RESUMO
PURPOSE: Benign Prostatic Hyperplasia (BPH) is a result of prostate inflammation, frequently occurring in metabolic syndrome (MetS). Low testosterone is common in MetS. A randomized clinical trial was designed to evaluate if 24 weeks of testosterone therapy (TTh) in BPH men with MetS and low testosterone improve urinary symptoms and prostate inflammation. METHODS: One-hundred-twenty men with MetS waitlisted for BPH surgery were enrolled. They were categorized into normal testosterone (TT ≥ 12 nmol/L and cFT ≥ 225 pmol/L; n = 48) and testosterone deficient (TD) (TT < 12 nmol/L and/or cFT < 225 pmol/L; n = 72) then randomized to testosterone gel 2% (5 g/daily) or placebo for 24 weeks. At baseline and follow-up, questionnaires for urinary symptoms and trans-rectal ultrasound were performed. Prostate tissue was collected for molecular and histopathological analyses. RESULTS: No differences in the improvement of urinary symptoms were found between TTh and placebo (OR [95% CI] 0.96 [0.39; 2.37]). In TD + TTh, increase in prostate but not adenoma volume was observed (2.64 mL [0.07; 5.20] and 1.82 mL [- 0.46; 0.41], respectively). Ultrasound markers of inflammation were improved. In a subset of 61 men, a hyper-expression of several pro-inflammatory genes was found in TD + placebo when compared with normal testosterone. TTh was able to counteract this effect. For 80 men, the inflammatory infiltrate was higher in TD + placebo than in normal testosterone (0.8 points [0.2; 1.4]) and TD + TTh men (0.9 points [0.2; 1.5]). CONCLUSIONS: Twenty-four weeks of TTh in TD men with BPH and MetS improves ultrasound, molecular and histological proxies of prostate inflammation. This does not result in symptom improvement.
Assuntos
Sintomas do Trato Urinário Inferior , Síndrome Metabólica , Hiperplasia Prostática , Prostatite , Biomarcadores , Humanos , Inflamação/tratamento farmacológico , Sintomas do Trato Urinário Inferior/tratamento farmacológico , Sintomas do Trato Urinário Inferior/etiologia , Masculino , Síndrome Metabólica/tratamento farmacológico , Hiperplasia Prostática/tratamento farmacológico , Hiperplasia Prostática/metabolismo , Testosterona/uso terapêuticoRESUMO
PURPOSE: Female sexual response involves a complex interplay between neurophysiological mechanisms and the nitric oxide (NO)-mediated relaxation of clitoris and vagina. The aim of this study was to evaluate sex steroids regulation of the relaxant pathway in vagina, using a validated animal model. METHODS: Subgroups of OVX Sprague-Dawley rats were treated with 17ß-estradiol, testosterone, or testosterone and letrozole, and compared with a group of intact animals. Masson's trichrome staining was performed for morphological evaluation of the distal vaginal wall, in vitro contractility studies investigated the effect of OVX and in vivo treatments on vaginal smooth muscle activity. RNA from vaginal tissue was analyzed by semi-quantitative RT-PCR. RESULTS: Immunohistochemical analysis showed that OVX induced epithelial and smooth muscle structural atrophy, testosterone and testo + letrozole increased the muscle bundles content and organization without affecting the epithelium while 17ß-estradiol mediated the opposite effects. In vitro contractility studies were performed on noradrenaline pre-contracted vaginal strips from each experimental group. Acetylcholine (0.001-10 µM) stimulation induced a concentration-dependent relaxation, significantly reduced by NO-synthase inhibitor L-NAME and by guanylate cyclase inhibitor ODQ. OVX resulted in a decreased responsiveness to acetylcholine, restored by testosterone, with or without letrozole, but not by 17ß-estradiol. OVX sensitivity to the NO-donor SNP was higher than in the control. Vardenafil, a PDE5 inhibitor, enhanced SNP effect in OVX + testosterone as well as in control, as supported by RNA expression analysis. CONCLUSIONS: Our study demonstrates that testosterone improves the NO-mediated smooth muscle vaginal cells relaxation confirming its role in maintaining the integrity of muscular relaxant machinery.
Assuntos
Acetilcolina , Óxido Nítrico , Animais , Estradiol/farmacologia , Feminino , Humanos , Letrozol/farmacologia , Óxido Nítrico/metabolismo , Ovariectomia , RNA , Ratos , Ratos Sprague-Dawley , Testosterona/farmacologia , Vagina/metabolismoRESUMO
Androgen deprivation therapy (ADT) has a deleterious effect on sexual functions and general well-being in men. Despite this evidence, however, patient and couple knowledge about ADT side effects as well as their management is poor. Similar considerations can be made for physician endorsement of management strategies. In this paper, we summarize and critically discuss available evidence regarding the possible associations between ADT and sexual dysfunction as well as the best therapeutical options. Preclinical data show that ADT is associated with penile contractility impairment as well as lower response to phosphodiesterase type 5 inhibitors (PDE5i). Available data indicate that ADT resulted in a five to sixfold increased risk of reduced libido and in a threefold increased risk of ED confirming the main role of testosterone in regulating sexual desire. Despite this evidence, sexuality remains an important aspect of health and well-being for men and their partner. The best therapeutical options depend on patient and couple desires and needs. When nonpenetrative erections are still possible, nonpenetrative activities should be encouraged to maintain sexual intimacy. A combined and personal educational program including the collaboration of different professional figures (including general physicians, oncologists, andrologists, sexologists, and psychologists) trained in sexual medicine is advisable in order to provide the best support to subjects undergoing ADT.
Assuntos
Neoplasias da Próstata , Disfunções Sexuais Fisiológicas , Antagonistas de Androgênios/efeitos adversos , Androgênios , Humanos , Libido , Masculino , Disfunções Sexuais Fisiológicas/induzido quimicamenteRESUMO
PURPOSE: In both preclinical and clinical settings, testosterone treatment (TTh) of hypogonadism has shown beneficial effects on insulin sensitivity and visceral and liver fat accumulation. This prospective, observational study was aimed at assessing the change in markers of fat and liver functioning in obese men scheduled for bariatric surgery. METHODS: Hypogonadal patients with consistent symptoms (n = 15) undergoing 27.63 ± 3.64 weeks of TTh were compared to untreated eugonadal (n = 17) or asymptomatic hypogonadal (n = 46) men. A cross-sectional analysis among the different groups was also performed, especially for data derived from liver and fat biopsies. Preadipocytes isolated from adipose tissue biopsies were used to evaluate insulin sensitivity, adipogenic potential and mitochondrial function. NAFLD was evaluated by triglyceride assay and by calculating NAFLD activity score in liver biopsies. RESULTS: In TTh-hypogonadal men, histopathological NAFLD activity and steatosis scores, as well as liver triglyceride content were lower than in untreated-hypogonadal men and comparable to eugonadal ones. TTh was also associated with a favorable hepatic expression of lipid handling-related genes. In visceral adipose tissue and preadipocytes, TTh was associated with an increased expression of lipid catabolism and mitochondrial bio-functionality markers. Preadipocytes from TTh men also exhibited a healthier morpho-functional phenotype of mitochondria and higher insulin-sensitivity compared to untreated-hypogonadal ones. CONCLUSIONS: The present data suggest that TTh in severely obese, hypogonadal individuals induces metabolically healthier preadipocytes, improving insulin sensitivity, mitochondrial functioning and lipid handling. A potentially protective role for testosterone on the progression of NAFLD, improving hepatic steatosis and reducing intrahepatic triglyceride content, was also envisaged. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02248467, September 25th 2014.
Assuntos
Hipogonadismo , Gordura Intra-Abdominal , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado , Hepatopatia Gordurosa não Alcoólica , Obesidade , Testosterona , Adulto , Biópsia/métodos , Estudos Transversais , Humanos , Hipogonadismo/diagnóstico , Hipogonadismo/tratamento farmacológico , Hipogonadismo/epidemiologia , Resistência à Insulina , Gordura Intra-Abdominal/efeitos dos fármacos , Gordura Intra-Abdominal/metabolismo , Gordura Intra-Abdominal/patologia , Itália/epidemiologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Obesidade/diagnóstico , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/farmacocinética , Testosterona/administração & dosagem , Testosterona/farmacocinética , Resultado do TratamentoRESUMO
BACKGROUND: Activation of the farnesoid X receptor (FXR), a member of the nuclear receptor steroid superfamily, leads to anti-inflammatory and anti-fibrotic effects in several tissues, including the lung. We have recently demonstrated a protective effect of the farnesoid X receptor (FXR) agonist obeticholic acid (OCA) in rat models of monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) and bleomycin-induced pulmonary fibrosis. The aim of the present study was to investigate whether the positive effects of OCA treatment could be exerted also in established MCT-induced PAH, i.e., starting treatment 2 weeks after MCT administration. METHODS: Rats with MCT-induced PAH were treated, 2 weeks after MCT administration, with OCA or tadalafil for two additional weeks. Pulmonary functional tests were performed at week 2 (before treatment) and four (end of treatment). At the same time points, lung morphological features and expression profile of genes related to smooth muscle relaxation/contraction and tissue remodeling were also assessed. RESULTS: 2 weeks after MCT-induced injury, the treadmill resistance (a functional parameter related to pulmonary hypertension) was significantly decreased. At the same time point, we observed right ventricular hypertrophy and vascular remodeling, with upregulation of genes related to inflammation. At week 4, we observed a further worsening of the functional and morphological parameters, accompanied by dysregulation of inflammatory and extracellular matrix markers mRNA expression. Administration of OCA (3 or 10 mg/kg/day), starting 2 weeks after MCT-induced injury, significantly improved pulmonary function, effectively normalizing the exercise capacity. OCA also reverted most of the lung alterations, with a significant reduction of lung vascular wall thickness, right ventricular hypertrophy, and restoration of the local balance between relaxant and contractile pathways. Markers of remodeling pathways were also normalized by OCA treatment. Notably, results with OCA treatment were similar, or even superior, to those obtained with tadalafil, a recently approved treatment for pulmonary hypertension. CONCLUSIONS: The results of this study demonstrate a significant therapeutic effect of OCA in established MCT-induced PAH, improving exercise capacity associated with reduction of right ventricular hypertrophy and lung vascular remodeling. Thus, OCA dosing in a therapeutic protocol restores the balance between relaxant and contractile pathways in the lung, promoting cardiopulmonary protective actions in MCT-induced PAH.
Assuntos
Ácido Quenodesoxicólico/análogos & derivados , Modelos Animais de Doenças , Hipertensão Pulmonar/tratamento farmacológico , Monocrotalina/toxicidade , Fibrose Pulmonar/tratamento farmacológico , Receptores Citoplasmáticos e Nucleares/agonistas , Animais , Antibióticos Antineoplásicos/toxicidade , Bleomicina/toxicidade , Ácido Quenodesoxicólico/farmacologia , Hipertensão Pulmonar/induzido quimicamente , Hipertensão Pulmonar/patologia , Masculino , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Ratos , Ratos Sprague-DawleyRESUMO
PURPOSE: We recently demonstrated a protective effect of the farnesoid X receptor agonist obeticholic acid (OCA) in rat models of bleomycin-induced pulmonary fibrosis (PF). Aim of the present study was to investigate whether the positive effects of OCA treatment are apparent also on ongoing bleomycin-induced PF, i.e., after 2 weeks of bleomycin administration. METHODS: Bleomycin-induced PF rats were treated 2 weeks after bleomycin administration with OCA or pirfenidone for two additional weeks. Pulmonary function test was performed at 2 and 4 weeks in all experimental groups. At the same time points, lung morphological features and mRNA expression profile of genes related to fibrosis, inflammation and epithelial-mesenchymal transition were also assessed. RESULTS: After 2 weeks, bleomycin significantly increased the pressure at the airway opening (PAO), a functional parameter related to fibrosis-induced lung stiffness, and induced diffuse lung interstitium fibrosis, with upregulation of inflammation (IL1ß, MCP1) and tissue remodeling (COL1A1, COL3A1, ET1, MMP7, PDGFa, αSMA, SNAI1) markers. At week four, a further increase of lung fibrosis and PAO was observed, accompanied by upregulation of extracellular matrix-related mRNA expression. OCA administration, even after the establishment of PF, significantly improved pulmonary function, normalizing PAO, and reverted the bleomycin-induced lung alterations, with significant reduction of markers of inflammation (CD206, COX2, HIF1, IL1ß, MCP1), epithelial proliferation (CTGF, PDGFa) and fibrosis (COL1A1, COL3A1, ET1, FN1, MMPs, αSMA, SNAIs, TGFß1, TIMPs). Results with OCA were similar or superior to those obtained with pirfenidone. CONCLUSIONS: In conclusion, our results demonstrate a significant therapeutic effect of OCA in already established PF.
Assuntos
Biomarcadores/metabolismo , Bleomicina/toxicidade , Ácido Quenodesoxicólico/análogos & derivados , Modelos Animais de Doenças , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Fibrose Pulmonar/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/toxicidade , Ácido Quenodesoxicólico/farmacologia , Masculino , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Ratos , Ratos Sprague-DawleyRESUMO
It has been shown, in animal models, that gastrointestinal tract (GIT) motility is influenced by temperature; nevertheless, the basic mechanism governing thermal GIT smooth muscle responses has not been fully investigated. Studies based on physiologically tuned mathematical models have predicted that thermal inhomogeneity may induce an electrochemical destabilization of peristaltic activity. In the present study, the effect of thermal cooling on human colonic muscle strip (HCMS) contractility was studied. HCMSs were obtained from disease-free margins of resected segments for cancer. After removal of the mucosa and serosa layers, strips were mounted in separate chambers. After 30 min, spontaneous contractions developed, which were measured using force displacement transducers. Temperature was changed every hour (37, 34, and 31°C). The effect of cooling was analyzed on mean contractile activity, oscillation amplitude, frequency, and contraction to ACh (10(-5) M). At 37°C, HCMSs developed a stable phasic contraction (~0.02 Hz) with a significant ACh-elicited mean contractile response (31% and 22% compared with baseline in the circular and longitudinal axis, respectively). At a lower bath temperature, higher mean contractile amplitude was observed, and it increased in the presence of ACh (78% and 43% higher than the basal tone in the circular and longitudinal axis, respectively, at 31°C). A simplified thermochemomechanical model was tuned on experimental data characterizing the stress state coupling the intracellular Ca(2+) concentration to tissue temperature. In conclusion, acute thermal cooling affects colonic muscular function. Further studies are needed to establish the exact mechanisms involved to better understand clinical consequences of hypothermia on intestinal contractile activity.
Assuntos
Temperatura Baixa , Colo/fisiologia , Motilidade Gastrointestinal , Modelos Biológicos , Contração Muscular , Músculo Liso/fisiologia , Acetilcolina/farmacologia , Idoso , Cálcio/metabolismo , Resposta ao Choque Frio , Colo/efeitos dos fármacos , Colo/metabolismo , Feminino , Motilidade Gastrointestinal/efeitos dos fármacos , Humanos , Técnicas In Vitro , Masculino , Mecanotransdução Celular , Pessoa de Meia-Idade , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/metabolismo , Fatores de TempoRESUMO
BACKGROUND: BPH and LUTS have been associated to obesity, hypogonadism, and metabolic syndrome (MetS). MetS-induced prostate and bladder alterations, including inflammation and tissue remodeling, have been related to a low-testosterone and high-estrogen milieu. In addition to ERs, GPR30/GPER is able to mediate several estrogenic non-genomic actions. METHODS: Supplementing a subgroup of MetS rabbits with tamoxifen, we analyzed the in vivo effects on MetS-induced prostate and bladder alterations. The effects of selective ER/GPER ligands and GPER silencing on prostate inflammation were also studied in vitro using hBPH cells. RESULTS: ERα, ERß, and PR expression was upregulated in MetS bladder, where tamoxifen decreased ERα and PR expression, further stimulating ERß. In addition, tamoxifen-dosing decreased MetS-induced overexpression of inflammatory and tissue remodeling genes. In prostate, sex steroid receptors, pro-inflammatory and pro-fibrotic genes were upregulated in MetS. However, tamoxifen did not affect them and even increased COX-2. In hBPH cells, 17ß-estradiol increased IL-8 secretion, an effect blunted by co-treatment with GPER antagonist G15 but not by ER antagonist ICI 182,780, which further increased it. GPER agonist G1 dose-dependently (IC50 = 1.6 nM) induced IL-8 secretion. In vitro analysis demonstrated that GPER silencing reverted these stimulatory effects. CONCLUSIONS: GPER can be considered the main mediator of estrogen action in prostate, whereas in bladder the mechanism appears to rely on ERα, as indicated by in vivo experiments with tamoxifen dosing. Limiting the effects of the MetS-induced estrogen action via GPER could offer new perspectives in the management of BPH/LUTS, whereas tamoxifen dosing showed potential benefits in bladder.
Assuntos
Síndrome Metabólica/metabolismo , Próstata/metabolismo , Receptores de Estrogênio/fisiologia , Receptores Acoplados a Proteínas G/fisiologia , Moduladores Seletivos de Receptor Estrogênico/farmacologia , Tamoxifeno/farmacologia , Bexiga Urinária/metabolismo , Animais , Linhagem Celular , Cromossomos Humanos Par 11/genética , Cromossomos Humanos Par 8/genética , Humanos , Masculino , Síndrome Metabólica/tratamento farmacológico , Ligação do Par , Próstata/efeitos dos fármacos , Coelhos , Moduladores Seletivos de Receptor Estrogênico/uso terapêutico , Tamoxifeno/uso terapêutico , Bexiga Urinária/efeitos dos fármacosRESUMO
Puberty is the developmental period during which rapid somatic changes and attainment of reproductive capacity take place. The sine qua non event for the onset of puberty is the increase in pulsatile GnRH release from GnRH neurons. GnRH neurons originate in the nasal placode and migrate, from the nasal compartment through the basal forebrain, before they attain their positions in the hypothalamus. Failure of GnRH neurons to migrate, mainly due to genetic alterations, leads to a clinical condition defined congenital hypogonadotropic hypogonadism. Other important factors demonstrated a permissive role for GnRH secretion at time of puberty, such as the kisspeptin G-protein-coupled receptor 54 (GPR54, now called KISS-1R) and its ligand, as well as neurokinin B (NKB) and the neurokinin 3 (NK3) receptor. Kisspeptin and neurokinin B colocalized in a subset of hypothalamic neurons that regulate GnRH secretion by GnRH neurons. Indeed, loss of function mutations in the above-mentioned and other genes result in hypogonadotrophic hypogonadism with absence of pubertal development. In males, pubertal increase of androgens levels seems to be required for the attainment of a normal bone density and male-specific body composition. However, also genetic variants of genes involved in bone metabolism as well as in osteoblast/osteoclast activation are associated to bone mineral density.
Assuntos
Androgênios/uso terapêutico , Composição Corporal/efeitos dos fármacos , Densidade Óssea/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/fisiologia , Neurônios/fisiologia , Puberdade/fisiologia , Animais , Movimento Celular , Hormônio Liberador de Gonadotropina/metabolismo , Humanos , Hipogonadismo/genética , Kisspeptinas , Leptina/fisiologia , Masculino , Receptores Acoplados a Proteínas G/fisiologia , Receptores de Kisspeptina-1 , Proteínas Supressoras de Tumor/fisiologiaRESUMO
Polycyclic aromatic hydrocarbons (PAH) such as dibenzo[a,l]pyrene (DBP) are wide-spread environmental pollutants most probably mutagenic and carcinogenic to humans. Detailed data on the cytogenetic effects of anti-11,12-dihydroxy-13,14-epoxy-11,12,13,14-tetrahydrodibenzo[a,l]pyrene (DBPDE) in mammalian cells are not available in the literature. The aim of this study is to elucidate the mechanisms involved in the induction of chromosomal aberrations and sister chromatid exchanges (SCEs) by DBPDE in mammalian cells. In order to achieve this a parental (AA8) and different DNA repair-deficient Chinese hamster ovary cell lines such as UV4, UV5, UV61 (nucleotide excision repair, NER), EM9 (base excision repair, BER), irs1SF (homologous recombination repair, HRR) and V3-3 (non-homologous end joining, NHEJ) were used. The most sensitive cell lines for DBPDE-induced chromosome aberrations were EM9 and irs1SF, while EM9 and V3-3 cell lines were the most sensitive in terms of SCEs induction. It can be suggested that the BER pathway plays an important role in the repair of lesions induced by DBPDE, affecting both chromosomal aberrations and SCEs induction. Moreover, the HRR pathway seems to play a role in cellular resistance to DBPDE mainly in terms of chromosomal aberration induction while the NHEJ pathway takes part affecting only the induction of SCEs.
Assuntos
Benzopirenos/farmacologia , Dano ao DNA/efeitos dos fármacos , Reparo do DNA , Poluentes Ambientais/farmacologia , Compostos de Epóxi/farmacologia , Animais , Células CHO , Aberrações Cromossômicas , Cricetinae , CricetulusRESUMO
BACKGROUND: Chronic inflammation is now considered a determinant of benign prostatic hyperplasia (BPH), promoting, together with the hormonal milieu, prostate overgrowth and lower urinary tract symptoms (LUTS). Prostatic urethra actively participates in determining progression of LUTS associated with BPH. AIM: To investigate the expression of the vitamin D receptor (VDR) and the ability of the VDR agonist elocalcitol to reduce inflammatory responses in human prostatic urethra (hPU) cells. MATERIALS AND METHODS: Human prostatic urethra, prostate and bladder neck were obtained from patients affected by BPH. Immunohistochemical studies for VDR expression were performed in tissue samples, from which primary cell cultures were also derived. In hPU cells, proliferation and chemiotaxis were studied, along with Rho kinase (ROCK) activity (MYPT-1 phosphorylation) by western blot. Quantitative RT-PCR was performed for VDR, cyclooxygenase (COX-2), and interleukin (IL)-8 expression. RESULTS: Urethra displays higher VDR expression compared to prostate and bladder neck tissues. The VDR agonist elocalcitol partially reverts COX-2 and IL-8 mRNA upregulation induced by a pro-inflammatory cytokine mixture (IL-17, interferon-γ, tumor necrosis factor-α) and inhibits cell migration in urethral cells. Elocalcitol prevents activation of ROCK, as previously demonstrated in bladder and prostate cell cultures. CONCLUSIONS: Our results suggest that prostatic urethra is, within the lower urinary tract, a novel target for VDR agonists, as shown by the capacity of elocalcitol to inhibit ROCK activity and to limit inflammatory responses in human primary urethra cells.
Assuntos
Calcitriol/análogos & derivados , Próstata/metabolismo , Receptores de Calcitriol/agonistas , Receptores de Calcitriol/genética , Uretra/metabolismo , Idoso , Calcitriol/farmacologia , Técnicas de Cultura de Células , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Avaliação Pré-Clínica de Medicamentos , Expressão Gênica , Humanos , Ligantes , Masculino , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Próstata/efeitos dos fármacos , Próstata/patologia , Hiperplasia Prostática/genética , Hiperplasia Prostática/metabolismo , Hiperplasia Prostática/patologia , Hiperplasia Prostática/cirurgia , Receptores de Calcitriol/metabolismo , Uretra/efeitos dos fármacos , Uretra/patologia , Bexiga Urinária/metabolismo , Bexiga Urinária/patologiaRESUMO
Paralytic ileus is a temporary syndrome with impairment of peristalsis and no passage of food through the intestine. Although improvements in supportive measures have been achieved, no therapy useful to specifically reduce or eliminate the motility disorder underlying postoperative ileus has been developed yet. In this paper, we draw a plausible, physiologically fine-tuned scenario, which explains a possible cause of paralytic ileus. To this aim we extend the existing 1D intestinal electrophysiological Aliev-Richards-Wikswo ionic model based on a double-layered structure in two and three dimensions. Thermal coupling is introduced here to study the influence of temperature gradients on intestine tissue which is an important external factor during surgery. Numerical simulations present electrical spiral waves similar to those experimentally observed already in the heart, brain and many other excitable tissues. This fact seems to suggest that such peculiar patterns, here electrically and thermally induced, may play an important role in clinically experienced disorders of the intestine, then requiring future experimental analyses in the search for possible implications for medical and physiological practice and bioengineering.
Assuntos
Pseudo-Obstrução Intestinal/fisiopatologia , Intestinos/fisiologia , Modelos Biológicos , Temperatura Corporal/fisiologia , Simulação por Computador , Eletrofisiologia , Humanos , Pseudo-Obstrução Intestinal/etiologia , Intestinos/anatomia & histologia , Intestinos/fisiopatologia , Modelos Anatômicos , Complicações Pós-Operatórias/fisiopatologiaRESUMO
Oxytocin (OT) is a neurohypophysial hormone with overall unclear physiological functions in the male. Several studies indicated that OT has a key role in the central regulation of penile erection. In this mini-review we summarize its possible involvement in another aspect of the male sexuality: the ejaculatory process. Because OT is released by posterior pituitary at the time of orgasm, we postulate that OT might help sperm progression during ejaculation. Our recent studies indicate that OT receptors (OTR) are present in rabbit and human epididymis and mediate contractility. Accordingly, they are immuno-localized in the smooth muscle cells of the epididymis. However, they are also present in the epithelial compartment of the tubules. In epididymal epithelial cells in culture, OT induces the release of another potent stimulator of epididymal contractility, endothelin-1 (ET-1), which most probably amplifies OT-induced contraction. Sex steroids regulate the density of OTR in epididymis. In fact, in an experimental model of hypogonadotropic hypogonadism (hypo) induced in rabbits, estrogens, but not androgens, fully restored OT-induced epididymal contractility, up-regulating OTR gene and protein expression. In addition, deprivation of endogenous estrogens, by blocking their formation using the aromatase inhibitor letrozole, induced OT hypo-responsiveness comparable to that observed in hypo rabbits. These findings suggest a new function of estrogens in the male: regulation of OT responsiveness in epididymis.
Assuntos
Ejaculação/fisiologia , Ocitocina/fisiologia , Animais , Estrogênios/fisiologia , Humanos , Masculino , Ereção Peniana/fisiologiaRESUMO
The nitric oxide synthase (NOS) pathway has been clearly demonstrated to regulate angiogenesis. Increased levels of NO correlate with tumour growth and spreading in different experimental and human cancers. Drugs interfering with the NOS pathway may be useful in angiogenesis-dependent tumours. The aim of this study was to pharmacologically characterise certain ruthenium-based compounds, namely NAMI-A, KP1339, and RuEDTA, as potential NO scavengers to be used as antiangiogenic/antitumour agents. NAMI-A, KP1339 and RuEDTA were able to bind tightly and inactivate free NO in solution. Formation of ruthenium-NO adducts was documented by electronic absorption, FT-IR spectroscopy and (1)H-NMR. Pretreatment of rabbit aorta rings with NAMI-A, KP1339 or RuEDTA reduced endothelium-dependent vasorelaxation elicited by acetylcholine. This effect was reversed by 8-Br-cGMP. The key steps of angiogenesis, endothelial cell proliferation and migration stimulated by vascular endothelial growth factor (VEGF) or NO donor drugs, were blocked by NAMI-A, KP1339 and RuEDTA, these compounds being devoid of any cytotoxic activity. When tested in vivo, NAMI-A inhibited angiogenesis induced by VEGF. It is likely that the antitumour properties previously observed for ruthenium-based NO scavengers, such as NAMI-A, are related to their NO-related antiangiogenic properties.
Assuntos
Inibidores da Angiogênese/farmacologia , Dimetil Sulfóxido/análogos & derivados , Dimetil Sulfóxido/farmacologia , Endotélio Vascular/fisiologia , Óxido Nítrico/fisiologia , Compostos Organometálicos/farmacologia , Compostos de Rutênio/farmacologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Vasos Coronários , Relação Dose-Resposta a Droga , Endotélio Vascular/efeitos dos fármacos , Sequestradores de Radicais Livres , Humanos , Óxido Nítrico Sintase/antagonistas & inibidores , Espectroscopia de Infravermelho com Transformada de Fourier , Vênulas/efeitos dos fármacos , Vênulas/fisiologiaRESUMO
Development and maintenance of penile erection requires the relaxation of the smooth muscle cells in the cavernous bodies and is essentially mediated by nitric oxide (NO). The penile flaccid state is conversely maintained by the alpha adrenergic neuroeffector system and by other vasoconstrictors, such as endothelin-1 (ET-1). In this study we examined the mechanisms involved in yohimbine-induced relaxation in human and rabbit corpora cavernosa (CC). We essentially found that yohimbine not only blocks contractions induced by adrenergic agonists, but also by non-adrenergic substances, such as ET-1. This effect was unrelated to antagonism at the level of ET receptors, because yohimbine did not affect ET-1-induced increase in intracellular calcium in isolated CC cells. Conversely, our data suggest that yohimbine counteracts ET-1-induced contractions by interfering with NO release from the endothelium. In fact, yohimbine-induced CC relaxation was inhibited by the mechanical removing of the endothelium and by blocking NO formation or signalling via guanylate cyclase and cGMP formation. Conversely, yohimbine activity was strongly increased by inhibiting cGMP degradation. In an experimental model of hypogonadism, performed on rabbits by chronic treatment with a long-lasting GnRH agonist, the relaxant yohimbine activity was also decreased, but completely restored by androgen supplementation. This effect was evident only in preparations in which the main source of NO was present (endothelium) or in which NO formation was not impaired by L-NAME. Our data indicate that the relaxant effect of yohimbine is both endothelium and androgen-dependent. This might justify the lack of efficacy of this drug in treatment of some form of organic erectile dysfunction.
Assuntos
Antagonistas Adrenérgicos alfa/farmacologia , Relaxamento Muscular/efeitos dos fármacos , Músculo Liso/fisiologia , Ereção Peniana/efeitos dos fármacos , Pênis/fisiologia , Ioimbina/farmacologia , Agonistas alfa-Adrenérgicos/farmacologia , Androgênios/deficiência , Animais , Antineoplásicos Hormonais/farmacologia , Células Cultivadas , Endotélio/fisiologia , Humanos , Hipogonadismo/induzido quimicamente , Hipogonadismo/fisiopatologia , Técnicas In Vitro , Masculino , Músculo Liso/citologia , Óxido Nítrico/metabolismo , Ereção Peniana/fisiologia , Pênis/citologia , Fenilefrina/farmacologia , Coelhos , Receptores Adrenérgicos alfa/metabolismo , Pamoato de Triptorrelina/farmacologiaRESUMO
Oxytocin (OT) is a neurohypophysial hormone with unclear physiological functions in the male. Several previous studies indicated that OT might have a role in the ejaculatory process, stimulating sperm release from the epididymal storage. In this study we investigated on the presence and function of OT receptor (OTR) in rabbit and human epididymis. By using RT-PCR, Western and binding studies, we found that OTR gene and protein is expressed in the human epididymis and stimulates in vitro contractility. The immunolocalization of OTR suggests that the receptor is not only present in the smooth muscle cells of the human epididymis but also in the epithelial compartment. Experiments performed in rabbit epididymal epithelial (rEE) cells in culture indicate that OT induces the release of an other potent stimulator of epididymal contractility, endothelin-1 (ET-1), Blocking the ET(A) subtype of the ET-1 receptors, by using a specific antagonist (BQ-123), partially counteracts the contractile effect of OT, suggesting positive interactions between the two peptides in regulating epididymal contractility. Finally, to investigate whether an acute OT administration increases sperm release also in humans, we treated oligozoospermic patients with an intravenous bolus of OT (2.5 IU), just before sperm collection. In a small, single blind study, we found that OT almost doubled sperm retrieval when compared with vehicle administration. Our results indicate that OT might have physiological functions also in the male, controlling epididymal motility and sperm progression through the male genital tract.
Assuntos
Epididimo/fisiologia , Receptores de Ocitocina/fisiologia , Adulto , Idoso , Animais , Ligação Competitiva , Endotelina-1/metabolismo , Endotelina-1/farmacologia , Epididimo/química , Epididimo/citologia , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Genitália Masculina/citologia , Genitália Masculina/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Contração Muscular/efeitos dos fármacos , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Oligospermia/tratamento farmacológico , Ocitocina/administração & dosagem , Ocitocina/farmacologia , RNA Mensageiro/metabolismo , Coelhos , Receptores de Ocitocina/análise , Receptores de Ocitocina/genética , Método Simples-Cego , Contagem de Espermatozoides , Distribuição TecidualRESUMO
We evaluated the synthesis of nitric oxide (NO) and of the neurotoxic kynurenine metabolites 3OH-kynurenine and quinolinic acid (QUIN) in interferon-gamma (IFN-gamma)-activated macrophages of the murine BACl.2F5 cell line with the aim of investigating the roles of mononuclear phagocytes in inflammatory neurological disorders. IFN-gamma induced indoleamine 2,3-dioxygenase (IDO) and NO synthase (NOS) and increased the synthesis of 3OH-kynurenine, QUIN, and NO that accumulated in the incubation medium where they reached neurotoxic levels. Macrophage exposure to norharmane, an IDO inhibitor, resulted in a decreased formation of not only the kynurenine metabolites but also NO. The inhibition of NO synthesis could not be ascribed to reduced NADPH availability or decreased NOS induction. Norharmane inhibited NOS activity also in coronary vascular endothelial cells and in isolated aortic rings. Our findings suggest that activated macrophages release large amounts of neurotoxic molecules and that norharmane may represent a prototype compound to study macrophage involvement in inflammatory brain damage.
Assuntos
Interferon gama/farmacologia , Cinurenina/biossíntese , Ativação de Macrófagos , Macrófagos/fisiologia , Óxido Nítrico Sintase/fisiologia , Ácido Quinolínico/metabolismo , Triptofano Oxigenase/fisiologia , Animais , Linhagem Celular , Indolamina-Pirrol 2,3,-Dioxigenase , Ativação de Macrófagos/efeitos dos fármacos , Camundongos , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II , Triptofano Oxigenase/antagonistas & inibidoresRESUMO
We have demonstrated that adenosine has potent relaxant activity on the rabbit corpus cavernosum, acting through the A2a subtype receptor for adenosine. We now report studies on the identification and functional characterization of adenosine receptors in human penile vessels. To identify A2 receptors in human corpora cavernosa (HCC) we performed binding studies using the selective radioligand [125I]PAPA-APEC in membranes from HCC. We found the presence of a single class of high affinity (Kd= 0.23 +/- 0.06 nM), low capacity (Bmax=134 +/- 37 fmoles/mg protein) binding sites. Adenosine and CGS 21680 completely displaced [125I]PAPA-APEC binding (Kd= 146.7 +/- 64 microM and 51.52 +/- 27 nM, respectively). Accordingly, in functional studies adenosine relaxed phenylephrine precontracted HCC with an IC50=2.28 +/- 0.17 mM. The effect of adenosine was independent from nitric oxide (NO), and was counteracted by the A2 antagonist CGS 15943. In order to evaluate the in vivo effect of adenosine, increasing concentrations (6, 60, 600 microg) of adenosine or prostaglandin E1 (PGE1) (10 microg) were injected into the corpora cavernosa of four healthy volunteers. Blood flow and erectile response were evaluated at different times by duplex sonography and visual inspection, respectively. It was found that adenosine increased cavernosal peak blood flow velocity in a time- and dose-dependent manner. The highest concentrations of injected adenosine elicited a response that was not statistically different from that of PGE1 (10 microg). However, in contrast to PGE1, a full or partial erection was never obtained. To further investigate the lack of effect of adenosine on penile tumescence (despite the substantial increase in cavernosal blood flow), in vitro experiments were performed on human deep dorsal penile veins (DDPV) obtained from surgical ligation for impotence. Adenosine did not affect basal tone, but it induced almost complete relaxation in noradrenaline-precontracted vein strips with an IC50=1.6 +/- 0.22 mM. Conversely, PGE1 stimulated a sustained increase in basal tone. Therefore, the lack of effect of adenosine on penile tumescence could be due to a simultaneous relaxing activity on penile corpora cavernosa and veins. In conclusion, our study indicates that adenosine relaxes HCC as well as penile veins without affecting erection, at least at the concentrations we have used. Conversely, PGE1 relaxes corpora cavernosa as well as adenosine but strongly stimulates vein contraction, allowing penile tumescence.
Assuntos
Pênis/fisiologia , Receptores Purinérgicos P1/fisiologia , Adenosina/farmacologia , Adulto , Idoso , Estimulação Elétrica , Humanos , Masculino , Pênis/efeitos dos fármacos , Pênis/metabolismo , Receptor A2A de Adenosina , Receptores Purinérgicos P1/metabolismoRESUMO
Previous studies have shown that the light emitted by halogen tungsten lamps contains UV radiation in the UV-A, UV-B and UV-C regions, induces mutations and irreparable DNA damage in bacteria, enhances the frequency of micronuclei in cultured human lymphocytes and is potently carcinogenic to the skin of hairless mice. The present study showed that the light emitted by an uncovered, traditional halogen lamp induces a significant, dose-related and time-related increase not only in micronuclei but also in chromosome-type aberrations, such as breaks, and even more in chromatid-type aberrations, such as isochromatid breaks, exchanges and isochromatid/chromatid interchanges, all including gaps or not, in cultured human lymphocytes. All these genotoxic effects were completely prevented by shielding the same lamp with a silica glass cover, blocking UV radiation. A new model of halogen lamp, having the quartz bulb treated in order to reduce the output of UV radiation, was considerably less genotoxic than the uncovered halogen lamp, yet induction of chromosomal alterations was observed at high illuminance levels.
Assuntos
Iluminação/efeitos adversos , Linfócitos/efeitos dos fármacos , Micronúcleos com Defeito Cromossômico/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Adulto , Células Cultivadas , Aberrações Cromossômicas/genética , Halogênios , Humanos , Linfócitos/ultraestrutura , Masculino , Micronúcleos com Defeito Cromossômico/genética , TungstênioRESUMO
The effect of ATP in human and rabbit corpus cavernosum (CC) smooth muscle was investigated. Strips of human CC were vertically mounted in an organ bath and the tonic tension was recorded. ATP (0.1-3 mM) induced a concentration-dependent relaxant effect, with a pD2 value of 3.01+/-0.3. The purine-induced relaxation was not affected by L-NAME (100 microM). In rabbit CC, ATP also induced a concentration-dependent relaxation, which was not influenced by L-NAME or by indomethacin (3 microM), with a pD2 value of 3.1 +/-0.4. The ATP-induced relaxant effect in rabbit CC was increased by both the inhibitor of adenosine reuptake, dipyridamole (3 microM) and by the inhibitor of adenosine deaminase, EHNA (0.3 microM). Moreover CGS 15943 (3 microM), an A2a adenosine antagonist, reduced the ATP-induced relaxation. UTP was not able to produce relaxation. The two ATP analogues 2-methylthioATP and alpha,beta-methylene ATP were able to induce relaxation in rabbit CC, with the following order of potency: 2-methylthioATP > ATP > alpha,beta-methylene ATP thus suggesting a role for P2y receptors. However, reactive blue (500 microM), an unspecific P2y antagonist, did not modify the ATP relaxant response. The inhibition of phospholipase C by U73122 (3 microM) and of the endoplasmic reticulum Ca2+ATPase by thapsigargin (1 microM) did not modify the ATP-induced relaxation. The P2x specific antagonist PPADS (30 microM) and suramine (500 microM) were not able to modify the ATP relaxation either in the absence or presence of CGS 15943 (3 microM). These results confirm that ATP acts as a potent and NO-independent relaxant agent of human and rabbit CC. Our findings also show that the ATP effect is partially attributable to the metabolic breakdown of ATP to adenosine, which acts through A2a receptor stimulation, but is also due to a direct stimulation of P2 receptors that are different from the classical P2y and P2X receptor subtypes for ATP.