RESUMO
Hemorrhagic cystitis (HC) is the major dose-limiting adverse effect of the clinical use ifosfamide (IFOS). The incidence of this side effect can be as high as 75%. Mesna has been used to reduce the risk of HC, although 5% of patients who get IFOS treatment may still suffer from HC. In previous studies, our group demonstrated that α-phellandrene (α-PHE) possesses anti-inflammatory activity, which opens the door for its study in the attenuation of HC. The objective of this study was to investigate the potential uroprotective effect of the α-PHE in the mouse model of IFOS-induced HC. In order to analyze the reduction of the urothelial damage, the bladder wet weight, hemoglobin content, and the Evans blue dye extravasation from the bladder matrix were evaluated. To investigate the involvement of neutrophil migration and lipid peroxidation and involvement of enzymatic and endogenous non-enzymatic antioxidants, the tissue markers myeloperoxidase (MPO), malondialdehyde, nitrite/nitrate (NOx), superoxide dismutase (SOD), and reduced glutathione (GSH) were evaluated. TNF-α and IL-1ß were measured by ELISA immunoassay technique. The results show that pretreatment with α-PHE significantly reduced urothelial damage that was accompanied by a decrease in the activity of MPO, MDA, and NOx levels and prevention of the depletion of SOD and GSH in bladder tissues. In the assessment of cytokines, α-PHE was able to significantly reduce TNF-α level. However, it does not affect the activities of IL-1ß. These data confirm that α-PHE exerts potent anti-inflammatory properties and demonstrates that α-PHE represents a promising therapeutic option for this pathological condition.
Assuntos
Monoterpenos Cicloexânicos/uso terapêutico , Cistite/prevenção & controle , Hemorragia/prevenção & controle , Ifosfamida/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Antineoplásicos Alquilantes/toxicidade , Monoterpenos Cicloexânicos/farmacologia , Cistite/induzido quimicamente , Cistite/metabolismo , Relação Dose-Resposta a Droga , Hemorragia/induzido quimicamente , Hemorragia/metabolismo , Masculino , Camundongos , Estresse Oxidativo/fisiologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Hemorrhagic cystitis (HC) is a common side effect of cyclophosphamide therapy, which deserves new therapeutic strategies, such as those based on natural products. The ethanol extract of the inner bark of Caesalpinia pyramidalis (Tul.) (EECp) possesses anti-inflammatory, antinociceptive, and antioxidant activities as previously showed by our group. We have investigated the effect of EECp on the cyclophosphamide-induced HC. Cystitis was induced in male Wistar rats by the injection of cyclophosphamide. These animals were pretreated with EECp (100-400 mg/kg), vehicle, or mesna. Myeloperoxidase activity and malondialdehyde formation were measured in urinary bladder and other tissues. Bladder edema and histopathological alterations and serum nitric oxide metabolites concentration NOx- were also evaluated. Treatment with EECp (100-400 mg/kg) or mesna impaired the increase of myeloperoxidase activity in urinary bladder and the serum NOx- induced by cyclophosphamide but did not reduce edema in this tissue, as did mesna. Total histological score was reduced by EECp (100 mg/kg). Lung myeloperoxidase activity, which was increased by cyclophosphamide, was decreased significantly by EECp (400 mg/kg). EECp also diminished the malondialdehyde formation in bladder, lung, and spleen, although these parameters were not affected by cyclophosphamide. These results indicate that EECp reduced urinary bladder damage during cyclophosphamide-induced HC in rats.
Assuntos
Anti-Inflamatórios/farmacologia , Caesalpinia/química , Cistite/patologia , Casca de Planta/química , Extratos Vegetais/farmacologia , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/patologia , Animais , Anti-Inflamatórios/administração & dosagem , Ciclofosfamida , Cistite/induzido quimicamente , Cistite/tratamento farmacológico , Cistite/metabolismo , Modelos Animais de Doenças , Ativação Enzimática/efeitos dos fármacos , Hemorragia/induzido quimicamente , Hemorragia/tratamento farmacológico , Hemorragia/patologia , Contagem de Leucócitos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Óxido Nítrico/metabolismo , Peroxidase/metabolismo , Extratos Vegetais/administração & dosagem , Ratos , Bexiga Urinária/metabolismoRESUMO
OBJECTIVE: To determine the concentration of nitrate/nitrite in the cerebrospinal fluid and in the dorsal horn interstice of the L6-S1 spinal cord boundary in rats with or without cystitis induced by cyclophosphamide. METHODS: All experiments were conducted using Wistar female rats. A microdialysis probe was implanted in the subarachnoid space or in the spinal cord tissue at the L6-S1 segments (confirmed histologically). Two days later, the microdialysis probe was perfused with artificial cerebrospinal fluid, containing or not NG-monomethyl-L-arginine. Samples were collected every 15 minutes and kept at -20ºC. Nitrite/nitrate concentrations were determined by chemiluminescence. RESULTS: In normal animals, the mean values of nitrite/nitrate concentrations in the first microdialysate sample of the cerebrospinal fluid and of the spinal cord interstice were similar (482.5±90.2pmol/75µL, n=20, and 505.7±11.5pmol/75µL, n=6, respectively), whereas, in the samples from rats with cystitis, these values were significantly greater (955.5±66.3pmol/75µL, n=8, and 926.5±131.7pmol/75µL, n=11, respectively). In both groups, NG-monomethyl-L- arginine caused a significant reduction in the nitrite/nitrate concentration. Interestingly, the maximal reduction of nitrite/nitrate concentration caused by NG-monomethyl-L- arginine was no greater than 30% of the initial values. CONCLUSIONS: These results constitute the first demonstration that nitrite/nitrate concentrations in the cerebrospinal fluid and spinal cord interstice are elevated between 20- and 22 hours after cyclophosphamide-induced cystitis, and indicate that cystitis is associated with changes in the production of nitric oxide in the spinal cord segments, where most primary bladder afferents end.
Assuntos
Cistite/induzido quimicamente , Óxido Nítrico/metabolismo , Medula Espinal/química , Animais , Ciclofosfamida , Cistite/metabolismo , Cistite/patologia , Feminino , Região Lombossacral , Luminescência , Microdiálise , Nitratos/líquido cefalorraquidiano , Nitratos/metabolismo , Óxido Nítrico/líquido cefalorraquidiano , Óxido Nítrico Sintase/antagonistas & inibidores , Nitritos/líquido cefalorraquidiano , Nitritos/metabolismo , Ratos , Ratos Wistar , Valores de Referência , Medula Espinal/metabolismo , Fatores de Tempo , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia , ômega-N-Metilarginina/farmacologiaRESUMO
The aim of this study was to investigate the involvement of the transient receptor potential ankyrin 1 (TRPA1) in haemorrhagic cystitis, the main side effect of cyclophosphamide-based chemotherapy. Hannover female rats received intraperitoneal (i.p.) injection of cyclophosphamide (three doses of 100 mg/kg, every other day, in a total of five days). This treatment was followed by the treatment with TRPA1 antagonist HC 030031 (50 mg/kg, p.o.). The threshold for hindpaw withdrawal or abdominal retraction to von Frey Hair and the locomotor activity were measured. The treatment with the TRPA1 antagonist HC 030031 significantly decreased mechanical hyperalgesia induced by cyclophosphamide without interfere with locomotor activity. Urodynamic parameters were performed by cystometry 24 h after a single treatment with cyclophosphamide (200 mg/kg, i.p.) in control and HC 030031 treated rats. Analyses of the urodynamic parameters showed that a single dose of cyclophosphamide was enough to significantly increase the number and amplitude of non-voiding contractions and to decrease the voided volume and voiding efficiency, without significantly altering basal, threshold or maximum pressure. The treatment with HC 030031 either before (100 mg/kg, p.o.) or after (30 mg/kg, i.v.) cyclophosphamide inhibited the non-voiding contractions but failed to counteract the loss in voiding efficiency. Our data demonstrates that nociceptive symptoms and urinary bladder overactivity caused by cyclophosphamide, in part, are dependent upon the activation of TRPA1. In this context, the antagonism of the receptor may be an alternative to minimise the urotoxic symptoms caused by this chemotherapeutic agent.
Assuntos
Ciclofosfamida/efeitos adversos , Cistite/induzido quimicamente , Cistite/complicações , Hemorragia/complicações , Hiperalgesia/tratamento farmacológico , Canais de Cátion TRPC/antagonistas & inibidores , Bexiga Urinária Hiperativa/tratamento farmacológico , Acetanilidas/farmacologia , Acetanilidas/uso terapêutico , Animais , Antineoplásicos Alquilantes/efeitos adversos , Cistite/metabolismo , Cistite/fisiopatologia , Feminino , Hiperalgesia/complicações , Purinas/farmacologia , Purinas/uso terapêutico , Ratos , Canal de Cátion TRPA1 , Bexiga Urinária Hiperativa/complicações , Urodinâmica/efeitos dos fármacosRESUMO
BACKGROUND AND PURPOSE: ATP is released in response to cellular damage, and P2X7 receptors have an essential role in the onset and maintenance of pathological changes. Haemorrhagic cystitis (HC) is a well-known adverse effect of therapy with cyclophosphamide used for the treatment of many solid tumours and autoimmune conditions. Here we have evaluated the role of P2X7 receptors in a model of HC induced by cyclophosphamide. EXPERIMENTAL APPROACH: Effects of pharmacological antagonism or genetic deletion of P2X7 receptor on cyclophosphamide-induced HC in mice was assessed by nociceptive and inflammatory measures. In addition, the presence of immunoreactive P2X7 receptors was assessed by immunohistochemistry. KEY RESULTS: Pretreatment with the selective P2X7 receptor antagonist A-438079 or genetic ablation of P2X7 receptors reduced nociceptive behaviour scores in the HC model. The same strategies decreased both oedema and haemorrhage indices, on macroscopic or histological evaluation. Treatment with A-438079 decreased the staining for c-Fos in the lumbar spinal cord and brain cortical areas. Treatment with A-438079 also prevented the increase of urinary bladder myeloperoxidase activity and macrophage migration induced by cyclophosphamide and reduced the tissue levels of IL-1ß and TNF-α. Finally, P2X7 receptors were markedly up-regulated in the bladders of mice with cyclophosphamide-induced HC. CONCLUSIONS AND IMPLICATIONS: P2X7 receptors were significantly involved in a model of HC induced by cyclophosphamide. Pharmacological inhibition of these receptors might represent a new therapeutic option for this pathological condition.
Assuntos
Ciclofosfamida/toxicidade , Cistite/induzido quimicamente , Hemorragia/induzido quimicamente , Inflamação/metabolismo , Nociceptividade/fisiologia , Receptores Purinérgicos P2X7/metabolismo , Animais , Movimento Celular , Cistite/metabolismo , Cistite/patologia , Citocinas/genética , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica , Genes fos , Hemorragia/metabolismo , Macrófagos/fisiologia , Masculino , Mesna/farmacologia , Camundongos , Camundongos Knockout , Antagonistas do Receptor Purinérgico P2X/farmacologia , Piridinas/administração & dosagem , Piridinas/farmacologia , Receptores Purinérgicos P2X7/genética , Tetrazóis/administração & dosagem , Tetrazóis/farmacologia , Bexiga Urinária/metabolismoRESUMO
PURPOSE: The underlying pathology of radiation cystitis is cellular and vascular damage followed by increased fibrosis and inflammation. This study was to determine if neovascular-promoting therapy could reduce the pathological changes in the bladder wall associated with pelvic irradiation. METHODS: Adult female Lewis inbred rats were irradiated with a single dose of 20 Gy directed at their bladder. Four weeks later, 30 rats were divided equally into one of three treatment groups for bladder wall injection of: (1) PBS (Control); (2) PBS containing 50 ng vascular endothelial growth factor (VEGF (165)); or (3) PBS containing 1 × 10(6) rat endothelial cells (EC). Age-matched non-irradiated rats (n = 10) served as untreated controls. At either 1.5 or 3 months following radiation, bladders were analyzed for collagen deposition using Masson's Trichrome staining of collagen and muscle and vascularization using Von Willebrand factor staining of ECs. Quantitative-PCR was used to examine markers of angiogenesis, hypoxia, and fibrosis. RESULTS: The collagen/muscle ratio was doubled in the control group 3 months post-irradiation (P < 0.05 vs. non-irradiated bladders). Both ECs and VEGF inhibited increases in collagen content (P < 0.05 vs. control). Similarly, irradiation reduced bladder wall vessel counts compared to non-irradiated controls (P < 0.05) and both ECs and VEGF maintained vessel counts similar to that of non-irradiated controls (P < 0.05). PCR analysis showed a higher expression of neovascular markers (CD31, KDR) in the EC and VEGF groups compared to non-irradiated controls (P < 0.05). CONCLUSIONS: Angiogenesis therapy may be useful in the prevention and/or treatment of the underlying pathology of radiation cystitis.
Assuntos
Indutores da Angiogênese/administração & dosagem , Cistite/terapia , Células Endoteliais/transplante , Neovascularização Fisiológica/efeitos dos fármacos , Lesões Experimentais por Radiação/terapia , Bexiga Urinária/irrigação sanguínea , Fator A de Crescimento do Endotélio Vascular/administração & dosagem , Administração Intravesical , Análise de Variância , Animais , Colágeno/metabolismo , Cistite/genética , Cistite/metabolismo , Cistite/patologia , Cistite/fisiopatologia , Células Endoteliais/metabolismo , Feminino , Fibrose , Regulação da Expressão Gênica , Neovascularização Fisiológica/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Lesões Experimentais por Radiação/genética , Lesões Experimentais por Radiação/metabolismo , Lesões Experimentais por Radiação/patologia , Lesões Experimentais por Radiação/fisiopatologia , Ratos , Ratos Endogâmicos Lew , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Fator de von Willebrand/metabolismoRESUMO
Introdução: Cistite glandular é um processo proliferativo benigno e infrequente da mucosa vesical, caracterizado por proliferação do epitélio e, em alguns casos, formação de glândulas intestinais. Alterações metaplásticas na cistite glandular são bem documentadas na literatura, embora sua etiologia não seja totalmente esclarecida. Relato do caso: Relatamos um caso de cistite glandular em um paciente de 55 anos, apresentando sintomas miccionais irritativos e obstrutivos persistentes sem resposta à terapia com alfabloqueadores. Ultrassonografia evidenciou lesão vegetante no trígono vesical e o paciente foi submetido à ressecção endoscópica por duas vezes e evoluiu com ureterohidronefrose bilateral. Dado o extenso acometimento vesical e a persistência dos sintomas, o paciente foi submetido a cistoprostatectomia e neobexiga ileal com boa evolução pós-operatória. Discussão: Há duas formas de cistite glandular: típica e intestinal. A forma típica é a mais comum e a intestinal é marcada pela produção de mucina, mais frequentemente associada ao adenocarcinoma de bexiga. A maioria dos casos de cistite glandular é assintomática, sendo que os pacientes sitomáticos normalmente apresentam hematúria, sintomas urinários irritativos e típicos de cistite crônica. Há controvérsias sobre o tratamento precoce agressivo, sendo que vários estudos propõem a ressecção transuretral e o acompanhamento com biópsias.
Introduction: glandular cystitis is a benign proliferative process and infrequent mucosal bladder, characterized by proliferation of the epithelium and in some cases, formation of intestinal glands. Metaplásticas changes in glandular cystitis are well documented in the literature, although its etiology is not fully understood. Case report: A case of glandular cystitis in one patient of 55 years, with irritative and obstructive urinary symptoms persisted without response to therapy with alpha blockers. Ultrasonography revealed a vegetative lesion in the trigonal and the patient underwent endoscopic resection and twice progressed to bilateral ureterohidronefrose. Given the extensive bladder involvement and persistence of symptoms, the patient underwent ileal neobladder cistoprostatectomia and with good postoperative evolution. Discussion: There are two types of glandular cystitis: typical and intestinal tract. The typical form is the most common and is characterized by intestinal mucin production, most often associated with adenocarcinoma of the bladder. Most cases of glandular cystitis is asymptomatic, and patients usually present sitomáticos hematuria, urinary symptoms and typical of chronic cystitis. There is controversy over early aggressive treatment, and several studies suggest transurethral resection and follow-up biopsies.
Assuntos
Humanos , Masculino , Adulto , Cistite/cirurgia , Cistite/metabolismo , Cistite/patologia , Cistite/terapia , Obstrução do Colo da Bexiga Urinária/diagnóstico , Obstrução do Colo da Bexiga Urinária/etiologia , Obstrução do Colo da Bexiga Urinária/patologia , Procedimentos Cirúrgicos Urológicos/métodos , Procedimentos Cirúrgicos UrológicosRESUMO
To investigate the role of substance P (sP), nitric oxide (ON) and prostaglandins (PGs) in acrolein (ACR)-induced cystitis, we studied the changes induced by ACR on bladder inducible nitric oxide synthase (iNOS) and mieloperoxidase (MPO) activities, along with PGs and NO metabolites levels. Sprague-Dawley male rats received i.p. ACR (5 mg/Kg) plus one of the following treatments: Group 1: saline 0.10 mL/100g i.p.; Group 2: Win-51.708 (WIN) 25 mg/Kg i.p.; Group 3: S-metilisothiourea (MITU) 35 mg/Kg i.p.; Group 4: Rofecoxib(ROF) 20 mg/Kg o.p.; Group 5: Meloxicam(MEL) 25 mg/Kg i.p.; Group 6: combination MITU+MEL. ACR-induced mortality was partially prevented by WIN (NK1 antagonist) and MITU (iNOS inhibitor). Animals that survived after 24h of ACR exposure, had histological inflammatory changes in bladder along with increased MPO activity. There was augmentation of nitrates+nitrites and of PGs. WIN didn't prevent any of these effects. ROF and MEL (COX-2 inhibitors) partially protected against bladder inflammation; MITU pre-treatment was able to prevent these changes and those of NO metabolites levels. The MITU+MEL combination produced the highest protection against ACR-induced damage. These results suggest that NO produced via iNOS and PGs produced by COX-1/COX-2, have an important role in the pathogenesis of cystitis induced by ACR. ACR could stimulate iNOS and COX-1/COX-2, producing lymphocyte migration and increases of NO and PGs.