The urothelium is a crucial stratified epithelial barrier that protects the urinary tract. It consists of basal cells in the lower layers and intermediate and superficial cells in the luminal layer. These urothelial cells can be identified by their distinct gene expression patterns. Superficial cells are terminally differentiated, binucleated, post-mitotic cells that are responsible for the barrier function of the urothelium via the production of uroplakin proteins. Intermediate cells act as the progenitor cells for superficial cells during development, homeostasis, and after acute injury. Basal cells consist of two populations, K14-basal cells and K5-basal cells. K14- basal cells have been shown to be progenitors that can repopulate the urothelium after chronic injury and are the cells of origin that produce bladder cancer. Bladder cancer can be classified as basal subtype or luminal subtype. The basal subtype is generally immune infiltrated, aggressive, and invasive with a poor prognosis. The luminal subtype is generally immune poor, less aggressive, and non-invasive with a better prognosis compared to basal tumors. Pparg is a nuclear hormone receptor that has been described as a master regulator of adipogenesis and cellular differentiation that also carries out important anti-inflammatory functions (in part by antagonizing the NFKB pathway). Pparg is downregulated in basal subtype muscle invasive bladder cancer and amplified in luminal subtype bladder cancer. In vivo we find that Pparg is a master regulator of cell specification during urothelial development, homeostasis, regeneration, and cancer. When Pparg is ablated in the entire urothelium, Pparg KO mutants lack mature superficial cells and undergo squamous differentiation, with an expansion of the K14-basal cell population. These Pparg KO mutants also display persistent inflammation and squamous metaplasia after injury by urinary tract infection (UTI), due to unregulated NFKB signaling. However, the squamous differentiation in the Pparg KO mutants did not progress to bladder cancer. Constitutive activation of Pparg in basal cells using a novel VP16;Pparg transgenic mouse line crossed to an Krt5CreERT2 driver induces basal cells to undergo a luminal differentiation program towards post-mitotic S-cells during homeostasis. Not surprisingly, these cells did not progress to form bladder cancer on their own. Interestingly, expression of VP16;Pparg in basal cells only drives tumor formation when the basal cells are in an “activated state,” induced by 1 month of BBN treatment. In a BBN mouse model which produces basal subtype bladder cancer in wild type animals, expression of the VP16;Pparg transgene in activated basal cells drives the formation of luminal tumors with papillary morphology, suggesting that this transcription factor is a master regulator of urothelial luminal differentiation, as has been suggested from previous in vitro studies. Like their human counterparts, these VP16;Pparg luminal tumors are immune cold. Additionally, these VP16;Pparg luminal tumors have different domains; a top domain that is “luminal,” and a bottom domain that is “basal”, suggesting the luminal tumors produced by activation of Pparg are not homogenous and undergo a phenotypic shift that mimics what has previously been reported in patient-derived organoids. Understanding the molecular mechanism that drives luminal bladder cancer provides critical information in bettering our approach in diagnosing and treating MIBCs.

Orientador: Luis Cuadrado Martin / Coorientador: Vanessa dos Santos Silva / Banca: Adriana Polachino do Vale / Banca: Maria Almerinda Vieira Fernandes / Resumo: Há discordância na literatura quanto à necessidade da realização da microscopia de fase para avaliação da origem da hematúria (glomerular ou não glomerular), isso mostra a necessidade de mais estudos para validar as modalidades de avaliação morfológica da hematúria. Os objetivos deste trabalho foram determinar o melhor ponto de corte da porcentagem dos diferentes padrões de células dismórficas na detecção da hematúria glomerular pelo microscópio óptico convencional e contraste de fase, verificar se a presença de proteinúria ou albuminúria pode auxiliar no diagnóstico da origem da hematúria e determinar o melhor ponto de corte para este parâmetro. Foram avaliadas de maneira cega 131 amostras de urina sendo 66 amostras de portadores de glomerulopatias e 65 amostras dos portadores de litíase renal da Faculdade de Medicina de Botucatu. Utilizaram-se amostras isoladas com densidade >1007 e com mais de 5 hemácias por campo de grande aumento. Verificou-se a presença e a porcentagem de codócitos e acantócitos tanto em microscópio óptico convencional com sedimento fresco e fixado submetidos à coloração de Papanicolaou e Panótico rápido LB e sedimento urinário fresco avaliado em microscópio de contraste de fase, além de determinar o índice de proteinúria e o índice de albuminúria. Os resultados desses métodos foram comparados entre si. Realizou-se regressão linear e diagrama de Bland-Altman das hemácias dismórficas para comparar os diferentes métodos. Curvas ROC foram traçadas para determinar a área sob a curva (ASC) e o melhor ponto de corte (PC) foi determinado pela maior soma de sensibilidade e especificidade. Nesse ponto foram calculadas a sensibilidade (S) e especificidade (Es), Valor Preditivo Positivo (VPP) e Valor Preditivo Negativo (VPN). No microscópio óptico convencional com sedimento urinário fresco, a ASC do dismorfismo eritrocitário total foi a que apresentou ... / Abstract: There is disagreement over the literature regarding the performance of phase microscopy to assess the origin f hematuria (glomerular or nonglomerular). This shows the need for further investigation in order to validate the best form of morphological evaluation of hematuria. The aims of this study were to determine the optimal cutoff point percentage for different patterns of dysmorphic cells in the detection of glomerular hematuria by conventional optical microscopy and phase contrast microscopy, to verify whether the presence of proteinuria or albuminuria may assist in the diagnosis of hematuria and also to establish an optimal cutoff point for this parameter. One hundred thirty-one urine samples were blinded evaluated at the Faculdade de Medicina de Botucatu, 66 samples of patients with glomerulopathies and 65 samples of patients with nephrolithiasis. Isolated samples with density greater than 1007 and with more than 5 erythrocytes per high-power field were used. The presence and percentage of codocytes and acanthocytes were verified by conventional optical microscopy using fresh and fixed urinary sediment subjected to Papanicolaou and Panótico Rápido LB staining and fresh urinary sediment was evaluated by phase contrast microscopy. Proteinuria and albuminuria rates were determined. The results of these methods were compared using linear regression analysis and Bland-Altman diagram of dysmorphic red blood cells. ROC curve plots were generated to determine the area under the ROC curve (AUC) and also an optimal cutoff point with the highest sum of sensitivity and specificity. At this point, it was possible to calculate sensitivity (TPR) and specificity (ES), positive predictive value (PPV) and negative predictive value (NPV). In conventional optical microscope with fresh urinary sediment, the AUC of total dysmorphic erythrocytes showed the best result for the diagnosis of hematuria with AUC (IC 95%) ... / Mestre

The purposes of this research are (1) to bridge this gap of deficiency by developing a simple, precise and reproducible routine analytical technique by using attenuated total reflectance, and (2) to serve as an atlas for identification of renal calculi.

by Lo, Kin Yin Anthony. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 168-195). / Abstracts in English and Chinese. / Abstract --- p.i / Declaration --- p.V / Publications and Conference Presentations --- p.vi / Scholarships and Awards --- p.vii / Acknowledgements --- p.viii / Table of contents --- p.X / Abbreviations --- p.xiv / List of Figures --- p.xvi / List of Tables --- p.xvii / Chapter 1. --- General Introduction --- p.1 / Chapter 2. --- Literature Review --- p.7 / Chapter 2.1 --- Introduction of nephrolithasis and surgical management --- p.9 / Chapter 2.1.1 --- Epidemology and physiochemistry --- p.9 / Chapter 2.1.2 --- Surgical management of nephrolithasis parallel with stone factors --- p.15 / Chapter 2.2 --- Extracorpoeral Shock Wave Lithotripsy in present study --- p.17 / Chapter 2.2.1 --- The 4th generation - Sonolith Vision electroconductive lithotripter --- p.18 / Chapter 2.2.2 --- The role of shock wave delivery rate in treatment outcome and its prediction --- p.23 / Chapter 2.2.3 --- Patient-controlled analgesia during Shock Wave Lithotripsy treatment and its pain management --- p.29 / Chapter 2.2.4 --- Shock wave induced renal injury & the use of urinary biomarker --- p.35 / Chapter 3. --- Materials and Methods --- p.62 / Chapter 3.1 --- Study Design --- p.63 / Chapter 3.2 --- Patient Selection --- p.64 / Chapter 3.3 --- Treatment Protocol --- p.63 / Chapter 3.4 --- Sample size calculation --- p.68 / Chapter 3.5 --- Statistical analysis --- p.68 / Chapter 4. --- The effect of shock wave delivery rate on treatment outcome and its prediction --- p.69 / Chapter 4.1 --- Introduction --- p.70 / Chapter 4.2 --- Materials and Methods --- p.72 / Chapter 4.2.1 --- ESWL treatment protocol --- p.72 / Chapter 4.2.2 --- Outcome Assessment --- p.73 / Chapter 4.2.3 --- Mathematical model development --- p.75 / Chapter 4.2.4 --- Statistical analysis --- p.76 / Chapter 4.3 --- Results --- p.77 / Chapter 4.3.1 --- Baseline characteristics and treatment modalities --- p.78 / Chapter 4.3.2 --- ESWL treatment outcome --- p.79 / Chapter 4.3.3 --- Mathematical model --- p.81 / Chapter 4.4 --- Discussion --- p.82 / Chapter 4.4.1 --- Overall treatment outcome improved by the use of slower rate --- p.82 / Chapter 4.4.2 --- When should we use fast/slow rate? --- p.86 / Chapter 4.4.3 --- Mathematical model to predict ESWL outcome --- p.88 / Chapter 4.5 --- Conclusion --- p.91 / Chapter 5. --- The role of shock wave delivery rate and patient-controlled analgesia in pain --- p.101 / Chapter 5.1 --- Introduction --- p.102 / Chapter 5.2 --- Materials and Methods --- p.104 / Chapter 5.2.1 --- ESWL treatment protocol and PCA settings --- p.104 / Chapter 5.2.2 --- Outcome Assessment --- p.105 / Chapter 5.2.3 --- Statistical analysis --- p.107 / Chapter 5.3 --- Results --- p.108 / Chapter 5.3.1 --- Baseline characteristics and treatment modalities --- p.108 / Chapter 5.3.2 --- Pain experience and satisfaction with PCA at different shock wave delivery rates --- p.108 / Chapter 5.3.3 --- Correlation between rate pain --- p.110 / Chapter 5.3.4 --- Vital signs --- p.110 / Chapter 5.4 --- Discussion --- p.111 / Chapter 5.4.1 --- Adverse complication was mild with PCA using alfentanil --- p.111 / Chapter 5.4.2 --- Less pain experience with 60 SWs/min --- p.112 / Chapter 5.4.3 --- Why PCA usage was the same in both groups? --- p.112 / Chapter 5.4.4 --- No correlation with treatment outcome --- p.114 / Chapter 5.5 --- Conclusion --- p.115 / Chapter 6. --- "The relations among rate of shock wave delivery, induced renal injury and acute complications" --- p.128 / Chapter 6.1 --- Introduction --- p.129 / Chapter 6.2 --- Materials and Methods --- p.130 / Chapter 6.2.1 --- ESWL treatment protocol --- p.130 / Chapter 6.2.2 --- Outcome Assessment --- p.131 / Chapter 6.2.3 --- Statistical analysis --- p.136 / Chapter 6.3 --- Results --- p.137 / Chapter 6.3.1 --- Baseline characteristics and treatment modalities --- p.137 / Chapter 6.3.2 --- Quality control of creatinine and NAG --- p.137 / Chapter 6.3.3 --- Standard curves ofIL-18 and NGAL --- p.137 / Chapter 6.3.4 --- Higher levels of urinary NAG and IL-18 in 60 SWs/min group --- p.138 / Chapter 6.3.5 --- Similar levels of urinary NGAL in both groups --- p.138 / Chapter 6.3.6 --- Unplanned hospital visits were similar in both groups --- p.139 / Chapter 6.4 --- Discussion --- p.140 / Chapter 6.4.1 --- More tubular damages caused by slower rate --- p.140 / Chapter 6.4.2 --- Escalated inflammatory activities in 60 SWs/min --- p.141 / Chapter 6.4.3 --- Vascular damage and ischemic insults were the same in both groups? --- p.142 / Chapter 6.4.4 --- Post-operative complications are similar in both groups --- p.142 / Chapter 6.4.5 --- 60 SWs/min vs. 120 SWs/min - What makes the difference in renal injury? --- p.143 / Chapter 6.5 --- Conclusion --- p.145 / Chapter 7. --- Discussion --- p.154 / Chapter 7.1 --- General discussion --- p.155 / Chapter 8. --- Conclusion --- p.158 / Chapter 8.1 --- General conclusion --- p.159 / Appendix --- p.160 / Appendix I --- p.161 / Appendix II --- p.163 / References --- p.167

As infecções do trato urinário (ITU) constituem uma das doenças mais comuns na prática médica geral, respondendo por grande parte dos processos infecciosos comunitários e hospitalares, ocorrendo em indivíduos de todas as faixas etárias, sendo as mulheres normalmente mais acometidas. No presente estudo foram analisados 295 isolados de Escherichia coli, provenientes de quatro grupos de mulheres, assim divididos: Grupo colonização; Grupo com ITU recorrente; Grupo com ITU comunitária e Grupo com ITU de internação hospitalar. Estes isolados foram analisados quanto aos fatores de virulência (produção de hemolisina e aerobactina, ligação do vermelho congo, pili 1 e P e mobilidade), a suscetibilidade a oito antimicrobianos e pela tipagem molecular (PFGE) para procurar estabelecer uma associação entre a E. coli de origem intestinal com a da ITU. Os resultados demonstraram que a expressão dos fatores de virulência dos isolados de E. coli das ITU e os de origem intestinal do grupo Colonização foi extremamente variada e a maioria dos perfis fenotípicos apresentou pelo menos duas características de urovirulência. Foi também verificada uma maior resistência aos antimicrobianos nos grupos de maior exposição a essas drogas, em ordem decrescente (Grupos com ITU de internação hospitalar; ITU recorrente e ITU comunitária). A comparação através da PFGE demonstrou uma concordância clonal entre os isolados de E. coli em nível uretral/periuretral e intestinal de 40,0% (6/15) dos casos do Grupo colonização e de 35,7% (5/14) dos pacientes do Grupo com ITU recorrente (urina e fezes). Comparando os resultados obtidos com a associação dos perfis fenotípicos (fatores de virulência) e de suscetibilidade com os genotípicos, também se obteve 40,0% de coincidência no Grupo colonização e de 42,9% (6/14) para o Grupo com ITU recorrente comparado com 35,7% obtido pela genotipagem. / Submitted by Marcelo Teixeira (mvteixeira@ucs.br) on 2014-06-05T16:45:00Z No. of bitstreams: 1 Dissertacao Fernando J Schreiner.pdf: 2493878 bytes, checksum: af52c8fa3356910bd52ceefbe045663e (MD5) / Made available in DSpace on 2014-06-05T16:45:00Z (GMT). No. of bitstreams: 1 Dissertacao Fernando J Schreiner.pdf: 2493878 bytes, checksum: af52c8fa3356910bd52ceefbe045663e (MD5) / The infections of the urinart tract (UTI) are one of the most common diseases in the general medicinal practice. They are responsables for an important number of the community and hospitalar infections affecting individuals of all ages, but with a higher frequency in women. In the present study 295 isolates of Escherichia coli were analysed. These isolates were obtained from four clinical groups of women: (1) Colonization group; (2) Recorrent UTI group; (3) Community UTI group; and (4) UTI in hospitalized group. In order to stablish a relation between UTI and intestinal E. coli the isolates were analysed for the presence of several putative virulence factors (hemolytic activity, aerobactin production, congo red absortion, motility, and the presence of pili 1 and pili P), the susceptibility against a panel of eight antibiotics, and further characterized by the comparison of their pulse field gel electrophoretic (PFGE) profiles. The results showed that the expression of virulence factor among UTI and intestinal isolates of the colonization group was variable, and most isolates exhibited at least two virulence factors. The antibiotic resistance was higher in the groups with exposed to these drugs, in decreasing order: UTI in hospitalized group, recorrent UTI group, and community UTI group. The comparison of PFGE profiles allowed to confirm the clonal origin of E. coli isolates obtained from urinary tract and intestinal samples in 40% (6/15) of the colonization group patients, and 35,7% (5/14) of recorrent UTI group patients. High coincidence was observed between the phenotypic (virulence factors and antibiotic susceptibility) and the genotypic (PFGE) characterization of clonal isolates.

Orientador: João Luiz Amaro / Banca: Hamilto Akihissa Yamamoto / Banca: Victor Augusto Sanguinetti Scherrer Leitão / Banca: Claudia Rucco Penteado Detregiachi / Banca: Aguinaldo Cesar Nardi / Resumo: A itíase urinária é a terceira causa mais comum de afeção do trato urinário. A orientação dietética e o tratamento medicamentoso específico são importantes na recorrência da litíase e, portanto, mudar a dieta e seguir o tratamento farmacológico específico pode prevenir a sua recorrência. Avaliar a influência da orientação nutricional e do tratamento medicamentoso na recorrência da litíase urinária. A partir do nosso registro de pacientes com litíase recorrente foram selecionados 57 que tiveram pelo menos 5 anos de seguimento. Intervenção: Durante o acompanhamento dos pacientes foram avaliados os seguintes parâmetros: questionário clínico, investigação metabólica e avaliação por imagem (ultra-sonografia e/ou raio-X simples de abdome). Todos os pacientes foram orientados a controlar a ingestão de proteína (entre 0,8 a 1g/Kg peso corporal/dia) e adequar a ingestão de cálcio (entre 800 a 1000 mg/dia), levando-se em consideração o registro alimentar de 3 dias. A restrição de sal (menor que 5 g/dia) foi realizada tendo como referência a variação do sódio excretado na urina de 24h. Durante o acompanhamento os pacientes receberam ainda orientações dietéticas e farmacológicas específicas de acordo com o distúrbio metabólico identificado. 56% dos pacientes eram sexo masculino e a média do IMC no pré-tratamento foi de 27,8 kg/m², observando-se uma correlação positiva entre o número médio de cálculos formados por ano no pré-tratamento e IMC (p=0,012) Notamos uma diminuição significativa do cálcio, sódio e ácido úrico na urina de 24 horas no pós tratamento em comparação ao período inicial. Observamos um aumento significativo do citrato na urina de 24 horas no pós-tratamento. O nº de cálculos formados durante seguimento de 5 anos, diminuiu significativamente em relação ao pré tratamento... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Urinary tract lithiasis is the third most common cause of urinary tract affection. Dietary factors have great importance in the formation of urolithiasis; therefore, changing the diet and specific pharmacological treatment can prevent its recurrence. Evaluate the influence of clinical therapy associated to nutritional orientation in the recurrence of urolithiasis. From our registry of patients with recurrent lithiasis we selected 57 who had at least 5-years of follow-up. We used the protocol composed by 2 non-consecutive urine samples of 24h and was performed: Ca, Na, uric acid, citrate, oxalate, Mg and urinary volume assessments. In the pre treatment the lithiasis clinical questionary and after five years was care out using abdomen x-ray and/or ultrasound. Individualized dietary orientation consisted of: to increase fluid intake (enough to form ≥ 2 L / day); sodium ingestion <5 g / day and protein intake range between 0.8 - 1 g / kg body weight / day, and the adequate calcium intake (800 to 1000 mg / day). During the follow-up of patients, specific and individualized dietary orientation was performed according to the diagnosed metabolic disorder. Patients received specific pharmacological treatment according to the metabolic alteration. 54% were male. In average the BMI was 27 kg / m2. It was observed that according to BMI classification, the patients were overweight. Urinary excretion of calcium, uric acid and sodium decreased significantly after 5 years of follow-up. The number of stones formed in the 5- year follow-up decreased significantly compare to pre treatment. Individualized dietary orientation and pharmacological... (Complete abstract click electronic access below) / Doutor

As infecções do trato urinário (ITU) constituem uma das doenças mais comuns na prática médica geral, respondendo por grande parte dos processos infecciosos comunitários e hospitalares, ocorrendo em indivíduos de todas as faixas etárias, sendo as mulheres normalmente mais acometidas. No presente estudo foram analisados 295 isolados de Escherichia coli, provenientes de quatro grupos de mulheres, assim divididos: Grupo colonização; Grupo com ITU recorrente; Grupo com ITU comunitária e Grupo com ITU de internação hospitalar. Estes isolados foram analisados quanto aos fatores de virulência (produção de hemolisina e aerobactina, ligação do vermelho congo, pili 1 e P e mobilidade), a suscetibilidade a oito antimicrobianos e pela tipagem molecular (PFGE) para procurar estabelecer uma associação entre a E. coli de origem intestinal com a da ITU. Os resultados demonstraram que a expressão dos fatores de virulência dos isolados de E. coli das ITU e os de origem intestinal do grupo Colonização foi extremamente variada e a maioria dos perfis fenotípicos apresentou pelo menos duas características de urovirulência. Foi também verificada uma maior resistência aos antimicrobianos nos grupos de maior exposição a essas drogas, em ordem decrescente (Grupos com ITU de internação hospitalar; ITU recorrente e ITU comunitária). A comparação através da PFGE demonstrou uma concordância clonal entre os isolados de E. coli em nível uretral/periuretral e intestinal de 40,0% (6/15) dos casos do Grupo colonização e de 35,7% (5/14) dos pacientes do Grupo com ITU recorrente (urina e fezes). Comparando os resultados obtidos com a associação dos perfis fenotípicos (fatores de virulência) e de suscetibilidade com os genotípicos, também se obteve 40,0% de coincidência no Grupo colonização e de 42,9% (6/14) para o Grupo com ITU recorrente comparado com 35,7% obtido pela genotipagem. / The infections of the urinart tract (UTI) are one of the most common diseases in the general medicinal practice. They are responsables for an important number of the community and hospitalar infections affecting individuals of all ages, but with a higher frequency in women. In the present study 295 isolates of Escherichia coli were analysed. These isolates were obtained from four clinical groups of women: (1) Colonization group; (2) Recorrent UTI group; (3) Community UTI group; and (4) UTI in hospitalized group. In order to stablish a relation between UTI and intestinal E. coli the isolates were analysed for the presence of several putative virulence factors (hemolytic activity, aerobactin production, congo red absortion, motility, and the presence of pili 1 and pili P), the susceptibility against a panel of eight antibiotics, and further characterized by the comparison of their pulse field gel electrophoretic (PFGE) profiles. The results showed that the expression of virulence factor among UTI and intestinal isolates of the colonization group was variable, and most isolates exhibited at least two virulence factors. The antibiotic resistance was higher in the groups with exposed to these drugs, in decreasing order: UTI in hospitalized group, recorrent UTI group, and community UTI group. The comparison of PFGE profiles allowed to confirm the clonal origin of E. coli isolates obtained from urinary tract and intestinal samples in 40% (6/15) of the colonization group patients, and 35,7% (5/14) of recorrent UTI group patients. High coincidence was observed between the phenotypic (virulence factors and antibiotic susceptibility) and the genotypic (PFGE) characterization of clonal isolates.

Orientador: Edson Antunes / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-17T00:06:55Z (GMT). No. of bitstreams: 1 RamosFilho_AntonioCelsoS._M.pdf: 1693116 bytes, checksum: 4526088a65a406983163ece45558e71c (MD5) Previous issue date: 2010 / Resumo: A hipertensão renovascular é uma forma secundária da hipertensão arterial, que corresponde de 1-5% dos casos de hipertensão. A associação entre hipertensão arterial e disfunções miccionais foi observada no modelo experimental de ratos espontaneamente hipertensos (SHR). Até o momento nenhum estudo avaliou as disfunções miccionais em animais hipertensos renovasculares. Dessa forma, neste estudo, caracterizamos a disfunção miccional em ratos hipertensos renovasculares através do modelo de dois rins, um clip (2K-1C). Em ratos Wistar (200-220 g) colocou-se um clip em torno da artéria renal. Depois de oito semanas, os ratos foram utilizados. Realizou-se estudo cistométrico em ratos anestesiados, assim como curvas concentração-resposta para agentes contráteis e relaxantes em detrusor isolado (DSM). Foram também realizados estudos histomorfométricos e da expressão de RNAm dos receptores muscarínicos M3 e M2 em DSM isolado. Os resultados histomorfométricos mostraram aumentos significantes na espessura da parede da bexiga, no volume intravesical, na densidade de musculatura lisa e na densidade de fibras neurais no grupo 2K-1C em comparação ao SHAM. O agonista muscarínico, carbacol, produziu contrações concentração-dependentes do DSM, as quais foram significantemente maiores no grupo 2K-1C. O inibidor da Rho-quinase, Y27-632 (10 µM), reduziu significantemente a contração induzida pelo carbacol nos ratos SHAM e 2K-1C; porém, no grupo 2K-1C, o DSM continuou hiperativo na presença do Y27-632. A estimulação elétrica (1 - 32 Hz) produziu contração freqüência-dependente do DSM as quais foram maiores no grupo 2K-1C. O agonista purinérgico P2X, ?,?-metileno-ATP (1 - 100 µM), o KCl (1 - 300 µM) e o Ca2+ extracelular (0,01-100 µM) produziram contrações concentração-dependente; porém, não observamos diferenças entre o grupo SHAM e 2K-1C. O agonista não seletivo ?-adrenérgico, isoproterenol, o agonista seletivo ?2-adrenérgico, metaproterenol, e o agonista seletivo ?3-adrenérgico, BRL37-344, produziram relaxamentos menores do DSM nos ratos 2K-1C, e também redução nos níveis intracelulares de AMPc nos detrusores. O efeito relaxante ao nitroprussiato de sódio e BAY41-2272 mantiveram-se iguais nos animais SHAM e 2K-1C. A expressão do RNAm do receptor muscarínico M3 (mas não do M2) no DSM foi significantemente maior nos ratos 2K-1C em comparação ao grupo controle. Os tratamentos crônicos com losartan e captopril normalizaram a pressão arterial sistólica dos animais 2K-1C, normalizaram a função miccional, e reduziram a hipercontratilidade do detrusor induzida pela estimulação elétrica e pelo carbacol, assim como restabeleceram o relaxamento induzido pelo isoproterenol ao nível do grupo SHAM. Concluimos que os ratos hipertensos renovasculares apresentam hiperatividade do detrusor, a qual envolve remodelamento tecidual e aumento da contração via receptor muscarínico M3 associado à redução no relaxamento ?-adrenérgico com redução da sinalização intracelular e produção de AMPc. Os tratamentos com losartan e captopril restauram a função miccional dos animais 2K-1C / Abstract: Renovascular hypertension is a secondary form of arterial hypertension, accounting for 1-5% of cases in unselected population. Association between arterial hypertension and urinary bladder dysfunction has been reported in spontaneously hypertensive rats, but no study evaluated the bladder dysfunction in renovascular hypertensive animals. Therefore, in this study, we explored the bladder dysfunction in renovascular hypertensive rats, using the two-kidney one-clip (2K-1C) model. A silver clip was placed around the renal artery of male Wistar Kyoto rats (200-220 g). After eight weeks, rats were used. Cystometric study in anesthetized rats, along with concentration-response curves to both contractile and relaxant agents in isolated detrusor smooth muscle (DSM) were performed. Histomorphometry and mRNA expression of muscarinic M3 and M2 receptors in DSM were also determined. The histomorphometric data showed significant increases in bladder wall thickness, intravesical volume and density of smooth muscle, as well as density of neural fibers in the 2K-1C group compared with SHAM. The muscarinic agonist carbachol produced concentration-dependent DSM contractions, which were markedly greater in 2K-1C rats. The Rho-kinase inhibitor Y27-632 (10 µM) significantly reduced the carbachol-induced contractions in sham and 2K-1C rats, but DSM in 2K-1C rats remained overactive in the presence of Y27632. Electrical-field stimulation (EFS; 1-32 Hz) produced frequency-dependent DSM contractions that were also greater in 2K-1C group. The P2X receptor agonist ?,?-methylene ATP (1-100 µM), KCl (1-300 mM) and extracellular Ca2+ (0.01-100 M) produced concentration-dependent DSM contractions, but no changes among sham and 2K-1C rats were seen. In 2K-1C rats, the non-selective ?-adrenoceptor agonist isoproterenol, the ?2-adrenoceptor agonist metaproterenol and the ?3-adrenoceptor agonist BRL 37-344 produced lower DSM relaxations, as well as decreased cAMP levels. The relaxant responses to sodium nitroprusside and BAY 41-2272 remained unchanged in 2K-1C rats. Expression of mRNA of muscarinic M3 (but not of M2) receptors in DSM was significantly increased in 2K-1C rats. The chronic treatment with losartan and captopril normalized the blood systolic pressure of 2K-1C animals, improved their urinary function by reducing DSM hypercontractility to EFS and carbacol stimulation, and restored the relaxation induced by the ?-adrenergic agonist isoproterenol to the level of SHAM group. In conclusion, renovascular hypertensive rats exhibit overactive DSM that involves tissue remodeling and enhanced muscarinic M3-mediated contractions associated with reduced ?-adrenoceptor-mediated signal transduction. The treatments with losartan and captopril improved urinary function of 2K-1C animals / Mestrado / Mestre em Farmacologia

Doctorat en Sciences médicales / info:eu-repo/semantics/nonPublished