RESUMO
This study explores low-intensity extracorporeal shock wave therapy (LiESWT)'s efficacy in alleviating detrusor hyperactivity with impaired contractility (DHIC) induced by ovarian hormone deficiency (OHD) in ovariectomized rats. The rats were categorized into the following four groups: sham group; OVX group, subjected to bilateral ovariectomy (OVX) for 12 months to induce OHD; OVX + SW4 group, underwent OHD for 12 months followed by 4 weeks of weekly LiESWT; and OVX + SW8 group, underwent OHD for 12 months followed by 8 weeks of weekly LiESWT. Cystometrogram studies and voiding behavior tracing were used to identify the symptoms of DHIC. Muscle strip contractility was evaluated through electrical-field, carbachol, ATP, and KCl stimulations. Western blot and immunofluorescence analyses were performed to assess the expressions of various markers related to bladder dysfunction. The OVX rats exhibited significant bladder deterioration and overactivity, alleviated by LiESWT. LiESWT modified transient receptor potential vanilloid (TRPV) channel expression, regulating calcium concentration and enhancing bladder capacity. It also elevated endoplasmic reticulum (ER) stress proteins, influencing ER-related Ca2+ channels and receptors to modulate detrusor muscle contractility. OHD after 12 months led to neuronal degeneration and reduced TRPV1 and TRPV4 channel activation. LiESWT demonstrated potential in enhancing angiogenic remodeling, neurogenesis, and receptor response, ameliorating DHIC via TRPV channels and cellular signaling in the OHD-induced DHIC rat model.
Assuntos
Modelos Animais de Doenças , Tratamento por Ondas de Choque Extracorpóreas , Contração Muscular , Canais de Cátion TRPV , Bexiga Urinária , Animais , Feminino , Ratos , Canais de Cátion TRPV/metabolismo , Canais de Cátion TRPV/genética , Tratamento por Ondas de Choque Extracorpóreas/métodos , Bexiga Urinária/fisiopatologia , Bexiga Urinária/metabolismo , Bexiga Urinária Hiperativa/terapia , Bexiga Urinária Hiperativa/metabolismo , Bexiga Urinária Hiperativa/fisiopatologia , Bexiga Urinária Hiperativa/etiologia , Ovariectomia , Ratos Sprague-Dawley , Ovário/metabolismoRESUMO
Autophagy is a lysosomal degradation process known as autophagic flux, involving the engulfment of damaged proteins and organelles by double-membrane autophagosomes. It comprises microautophagy, chaperone-mediated autophagy (CMA), and macroautophagy. Macroautophagy consists of three stages: induction, autophagosome formation, and autolysosome formation. Atg8-family proteins are valuable for tracking autophagic structures and have been widely utilized for monitoring autophagy. The conversion of LC3 to its lipidated form, LC3-II, served as an indicator of autophagy. Autophagy is implicated in human pathophysiology, such as neurodegeneration, cancer, and immune disorders. Moreover, autophagy impacts urological diseases, such as interstitial cystitis /bladder pain syndrome (IC/BPS), ketamine-induced ulcerative cystitis (KIC), chemotherapy-induced cystitis (CIC), radiation cystitis (RC), erectile dysfunction (ED), bladder outlet obstruction (BOO), prostate cancer, bladder cancer, renal cancer, testicular cancer, and penile cancer. Autophagy plays a dual role in the management of urologic diseases, and the identification of potential biomarkers associated with autophagy is a crucial step towards a deeper understanding of its role in these diseases. Methods for monitoring autophagy include TEM, Western blot, immunofluorescence, flow cytometry, and genetic tools. Autophagosome and autolysosome structures are discerned via TEM. Western blot, immunofluorescence, northern blot, and RT-PCR assess protein/mRNA levels. Luciferase assay tracks flux; GFP-LC3 transgenic mice aid study. Knockdown methods (miRNA and RNAi) offer insights. This article extensively examines autophagy's molecular mechanism, pharmacological regulation, and therapeutic application involvement in urological diseases.
Assuntos
Cistite , Neoplasias Testiculares , Animais , Masculino , Camundongos , Humanos , Neoplasias Testiculares/metabolismo , Autofagia/genética , Autofagossomos/metabolismo , Família da Proteína 8 Relacionada à Autofagia/metabolismo , Camundongos Transgênicos , Cistite/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Lisossomos/metabolismoRESUMO
The present study attempted to elucidate whether intravesical instillation of platelet-rich plasma (PRP) could decrease bladder inflammation and ameliorate bladder hyperactivity in ketamine ulcerative cystitis (KIC) rat model. Female Sprague Dawley (S-D) rats were randomly divided into control group, ketamine-treated group, ketamine with PRP treated group, and ketamine with platelet-poor plasma (PPP) treated group. Cystometry and micturition frequency/volume studies were performed to investigate bladder function. The morphological change of bladder was investigated by Mason's trichrome staining. Western blotting analysis were carried out to examine the protein expressions of inflammation, urothelial differentiation, proliferation, urothelial barrier function, angiogenesis and neurogenesis related proteins. The results revealed that treatment with ketamine significantly deteriorated bladder capacity, decreased voiding function and enhanced bladder overactivity. These pathological damage and interstitial fibrosis may via NF-κB/COX-2 signaling pathways and muscarinic receptor overexpression. PRP treatment decreased inflammatory fibrotic biosynthesis, attenuated oxidative stress, promoted urothelial cell regeneration, and enhanced angiogenesis and neurogenesis, thereafter recovered bladder dysfunction and ameliorate the bladder hyperactivity in KIC rat model. These findings suggested that the PRP therapy may offer new treatment options for those clinical KIC patients.
Assuntos
Cistite , Ketamina , Plasma Rico em Plaquetas , Animais , Cistite/induzido quimicamente , Cistite/terapia , Feminino , Humanos , Ketamina/farmacologia , Plasma Rico em Plaquetas/metabolismo , Agitação Psicomotora , Ratos , Ratos Sprague-Dawley , Bexiga Urinária/patologiaRESUMO
Background and Objectives: To evaluate the effects of low intensity extracorporeal shock wave therapy (LiESWT) on stress urinary incontinence (SUI). Materials and Methods: This investigation was a multicenter, single-blind, randomized-controlled trial study. Sixty female SUI patients were randomly assigned to receive LiESWT with 0.25 mJ/mm2 intensity, 3000 pulses, and 3 pulses/s, once weekly for a 4-week (W4) and 8-week (W8) period, or an identical sham LiESWT treatment without energy transmission. The primary endpoint was the changes in urine leakage as measured by a pad test and validated standardized questionnaires, while the secondary endpoint was the changes in a 3-day urinary diary among the baseline (W0), the W4 and W8 of LiESWT, and 1-month (F1), 3-month (F3), and 6-month (F6) follow-up after LiESWT. Results: The results showed that 4 weeks of LiESWT could significantly decrease urine leakage based on the pad test and validated standardized questionnaire scores, as compared to the sham group. Moreover, 8 weeks of LiESWT could significantly reduce urine leakage but increase urine volume and attenuate urgency symptoms, which showed meaningful and persistent improvement at W8, F1, F3, and F6. Furthermore, validated standardized questionnaire scores were significantly improved at W8, F1, F3, and F6 as compared to the baseline (W0). Conclusions: Eight weeks of LiESWT attenuated SUI symptoms upon physical activity, reduced urine leakage, and ameliorated overactive bladder symptoms, which implied that LiESWT significantly improved the quality of life. Our findings suggested that LiESWT could serve as a potentially novel and non-invasive treatment for SUI.
Assuntos
Tratamento por Ondas de Choque Extracorpóreas , Bexiga Urinária Hiperativa , Incontinência Urinária por Estresse , Feminino , Humanos , Qualidade de Vida , Método Simples-Cego , Resultado do Tratamento , Bexiga Urinária Hiperativa/terapia , Incontinência Urinária por Estresse/terapiaRESUMO
Postmenopausal women with ovary hormone deficiency (OHD) are subject to overactive bladder (OAB) symptoms. The present study attempted to elucidate whether low-intensity extracorporeal shock wave therapy (LiESWT) alters bladder angiogenesis, decreases inflammatory response, and ameliorates bladder hyperactivity to influence bladder function in OHD-induced OAB in human clinical trial and rat model. The ovariectomized (OVX) for 12 months Sprague-Dawley rat model mimicking the physiological condition of menopause was utilized to induce OAB and assess the potential therapeutic mechanism of LiESWT (0.12 mJ/mm2, 300 pulses, and 3 pulses/second). The randomized, single-blinded clinical trial was enrolled 58 participants to investigate the therapeutic efficacy of LiESWT (0.25 mJ/mm2, 3000 pulses, 3 pulses/second) on postmenopausal women with OAB. The results revealed that 8 weeks' LiESWT inhibited interstitial fibrosis, promoted cell proliferation, enhanced angiogenesis protein expression, and elevated the protein phosphorylation of ErK1/2, P38, and Akt, leading to decreased urinary frequency, nocturia, urgency, urgency incontinence, and post-voided residual urine volume, but increased voided urine volume and the maximal flow rate of postmenopausal participants. In conclusion, LiESWT attenuated inflammatory responses, increased angiogenesis, and promoted proliferation and differentiation, thereby improved OAB symptoms, thereafter promoting social activity and the quality of life of postmenopausal participants.
Assuntos
Modelos Animais de Doenças , Tratamento por Ondas de Choque Extracorpóreas/métodos , Insuficiência Ovariana Primária/complicações , Qualidade de Vida , Regeneração , Bexiga Urinária Hiperativa/terapia , Bexiga Urinária/citologia , Adulto , Idoso , Animais , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Ratos , Ratos Sprague-Dawley , Método Simples-Cego , Bexiga Urinária Hiperativa/etiologia , Bexiga Urinária Hiperativa/patologiaRESUMO
Overactive bladder (OAB) syndrome, including frequency, urgency, nocturia and urgency incontinence, has a significantly negative impact on the quality-of-life scale (QoL) and can cause sufferer withdrawal from social activities. The occurrence of OAB can result from an imbalance between the production of pro-oxidants, such as free radicals and reactive species, and their elimination through protective mechanisms of antioxidant-induced oxidative stress. Several animal models, such as bladder ischemia/reperfusion (I/R), partial bladder outlet obstruction (PBOO) and ovarian hormone deficiency (OHD), have suggested that cyclic I/R during the micturition cycle induces oxidative stress, leading to bladder denervation, bladder afferent pathway sensitization and overexpression of bladder-damaging molecules, and finally resulting in bladder hyperactivity. Based on the results of previous animal experiments, the present review specifically focuses on four issues: (1) oxidative stress and antioxidant defense system; (2) oxidative stress in OAB and biomarkers of OAB; (3) OAB animal model; (4) potential nature/plant antioxidant treatment strategies for urinary dysfunction with OAB. Moreover, we organized the relationships between urinary dysfunction and oxidative stress biomarkers in urine, blood and bladder tissue. Reviewed information also revealed the summary of research findings for the effects of various antioxidants for treatment strategies for OAB.
Assuntos
Antioxidantes/uso terapêutico , Isquemia/tratamento farmacológico , Bexiga Urinária Hiperativa/tratamento farmacológico , Incontinência Urinária/tratamento farmacológico , Humanos , Isquemia/patologia , Estresse Oxidativo/genética , Estresse Oxidativo/fisiologia , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/patologia , Incontinência Urinária/patologiaRESUMO
BACKGROUND: The present study attempted to investigate the therapeutic effect and duration of low intensity extracorporeal shock wave therapy (LiESWT) on overactive bladder (OAB) symptoms, including social activity and the quality of life (QoL). METHODS: In this prospective, randomized, single-blinded clinical trial, 65 participants with OAB symptom were randomly divided into receive LiESWT (0.25 mJ/mm2, 3000 pulses, 3 pulses/second) once a week for 8 weeks, or an identical sham LiESWT treatment without the energy transmission. We analyzed the difference in overactive bladder symptom score (OABSS) and 3-day urinary diary as the primary end. The secondary endpoint consisted of the change in uroflowmetry, post-voided residual (PVR) urine, and validated standardized questionnaires at the baseline (W0), 4-week (W4) and 8-week (W8) of LiESWT, and 1-month (F1), 3-month (F3) and 6-month (F6) follow-up after LiESWT. RESULTS: 8-week LiESWT could significantly decrease urinary frequency, nocturia, urgency, and PVR volume, but meaningfully increase functional bladder capacity, average voided volume and maximal flow rate (Qmax) as compared with the W0 in the LiESWT group. In addition, the scores calculated from questionnaires were meaningfully reduced at W4, W8, F1, F3, and F6 in the LiESWT group. CONCLUSIONS: Our results revealed that the therapeutic efficacy of LiESWT could improve voided volume and ameliorate OAB symptoms, such as urgency, frequency, nocturia, and urinary incontinence, and lasted up to 6 month of follow-up. Moreover, LiESWT treatment brought statistically significant and clinically meaningful improvements in social activity and QoL of patients. These findings suggested that LiESWT could serve as an alternative non-invasive therapy for OAB patients.
RESUMO
Interstitial cystitis/bladder pain syndrome (IC/BPS) is defined as a chronic bladder disorder with suprapubic pain (pelvic pain) and pressure and/or discomfort related to bladder filling accompanied by lower urinary tract symptoms, such as urinary frequency and urgency without urinary tract infection (UTI) lasting for at least 6 weeks. IC/BPS presents significant bladder pain and frequency urgency symptoms with unknown etiology, and it is without a widely accepted standard in diagnosis. Patients' pathological features through cystoscopy and histologic features of bladder biopsy determine the presence or absence of Hunner lesions. IC/PBS is categorized into Hunner (ulcerative) type IC/BPS (HIC/BPS) or non-Hunner (nonulcerative) type IC/BPS (NHIC/BPS). The pathophysiology of IC/BPS is composed of multiple possible factors, such as chronic inflammation, autoimmune disorders, neurogenic hyperactivity, urothelial defects, abnormal angiogenesis, oxidative stress, and exogenous urine substances, which play a crucial role in the pathophysiology of IC/BPS. Abnormal expressions of several urine and serum specimens, including growth factor, methylhistamine, glycoprotein, chemokine and cytokines, might be useful as biomarkers for IC/BPS diagnosis. Further studies to identify the key molecules in IC/BPS will help to improve the efficacy of treatment and identify biomarkers of the disease. In this review, we discuss the potential medical therapy and assessment of therapeutic outcome with urinary biomarkers for IC/BPS.
RESUMO
OBJECTIVE: In the present clinical trial, we evaluated the therapeutic effects of low-intensity extracorporeal shockwave therapy (LiESWT) on overactive bladder (OAB). METHODS: Female subjects with ages of 20-75 years and who have been clinically diagnosed with OAB were included in the study. The LiESWT (DUOLITH SD1 T-TOP, AG) applicator was placed on the suprapubic skin area and applied with an intensity of 0.25 mJ/mm2, 3000 pulses, and 3 pulses/second. To assess the therapeutic efficacy, all subjects were required to complete the validated OAB symptoms and life bothersome questionnaires, 3-day urinary diary, uroflowmetry, and post-voided residual urine (PVR) measurement at 4 weeks of LiESWT (W4), 8 weeks of LiESWT (W8), 1-month follow-up (F1), and 3-month follow-up (F3) after LiESWT. RESULT: 82 subjects with the mean age of 56.5 ± 1.2 years were enrolled. The questionnaire scores were significantly improved at W4, W8, F1, and F3 as compared to baseline data (W0). At W8, the mean values of functional bladder capacity were meaningfully increased. According to the 3-day urinary diary, daytime frequency, urgency, and nocturia were significantly decreased. The uroflowmetry results showed that the mean voided urine volume and the maximal flow rate (Q max) were noticeably increased. PVR volume was also significantly decreased. CONCLUSIONS: The data demonstrated that 8-week LiESWT ameliorated the OAB symptoms, promoted the uroflow parameters, and improved the quality of life (QoL) in OAB patients, suggesting that LiESWT might serve as an alternative noninvasive therapy for OAB.
Assuntos
Bexiga Urinária Hiperativa/terapia , Bexiga Urinária/fisiopatologia , Adulto , Idoso , Tratamento por Ondas de Choque Extracorpóreas/métodos , Feminino , Humanos , Pessoa de Meia-Idade , Noctúria/terapia , Projetos Piloto , Estudos Prospectivos , Qualidade de Vida , Inquéritos e Questionários , Micção/fisiologia , Adulto JovemRESUMO
This study aimed to evaluate the therapeutic effects of Low intensity extracorporeal low energy shock wave therapy (LiESWT) on stress urinary incontinence (SUI). The investigation was a single-arm, open-label, multicentre study conducted in Taiwan. 50 female patients with SUI received LiESWT-treated with 0.25 mJ/mm2 intensity, 3000 pulses, and 3 pulses/second, once weekly for 4-weeks (W4) and 8-weeks (W8). The pad test, uroflowmetry, life quality questionnaires, and 3-day urinary diary measurement were performed before and after LiESWT intervention. The results revealed that 8-week of LiESWT treatment meaningfully improved urine leakage (pad test), maximum flow rate, post-voided residual urine, average urine volume, functional bladder capacity, urinary frequency, urgency symptom, and nocturia, which also persisted to show significant improvements at 1-month follow up (F1). Moreover, bothersome questionnaires scores were significantly improved at W4, W8, and F1 as compared to the baseline (W0). These results indicated that 8 weeks of LiESWT attenuated SUI symptoms on physical activity, reduced bladder leaks and overactive bladder (OAB), implying that LiESWT brought significant improvement in the quality of life. (ClinicalTrials.gov number, NCT04059133).
Assuntos
Incontinência Urinária por Estresse/terapia , Adulto , Idoso , Tratamento por Ondas de Choque Extracorpóreas/métodos , Feminino , Humanos , Pessoa de Meia-Idade , Noctúria/fisiopatologia , Qualidade de Vida , Inquéritos e Questionários , Taiwan , Resultado do Tratamento , Bexiga Urinária/fisiopatologia , Bexiga Urinária Hiperativa/fisiopatologia , Incontinência Urinária por Estresse/fisiopatologia , Adulto JovemRESUMO
The present study investigated the methylation of CpG sites in the cyclooxygenase (COX)2 promoter via nuclear factor (NF)κB transcriptional regulation and elucidated its effect on the COX2 transcriptional expression in a ketamineinduced ulcerative cystitis (KIC) animal model. The results of the present study revealed that ketamine treatment induced NFκB p65 translocation to nuclei and activated COX2 expression and prostaglandin (PGE)2 production in bladder tissue, whereas COX2 inhibitor suppressed the inflammatory effect. Moreover, DNA hypomethylation of the COX2 promoter region located from 1,522 to 829 bp might contribute to transcriptional regulation of COX2 expression and induce a proinflammatory response in KIC. Ketamine treatment increased the binding of NFκB and permissive histone H3 lysine4 (H3K4)m3, but caused a decrease in the repressive histone H3K27m3 and H3K36m3 on the COX2 promoter ranging from 1,522 to 1,331 bp as determined by a chromatin immunoprecipitation assay. Moreover, in the ketamine group, the level of TenElevenTranslocation methylcytosine dioxygenase for demethylation as determined by reverse transcriptionquantitative PCR assay was increased in comparison with the control group, but that was not the case for the level of DNA methyltransferases for methylation. The present findings revealed that there was a hypomethylation pattern of the COX2 promoter in association with the level of COX2 transcription in KIC.
Assuntos
Ciclo-Oxigenase 2/genética , Cistite/etiologia , Cistite/metabolismo , Metilação de DNA , Epigênese Genética , Regulação da Expressão Gênica , NF-kappa B/metabolismo , Animais , Imunoprecipitação da Cromatina , Ilhas de CpG , Cistite/patologia , Modelos Animais de Doenças , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Ketamina/efeitos adversos , Modelos Biológicos , Regiões Promotoras Genéticas , Transporte Proteico , RatosRESUMO
Metabolic syndrome (MetS) and ovarian hormone deficiency could affect bladder storage dysfunction. Epigallocatechin-3-gallate (EGCG), a polyphenolic compound in green tea, has been shown to protect against ovarian hormone deficiency induced overactive bladder (OAB). The present study investigated oxidative stress induced by MetS and bilateral ovariectomy (OVX), and elucidated the mechanism underlying the protective effect of EGCG (10 umol/kg/day) on bladder overactivity. Rats were fed with high fat high sugar (HFHS) diet to induce MetS and received ovariectomy surgery to deprive ovarian hormone. By dieting with HFHS for 6 months, rats developed MetS and OAB. MetS + OVX deteriorated bladder storage dysfunction more profound than MetS alone. MetS and MetS + OVX rats showed over-expression of inflammatory and fibrosis markers (1.7~3.8-fold of control). EGCG pretreatment alleviated storage dysfunction, and protected the bladders from MetS and OVX - induced interstitial fibrosis changes. Moreover, OVX exacerbated MetS related bladder apoptosis (2.3~4.5-fold of control; 1.8~2.6-fold of Mets group), enhances oxidative stress markers (3.6~4.3-fold of control; 1.8~2.2-fold of Mets group) and mitochondrial enzyme complexes subunits (1.8~3.7-fold of control; 1.5~3.4-fold of Mets group). EGCG pretreatment alleviated bladder apoptosis, attenuated oxidative stress, and reduced the mitochondrial and endoplasmic reticulum apoptotic signals. In conclusions, HFHS feeding and ovarian hormone deficiency enhances the generation of oxidative stress mediated through mitochondrial pathway. EGCG reduced the generation of oxidative stress and lessened bladder overactivity.
Assuntos
Apoptose/efeitos dos fármacos , Catequina/análogos & derivados , Síndrome Metabólica/complicações , Mitocôndrias/efeitos dos fármacos , Insuficiência Ovariana Primária/complicações , Bexiga Urinária Hiperativa/tratamento farmacológico , Bexiga Urinária Hiperativa/etiologia , Animais , Biomarcadores/sangue , Catequina/farmacologia , Catequina/uso terapêutico , Dieta da Carga de Carboidratos/métodos , Dieta Hiperlipídica , Modelos Animais de Doenças , Feminino , Mitocôndrias/metabolismo , Ovariectomia/métodos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Ratos , Ratos Sprague-Dawley , CháRESUMO
Ketamine-induced ulcerative cystitis (KIC) initially damaged the bladder mucosa and induced contracted bladder thereafter. Hyaluronan (hyaluronic acid; HA) instillation to the bladder has been used to treat KIC. The present study investigated bladder injury by urothelial defect and HA degeneration and bladder repair by urothelium proliferation and differentiation. This work was based on the hypothesis that HA treatment altered the bladder urothelial layer and the expression of hyaluronan-metabolizing enzymes and/or HA receptors in KIC. Cystometrogram study and tracing analysis of voiding behavior revealed that the ketamine-treated rats exhibited significant bladder hyperactivity with an increase in micturition frequency and a decrease in bladder capacity. The expression of inflammatory and fibrosis markers was also increased in the ketamine-treated group. Moreover, ketamine administration decreased the expression of urothelial barrier-associated protein, altered HA production, and induced abnormal urothelial differentiation, which might attribute to urothelial lining defects. However, HA instillation ameliorated bladder hyperactivity, lessened bladder mucosa damage, and decreased interstitial fibrosis. HA instillation also improved the level of HA receptors (CD44, Toll-like receptor-4, and receptor for HA-mediated motility) and HA synthases 1 to 3 and decreased the expression of hyaluronidases in the urothelial layer of bladder, resulting in enhanced mucosal regeneration. These findings suggested that HA could modulate inflammatory responses, enhance mucosal regeneration, and improve urothelial lining defects in KIC.
Assuntos
Cistite/fisiopatologia , Ácido Hialurônico/uso terapêutico , Bexiga Urinária/efeitos dos fármacos , Animais , Cistite/induzido quimicamente , Cistite/metabolismo , Modelos Animais de Doenças , Feminino , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/metabolismo , Ácido Hialurônico/farmacologia , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/fisiopatologia , Ketamina , Ratos , Ratos Sprague-Dawley , Receptor 4 Toll-Like/metabolismo , Bexiga Urinária/metabolismo , Bexiga Urinária/fisiopatologia , Urotélio/metabolismo , Urotélio/fisiopatologiaRESUMO
The number of ketamine abusers has increased significantly recently. Ketamine abusers exhibit urinary frequency, urgency, and at times urinary incontinence. Our aim was to investigate the role of transcription factor NF-κB and cyclooxygenase (COX)-2 in ketamine-induced cystitis. Sprague-Dawley rats were distributed into three groups, which received saline or treatment with ketamine or ketamine combined with a Cox-2 inhibitor (parecoxib). In addition, the toxic effect of ketamine and its metabolites were examined by primary urothelial cell culture. The ketamine-treated group displayed bladder hyperactivity and decreased bladder capacity. Treatment with ketamine + COX-2 inhibitor prevented these bladder dysfunctions. These bladder dysfunctions were accompanied by increases in the expression of NF-κB and COX-2 at the protein and mRNA levels. Ketamine treatment also enhanced bladder interstitial fibrosis, whereas ketamine + Cox-2 inhibitor decreased the intensity of fibrosis. Treatment of primary urothelial cells in vitro with ketamine or urine obtained from ketamine-treated rats stimulated the expression of NF-κB p65 and COX-2. Ketamine also initiated NF-κB translocation from cell cytoplasm to nucleus. Treatment with NF-κB inhibitor suppressed Cox-2 mRNA expression. Promoter-deletion analysis revealed that NF-κB was a necessary transcription factor for COX-2 gene (Ptgs2) activation. These results demonstrate that the regulation of COX-2 via the NF-κB pathway is involved in the inflammatory signaling of ketamine-induced cystitis in rat urinary bladder.
Assuntos
Ciclo-Oxigenase 2/metabolismo , Cistite/metabolismo , NF-kappa B/metabolismo , Bexiga Urinária/metabolismo , Animais , Linhagem Celular , Cistite/induzido quimicamente , Feminino , Humanos , Ketamina , Transporte Proteico , Ratos , Ratos Sprague-DawleyRESUMO
Ketamine abusers develop severe lower urinary tract symptoms. The major aims of the present study were to elucidate ketamine-induced ulcerative cystitis and bladder apoptosis in association with oxidative stress mediated by mitochondria and the endoplasmic reticulum (ER). Sprague-Dawley rats were distributed into three different groups, which received normal saline or ketamine for a period of 14 or 28 days, respectively. Double-labeled immunofluorescence experiments were performed to investigate tight junction proteins for urothelial barrier functions. A TUNEL assay was performed to evaluate the distribution of apoptotic cells. Western blot analysis was carried out to examine the expressions of urothelial tight junction proteins, ER stress markers, and apoptosis-associated proteins. Antioxidant enzymes, including SOD and catalase, were investigated by real-time PCR and immunofluorescence experiments. Ketamine-treated rats were found to display bladder hyperactivity. This bladder dysfunction was accompanied by disruptions of epithelial cadherin- and tight junction-associated proteins as well as increases in the expressions of apoptosis-associated proteins, which displayed features of mitochondria-dependent apoptotic signals and ER stress markers. Meanwhile, expressions of mitochondria respiratory subunit enzymes were significantly increased in ketamine-treated bladders. Conversely, mRNA expressions of the antioxidant enzymes Mn-SOD (SOD2), Cu/Zn-SOD (SOD1), and catalase were decreased after 28 days of ketamine treatment. These results demonstrate that ketamine enhanced the generation of oxidative stress mediated by mitochondria- and ER-dependent pathways and consequently contributed to bladder apoptosis and urothelial lining defects. Such oxidative stress-enhanced bladder cell apoptosis and urothelial barrier defects are potential factors that may play a crucial role in bladder overactivity and ulceration.
Assuntos
Apoptose , Cistite/metabolismo , Retículo Endoplasmático/metabolismo , Ketamina , Mitocôndrias/metabolismo , Estresse Oxidativo , Úlcera/metabolismo , Bexiga Urinária/metabolismo , Urotélio/metabolismo , Animais , Antioxidantes/metabolismo , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Biomarcadores/metabolismo , Cistite/induzido quimicamente , Cistite/genética , Cistite/patologia , Cistite/fisiopatologia , Modelos Animais de Doenças , Retículo Endoplasmático/patologia , Feminino , Fibrose , Regulação da Expressão Gênica , Mitocôndrias/patologia , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Fatores de Tempo , Úlcera/induzido quimicamente , Úlcera/genética , Úlcera/patologia , Úlcera/fisiopatologia , Bexiga Urinária/patologia , Bexiga Urinária/fisiopatologia , Urodinâmica , Urotélio/patologia , Urotélio/fisiopatologiaRESUMO
INTRODUCTION: The roles of testosterone and orchiectomy on male bladder subjected to ischemic/reperfusion (I/R) injuries received little attention. To fill this gap, the present study intended to examine testosterone and orchiectomy effects on male rabbits subjected to I/R damages. AIM: To elucidate the effects of testosterone and orchiectomy on contractile response, bladder morphology, interstitial fibrosis, and oxidative stress in male rabbit bladder subjected to I/R surgery. METHODS: Male New Zealand rabbits were distributed into five groups as follows: Group 1 received sham surgical procedure. In group 2, I/R surgery was performed. In group 3, testosterone (100 µg/kg/day) was intramuscularly injected prior to I/R surgery. In group 4, orchiectomy was performed prior to I/R surgery. In group 5, orchiectomy was performed with subsequent testosterone administration, followed by I/R surgery. All the rabbits were euthanized 7 days after I/R. Comparative studies were analyzed to elucidate the effects of testosterone and orchiectomy on bladder dysfunction subjected to I/R injuries. MAIN OUTCOME MEASURES: Bladder contractile function was evaluated. Masson's trichrome staining and immunohistochemical studies were performed to evaluate bladder morphology and intramural nerve terminals. Western blotting was examined to investigate the expressions of fibrosis and oxidative stress markers. RESULTS: I/R surgery significantly decreased bladder contractility in response to various stimulations with and without testosterone treatment. I/R damages decreased bladder nerve density with and without testosterone. The expressions of fibrosis and oxidative stress-related proteins were increased by I/R injuries with or without testosterone treatment. Testosterone depletion significantly decreased the expressions of transforming growth factor-ß and fibronectin expressions after I/R injury. Supraphysiological testosterone treatment after orchiectomy greatly increased the expressions of these fibrosis proteins; however, orchiectomy alone ameliorated I/R injuries. CONCLUSIONS: Testosterone treatment or orchiectomy affected I/R-induced bladder damages in male rabbits. Orchiectomy decreased the level of fibrosis and oxidative stress markers and increased neurofilament densities. Supraphysiological exogenous testosterone administration after orchiectomy further exacerbated such detrimental effects of I/R.
Assuntos
Contração Muscular/efeitos dos fármacos , Orquiectomia , Traumatismo por Reperfusão/fisiopatologia , Testosterona/farmacologia , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/fisiopatologia , Animais , Humanos , Masculino , Estresse Oxidativo/efeitos dos fármacos , Coelhos , Bexiga Urinária/irrigação sanguíneaRESUMO
AIMS: The aims of the present study were to investigate voiding patterns, tissue constituents and the expressions of cyclooxygenase-2 (COX-2) and nitric oxide synthase (NOS) involved in ketamine-induced ulcerative cystitis in rat urinary bladder. METHODS: Thirty Sprague-Dawley rats were distributed into three groups which received saline or ketamine (25 mg/kg/day) for a period of 14 and 28 days. In each group, cystometry was performed weekly and the concentration of ketamine and its metabolites (norketamine) was assayed. Paraffin-embedded sections were stained with Masson's trichrome stain, and ketamine-induced morphological changes were examined. Western blot analyses were carried out to examine the expressions of COX-2 and different NOS isoforms in bladder tissues. Immunofluorescence study was done to evaluate the expressions of COX-2 and macrophage infiltration (stained with ED-1 macrophage cell surface antigen) within the bladder. RESULTS: Ketamine treatment resulted in bladder hyperactivity and the non-voiding contractions were significantly increased. The urine concentrations of ketamine and norketamine were much higher in ketamine-treated group. Moreover, ulcerated urothelium and mononuclear cell infiltration were noted in ketamine-treated group. These alterations in urodynamic functions and tissue constituents were accompanied by increases in the expression of COX-2. Two NOS isoforms (iNOS and eNOS) were also overexpressed, but no significant change was observed for nNOS. COX-2 positive stained cells were significantly increased. Meanwhile, increased amounts of ED-1 positive stained macrophages were present and most of COX-2 expressed cells were co-stained with ED-1 in the early stage of ketamine treatment. CONCLUSIONS: Ketamine treatment affected bladder tissues by enhancing interstitial fibrosis and accelerating macrophages infiltration. Ketamine also initiated the up-regulations of COX-2 and iNOS and eNOS expressions. These up-regulated enzymes might play an important role in contributing to ketamine-induced alterations in micturition patterns and ulcerative cystitis.
Assuntos
Cistite/enzimologia , Óxido Nítrico Sintase/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Bexiga Urinária/enzimologia , Animais , Cistite/induzido quimicamente , Cistite/fisiopatologia , Ketamina , Ratos , Ratos Sprague-Dawley , Regulação para Cima , Bexiga Urinária/fisiopatologia , Micção/fisiologia , Urodinâmica/fisiologiaRESUMO
OBJECTIVE: The present study investigated the effects of ovarian hormone depletion and estrogen administration on ischemia/reperfusion (I/R)-induced bladder damage in female rabbits. METHODS: Female New Zealand white rabbits were divided into five groups. A sham surgical procedure was performed on rabbits in group 1. In group 2, both vesical arteries were clamped for 2 hours and then released (I/R surgical procedure). In group 3, 17ß-estradiol (100 µg/kg/d) was injected intramuscularly before I/R surgical procedure. In group 4, ovariectomies were performed before I/R surgical procedure. Group 5 had ovariectomy, recovered for 2 weeks, and then received 17ß-estradiol for 2 weeks. I/R surgical procedure was performed thereafter. Rabbits were killed 7 days after I/R surgical procedure. Masson's trichrome stain was used, and immunohistochemical experiments were performed to evaluate interstitial fibrosis and intramural nerve changes. Western immunoblots were examined to determine the expressions of markers for inflammation, fibrosis, and oxidative stress. RESULTS: I/R surgical procedure decreased bladder contractile responses by 30% to 50%. Ovarian hormone depletion further reduced bladder contractile function by 45% to 55% compared with the I/R group members that retained their ovaries. Moreover, I/R surgical procedure significantly decreased intramural neurofilament staining by two thirds compared with the control group. Estrogen replacement after ovariectomy significantly increased the density of nerve terminals. In addition, the expression of transforming growth factor-ß and fibronectin increased twofold and fivefold after I/R, respectively. Ovarian hormone depletion further increased the expression of these inflammatory and fibrosis markers. Ovariectomy significantly exacerbated oxidative damage, whereas estrogen replacement diminished oxidative stress to a level approaching that of the control group. CONCLUSIONS: I/R surgical procedure increases oxidative damage, enhances interstitial fibrosis, and results in bladder denervation. Ovarian hormone deficiency exacerbates this I/R-induced bladder damage, whereas estrogen therapy after ovariectomy attenuates this injury. These results reveal estrogen's protective effects on bladders subjected to I/R injury and the potential benefits of estrogen therapy on I/R-induced bladder damage.
Assuntos
Estradiol/administração & dosagem , Ovariectomia , Traumatismo por Reperfusão/complicações , Doenças da Bexiga Urinária/prevenção & controle , Bexiga Urinária/irrigação sanguínea , Animais , Terapia de Reposição de Estrogênios , Feminino , Fibronectinas/genética , Expressão Gênica/efeitos dos fármacos , Injeções Intramusculares , Contração Muscular/efeitos dos fármacos , Coelhos , Fator de Crescimento Transformador beta/genética , Doenças da Bexiga Urinária/etiologia , Doenças da Bexiga Urinária/patologiaRESUMO
UNLABELLED: What's known on the subject? and What does the study add? Ovary hormone deficiency and the age-related changes in post-menopausal women are subjected to a number of urological dysfunctions, including overactive bladder syndrome. Green tea is a popular healthy drink worldwide and its extract catechin has strong anti-inflammatory and antioxidant properties. EGCG, the major type of catechin, is an antioxidant polyphenol flavonoid isolated from green tea. EGCG supplement could prevent ovariectomy-induced bladder dysfunction in a dose-related manner through its anti-oxidant, anti-fibrosis and anti-apoptosis effects. OBJECTIVE: To evaluate whether green tea extract, epigallocatechin gallate (EGCG), could prevent ovariectomy-induced overactive bladder (OAB) and to investigate its antioxidant, anti-apoptotic and anti-fibrosis effects. MATERIALS AND METHODS: In all, 48 female Sprague-Dawley rats were divided into four groups. After bilateral ovariectomy, the first group served as the ovariectomy control, the second group received EGCG 1 µM/kg daily i.p. injection after ovariectomy surgery, and the third group received EGCG 10 µM/kg daily i.p. injection. The fourth group was taken as the sham without ovariectomy surgery. The rats were killed after 6 months after ovariectomy surgery. Cystometrograms were performed for the measure of bladder overactivity. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) assay was used to evaluate apoptotic cells. Western immunoblots were performed to determine the expressions of inflammatory markers, apoptosis-associated proteins and oxidative stress markers. RESULTS: Long-term ovariectomy significantly increased non-voiding contractions and decreased bladder compliance. Treatment with EGCG significantly increased bladder compliance and diminished non-voiding contractions. Ovariectomy significantly increased apoptotic cells and enhanced interstitial fibrosis in bladders. The expression of caspase-3 significantly increased, while that of Bcl-2 notably decreased after ovariectomy. Inflammatory and fibrosis markers, TGF-ß, fibronectin and type I collagen expressions were significantly increased after 6 months of ovariectomy surgery. Treatment with EGCG significantly decreased TGF-ß and type I collagen expressions. Oxidative stress markers, nitrotyrosine and protein carbonylation levels were significantly increased in the ovariectomy group. EGCG could attenuate this oxidative damage in dose-dependent fashion. CONCLUSIONS: Ovariectomy increased oxidative damage, enhanced voiding frequency and decreased bladder compliance. EGCG could restore ovariectomy-induced bladder dysfunction in a dose-dependent fashion through antioxidant, anti-fibrosis and anti-apoptosis effects.
Assuntos
Antioxidantes/farmacologia , Catequina/análogos & derivados , Menopausa/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Chá , Bexiga Urinária Hiperativa/metabolismo , Animais , Catequina/farmacologia , Modelos Animais de Doenças , Feminino , Ovariectomia , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVES: To assess the effects of abdominal fat on shock wave lithotripsy (SWL). We used pre-SWL unenhanced computed tomography (CT) to evaluate the impact of abdominal fat distribution and calculus characteristics on the outcome of SWL. METHODS: One hundred and eighty-five patients with a solitary ureteric calculus treated with SWL were retrospectively reviewed. Each patient underwent unenhanced CT within 1 month before SWL treatment. Treatment outcomes were evaluated 1 month later. Unenhanced CT parameters, including calculus surface area, Hounsfield unit (HU) density, abdominal fat area and skin to calculus distance (SSD) were analysed. RESULTS: One hundred and twenty-eight of the 185 patients were found to be calculus-free following treatment. HU density, total fat area, visceral fat area and SSD were identified as significant variables on multivariate logistic regression analysis. The receiver-operating characteristic analyses showed that total fat area, para/perirenal fat area and visceral fat area were sensitive predictors of SWL outcomes. CONCLUSION: This study revealed that higher quantities of abdominal fat, especially visceral fat, are associated with a lower calculus-free rate following SWL treatment. Unenhanced CT is a convenient technique for diagnosing the presence of a calculus, assessing the intra-abdominal fat distribution and thereby helping to predict the outcome of SWL. KEY POINTS: ⢠Unenhanced CT is now widely used to assess ureteric calculi. ⢠The same CT protocol can provide measurements of abdominal fat distribution. ⢠Ureteric calculi are usually treated by shock wave lithotripsy (SWL). ⢠Greater intra-abdominal fat stores are generally associated with poorer SWL results.