RESUMEN
We present a scientific investigation into the pathogenesis of a urinary bladder disease. The disease in question is called urofacial syndrome (UFS), a congenital condition inherited in an autosomal recessive manner. UFS features incomplete urinary bladder emptying and vesicoureteric reflux, with a high risk of recurrent urosepsis and end-stage renal disease. The story starts from a human genomic perspective, then proceeds through experiments that seek to determine the roles of the implicated molecules in embryonic frogs and newborn mice. A future aim would be to use such biological knowledge to intelligently choose novel therapies for UFS. We focus on heparanase proteins and the peripheral nervous system, molecules and tissues that appear to be key players in the pathogenesis of UFS and therefore must also be critical for functional differentiation of healthy bladders. These considerations allow the envisioning of novel biological treatments, although the potential difficulties of targeting the developing bladder in vivo should not be underestimated.
Asunto(s)
Estudios de Asociación Genética , Glucuronidasa/genética , Glucuronidasa/metabolismo , Mutación/genética , Enfermedades de la Vejiga Urinaria/congénito , Enfermedades de la Vejiga Urinaria/genética , Animales , Humanos , Ratones , Enfermedades de la Vejiga Urinaria/enzimologíaRESUMEN
Glandular lesions of the urinary bladder include a broad spectrum of entities ranging from completely benign to primary and secondary malignancies. The accurate diagnosis of these lesions is both important and challenging. Recently, studies suggest that telomerase reverse transcriptase (TERT) promoter mutations could be a biomarker for urothelial carcinoma (UC). We hypothesized that these mutations can distinguish UC with glandular differentiation from nephrogenic adenoma, primary adenocarcinoma of the urinary bladder (PAUB), or secondary malignancies. Twenty-five cases of benign glandular lesions (including nephrogenic adenoma); 29 cases of UC with glandular differentiation; 10 cases of PAUB; and 10 cases each of metastatic colon cancer, prostatic carcinoma, and carcinoma from Mullerian origin were collected. Slides were reviewed and selected to make sure the lesion was at least 10% to 20% of all tissue. Macrodissection was performed in some of cases, and genomic DNA was extracted from the tissue. Telomerase reverse transcriptase promoter mutations were determined by standard polymerase chain reaction sequencing. Twenty-one cases (72%) of UC with glandular differentiation were positive for TERT promoter mutations. However, none of the remaining cases (total 65 cases of benign lesions, PAUB, and metastatic carcinomas) was positive for TERT promoter mutation. Telomerase reverse transcriptase promoter mutations were highly associated with UC including UC with glandular differentiation but not other glandular lesions of bladder. Therefore, in conjunction with morphologic features, Immunohistochemistry stain profile, and clinical information, TERT promoter mutations could distinguish UC with glandular differentiation from other bladder glandular lesions. In addition, lack of TERT promoter mutations in primary adenocarcinoma of bladder suggests that this entity may have different origin or carcinogenesis from those of UC.
Asunto(s)
Mutación , Neoplasias Glandulares y Epiteliales/genética , Telomerasa/genética , Enfermedades de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/genética , Adenoma/enzimología , Adenoma/genética , Adenoma/patología , Biomarcadores de Tumor/genética , Cistitis/enzimología , Cistitis/genética , Cistitis/patología , Femenino , Humanos , Masculino , Neoplasias Glandulares y Epiteliales/enzimología , Neoplasias Glandulares y Epiteliales/patología , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/patología , Neoplasias de la Vejiga Urinaria/enzimología , Neoplasias de la Vejiga Urinaria/patología , Neoplasias de la Vejiga Urinaria/secundarioRESUMEN
BACKGROUND: Bladder dysfunction has one of the highest prevalences as a comorbidity of obesity in industrialized countries. The aetiopathogenesis of obesity-associated bladder dysfunction is still obscure, but there is growing evidence that general metabolic changes in obese patients may be in part responsible. As demonstrated recently, high fat diet (HFD) significantly alters the protein expression in the urinary bladder, activates multiple signalling pathways associated with cell survival and inflammation and ultimately provokes bladder fibrosis in an obese rat model. The study aimed to elucidate the role of matrix metalloproteases (MMPs) and their specific tissue inhibitors of metalloproteases (TIMPs) in obesity-related bladder extracellular matrix (ECM) remodelling and the effect of weight loss surgery via sleeve gastrectomy (SG) on phenotype and molecular parameters. METHODS: Twenty-four male Sprague-Dawley rats were used for (i) characterization of the HFD phenotype and (ii) evaluation of alterations following SG. Metabolic status, the degree of bladder fibrosis and tissue expression and activity of MMP2, MMP9, MMP14, TIMP1 and TIMP2 were analysed by immunohistochemistry, enzyme-linked immunosorbent assay and activity assays. Statistical differences were calculated by analysis of variance or independent Student's t-test. A P-value <0.05 was considered statistically significant. RESULTS: In HFD rats, we found significant alterations in lipid metabolism, fat mass, free fatty acid profile, insulin resistance and inflammatory markers. Voided volume was significantly decreased, and bladder showed marked fibrosis. MMPs and TIMPs were differentially regulated depending on animal status (controls, chow diet, HFD, and SG- and sham-operated animals) in both urothelium and detrusor smooth muscle. Although animal weight and most metabolic parameters were positively affected by SG, bladder fibrosis persisted. The limitations of this study were 1 month follow-up and lack of direct measurement of bladder function. CONCLUSIONS: Early diagnosis of the bladder dysfunction associated with obesity is essential to allow targeted early intervention, that is, before manifestation of potentially irreversible ECM fibrotic alterations.
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Metaloproteinasas de la Matriz/metabolismo , Obesidad/metabolismo , Obesidad/cirugía , Enfermedades de la Vejiga Urinaria/patología , Vejiga Urinaria/patología , Pérdida de Peso , Animales , Cirugía Bariátrica , Células Cultivadas , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Fibrosis , Metabolismo de los Lípidos , Masculino , Obesidad/complicaciones , Obesidad/enzimología , Obesidad/patología , Ratas , Ratas Sprague-Dawley , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/etiología , Enfermedades de la Vejiga Urinaria/cirugíaRESUMEN
Nitric oxide and the cyclic nucleotide monophosphates cAMP and cGMP have a role in control of the micturition process and hence, are suggested to be involved in the pathophysiology of storage and voiding disorders. Phosphodiesterase enzymes (PDEs) hydrolyse cAMP and cGMP. Inhibition of PDEs increases cAMP and cGMP levels and relaxes urinary bladder smooth musculature. Although many preclinical studies have been conducted, to date, only PDE1 and PDE5 inhibitors have been tested clinically for the management of storage and voiding disorders. Treatment with PDE1 inhibitors might improve micturition frequency in patients with overactive bladder, whereas inhibition of PDE5 improves lower urinary tract symptoms in men, either with or without BPH and erectile dysfunction (ED). Furthermore, the combination of a PDE5 inhibitor and an α-adrenoceptor antagonist has superior efficacy to monotherapy with either agent. However, the role of PDE5 inhibitors in the treatment of women with detrusor overactivity remains unclear. The clinical application of agents that inhibit other PDEs, including PDE4, also certainly merits scientific attention. PDE inhibitors seem likely to become a valuable alternative treatment for patients with storage and voiding disorders in the future.
Asunto(s)
Hidrolasas Diéster Fosfóricas/fisiología , Enfermedades de la Vejiga Urinaria/enzimología , Vejiga Urinaria/enzimología , Vejiga Urinaria/fisiopatología , Animales , Disfunción Eréctil/enzimología , Disfunción Eréctil/fisiopatología , Disfunción Eréctil/terapia , Femenino , Humanos , Síntomas del Sistema Urinario Inferior/enzimología , Síntomas del Sistema Urinario Inferior/fisiopatología , Síntomas del Sistema Urinario Inferior/terapia , Masculino , Inhibidores de Fosfodiesterasa/farmacología , Inhibidores de Fosfodiesterasa/uso terapéutico , Hidrolasas Diéster Fosfóricas/metabolismo , Vejiga Urinaria/patología , Enfermedades de la Vejiga Urinaria/fisiopatología , Enfermedades de la Vejiga Urinaria/terapia , Vejiga Urinaria Hiperactiva/enzimología , Vejiga Urinaria Hiperactiva/fisiopatología , Vejiga Urinaria Hiperactiva/terapiaRESUMEN
OBJECTIVES: To investigate whether overactivated Poly (ADP-ribose) polymerase (PARP) and subsequent activation of nuclear factor kappa B (NF-κB) correlate with the development of diabetic cystopathy via induction of bladder apoptosis. Diabetic cystopathy as a common complication of diabetes is frequently associated with increased oxidative stress and apoptosis of the bladder. PARP is activated by hyperglycemia-induced oxidative stress and plays a critical role in cell apoptosis and the development of diabetic complications, such as retinopathy and nephropathy. METHODS: Sprague-Dawley rats were divided into 3 groups: control, diabetic, and diabetic treated with PARP inhibitor (DM+Vit-B3). Four weeks after induction of diabetes, the DM+Vit-B3 group was treated with PARP inhibitor (nicotinamide, 400 mg/kg/d) for 3 weeks. Bladder function was then assessed by conscious cystometry. The extent of oxidative stress and apoptosis, expression of poly(ADP-ribose) (PAR), NF-κB, phosphorylated inhibitor of NF-κB (IκB)-α, Bcl-2, and Bax in the bladder were also investigated. RESULTS: Bladder dysfunction was strongly associated with increased oxidative stress and bladder apoptosis. In addition, the amount of PAR, phosphorylated IκB-α, expression of NF-κB, and Bax were significantly increased in diabetic rat bladder. Inhibition of PARP significantly reduced PARP activation and expression of NF-κB and Bax. As a result, bladder apoptosis was attenuated and bladder function was improved. CONCLUSIONS: These results indicate that overactivated PARP and subsequent activation of NF-κB play important roles in the development of diabetic cystopathy via induction of bladder apoptosis. These findings may be applied in the development of novel therapies for patients with diabetic cystopathy.
Asunto(s)
Complicaciones de la Diabetes/enzimología , FN-kappa B/fisiología , Poli(ADP-Ribosa) Polimerasas/fisiología , Enfermedades de la Vejiga Urinaria/enzimología , Animales , Masculino , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Ratas , Ratas Sprague-DawleyRESUMEN
AIMS: Diabetic cystopathy, a common complication of diabetes, is frequently associated with an increase in oxidative stress and apoptosis of the bladder. Poly(ADP-ribose) polymerase (PARP) is activated under such conditions of oxidative stress, and plays a critical role in cell apoptosis. The aim of this study was to investigate whether the activation of PARP and subsequent activation of c-Jun N-terminal kinase (JNK) and the mitochondrial apoptotic pathway are involved in the development of diabetic cystopathy. METHODS: Bladder function was assessed in a streptozotocin (STZ)-induced diabetic rat model with or without 3-aminobenzamide treatment, a PARP inhibitor. The degree of bladder apoptosis, expression of poly(ADP-ribose) (PAR) in the bladder, phosphorylated JNK, the levels of Bcl-2 and Bax, caspase 3 activity and nuclear translocation of the apoptotic inducing factor (AIF) from mitochondria were investigated. RESULTS: Bladder dysfunction was significantly associated with an increase of bladder apoptosis, and a reduction of the Bcl-2/Bax ratio. In addition, the amount of PAR, phosphorylated JNK, caspase 3 activity, and nuclear translocation of AIF were significantly increased in the diabetic rats. Inhibition of PARP significantly suppressed activation of PARP, JNK and restored the Bcl-2/Bax ratio. Activation of caspase 3 and nuclear translocation of AIF were also significantly reduced by PARP inhibition. As a result, the bladder apoptosis was attenuated and the bladder function improved. CONCLUSIONS: These results indicate that bladder apoptosis is involved in diabetic cystopathy via activation of the PARP/JNK/mitochondrial apoptotic pathway. These findings may be used to develop novel therapies for patients with diabetic bladder dysfunction.
Asunto(s)
Apoptosis , Complicaciones de la Diabetes/enzimología , Diabetes Mellitus Experimental/complicaciones , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Mitocondrias/enzimología , Poli(ADP-Ribosa) Polimerasas/metabolismo , Enfermedades de la Vejiga Urinaria/enzimología , Vejiga Urinaria/enzimología , Transporte Activo de Núcleo Celular , Animales , Factor Inductor de la Apoptosis/metabolismo , Benzamidas/farmacología , Caspasa 3/metabolismo , Complicaciones de la Diabetes/etiología , Complicaciones de la Diabetes/patología , Complicaciones de la Diabetes/prevención & control , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/enzimología , Diabetes Mellitus Experimental/patología , Inhibidores Enzimáticos/farmacología , Masculino , Mitocondrias/efectos de los fármacos , Mitocondrias/patología , Estrés Oxidativo , Fosforilación , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Vejiga Urinaria/efectos de los fármacos , Vejiga Urinaria/patología , Vejiga Urinaria/fisiopatología , Enfermedades de la Vejiga Urinaria/etiología , Enfermedades de la Vejiga Urinaria/patología , Enfermedades de la Vejiga Urinaria/prevención & control , Urodinámica , Proteína X Asociada a bcl-2/metabolismoRESUMEN
OBJECTIVES: Nitric oxide is a neurotransmitter that is related to overactive bladder. This study was undertaken to evaluate the expression of nitric oxide synthases (NOS) in relation to bladder function after the removal of the bladder outlet obstruction in rats. MATERIALS AND METHODS: 50 rats were divided into a control group (n = 10) and an experimental group (n = 40). The bladders in the experimental group were partially obstructed for 3 weeks. 3 weeks after removal, cystometrograms (CMG) were performed. On the basis of CMG results, the experimental group was subdivided into 'normalized' and 'overactive' groups. Expression of neuronal, inducible, and endothelial NOS mRNA in bladders of each group was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: On the CMG, the contraction interval of the overactive group was markedly decreased. RT-PCR results showed significantly decreased expression of nNOS and iNOS mRNA in the normalized group compared to the control group, and increased nNOS levels in the overactive group, compared to the normalized group (p < 0.05). A marked increase in eNOS mRNA expression was evident in the normalized and overactive groups compared to the control group (p < 0.05). CONCLUSIONS: Our results clearly show increases in nNOS and eNOS expression according to bladder function, which may be related to a persistently overactive bladder or irritative symptoms, following the removal of a bladder outlet obstruction.
Asunto(s)
Músculo Liso/enzimología , Óxido Nítrico Sintasa de Tipo III/biosíntesis , Óxido Nítrico Sintasa de Tipo I/biosíntesis , Obstrucción del Cuello de la Vejiga Urinaria/cirugía , Vejiga Urinaria/enzimología , Animales , Cartilla de ADN/química , Masculino , Óxido Nítrico/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Transducción de Señal , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/terapia , Obstrucción del Cuello de la Vejiga Urinaria/fisiopatologíaRESUMEN
It was previously shown that nitric oxide produced by inducible nitric oxide synthase (iNOS) and peroxynitrite are responsible for cyclophosphamide (CP)-induced cystitis. Since endogenous production of peroxynitrite is known to lead to poly(ADP-ribose) polymerase (PARP) activation, in this study, the aim was to evaluate whether the PARP activation pathway is also included in the pathogenesis of CP-induced bladder ulceration in rats. A total of 48 male albino Wistar rats were divided into 5 groups. Group 1 served as control and was given 2 ml saline; four groups received a single dose of CP (200 mg/kg) with the same time intervals. Group 2 received CP only; Group 3, selective iNOS inhibitor 1400W (20 mg/kg); Group 4, peroxynitrite scavenger ebselen (30 mg/kg); and Group 5, PARP inhibitor 3-aminobenzamide (20 mg/kg). CP injection resulted in severe cystitis with continuous macroscopic hemorrhage, strong edema, inflammation, and ulceration. Moreover, bladder iNOS activation and urine nitrite-nitrate levels were dramatically increased. Histologically, 1400W protected bladder against CP damage and decreased urine nitrite-nitrate levels and bladder iNOS induction. Ebselen has shown similar histologic results with 1400W without changing urinary nitrite-nitrate level and iNOS activity. Furthermore in the 3-aminobenzamide group, beneficial effects had also occurred including decreased ulceration. These results suggest that PARP activation involves pathogenesis of CP-induced bladder ulceration. Furthermore, PARP is not only important for ulceration but also for bladder edema, hemorrhage, and inflammation because of broken uroepithelial cellular integrity.
Asunto(s)
Ciclofosfamida/farmacología , Inhibidores Enzimáticos/farmacología , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/prevención & control , Animales , Biomarcadores/metabolismo , Peso Corporal/efectos de los fármacos , Masculino , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/metabolismo , Nitrosación , Tamaño de los Órganos/efectos de los fármacos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Ratas , Ratas Wistar , Enfermedades de la Vejiga Urinaria/inducido químicamente , Enfermedades de la Vejiga Urinaria/patologíaAsunto(s)
3',5'-GMP Cíclico Fosfodiesterasas/metabolismo , Disfunción Eréctil/enzimología , Pelvis/fisiopatología , Inhibidores de Fosfodiesterasa/uso terapéutico , Hiperplasia Prostática/enzimología , Enfermedades de la Vejiga Urinaria/enzimología , 3',5'-GMP Cíclico Fosfodiesterasas/antagonistas & inhibidores , Anciano , Anciano de 80 o más Años , Disfunción Eréctil/complicaciones , Disfunción Eréctil/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Hiperplasia Prostática/complicaciones , Hiperplasia Prostática/tratamiento farmacológico , Enfermedades de la Vejiga Urinaria/complicaciones , Enfermedades de la Vejiga Urinaria/tratamiento farmacológicoRESUMEN
The role of dendritic cells (DC) in urinary tract infections (UTI) is unknown. These cells contribute directly to the innate defense against various viral and bacterial infections. Here, we studied their role in UTI using an experimental model induced by transurethral instillation of the uropathogenic Escherichia coli (UPEC) strain 536 into C57BL/6 mice. While few DC were found in the uninfected bladder, many had been recruited after 24 h, mostly to the submucosa and uroepithelium. They expressed markers of activation and maturation and exhibited the CD11b+ F4/80+ CD8- Gr-1- myeloid subtype. Also, tumor necrosis factor alpha (TNF-alpha)- and inducible nitric oxide synthase (iNOS)-producing CD11bINT DC (Tip-DC) were detected, which recently were proposed to be critical in the defense against bacterial infections. However, Tip-DC-deficient CCR2-/- mice did not show reduced clearance of UPEC from the infected bladder. Moreover, clearance was also unimpaired in CD11c-DTR mice depleted of all DC by injection of diphtheria toxin. This may be explained by the abundance of granulocytes and of iNOS- and TNF-alpha-producing non-DC that were able to replace Tip-DC functionality. These findings demonstrate that some of the abundant DC recruited in UTI contributed innate immune effector functions, which were, however, dispensable in the microenvironment of the bladder.
Asunto(s)
Movimiento Celular/inmunología , Células Dendríticas/enzimología , Células Dendríticas/inmunología , Infecciones por Escherichia coli/microbiología , Óxido Nítrico Sintasa de Tipo II/fisiología , Factor de Necrosis Tumoral alfa/fisiología , Enfermedades de la Vejiga Urinaria/microbiología , Animales , Células Dendríticas/microbiología , Modelos Animales de Enfermedad , Infecciones por Escherichia coli/enzimología , Infecciones por Escherichia coli/inmunología , Inmunidad Celular , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Factor de Necrosis Tumoral alfa/biosíntesis , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/inmunologíaRESUMEN
Cyclophosphamide (CP) is a commonly used anti-cancer drug which causes toxicity by its reactive metabolites such as acrolein and phosphoramide mustard. In the present study modulation of toxicity caused by concomitant exposure to CP and l-buthionine-SR-sulfoximine (BSO) by fenugreek (Trigonella foenum-graecum L.) extract was evaluated by measuring lipid peroxidation (LPO) and anti-oxidants in urinary bladder in mice. Fenugreek, a common dietary and medicinal herb, showed protective effect not only on LPO but also on the enzymatic anti-oxidants. CP-treated animals exhibited a significant decrease in the activities of glutathione S-transferase (GST), glutathione reductase (GR), glutathione peroxidase (GP) and catalase (CAT) when compared to the controls. Level of reduced glutathione (GSH) was also reduced with an increase in LPO in CP-treated animals. BSO treatment depicted an additive toxic effect in CP-treated animals. Pre-treatment of herbal extract restored activities of all the enzymes and thus showed an overall protective effect on additive effect of CP and BSO. Restoration of GSH by extract treatment may play an important role in reversing CP-induced apoptosis and free radical-mediated LPO in urinary bladder. Fenugreek, known for its hypoglycemic, anti-inflammatory and immunomodulatory activity, may be a promising protective medicinal herb for consideration in complementary therapy in cancer patients under chemotherapeutic interventions.
Asunto(s)
Butionina Sulfoximina/toxicidad , Ciclofosfamida/toxicidad , Extractos Vegetales/farmacología , Trigonella/química , Enfermedades de la Vejiga Urinaria/inducido químicamente , Enfermedades de la Vejiga Urinaria/prevención & control , Animales , Antioxidantes/metabolismo , Interacciones Farmacológicas , Glutatión/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Vejiga Urinaria/efectos de los fármacos , Vejiga Urinaria/enzimología , Vejiga Urinaria/metabolismo , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/metabolismo , Agua/químicaRESUMEN
The extracellular signal-regulated kinase 1 and 2 (ERK) pathway, regulated by phosphorylation on specific amino acids, is emerging as an important signaling cascade in neurones, transducing sensory input into cellular responses. In the mammalian nervous system, the ERK pathway has been found to mediate plasticity events. Particularly, in the spinal cord, ERK play an important role in allodynia and hyperalgesia. Recently, it was demonstrated that ERK activation is upregulated in the spinal cord of rats with chronic bladder inflammation and contributes to bladder overactivity. Thus, in this study we sought to assess the involvement of ERK in micturition reflexes associated to spinal cord injury (SCI) in the rat. Bladder function in chronic SCI rats was altered compared to spinal intact rats. PhosphoERK levels were upregulated in the L6 spinal cord segment, particularly after saline infusion for 2 h. The increase in spinal ERK phosphorylation was specifically restricted to L6 spinal segment. No variation in the levels of total ERK protein was observed. Intrathecal administration of PD98059, a specific inhibitor of ERK phosphorylation, reduced the frequency and amplitude of bladder contractions in SCI animals but not in spinal intact ones. Overall, our results demonstrate increased activation of the ERK pathway in the spinal cord from SCI rats, restricted to spinal segments that receive sensory input arising from the bladder. Since the use of PD98059 reduced the frequency and amplitude of bladder contractions, ERK inhibitors may provide a new therapeutic approach to the treatment of bladder overactivity after spinal injuries.
Asunto(s)
Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Traumatismos de la Médula Espinal/enzimología , Médula Espinal/enzimología , Vejiga Urinaria/fisiología , Animales , Activación Enzimática/fisiología , Femenino , Fosforilación , Ratas , Ratas Wistar , Traumatismos de la Médula Espinal/fisiopatología , Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/fisiopatologíaRESUMEN
OBJECTIVES: To evaluate the oxidative status of the bladder 8 weeks after diabetes induction. Oxidative stress has recently been implicated in the pathogenesis of diabetes complications, but its role in diabetic cystopathy has not been studied. METHODS: Sprague-Dawley rats were divided into three groups: control (n = 11), diuretic control (5% sucrose drink; n = 6), and streptozotocin-induced diabetic group (n = 14). Eight weeks later, the bladders were dissected. We measured the antioxidant scavenging enzymes (catalase and superoxide dismutase)-like activity and the levels of the thiobarbituric acid reactive substances, as a marker of lipid peroxidation. We also examined the levels of inducible nitric oxide synthase and apoptosis in the bladders. RESULTS: We found a statistically significant reduction in the catalase-like activity in the bladders from the diabetic group compared with the other groups (P = 0.017, diabetic versus control); the difference in the superoxide dismutase-like activity was not statistically significant among the groups. The thiobarbituric acid reactive substances levels were significantly greater in the diabetic compared with other groups (131.9 +/- 47.5, 46.7 +/- 17.9, and 60.9 +/- 25.4 nmol/mg protein in the diabetic, control, and diuretic group, respectively, P = 0.006, diabetic versus control). Immunohistochemical and apoptosis studies showed a statistically significant increased number of inducible nitric oxide synthase-positive cells and apoptotic cells in the diabetic bladder smooth muscle cells (P <0.001). CONCLUSIONS: Our findings showed that oxidative stress occurred in the bladders of the STZ-diabetic rats and was not mediated by diuresis. The oxidative damage of the smooth muscle cells may be a contributory factor in diabetic cystopathy.
Asunto(s)
Diabetes Mellitus Experimental/fisiopatología , Nefropatías Diabéticas/fisiopatología , Estrés Oxidativo , Enfermedades de la Vejiga Urinaria/etiología , Animales , Apoptosis , Catalasa/análisis , Nefropatías Diabéticas/etiología , Modelos Animales de Enfermedad , Inmunohistoquímica , Peroxidación de Lípido , Masculino , Músculo Liso/fisiopatología , Óxido Nítrico Sintasa/análisis , Óxido Nítrico Sintasa de Tipo II , Ratas , Ratas Sprague-Dawley , Superóxido Dismutasa/análisis , Sustancias Reactivas al Ácido Tiobarbitúrico/análisis , Factores de Tiempo , Vejiga Urinaria/fisiopatología , Enfermedades de la Vejiga Urinaria/enzimologíaRESUMEN
Inflammatory myofibroblastic tumours (IMFT) may arise at any anatomical site, including lung, soft tissues, retroperitoneum and bladder. Although morphologically similar, these lesions encompass a spectrum of entities with differing aetiology, ranging from reactive/regenerative proliferations to low-grade neoplasms with a risk of local recurrence, but no significant metastatic potential. Vesical IMFT usually presents as a polypoid mass with a pale firm cut surface and can be of considerable size, mimicking a malignant tumour clinically and radiologically. Its good outcome, however, warrants conservative surgical excision, emphasising the importance of identification and distinction from malignant tumours of the bladder that may require more radical surgery and/or adjuvant therapy. We conducted a preliminary retrospective, comparative immunocytochemical study of 20 bladder tumours, including nine IMFTs, five spindle cell (sarcomatoid) carcinomas, two rhabdomyosarcomas, two leiomyosarcomas and two neurofibromas. The results confirmed IMFT positivity for smooth muscle actin, desmin and cytokeratin in 78-89% cases, resulting in potential confusion with sarcomatoid carcinoma or leiomyosarcoma. In contrast, cytoplasmic anaplastic lymphoma kinase (ALK 1) staining was present in eight IMFT (89%), but was not seen in any other lesion examined. The ALK 1 staining was confirmed by fluorescence in situ hybridisation, with translocation of the ALK gene present in 15-60% tumour cells in four of six IMFT examined, but not in four cases of sarcomatoid carcinoma or three of leiomyosarcoma. In conclusion, ALK 1 staining may be of value in the distinction of vesical IMFT from morphologically similar entities, and often reflects ALK gene translocations in these lesions.
Asunto(s)
Granuloma de Células Plasmáticas/patología , Proteínas Tirosina Quinasas/análisis , Enfermedades de la Vejiga Urinaria/patología , Actinas/análisis , Adolescente , Adulto , Quinasa de Linfoma Anaplásico , Proteínas de Unión al Calcio/análisis , Proteínas de Unión a Calmodulina/análisis , Desmina/análisis , Diagnóstico Diferencial , Femenino , Reordenamiento Génico , Granuloma de Células Plasmáticas/enzimología , Granuloma de Células Plasmáticas/genética , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Queratinas/análisis , Masculino , Proteínas de Microfilamentos , Persona de Mediana Edad , Músculo Liso/química , Proteínas Tirosina Quinasas/genética , Proteínas Tirosina Quinasas Receptoras , Coloración y Etiquetado , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/enzimología , Neoplasias de la Vejiga Urinaria/patología , Vimentina/análisis , CalponinasRESUMEN
AIMS: To study the expression of the endothelial and inducible isoforms of nitric oxide synthase (eNOS and iNOS, respectively) in human bladder carcinoma and schistosomal bladder disease, and to compare it with normal adult and fetal urothelium. Nitric oxide is thought to play a complex role in human carcinogenesis, but has only recently been investigated in bladder cancer. METHODS: Immunohistochemistry was performed on paraffin wax embedded sections of 33 human bladder carcinomas and five bladder carcinoma cell lines; in addition, seven schistosomal bladder cases and normal and fetal urothelium were investigated. In the cell lines enzymatic activity was examined by the NADPH diaphorase reaction. RESULTS: Immunoreactivity for eNOS was present in most cells of all 31 cases examined. Immunoreactivity for iNOS was less abundant and was seen in 23 of 25 cases. Similar findings were noted in schistosomal bladder cancer. In the normal bladder mucosa, eNOS immunoreactivity was found only in the superficial cell layer and iNOS was not expressed, whereas in the fetal urothelium immunoreactivity for both isoforms was seen in all cell layers. Enzymatic activity and immunoreactivity for eNOS and iNOS were evident in the five bladder carcinoma cell lines. CONCLUSIONS: It is possible that NOS plays a role in the differentiation of the transitional epithelium in fetal life, has a biological function in the adult bladder mucosa, and is involved in bladder carcinogenesis. eNOS and iNOS immunoreactivity do not differ in schistosomal and non-schistosomal bladder carcinoma, but resemble the pattern of expression typical of fetal urothelium.
Asunto(s)
Carcinoma de Células Transicionales/enzimología , Óxido Nítrico Sintasa/análisis , Neoplasias de la Vejiga Urinaria/enzimología , Adulto , Carcinoma de Células Transicionales/parasitología , Humanos , Inmunohistoquímica , Óxido Nítrico Sintasa de Tipo II , Óxido Nítrico Sintasa de Tipo III , Esquistosomiasis/enzimología , Células Tumorales Cultivadas/enzimología , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/parasitología , Neoplasias de la Vejiga Urinaria/parasitología , Urotelio/embriología , Urotelio/enzimologíaRESUMEN
PURPOSE: To investigate the effect of diabetes mellitus (DM) on the density and distribution of nitric oxide synthase (NOS) and the smooth muscle responses to non-adrenergic, non-cholinergic (NANC) nerve stimulation and exogenous nitric oxide (NO) in the rabbit lower urinary tract. MATERIALS AND METHODS: Transverse sections of detrusor, bladder neck and urethra, from control and six months alloxan-induced DM New Zealand White rabbits were incubated with a radioligand for NOS ([3H]-L-N(G)-nitroarginine). Densitometric analysis was performed on the autoradiographs. NADPH diaphorase histochemistry was also used as a marker for NOS activity. Responses to NANC nerve stimulation (5 to 20 Hz) and to NO (10(-6) to 3x10(-4) M.) on smooth muscle strips from detrusor, bladder neck and urethra were measured in organ baths. RESULTS: NOS binding sites were significantly (p<0.03) more dense in the bladder neck than in the detrusor in both DM and control groups. In DM bladder neck, NOS binding sites were significantly (p<0.04) increased compared with the controls. NADPH diaphorase activity appeared markedly increased in the detrusor, bladder neck and urethra of DM animals compared with controls. The mean IC50 for exogenous NO in control versus DM were not statistically different in the bladder neck (1.03x10(-4) M versus 9.8x10(-5) M) and urethra (8.1x10(-5) M versus 8.8x10(-5) M), but the relaxations to 5x10(-6) M of NO were significantly impaired (p<0.04) in the DM urethral smooth muscle. NANC nerve-mediated relaxations were significantly impaired (p<0.001) in the DM urethral smooth muscle. CONCLUSIONS: Alterations of both the NOS binding sites and functional responses to NANC nerve stimulation suggest that NO may have a pathophysiological role in the urinary bladder dysfunction associated with DM.
Asunto(s)
Diabetes Mellitus Experimental/enzimología , Diabetes Mellitus Experimental/fisiopatología , Relajación Muscular/fisiología , Músculo Liso/enzimología , Músculo Liso/fisiopatología , Óxido Nítrico Sintasa/metabolismo , Enfermedades de la Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/fisiopatología , Vejiga Urinaria/enzimología , Vejiga Urinaria/fisiopatología , Animales , Autorradiografía , Sitios de Unión , Diabetes Mellitus Experimental/complicaciones , Masculino , NADPH Deshidrogenasa/metabolismo , Nitroarginina , Conejos , Uretra/enzimología , Uretra/fisiopatología , Enfermedades de la Vejiga Urinaria/etiologíaRESUMEN
This study investigated whether the enzyme telomerase is active in bladder tumours, whether there is any correlation between activity and grade, and whether the enzyme is expressed in non-malignant conditions. Fifty-two patients undergoing cystoscopy or TURBT at a district general hospital were included, 25 with current bladder tumours, 13 with previous but no current tumours, and 14 with non-malignant pathology. Specimens were analysed by the telomerase repeat amplification protocol (TRAP assay), a highly-sensitive polymerase chain reaction (PCR)-based assay, and a commercially-available ELISA kit. Telomerase activity was detected in 80% of bladder tumours, more frequently in moderate- or poorly-differentiated (93%) than well-differentiated (56%) tumours. Activity was not uniform across individual tumours. Telomerase was also frequently (61%) detected in inflammatory lesions found in patients being followed up for previous bladder tumours, and in two (14%) patients with benign pathologies. In conclusion, telomerase was frequently but not uniformly detected in bladder tumours; its presence was not specific to malignancy. There is a possible correlation between tumour grade and telomerase activity.
Asunto(s)
Telomerasa/metabolismo , Neoplasias de la Vejiga Urinaria/enzimología , Biopsia , Activación Enzimática , Ensayo de Inmunoadsorción Enzimática , Humanos , Inflamación/enzimología , Reacción en Cadena de la Polimerasa , Vejiga Urinaria/enzimología , Enfermedades de la Vejiga Urinaria/enzimología , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
OBJECTIVE: To assess whether there is a reduction in serum antioxidant activity in patients who have undergone a clam enterocystoplasty procedure. PATIENTS AND METHODS: Serum glutathione peroxidase (GPase) activity was measured in 20 patients who had undergone clam enterocystoplasty. Serum selenium concentration was also measured in 62 similar patients and compared with 56 healthy controls and 44 patients with a neuropathic bladder, mainly patients with spinal cord injuries who had not undergone surgery. RESULTS: GPase activity correlated well with serum selenium measurement. There was a significant reduction (P < 0.001) in serum selenium level in young (< 50 years old) non-neuropathic bladder patients following clam enterocystoplasty. This reduction in serum selenium was similar to that found in both unoperated patients with a neuropathic bladder (who are known to have an increased risk of developing bladder cancer) and those patients with a neuropathic bladder who had undergone augmentation. This reduction was not related to urinary tract infection nor the time since surgery. CONCLUSION: A reduction in serum selenium has been shown to increase susceptibility to bladder cancer following carcinogenic exposure to compounds such as nitrosamines. This study suggests that patients with idiopathic and congenital instability may be at an equally high risk as a result of undergoing this procedure.
Asunto(s)
Antioxidantes/metabolismo , Glutatión Peroxidasa/sangre , Selenio/deficiencia , Enfermedades de la Vejiga Urinaria/enzimología , Adolescente , Adulto , Anciano , Niño , Femenino , Humanos , Intestinos/cirugía , Masculino , Persona de Mediana Edad , Vejiga Urinaria/cirugía , Enfermedades de la Vejiga Urinaria/cirugía , Vejiga Urinaria Neurogénica/enzimología , Vejiga Urinaria Neurogénica/cirugíaRESUMEN
OBJECTIVES: The objective of this study was to establish the identity of a protein found in high concentrations in squamous metaplasia of the bladder. DESIGN AND METHODS: The protein was isolated and subjected to a series of physical, chemical, and catalytic studies. RESULTS: In the normal urothelium the protein was confined to a juxtanuclear pattern on the luminal side of the umbrella cells; in squamous metaplasia and squamous cell carcinoma the protein was increased and exhibited a more diffuse intracellular distribution. The protein was found to be identical to triosephosphate isomerase (EC 5.3.1.1; TPI) with respect to its immunological properties, native and subunit molecular weights, electrophoretic mobility, catalytic activity, and amino acid sequence. CONCLUSIONS: While the basis for the altered distribution of TPI remains to be established, the increased amounts of the protein in urine or bladder tissue may be indicative of squamous metaplasia, squamous cell carcinoma, or other bladder injuries.