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1.
Artigo em Inglês | MEDLINE | ID: mdl-31807839

RESUMO

To examine the role of the transcription factor nuclear factor-erythroid 2 (NF-E2)-related factor 2 (Nrf2) and the SAFE pathway (JAK/STAT) in the induction of germ cell apoptosis (GCA) and the protective role of epigallocatechin-3-gallate (EGCG) during testicular ischemia reperfusion injury (tIRI). Male Sprague-Dawley rats (n = 18) were divided into three groups: sham, unilateral tIRI, tIRI + epigallocatechin-3-gallate (EGCG, 50 mg/Kg). Unilateral tIRI was induced by 1-h ischemia followed by 4-h reperfusion, and EGCG was injected i.p. 30-min post ischemia. Immuno-histological analyses were used to detect spermatogenesis, oxidative DNA damage, and the immuno-expression of the JAK2, STAT3, and STAT1. Biochemical assays were used to investigate the oxidative, apoptosis proteins and enzymes, while Western blot was used to detect the expression of NOX and Nrf2. Expression of apoptosis-related genes was measured by real-time PCR. During tIRI, there was a clear damage to spermatogenesis associated with decreased activities of SOD, CAT, and GPx and increased levels of lipid peroxidation and oxidative DNA damage. In addition, GCA was indicated by the activation of caspase1, PARP, decreased gene expression of survivin and increased Bax to Bcl2 ratio. In contrast to lowered Nrf2 levels, NOX levels were augmented and phosphorylation of JAK2, STAT3, and STAT1 was increased. Pre-perfusion treatment with EGCG prevented the above modulations. The coordinated activation of the SAFE pathway and suppression of Nrf2 contribute to the tIRI-induced oxidative damages and GCA, which were modulated by EGCG.

2.
Molecules ; 24(18)2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31547465

RESUMO

The aim is to explore the mechanism of the apoptosis signal-regulating kinase-1 (ASK-1) signaling pathway and the involvement of the thioredoxin (Trx) system during testicular ischemia reperfusion injury (tIRI) by using ASK-1 specific inhibitor, NQDI-1. Male Sprague-Dawley rats (n = 36, 250-300 g) were equally divided into 3 groups: sham, tIRI, and tIRI + NQDI-1 (10 mg/kg, i.p, pre-reperfusion). For tIRI induction, the testicular cord and artery were occluded for 1 h followed by 4 h of reperfusion. Histological analyses, protein immunoexpression, biochemical assays, and real-time PCR were used to evaluate spermatogenesis, ASK-1/Trx axis expression, enzyme activities, and relative mRNA expression, respectively. During tIRI, ipsilateral testes underwent oxidative stress indicated by low levels of superoxide dismutase (SOD) and Glutathione (GSH), increased oxidative damage to lipids and DNA, and spermatogenic damage. This was associated with induced mRNA expression of pro-apoptosis genes, downregulation of antiapoptosis genes, increased caspase 3 activity and activation of the ASK-1/JNK/p38/survivin apoptosis pathway. In parallel, the expression of Trx, Trx reductase were significantly reduced, while the expression of Trx interacting protein (TXNIP) and the NADP+/ nicotinamide Adenine Dinucleotide phosphate (NADPH) ratio were increased. These modulations were attenuated by NQDI-1 treatment. In conclusion, the Trx system is regulated by the ASK-1/Trx/TXNIP axis to maintain cellular redox homeostasis and is linked to tIRI-induced germ cell apoptosis via the ASK-1/JNK/p38/survivin apoptosis pathway.


Assuntos
Aporfinas/farmacologia , MAP Quinase Quinase Quinase 5/metabolismo , Quinolinas/farmacologia , Traumatismo por Reperfusão/metabolismo , Espermatozoides/metabolismo , Tiorredoxinas/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Proteínas de Ciclo Celular/metabolismo , MAP Quinase Quinase 4/metabolismo , MAP Quinase Quinase Quinase 5/antagonistas & inibidores , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/patologia , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/patologia , Survivina/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo , Testículo/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
3.
PeerJ ; 4: e2195, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27441124

RESUMO

Background. Testicular ischemia reperfusion injury (tIRI) is considered the mechanism underlying the pathology of testicular torsion and detorsion. Left untreated, tIRI can induce testis dysfunction, damage to spermatogenesis and possible infertility. In this study, we aimed to assess the activities and expression of glycolytic enzymes (GEs) in the testis and their possible modulation during tIRI. The effect of fructose 1,6-diphosphate (FDP), a glycolytic intermediate, on tIRI was also investigated. Methods. Male Sprague-Dawley rats were divided into three groups: sham, unilateral tIRI, and tIRI + FDP (2 mg/kg). tIRI was induced by occlusion of the testicular artery for 1 h followed by 4 h of reperfusion. FDP was injected peritoneally 30 min prior to reperfusion. Histological and biochemical analyses were used to assess damage to spermatogenesis, activities of major GEs, and energy and oxidative stress markers. The relative mRNA expression of GEs was evaluated by real-time PCR. ELISA and immunohistochemistry were used to evaluate the expression of p53 and TP53-induced glycolysis and apoptosis regulator (TIGAR). Results. Histological analysis revealed tIRI-induced spermatogenic damage as represented by a significant decrease in the Johnsen biopsy score. In addition, tIRI reduced the activities of hexokinase 1, phosphofructokinase-1, glyceraldehyde 3-phosphate dehydrogenase, and lactate dehydrogenase C. However, mRNA expression downregulation was detected only for hexokinase 1, phosphoglycerate kinase 2, and lactate dehydrogenase C. ATP and NADPH depletion was also induced by tIRI and was accompanied by an increased Malondialdehyde concentration, reduced glutathione level, and reduced superoxide dismutase and catalase enzyme activities. The immunoexpression of p53 and TIGAR was markedly increased after tIRI. The above tIRI-induced alterations were attenuated by FDP treatment. Discussion. Our findings indicate that tIRI-induced spermatogenic damage is associated with dysregulation of GE activity and gene expression, which were associated with activation of the TIGAR/p53 pathway. FDP treatment had a beneficial effect on alleviating the damaging effects of tIRI. This study further emphasizes the importance of metabolic regulation for proper spermatogenesis.

4.
Urology ; 94: 312.e1-8, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27125877

RESUMO

OBJECTIVE: To investigate whether exogenous angiotensin (Ang)-(1-7) administration can protect against the damaging consequences of testicular ischemia reperfusion (tIR) injury. MATERIALS AND METHODS: Eighteen male Sprague-Dawley rats were divided equally among the following 3 groups: sham, unilateral tIR injury (1 hour of ischemic treatment and 4 hours of reperfusion), and tIR + Ang-(1-7) (0.3 mg/kg). Testicular tissues obtained from the rats were evaluated for the expression of testicular angiotensin-converting enzyme (tACE), Ang-(1-7), and the Ang-(1-7)-specific receptor Mas by immunohistochemistry and enzyme-linked immunosorbent assay. Reduced spermatogenesis, induction of the caspase-8 pathway, and nitric oxide (NO) generation were assessed. The effects of tIR and Ang-(1-7) treatment on the PI3K/Akt antiapoptosis pathway were also investigated. RESULTS: Testicular morphological changes and reduced spermatogenesis associated with decreased expression of the tACE/Ang-(1-7)/Mas axis were observed during tIR. These effects were also accompanied by increased activity of caspase-3 and -8, downregulation of the survivin and BAD transcripts, and decreased NO formation. During tIR, PTEN expression was increased, leading to inactivation of the PI3K/Akt pathway. Acute treatment with Ang-(1-7) prior to reperfusion attenuated the tIR-induced damage described above. CONCLUSION: Expression of the tACE/Ang-(1-7)/Mas axis was downregulated during tIR. Administration of exogenous Ang-(1-7) prior to reperfusion rescued tACE and Mas expression and protected against germ cell apoptosis and oxidative stress. Increased NO generation and activation of the PI3K/Akt signaling pathway may have partially contributed to these effects. The tACE/Ang-(1-7)/Mas axis likely plays a role in the maintenance of normal testis physiology and spermatogenesis.


Assuntos
Angiotensina I/fisiologia , Angiotensina I/uso terapêutico , Fragmentos de Peptídeos/fisiologia , Fragmentos de Peptídeos/uso terapêutico , Peptidil Dipeptidase A/biossíntese , Traumatismo por Reperfusão/prevenção & controle , Testículo/irrigação sanguínea , Testículo/metabolismo , Animais , Masculino , Peptidil Dipeptidase A/fisiologia , Proteínas Proto-Oncogênicas/fisiologia , Ratos , Ratos Sprague-Dawley , Receptores Acoplados a Proteínas-G/fisiologia , Traumatismo por Reperfusão/etiologia
5.
Naunyn Schmiedebergs Arch Pharmacol ; 389(5): 539-51, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26915501

RESUMO

Testicular ischemia reperfusion injury (tIRI) is considered as the underlying mechanism of testicular torsion, which can cause male infertility. tIRI-induced damage was investigated by assessing the gene expression of spermatogenesis master transcription factors (TFs) and that of major adhesion molecules of the blood-testis barrier. The effect of lutein, a hydroxyl carotenoid, in alleviating tIRI-induced damage was also studied. Male Sprague-Dawley rats were divided into three groups: sham, unilateral tIRI, and tIRI + lutein (0.2 mg/kg). tIRI was induced by occlusion of the testicular artery for 1 h, followed by 4 h of reperfusion. Lutein was injected 15 min after the start of ischemia. Histological analysis and real-time polymerase chain reaction revealed significant decreases in tissue biopsy scores, reduced seminiferous tubule diameters, and downregulated the mRNA expression of the TFs cAMP-responsive element modulator (CREM), TATA box-binding protein-related factor 2 (TRF2), and regulatory factor X 2 (RFX2) compared with the sham group. Lutein treatment reversed these effects. The mRNA expression of the adhesion molecules N-cadherin, nectin-2, claudin-11, occludin, and connexin-43 was significantly downregulated during tIRI, but this change was prevented by lutein treatment. In addition, lutein normalized the tIRI-induced increase in total antioxidant capacity, increased malondialdehyde (MDA) levels, augmented number of TdT-mediated dUTP-X nick-end labeling (TUNEL)-positive nuclei, and activated caspase-8 pathway. The components of survivor activating factor enhancement (SAFE) were also activated during tIRI. Increased tissue expression of TNF-α and its receptor, TNFR1, was accompanied by increased phosphorylation of Janus kinase (JAK) and STAT3, which was prevented by lutein treatment. Our findings suggested that tIRI-induced spermatogenic damage may involve modulation of the SAFE pathway and could benefit from lutein treatment.


Assuntos
Luteína/farmacologia , Traumatismo por Reperfusão/metabolismo , Testículo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 8/metabolismo , Marcação In Situ das Extremidades Cortadas , Janus Quinase 1/metabolismo , Masculino , Malondialdeído/metabolismo , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Fator de Transcrição STAT3/metabolismo , Espermatogênese/efeitos dos fármacos , Testículo/metabolismo , Fatores de Transcrição/genética , Transcrição Genética/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo
6.
Indian J Urol ; 31(1): 57-64, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25624578

RESUMO

INTRODUCTION: The objective of this study is to investigate the molecular mechanisms underlying the effects of zinc deficiency on spermatogenesis in the Sprague-Dawley (SD) rat. MATERIALS AND METHODS: Three groups of eight adult male SD rats were maintained for 4 weeks on a normal diet as control, zinc deficient diet and zinc deficient diet with zinc supplementation of 28 mg zinc/kg body weight respectively. Using standard techniques, the following parameters were compared between the three groups of experimental animals at the end of 4 weeks: (a) Serum zinc, magnesium (Mg), copper (Cu), selenium (Se) and cadmium (Cd), (b) serum sex hormones, malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPX), (c) interleukin-4 (IL-4), tumor necrosis factor-alpha (TNF-α), Bcl-2, Bax and caspase-3 expression in the testes, (d) assessment of apoptosis of testicular cells using electron microscopy and (e) testicular volume and histology using the orchidometer and Johnsen score, respectively. RESULTS: The zinc deficient group showed a reduction of testicular volume, serum concentrations of Zn, Cu, Se, Mg, SOD, GPX, IL-4, Bcl-2 and testosterone (P < 0.05), as well as increased levels of serum Cd, MDA and tissue TNF-α, Bax, caspase-3 and apoptosis of the germ cells (P < 0.05) compared with control and zinc supplementation groups. CONCLUSION: Zinc deficiency is associated with impaired spermatogenesis because of reduced testosterone production, increased oxidative stress and apoptosis. These findings suggest that zinc has a role in male reproduction.

7.
J Neurotrauma ; 32(3): 170-84, 2015 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-25025489

RESUMO

Our previous studies have established that (-)-epigallocatechin-3-gallate (EGCG) has both neuroprotective and -regenerative capacity after sciatic nerve injury. Moreover, this improvement was evident on the behavioral level. The aim of this study was to investigate the central effects of ECGC on spinal cord motor neurons after sciatic nerve injury. Our study showed that administering 50 mg/kg intraperitoneally i.p. of EGCG to sciatic nerve-injured rats improved their performance on different motor functions and mechanical hyperesthesia neurobehavioral tests. Histological analysis of spinal cords of EGCG-treated sciatic nerve-injured (CRUSH+ECGC) animals showed an increase in the number of neurons in the anterior horn, when compared to the naïve, sham, and saline-treated sciatic nerve-injured (CRUSH) control groups. Additionally, immunohistochemical study of spinal cord sections revealed that EGCG reduced the expression of glial fibrillary acidic protein and increased the expression of growth-associated protein 43, a marker of regenerating axons. Finally, EGCG reduced the ratio of B-cell lymphoma 2 (Bcl-2)-associated X protein/Bcl-2 and increased the expression of survivin gene. This study may shed some light on the future clinical use of EGCG and its constituents in the treatment of peripheral nerve injury.


Assuntos
Catequina/análogos & derivados , Fármacos Neuroprotetores/farmacologia , Traumatismos dos Nervos Periféricos/patologia , Recuperação de Função Fisiológica/efeitos dos fármacos , Medula Espinal/efeitos dos fármacos , Animais , Catequina/farmacologia , Modelos Animais de Doenças , Proteína GAP-43/biossíntese , Imuno-Histoquímica , Masculino , Compressão Nervosa , Degeneração Neural/patologia , Traumatismos dos Nervos Periféricos/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Nervo Isquiático/lesões , Medula Espinal/patologia , Proteína bcl-X/biossíntese
8.
Korean J Physiol Pharmacol ; 17(4): 259-65, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23946684

RESUMO

The anti-apoptotic effect of (-)-epigallocatechin-3-gallate (EGCG) during unilateral testicular torsion and detorsion (TT/D) was established in our previous study. In mice, the smallest inhibitor of apoptosis, survivin, is alternatively spliced into three variants, each suggested to have a unique function. Here, we assessed how EGCG exerts its protective effect through the expression of the different survivin splice variants and determined its effect on the morphology of the seminiferous tubules during TT/D. Three mouse groups were used: sham, TT/D+vehicle and TT/D treated with EGCG. The expression of the survivin variants (140 and 40) and other apoptosis genes (p53, Bax and Bcl-2) was measured with semi-quantitative RT-PCR. Histological analysis was performed to assess DNA fragmentation, damage to spermatogenesis and morphometric changes in the seminiferous tubules. In the TT/D+vehicle group, survivin 140 expression was markedly decreased, whereas survivin 40 expression was not significantly different. In parallel, there was an increase in the mRNA level of p53 and the Bax to Bcl-2 ratio in support of apoptosis induction. Histological analyses revealed increased DNA fragmentation and increased damage to spermatogenesis associated with decreased seminiferous tubular diameter and decreased germinal epithelial cell thickness in the TT/D+vehicle group. These changes were reversed to almost sham levels upon EGCG treatment. Our data indicate that EGCG protects the testis from TT/D-induced damage by protecting the morphology of the seminiferous tubules and modulating survivin 140 expression.

9.
Neurochem Int ; 62(3): 221-31, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23313191

RESUMO

Recently, we have shown that green tea (GT) consumption improves both reflexes and sensation in unilateral chronic constriction injury to the sciatic nerve. Considering the substantial neuroprotective properties of GT polyphenols, we sought to investigate whether (-)-epigallocatechin-3-gallate (EGCG) could protect the sciatic nerve and improve functional impairments induced by a crushing injury. We also examined whether neuronal cell apoptosis induced by the crushing injury is affected by EGCG treatment. Histological examination of sciatic nerves from EGCG-treated (50mg/kg; i.p.) showed that axonotmized rats had a remarkable axonal and myelin regeneration with significant decrease in the number of myelinated axonal fibers compared to vehicle-treated crush group. Similarly, ultrastructural evaluation of EGCG-treated nerves displayed normal unmyelinated and myelinated axons with regular myelin sheath thickness and normalized appearance of Schmidt-Lantermann clefts. Extracellular matrix displayed normal collagen fibers appearance with distinctively organized distribution similar to sham animals. Analysis of foot position and extensor postural thrust test showed a progressive and faster recovery in the EGCG-treated group compared to vehicle-treated animals. EGCG-treated rats showed significant increase in paw withdrawal thresholds to mechanical stimulation compared to vehicle-treated crush group. EGCG treatment also restored the mRNA expression of Bax, Bcl-2 and survivin but not that of p53 to sham levels on days 3 and 7 post-injury. Our results demonstrate that EGCG treatment enhanced functional recovery, advanced morphological nerve rescue and accelerated nerve regeneration following crush injury partly due to the down regulation of apoptosis related genes.


Assuntos
Catequina/análogos & derivados , Síndrome de Esmagamento/tratamento farmacológico , Nervo Isquiático/lesões , Animais , Catequina/uso terapêutico , Masculino , Microscopia Eletrônica de Transmissão , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Transcrição Genética
10.
J Urol ; 189(3): 911-5, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23009873

RESUMO

PURPOSE: A steady increase in the incidence of septicemia after prostate biopsy in our unit between 2001 and 2005 prompted us to review our prophylactic antibiotic regimen. We compared the incidence of septicemia in patients undergoing prostate biopsy between 2001 and 2005 when only oral ciprofloxacin was used prophylactically (group 1) to the incidence among patients undergoing biopsy between 2006 and 2010 when a single dose of intravenous amikacin was added to ciprofloxacin (group 2). MATERIALS AND METHODS: In group 1 the 300 patients were given 500 mg oral ciprofloxacin twice daily 1 day before and for 2 days after the biopsy while in group 2 the 897 patients, in addition to the ciprofloxacin previously mentioned, received 500 mg intravenous amikacin 30 minutes before the biopsy. Patients admitted to the hospital with septicemia after prostate biopsy had urine and blood culture and sensitivity tests. The number of patients in whom septicemia developed in each group after prostate biopsy and the microorganisms isolated from the urine and blood of such patients were compared using the chi-square test. RESULTS: Septicemia was seen in 24 of 300 (8%) and 15 of 897 (1.7%) patients in groups 1 and 2, respectively (p <0.001). In group 1 the rate of septicemia after prostate biopsy was 2.1% and 13% in 2001 and 2005, respectively (p <0.001). In group 2 the rate of septicemia was 1.5% in 2006 and 1.6% in 2010 (p <0.25). Escherichia coli resistant to quinolones was responsible for 33 of 39 (84.6%) septicemic cases. CONCLUSIONS: The addition of amikacin to ciprofloxacin prophylaxis significantly reduces the incidence of septicemia after prostate biopsy.


Assuntos
Amicacina/administração & dosagem , Antibioticoprofilaxia/métodos , Biópsia por Agulha/efeitos adversos , Ciprofloxacino/administração & dosagem , Endossonografia/métodos , Próstata/patologia , Sepse/prevenção & controle , Idoso , Idoso de 80 Anos ou mais , Anti-Infecciosos/administração & dosagem , Relação Dose-Resposta a Droga , Quimioterapia Combinada , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Próstata/diagnóstico por imagem , Reto , Sepse/etiologia , Resultado do Tratamento
11.
Int Urol Nephrol ; 44(6): 1681-9, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22718029

RESUMO

AIM: To determine whether the measurement of tissue mRNA levels of AMACR and survivin has a role in distinguishing prostate cancer (PCa) from benign lesions and high risk from low-risk PCa in men with suspected PCa. METHODS: TRUS prostate biopsies from 170 patients with suspected PCa were used to measure the mRNA levels of AMACR and survivin using semi-quantitative RT-PCR technique. The diagnosis, staging and risk stratification of PCa was based on established clinical criteria. The ability of tissue mRNA levels to distinguish benign from malignant prostate and high- and low-risk PCa was assessed. The diagnostic value for the two genes was evaluated by calculating their individual and combined sensitivity and specificity, which were compared with that of PSA. RESULTS: Histological examination showed 90/170 (53%) of patients had benign prostate pathology, while 80/170 (47%) had PCa. Tissue mRNA levels of both AMACR and survivin were able to distinguish benign from PCa biopsies (p<0.0001) and also high-risk from low-risk PCa cases (p<0.02, p<0.05, respectively). Compared with serum PSA levels, the combined use of tissue mRNA levels of AMACR and survivin yielded a higher detection specificity (84 vs. 22%). CONCLUSION: Based on AMACR and survivin combined sensitivity and specificity, these mRNA markers can be used as an adjunct to distinguish patients with and without PCa and in men with PCa may help to identify those with low- or high-risk PCa.


Assuntos
Proteínas Inibidoras de Apoptose/genética , Próstata/química , Neoplasias da Próstata/química , Neoplasias da Próstata/diagnóstico , RNA Mensageiro/análise , Racemases e Epimerases/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Próstata/patologia , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Medição de Risco/métodos , Survivina
12.
Ann Saudi Med ; 32(3): 262-8, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22588437

RESUMO

BACKGROUND AND OBJECTIVES: Enhancer of zeste homolog 2 (EZH2) has been recently found to regulate several genes involved in immunoresponse and autocrine inflammation network. The aim of the study was to quantitate EZH2 messenger ribonucleic acid (mRNA) expression, evaluate its relation to conditions of prostatitis associated with benign prostatic hyperplasia (BPH), and correlate it with the levels of the inflammatory marker interlukin 6 (IL-6). DESIGN AND SETTING: Cross-sectional study in Middle Eastern men with BPH and prostatitis or BPH only. PATIENTS AND METHODS: Transrectal ultrasound-guided prostate biopsies were collected from 106 patients suspected of having prostate cancer; however, the histology revealed BPH. Upon further pathological examination, 56 of these cases were identified as BPH with prostatitis and classified as: acute prostatitis (n=13); active chronic prostatitis (n=32); and, chronic inactive prostatitis (n=12). Serum IL-6 levels and EZH2 mRNA expression were measured and compared between patient groups. RESULTS: EZH2 mRNA was overexpressed in BPH with prostatitis patients compared to BPH only patients (P<.0001). BPH with active chronic prostatitis had higher EZH2 expression than BPH with acute or chronic inactive prostatitis compared to BPH only (P=.05 and .73, respectively). EZH2 mRNA expression showed a negative correlation with IL-6 concentrations in BPH with prostatitis patients (rs=-0.31, P=.02). EZH2 overexpression was associated with an increased risk of having BPH with prostatitis (crude odds ratio 0.20, 95% CI 0.06-0.65, P=.0076). CONCLUSIONS: EZH2 mRNA expression correlates positively with prostatitis conditions associated with BPH and negatively with serum IL-6 levels. This supports the possible involvement of EZH2 mRNA overexpression in the development of prostate inflammation, and its new regulatory role in suppressing the expression of some inflammatory network genes.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Próstata/patologia , Hiperplasia Prostática/metabolismo , Prostatite/metabolismo , RNA Mensageiro/análise , Fatores de Transcrição/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Biópsia , Estudos Transversais , Proteínas de Ligação a DNA/genética , Proteína Potenciadora do Homólogo 2 de Zeste , Humanos , Interleucina-6/sangue , Masculino , Pessoa de Meia-Idade , Complexo Repressor Polycomb 2 , Antígeno Prostático Específico/sangue , Hiperplasia Prostática/complicações , Hiperplasia Prostática/patologia , Prostatite/complicações , Prostatite/patologia , Fatores de Transcrição/genética
13.
Naunyn Schmiedebergs Arch Pharmacol ; 385(8): 807-22, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22573016

RESUMO

Muscle degeneration and impairment following nerve injury could lead to apoptosis as a result of increased levels of reactive oxygen species. This activates the apoptotic cascade through mitochondrial dysfunction and damage to lipids, proteins, and DNA. In considering of the multifactorial protective properties of green tea polyphenols (-)-epigallocatechin-3-gallate (EGCG), this study investigates whether EGCG treatment does improve skeletal muscle function impairments, induced by crushing of the sciatic nerve. Compared to the saline-treated injured group of animals, EGCG treatment of axonotomized animals showed significant motor enhancement in the toe spread and foot positioning analysis and gain in the percentage motor deficit. The proprioceptive function expressed by the hopping response showed significant progression in the EGCG-treated group. Recovery of sensory innervation was followed by a slowly retreating neuropathic pain-like syndrome in the EGCG-treated animals. Muscle tissues from injured limb showed severe histopathological alterations that were significantly attenuated by EGCG treatment at the end of week 3 post-surgery. Semi-quantitative desmin immunohistochemistry revealed intense staining in the saline-treated injured animals, whereas EGCG treatment decreased the desmin immunoreactivity back to sham control levels. Using RT-PCR, EGCG treatment induced a significant anti-apoptotic effect in injured muscle tissues by normalizing the Bax/Bcl-2 ratio back to baseline levels and inhibiting overexpression of the p53 apoptotic gene at days 3 and 7 post-surgery. In conclusion, our results demonstrate that EGCG enhances functional recovery, protects muscle fibers from cellular death by activating anti-apoptotic signaling pathway, and improves morphological recovery in skeletal muscle after nerve injuries.


Assuntos
Proteínas Reguladoras de Apoptose/biossíntese , Proteínas Reguladoras de Apoptose/genética , Catequina/análogos & derivados , Músculo Esquelético/metabolismo , Compressão Nervosa , Fármacos Neuroprotetores/farmacologia , Neuropatia Ciática/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Catequina/farmacologia , Desmina/metabolismo , Pé/fisiologia , Temperatura Alta , Hiperalgesia/tratamento farmacológico , Hiperalgesia/etiologia , Imuno-Histoquímica , Masculino , Músculo Esquelético/patologia , Medição da Dor/efeitos dos fármacos , Postura/fisiologia , RNA/biossíntese , RNA/genética , Ratos , Ratos Wistar , Tempo de Reação/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Recuperação de Função Fisiológica , Neuropatia Ciática/metabolismo , Neuropatia Ciática/patologia , Transdução de Sinais/efeitos dos fármacos , Dedos do Pé/fisiologia
14.
Biochem Biophys Res Commun ; 420(2): 434-9, 2012 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-22426481

RESUMO

Testicular torsion (TT) is a urologic emergency that may result in future infertility problems. The pathologic process of TT is similar to an ischemia reperfusion injury (IRI). The purpose of this study was to evaluate the effect of epigallocatechin-3-gallate (EGCG) on reversing the damaging consequences of TT-induced IRI by examining its inhibitory effects on the expression of inflammatory and apoptosis mediators in a unilateral TT rat model. Eighteen male Sprague-Dawley rats were divided into 3 groups. Group 1 underwent a sham operation of the left testis under general anesthesia. Group 2 underwent ischemia for 1h followed by 4h reperfusion in the presence of saline. The third group was similar to group 2, however, EGCG (50 mg/kg) was injected i.p. 30 min after ischemia induction. The in vivo protective effect of EGCG was tested by measuring testicular levels of TNF-α, IL-6 and IL-1ß by ELISA and mRNA expression of iNOS, MCP-1, p53, Bax, Bcl-2 and survivin by real-time PCR. Also, testicular morphological changes and damage to spermatogenesis were evaluated using H&E staining and Johnsen's scoring system, respectively. EGCG treatment improved testicular structures in the ipsilateral testis, markedly inhibited germ cell apoptosis (GCA) and significantly decreased testicular cytokine levels. In addition, EGCG was able to down regulate the mRNA expression of iNOS, MCP-1 and pro-apoptosis genes in favor of cell survival. For the first time we show that in vivo EGCG treatment rescued the torsed testes from IRI-induced inflammation, GCA and damage to spermatogenesis thus suggesting a new preventive approach to inhibiting the inflammatory and apoptotic consequences of TT-induced IRI.


Assuntos
Apoptose/efeitos dos fármacos , Catequina/análogos & derivados , Orquite/prevenção & controle , Traumatismo por Reperfusão/prevenção & controle , Túbulos Seminíferos/irrigação sanguínea , Torção do Cordão Espermático/complicações , Animais , Catequina/uso terapêutico , Ensaio de Imunoadsorção Enzimática , Interleucina-1beta/análise , Interleucina-6/análise , Masculino , Óxido Nítrico Sintase Tipo II , Orquite/etiologia , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Receptores CCR2/biossíntese , Traumatismo por Reperfusão/etiologia , Fator de Necrose Tumoral alfa/análise , Proteína Supressora de Tumor p53/biossíntese , Proteína X Associada a bcl-2/biossíntese
15.
Med Princ Pract ; 21(3): 295-7, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22236881

RESUMO

OBJECTIVE: To compare the diagnostic performance of urine cytology (UC), survivin mRNA expression, and the NMP22 BladderChek® (NMP22BC) test for the detection, grading and staging of transitional cell carcinoma (TCC) of the bladder. MATERIALS AND METHODS: Voided urine samples collected from 25 healthy controls and 80 patients diagnosed with TCC of the bladder were subjected to UC, the NMP22BC test and reverse-transcription real-time PCR for survivin mRNA expression. RESULTS: Survivin mRNA expression showed the highest sensitivity (87.5%) followed by the NMP22BC test (61.3%) while UC exhibited the lowest sensitivity (40%). All three urine markers had a similar specificity of 96% (95% CI 80.5-99.3%). Survivin mRNA expression was the only urine marker that showed a significant difference in relation to tumour histological grade (χ(2) 8.5, p = 0.015). None of the three urine markers was significantly related to tumour pathological stages. CONCLUSION: The diagnostic sensitivity of urinary survivin mRNA expression was superior to that of UC and the NMP22BC test and correlates with tumour pathological grade but not stage.


Assuntos
Biomarcadores Tumorais/análise , Carcinoma de Células de Transição/urina , Proteínas Nucleares/metabolismo , Neoplasias da Bexiga Urinária/urina , Bexiga Urinária/patologia , Adolescente , Adulto , Idoso , Antígenos de Neoplasias/análise , Antígenos de Neoplasias/metabolismo , Carcinoma de Células de Transição/diagnóstico , Carcinoma de Células de Transição/patologia , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Intervalos de Confiança , Citodiagnóstico , Estudos de Viabilidade , Feminino , Humanos , Masculino , Proteínas Associadas aos Microtúbulos , Pessoa de Meia-Idade , Proteínas Nucleares/análise , Valor Preditivo dos Testes , RNA Mensageiro/biossíntese , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/patologia , Adulto Jovem
16.
Med Princ Pract ; 20(5): 449-54, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21757935

RESUMO

OBJECTIVE: To evaluate the expression of the apoptotic genes survivin, Bax and Bcl-2 in vasectomized rabbits and to determine their relation with vasectomy-induced spermatogenic impairment and germ cell apoptosis. MATERIALS AND METHODS: Twelve adult rabbits (6-12 months old) were divided into three groups: sham control, unilateral vasectomy or bilateral vasectomy. Six months after vasectomy, testicular tissue was analyzed for germ cell apoptosis and DNA fragmentation by the TUNEL assay and gel electrophoresis, respectively. Spermatogenesis was assessed using the Johnsen score. The relative gene expression of survivin, Bax and Bcl-2 was measured using reverse transcription followed by real-time PCR. RESULTS: Compared to sham animals, a significant decrease in testicular survivin mRNA levels was measured in the two vasectomy animal groups (p < 0.05). This was accompanied by a significant increase in the Bax:Bcl-2 ratio in the vasectomized animals (p < 0.05). In addition, these data showed positive correlation with enhanced apoptotic index, damage to spermatogenesis and DNA fragmentation after vasectomy. CONCLUSION: These findings demonstrate that vasectomy-induced damage to spermatogenesis due to testicular apoptosis may be associated with survivin downregulation and Bax overexpression.


Assuntos
Inibidores de Cisteína Proteinase , Genes bcl-2/genética , Células Germinativas , Proteínas Inibidoras de Apoptose/biossíntese , Espermatogênese , Vasectomia/efeitos adversos , Animais , Expressão Gênica , Proteínas Inibidoras de Apoptose/antagonistas & inibidores , Proteínas Inibidoras de Apoptose/metabolismo , Masculino , RNA Mensageiro , Coelhos , Survivina , Testículo/citologia
17.
Cancer Res ; 71(4): 1334-43, 2011 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-21303975

RESUMO

Raf kinase inhibitory protein (RKIP) is a physiologic inhibitor of c-RAF kinase and nuclear factor κB signaling that represses tumor invasion and metastasis. Glycogen synthase kinase-3ß (GSK3ß) suppresses tumor progression by downregulating multiple oncogenic pathways including Wnt signaling and cyclin D1 activation. Here, we show that RKIP binds GSK3 proteins and maintains GSK3ß protein levels and its active form. Depletion of RKIP augments oxidative stress-mediated activation of the p38 mitogen activated protein kinase, which, in turn, inactivates GSK3ß by phosphorylating it at the inhibitory T390 residue. This pathway de-represses GSK3ß inhibition of oncogenic substrates causing stabilization of cyclin D, which induces cell-cycle progression and ß-catenin, SNAIL, and SLUG, which promote epithelial to mesenchymal transition. RKIP levels in human colorectal cancer positively correlate with GSK3ß expression. These findings reveal the RKIP/GSK3 axis as both a potential therapeutic target and a prognosis-based predictor of cancer progression.


Assuntos
Quinase 3 da Glicogênio Sintase/fisiologia , Proteína de Ligação a Fosfatidiletanolamina/fisiologia , Animais , Carcinoma/genética , Carcinoma/metabolismo , Carcinoma/patologia , Células Cultivadas , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Progressão da Doença , Estabilidade Enzimática , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Camundongos , Camundongos Knockout , Estresse Oxidativo/genética , Estresse Oxidativo/fisiologia , Proteína de Ligação a Fosfatidiletanolamina/genética , Proteína de Ligação a Fosfatidiletanolamina/metabolismo , Fosforilação , Ligação Proteica/fisiologia , Transdução de Sinais/genética , Regulação para Cima/fisiologia
18.
Biochem Biophys Res Commun ; 395(3): 342-7, 2010 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-20380811

RESUMO

Testicular torsion is associated with damage to the testicular tissue as a result of ischemia-reperfusion injury (IRI) and induction of apoptosis leading to progressive damage to spermatogenesis. Survivin is suggested to be an important regulator in the control of the mitochondrial apoptotic pathway, although its role in torsion-induced IRI is unknown. Therefore, we sought to evaluate testicular survivin expression after long term IRI induced by testicular torsion. Survivin expression was measured by real-time PCR in 6-12 month old New Zealand white rabbits divided into three groups (4 animals/group): group (A) sham control, group (B) ischemia alone for 60 min and group (C) ischemia for 60 min followed by reperfusion for 6 months. Germ cell apoptosis was evaluated by TUNEL assay, Bax/Bcl-2 ratio and DNA fragmentation. The Johnsen score was used to assess testicular morphological damage, while lipid peroxidation was used as an indicator for oxidative stress. Survivin expression was detected in all testicular tissue samples. The rate of survivin expression after IRI was significantly higher (p<0.05) compared with ischemic only and sham control testes. Its expression in IRI samples was inversely correlated with the significant increase (p<0.05) in apoptosis, oxidative levels and spermatogenic damage. In conclusion, down-regulation of testicular survivin expression after long term IRI to the testis and its association with apoptosis induction suggests its involvement in the regulation of this apoptotic pathway. These findings also identify survivin as a potential new target for the prevention of germ cell death during testicular torsion.


Assuntos
Apoptose , Proteínas Associadas aos Microtúbulos/metabolismo , Traumatismo por Reperfusão/patologia , Torção do Cordão Espermático/patologia , Espermatozoides/patologia , Testículo/irrigação sanguínea , Animais , Regulação para Baixo , Masculino , Proteínas Associadas aos Microtúbulos/genética , Coelhos , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/metabolismo , Torção do Cordão Espermático/complicações , Torção do Cordão Espermático/metabolismo , Espermatozoides/metabolismo
19.
Pharmacol Res ; 59(4): 263-8, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19166939

RESUMO

Angiotensin-(1-7) [Ang-(1-7)] is a vasodilator peptide with cardiac and vascular protective properties. We examined the influence of Ang-(1-7), both endogenous and after chronic treatment with the peptide (576microg/(kgday)), on ischemia/reperfusion (I/R)-induced cardiac dysfunction in streptozotocin-treated spontaneously hypertensive rats (diabetic SHR). In isolated perfused hearts, recovery of left ventricular function from 40min of global ischemia was improved significantly in Ang-(1-7)- or captopril-treated diabetic SHR and worsened in animals treated with A779, an Ang-(1-7) receptor (AT((1-7))) antagonist. The beneficial effect of captopril on cardiac recovery was reduced when co-administered with A779. Cardiac NF-kappaB activity appears to be higher in diabetic SHR and treatment with Ang-(1-7) or captopril decreased NF-kappaB activity in diabetic SHR, an effect partially reversed by co-administration of A779. Real-time PCR-based gene array analysis of cardiac tissue revealed that Ang-(1-7) or captopril treatment may reduce expression of several genes of inflammation involved in the NF-kappaB signalling pathway. The data provide for the first time a role for endogenous Ang-(1-7) as well as confirmation that exogenous treatment with the peptide produces cardioprotection. Whether potential anti-inflammatory and transcriptional factor changes are directly linked to the cardioprotection produced by Ang-(1-7) in diabetic SHR remains to be determined.


Assuntos
Angiotensina I/fisiologia , Angiotensina I/uso terapêutico , Coração/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Fragmentos de Peptídeos/fisiologia , Fragmentos de Peptídeos/uso terapêutico , Angiotensina I/administração & dosagem , Angiotensina I/antagonistas & inibidores , Angiotensina II/administração & dosagem , Angiotensina II/análogos & derivados , Angiotensina II/uso terapêutico , Antagonistas de Receptores de Angiotensina , Animais , Pressão Sanguínea/efeitos dos fármacos , Captopril/administração & dosagem , Captopril/uso terapêutico , Diabetes Mellitus Experimental/fisiopatologia , Quimioterapia Combinada , Coração/fisiopatologia , Técnicas In Vitro , Masculino , Miocárdio/enzimologia , NF-kappa B/metabolismo , Proteínas Nucleares/metabolismo , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/antagonistas & inibidores , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Receptores de Angiotensina/agonistas , Transdução de Sinais/efeitos dos fármacos
20.
Am J Physiol Heart Circ Physiol ; 292(1): H666-72, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17213482

RESUMO

The aim of this study was to test the hypothesis that treatment with angiotensin-(1-7) [ANG-(1-7)] or ANG-(1-7) nonpeptide analog AVE-0991 can produce protection against diabetes-induced cardiovascular dysfunction. We examined the influence of chronic treatment (4 wk) with ANG-(1-7) (576 microg.kg(-1).day(-1) ip) or AVE-0991 (576 microg.kg(-1).day(-1) ip) on proteinuria, vascular responsiveness of isolated carotid and renal artery ring segments and mesenteric bed to vasoactive agonists, and cardiac recovery from ischemia-reperfusion in streptozotocin-treated rats (diabetes). Animals were killed 4 wk after induction of diabetes and/or treatment with ANG-(1-7) or AVE-0991. There was a significant increase in urine protein (231 +/- 2 mg/24 h) in diabetic animals compared with controls (88 +/- 6 mg/24 h). Treatment of diabetic animals with ANG-(1-7) or AVE-0991 resulted in a significant reduction in urine protein compared with vehicle-treated diabetic animals (183 +/- 16 and 149 +/- 15 mg/24 h, respectively). Treatment with ANG-(1-7) or AVE-0991 also prevented the diabetes-induced abnormal vascular responsiveness to norepinephrine, endothelin-1, angiotensin II, carbachol, and histamine in the perfused mesenteric bed and isolated carotid and renal arteries. In isolated perfused hearts, recovery of left ventricular function from 40 min of global ischemia was significantly better in ANG-(1-7)- or AVE-0991-treated animals. These results suggest that activation of ANG-(1-7)-mediated signal transduction could be an important therapeutic strategy to reduce cardiovascular events in diabetic patients.


Assuntos
Angiotensina I/administração & dosagem , Diabetes Mellitus Experimental/tratamento farmacológico , Angiopatias Diabéticas/prevenção & controle , Angiopatias Diabéticas/fisiopatologia , Fragmentos de Peptídeos/administração & dosagem , Disfunção Ventricular Esquerda/prevenção & controle , Disfunção Ventricular Esquerda/fisiopatologia , Animais , Pressão Sanguínea/efeitos dos fármacos , Cardiotônicos/administração & dosagem , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/fisiopatologia , Angiopatias Diabéticas/etiologia , Relação Dose-Resposta a Droga , Masculino , Ratos , Ratos Wistar , Estreptozocina , Vasoconstrição/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Disfunção Ventricular Esquerda/etiologia
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