF2). MATERIALS AND METHODS: The median third of sixteen kidneys were submitted to radiofrequency (exposition of 1 cm) controlled by intra-surgical ultrasound, with eight minutes cycles and median temperature of 90°C in eight female pigs. One excretory renal system was cooled with SF2, at a 30mL/min rate, and the other kidney was not. After 14 days of post-operatory, the biggest diameters of the lesions and the radiological aspects of the excretory system were compared by bilateral ascending pyelogram and the animals were sacrificed in order to perform histological analysis. RESULTS: There were no significant differences between the diameters of the kidney lesions whether or not exposed to cooling of the excretory system. Median diameter of the cooled kidneys and not cooled kidneys were respectively (in mm): anteroposterior: 11.46 vs. 12.5 (p = 0.23); longitudinal: 17.94 vs. 18.84 (p = 0.62); depth: 11.38 vs. 12.25 (p = 0.47). There was no lesion of the excretory system or signs of leakage of contrast media or hydronephrosis at ascending pyelogram. CONCLUSION: Cooling of excretory system during radiofrequency ablation does not sig¬nificantly alter generated coagulation necrosis or affect the integrity of the excretory system in the studied model.
Asunto(s)
Ablación por Catéter/métodos , Frío , Riñón/cirugía , Modelos Animales , Solución Salina Hipertónica/farmacología , Animales , Femenino , Riñón/patología , Necrosis , Tamaño de los Órganos , Valores de Referencia , Reproducibilidad de los Resultados , Porcinos , Irrigación Terapéutica , Factores de Tiempo , Ultrasonografía Intervencional/métodos , Urotelio/lesionesRESUMEN
Objective: Analysis of renal excretory system integrity and efficacy of radiofrequency ablation with and without irrigation with saline at 2 o C (SF2). Materials and Methods: The median third of sixteen kidneys were submitted to radiofrequency (exposition of 1 cm) controlled by intra-surgical ultrasound, with eight minutes cycles and median temperature of 90 o C in eight female pigs. One excretory renal system was cooled with SF2, at a 30ml/min rate, and the other kidney was not. After 14 days of post-operatory, the biggest diameters of the lesions and the radiological aspects of the excretory system were compared by bilateral ascending pyelogram and the animals were sacrificed in order to perform histological analysis. Results: There were no significant differences between the diameters of the kidney lesions whether or not exposed to cooling of the excretory system. Median diameter of the cooled kidneys and not cooled kidneys were respectively (in mm): anteroposterior: 11.46 vs. 12.5 (p = 0.23); longitudinal: 17.94 vs. 18.84 (p = 0.62); depth: 11.38 vs. 12.25 (p = 0.47). There was no lesion of the excretory system or signs of leakage of contrast media or hydronephrosis at ascending pyelogram. Conclusion: Cooling of excretory system during radiofrequency ablation does not significantly alter generated coagulation necrosis or affect the integrity of the excretory system in the studied model. .
Asunto(s)
Animales , Femenino , Frío , Ablación por Catéter/métodos , Riñón/cirugía , Modelos Animales , Solución Salina Hipertónica/farmacología , Riñón/patología , Necrosis , Tamaño de los Órganos , Valores de Referencia , Reproducibilidad de los Resultados , Porcinos , Irrigación Terapéutica , Factores de Tiempo , Ultrasonografía Intervencional/métodos , Urotelio/lesionesRESUMEN
In addition to its role as a physical barrier, the urothelium is considered to play an active role in mechanosensation. A key mechanism is the release of transient mediators that activate purinergic P2 receptors and transient receptor potential (TRP) channels to effect changes in intracellular Ca²âº. Despite the implied importance of these receptors and channels in urothelial tissue homeostasis and dysfunctional bladder disease, little is known about their functional expression by the human urothelium. To evaluate the expression and function of P2X and P2Y receptors and TRP channels, the human ureter and bladder were used to separate urothelial and stromal tissues for RNA isolation and cell culture. RT-PCR using stringently designed primer sets was used to establish which P2 and TRP species were expressed at the transcript level, and selective agonists/antagonists were used to confirm functional expression by monitoring changes in intracellular Ca²âº and in a scratch repair assay. The results confirmed the functional expression of P2Y4 receptors and excluded nonexpressed receptors/channels (P2X1, P2X3, P2X6, P2Y6, P2Y11, TRPV5, and TRPM8), while a dearth of specific agonists confounded the functional validation of expressed P2X2, P2X4, P2Y1, P2Y2, TRPV2, TRPV3, TRPV6 and TRPM7 receptors/channels. Although a conventional response was elicited in control stromal-derived cells, the urothelial cell response to well-characterized TRPV1 and TRPV4 agonists/antagonists revealed unexpected anomalies. In addition, agonists that invoked an increase in intracellular Ca²âº promoted urothelial scratch repair, presumably through the release of ATP. The study raises important questions about the ligand selectivity of receptor/channel targets expressed by the urothelium. These pathways are important in urothelial tissue homeostasis, and this opens the possibility of selective drug targeting.
Asunto(s)
Receptores Purinérgicos P2/biosíntesis , Canales de Potencial de Receptor Transitorio/biosíntesis , Urotelio/metabolismo , Adenosina Trifosfato/farmacología , Adulto , Anciano , Calcio/metabolismo , Capsaicina/farmacología , Células Cultivadas , Cartilla de ADN , Homeostasis/fisiología , Humanos , Persona de Mediana Edad , Antagonistas del Receptor Purinérgico P2/farmacología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismo , Canales de Potencial de Receptor Transitorio/agonistas , Canales de Potencial de Receptor Transitorio/antagonistas & inhibidores , Uréter/patología , Vejiga Urinaria/patología , Urotelio/lesiones , Cicatrización de Heridas/fisiologíaRESUMEN
Oxytocin (OXY), a well-known nonapeptide, plays a crucial role in reproduction, and has effects on modulating the immune and inflammatory processes in living organisms as well. Recently it is also known as an antioxidant in several organs. The present study aims to demonstrate the protective effect of OXY against ischemia/reperfusion (I/R) injury in urinary bladder tissue. Abdominal aorta of rats, were clamped to perform urinary bladder ischemia. OXY (0.5 µg/kg) was injected intraperitoneally before ischemia in I/R+OXY group, whereas the vehicle solution was injected to I/R group. At the end of reperfusion, tissue samples from urinary bladder were processed for histochemical, ultrastructural and biochemical analysis. Tissue sections were stained by toluidine blue for mast cell counting and hematoxylin-eosin for histopathology. In addition, malondialdehyde (MDA) and glutathione (GSH) levels were determined biochemically. The results demonstrated that there was an extreme damage at urothelium, dilatation of intercellular junctions, inflammatory cell infiltration in I/R group. I/R+OXY group demonstrated a reduction in the severity of urinary bladder damage. According to mast cell counting results, both granulated and degranulated mast cells were decreased in I/R+OXY group compared to I/R group. The mean MDA level was higher in I/R group compared to control and lower in I/R+OXY group compared to I/R group. GSH level reduced in I/R group compared to the control and increased in I/R+OXY group compared to I/R group. In conclusion, oxytocin, as confirmed by histological evaluation and biochemical assays has a potential protective effect in the urinary bladder tissue against ischemia/reperfusion injury.
Asunto(s)
Oxitocina/administración & dosificación , Daño por Reperfusión , Vejiga Urinaria/efectos de los fármacos , Urotelio/efectos de los fármacos , Animales , Aorta Abdominal/efectos de los fármacos , Aorta Abdominal/patología , Glutatión/metabolismo , Malondialdehído/metabolismo , Ratas , Vejiga Urinaria/patología , Urotelio/lesionesRESUMEN
Catheter-related bladder discomfort (CRBD) is an unrecognized clinical event. Symptoms of CRBD secondary to an indwelling urinary catheter mimic those of an overactive bladder, i.e. urinary frequency and urgency with or without urge incontinence. Stimulation of muscarinic receptors located in the bladder wall by the catheter is the triggering factor. Postoperative pain may be increased by the CRBD. Antimuscarinic drugs, as oxybutynin, are today the main treatment. Further studies are warranted to confirm efficacy of ketamine, tramadol and gabapentin in this situation.
Asunto(s)
Disuria/etiología , Dolor Postoperatorio/etiología , Sala de Recuperación , Cateterismo Urinario/efectos adversos , Aminas/uso terapéutico , Periodo de Recuperación de la Anestesia , Ácidos Ciclohexanocarboxílicos/uso terapéutico , Disuria/tratamiento farmacológico , Disuria/fisiopatología , Femenino , Gabapentina , Humanos , Ketamina/uso terapéutico , Masculino , Ácidos Mandélicos/uso terapéutico , Antagonistas Muscarínicos/uso terapéutico , Dolor Postoperatorio/tratamiento farmacológico , Dolor Postoperatorio/fisiopatología , Receptores Muscarínicos/efectos de los fármacos , Receptores Muscarínicos/fisiología , Factores de Riesgo , Índice de Severidad de la Enfermedad , Evaluación de Síntomas , Síndrome , Tramadol/uso terapéutico , Urotelio/lesiones , Urotelio/fisiopatología , Ácido gamma-Aminobutírico/uso terapéuticoRESUMEN
When the urothelial barrier, i.e., the blood-urine barrier, is injured, rapid resealing of the injury is crucial for the normal functioning of the organism. In order to investigate the mechanisms required for rapid resealing of the barrier, we established in vitro models of hyperplastic and normoplastic urothelia. We found that hyperplastic urothelia achieve significantly higher transepithelial resistance (TER) than normoplastic urothelia. However, the expression of cell junctional (claudin-8, occludin, E-cadherin) and differentiation-related proteins (cytokeratin 20 and uroplakins) is weaker in hyperplastic urothelia. Further investigation of cell differentiation status at the ultrastructural level confirmed that superficial urothelial cells (UCs) in hyperplastic urothelial models achieve a lower differentiation stage than superficial UCs in normoplastic urothelial models. With the establishment of such in vitro models and the aid of TER measurements, flow cytometry, molecular and ultrastructural analysis, we here provide unequivocal evidence that the specific cell-cycle distribution and, consequently, the number of cell layers have a significant influence on the barrier function of urothelia. We demonstrate the importance of hyperplasia for the rapid restoration of the urothelial barrier and the maintenance of high TER until the UCs reach a highly differentiated stage and restoration of the urothelial barrier after injury is complete. The information that this approach provides is unique and we expect that further exploitation of hyperplastic and normoplastic urothelial models in future studies may advance our understanding of blood-urine barrier development and functionality.
Asunto(s)
Ciclo Celular/fisiología , Vejiga Urinaria/metabolismo , Vejiga Urinaria/patología , Urotelio/metabolismo , Urotelio/ultraestructura , Cicatrización de Heridas/fisiología , Animales , Cadherinas/metabolismo , Diferenciación Celular/fisiología , Células Cultivadas , Claudinas/metabolismo , Células Epiteliales/metabolismo , Células Epiteliales/patología , Hiperplasia/metabolismo , Queratina-20/metabolismo , Proteínas de la Membrana/metabolismo , Ocludina , Porcinos , Vejiga Urinaria/lesiones , Uroplaquinas/metabolismo , Urotelio/lesiones , Urotelio/patologíaRESUMEN
Our aim is to study the prospective trial where urine cytology was used to detect the acute urothelial mucosal damage in patients who undergo extracorporeal shock waves lithotripsy (SWL). The study included 48 consecutive patients (28 male, 20 female) with mean age of 49.02 years (range 18-66) who were treated with SWL due to renal stones (30 patients) or upper ureter stones (18 patients). The mean calculi diameter was 12.44 mm (range 5-20). Urinary cytologic examinations were done for all patients immediately before and after SWL therapy and 10 days latter. The average numbers of transitional cells, red blood cells and myocytes were counted under 40 × magnification. In overall patients the average numbers of transitional cells at the cytologic examinations done immediately before and after SWL therapy were 1.6 and 7.53 cell/field, respectively (p = 0.001). The increment in transitional cells at cytologic examination after SWL was significantly influenced only by number of shock waves applied (p = 0.003). No muscle cell was detected in all cytologic examinations. The cytologic examinations which were done after 10 days of SWL therapy showed recovery from all cytologic abnormalities. The acute increment in number of transitional cells after the SWL is not clinically important and it is a temporary change. Urothelial lesion is limited to mucosal layer and there is no evidence of damage to basal membrane or deeper muscle layer. SWL safety on urothelial and muscular layer was demonstrated. However, evaluation of larger series with use of other lithotripters is necessary before reaching any definitive conclusions.