Contractility Of Vas Deferens Research Articles

6‐Nitrodopamine potentiates catecholamine‐induced contractions of human isolated vas deferens

Abstract6‐Nitrodopamine (6‐ND) is the main catecholamine released from human isolated vas deferens and the adrenergic nervous system is known to play a major role in the contractions of the epididymal portion of the vas deferens. Here it was investigated the interactions of 6‐ND on the contractions of the human isolated vas deferens induced by either classical catecholamines or electric‐field stimulation (EFS). The vas deferens obtained from 106 patients who underwent vasectomy surgery were mounted in a 10‐mL glass chamber filled with warmed (37°C) and oxygenated Krebs–Henseleit's solution. The strips were pretreated (30 min) with 6‐ND (0.1–100 nM) and exposed to increasing concentrations of noradrenaline (0.01–300 M), dopamine (0.00001–10 mM), or adrenaline (0.01–300 M). The strips were also submitted to EFS in tissues pre‐incubated or not with 6‐ND (1–100 nM), noradrenaline (100 nM), adrenaline (100 nM), or dopamine (100 nM). Catecholamine basal release was evaluated by LC–MS/MS and expression of tyrosine hydroxylase by both immunohistochemistry (IC) and fluorescence in‐situ hybridization (FISH). Pre‐incubation of the vas deferens with 6‐ND caused marked potentiation of the contractions induced by noradrenaline, adrenaline, and dopamine, as characterized by significant increases in Emax, without changes in pEC50 values. 6‐nitrodopamine also caused significant increases in the EFS‐induced contractions. The basal release of 6‐ND was not affected by pre‐treatment of the tissues with tetrodotoxin. Tyrosine hydroxylase was detected in epithelial cells of human vas deferens samples by both IC and FISH. The results clearly demonstrate that epithelium‐derived 6‐ND is a major modulator of human vas deferens contractility.

6-Nitrodopamine is an endogenous mediator of rat seminal vesicles contractility.

6-Nitrodopamine (6-ND) released from rat vas deferens acts an endogenous modulator of vas deferens contractility. To investigate whether rat isolated seminal vesicles (RISV) releases 6-ND, the mechanisms involved in the release, and the modulatory role of 6-ND on tissue contractility. Rat seminal vesicles were removed and placed in Krebs-Henseleit's solution at 37°C for 30 min, and an aliquot was used to analyze the concentrations of 6-ND, dopamine, noradrenaline, and adrenaline by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The effect of mechanical removal of the epithelium and the effects of pre-incubation of RISV strips with Nω-nitro-L-arginine methyl ester (L-NAME; in combination or not with L-arginine), tetrodotoxin (TTX), GKT137831, and hydrogen peroxide (H2O2) on 6-ND release were evaluated. For functional studies, RISV strips were mounted in an organ bath and tied to an isometric force transducer. The expressions of tyrosine hydroxylase and endothelial nitric oxide synthase (eNOS, type III NOS) were investigated by immunohistochemistry and/or fluorescence in situ hybridization (FISH). 6-ND was the major released catecholamine from RISV strips compared with noradrenaline, adrenaline, and dopamine. Epithelium removal significantly reduced the release of 6-ND, noradrenaline and dopamine. In RISV strips obtained from animals chronically treated with L-NAME, the 6-ND release significantly reduced. Pre-incubation with L-NAME reduced 6-ND release, which was partly restored by co-incubation with L-arginine. Pre-incubation with TTX had no effect on the release of any catecholamine, whereas GKT137831 and H2O2 significantly increased 6-ND release. All catecholamines produced concentration-dependent RISV contractions, but 6-ND was approximately 30× less potent than the others. 6-ND (0.1 nM) significantly potentiated noradrenaline-, adrenaline-, and dopamine-induced contractions, with such potentiations inhibited by TTX. Immunohistochemistry and FISH assays in RISV tissues identified tyrosine hydroxylase in epithelial cells and eNOS expression in both epithelial and endothelial cells. This is the first demonstration that epithelial-derived 6-ND modulates rat seminal vesicle contractility.

Effect of Atomoxetine on Mouse Isolated Vas Deferens Contractility

Objective: Atomoxetine (ATX), a selective noradrenaline re-uptake inhibitor, is a preferred drug with sufficient efficacy and favorable safety profile for the treatment of attention-deficit hyperactivity disorder. Ejaculatory dysfunctions have been reported in the patients receiving ATX as sexual side effect, of which underlying mechanisms are largely unknown. The present study aimed to investigate the effect of ATX on mouse isolated vas deferens (VD) contractility as a potential mechanism of ATX-induced ejaculatory dysfunction. Material and Methods: Isolated organ bath studies were performed on prostatic parts of VD obtained from adult male Balb/c mice. The effect of ATX (10-6, 10-5, 3x10-5 and 10-4 M) on KCl (80 mM), phenylephrine (PhE, 3x10-4 M), adenosine 5’-triphosphate (ATP, 10-2 M) and electrical field stimulation (EFS; 100 V, 64 Hz)-induced contractions of VD strips were evaluated in concentration dependent manner. Results: ATX at 10-6 and 10-5 did not alter the contractile responses (p>0.05), however, higher concentrations of ATX (3x10-5 or 10-4 M) significantly inhibited the KCl, PhE, ATP and EFS-induced contractions of VD strips (p<0.05). Conclusion: The present study demonstrated for the first time that ATX decreased the contractile responses of mouse isolated VD concentration-dependently. Our results suggest that ejaculatory dysfunction might be related to the inhibitory effect on ATX on VD.

Extracellular cAMP elicits contraction of rat vas deferens: Involvement of ecto-5′-nucleotidase and adenosine A1 receptors

AimsIt is well established that intracellular cAMP contributes to the relaxation of vas deferens smooth muscle. In many tissues, intracellular cAMP is actively transported to the extracellular space, where it exerts regulatory functions, via its metabolite adenosine. These actions take place through the cAMP conversion to adenosine by ectoenzymes, a process called “extracellular cAMP-adenosine pathway”. Herein, we investigated whether, in addition to ATP, extracellular cAMP might be an alternative source of adenosine, influencing the contraction of vas deferens smooth muscle. Main methodsThe effects of cAMP, 8-Br-cAMP and adenosine were analyzed in the isometric contractions of rat vas deferens. cAMP efflux was analyzed by measuring extracellular cAMP levels after exposure of vas deferens segments to isoproterenol and forskolin in the presence or absence of MK-571, an inhibitor of MRP/ABCC transporters. Key findingsWhile 8-Br-cAMP, a cell-permeable cAMP analog, induced relaxation of KCl-precontracted vas deferens, the non-permeant cAMP increased the KCl-induced contractile response, which was mimicked by adenosine, but prevented by inhibitors of ecto-5′-nucleotidase or A1 receptors. Our results also showed that isoproterenol and forskolin increases cAMP efflux via an MRP/ABCC transporter-dependent mechanism, since it is inhibited by MK-571. SignificanceOur data show that activation of β-adrenoceptors and adenylyl cyclase increases cAMP efflux from vas deferens tissue, which modulates the vas deferens contractile response via activation of adenosine A1 receptors. Assuming that inhibition of vas deferens contractility has been proposed as a strategy for male contraception, the extracellular cAMP-adenosine pathway emerges as a potential pharmacological target that should be considered in studies of male fertility.

(125) BASAL RELEASE OF 6-CYANODOPAMINE FROM RAT ISOLATED VAS DEFERENS AND ITS ROLE ON THE VAS DEFERENS CONTRACTILITY

Abstract Introduction 6-Cyanodopamine is released from isolated rabbit heart and is a novel endothelial catecholamine similar to 6-nitrodopamine described by our research group. 6-Nitrodopamine is a major endogenous modulator of human vas deferens contractility. However, it is not known whether 6-cyanodopamine presents any biological activity. Objective Here it was evaluated whether 6-cyanodopamine was released from rat isolated vas deferens and wether, like 6-nitrodopamine, modulates vas deferens contractility. Methods The basal release of 6-bromodopamine, 6-nitrodopa, 6-nitrodopamine, 6-cyanodopamine, and 6-nitroadrenaline from the vas deferens was quantified using LC–MS/MS. Electric-field stimulation (EFS) and concentration-response curves for noradrenaline, adrenaline, and dopamine were performed on the rat isolated epididymal vas deferens (RIEVD) in the absence and presence of 6-cyanodopamine. The expression of tyrosine hydroxylase and S100 was assessed by immunohistochemistry. Additionally, we evaluated the effect of tetrodotoxin on the release and effect of 6-cyanodopamine in the rat vas deferens. The synergism between 6-cyanodopamine and 6-nitrodopamine was analyzed in the RIEVD. Results The rat isolated vas deferens release both 6-cyanodopamine and 6-nitrodopamine. Pre-incubation with tetrodotoxin, had no effect on the release of 6-cyanodopamine, but it virtually abolished 6-nitrodopamine release. 6-Cyanodopamine contracted RIEVD only at 1 mM, but significantly potentiated the contractions induced by both noradrenaline and EFS at 1 nM. At 10 and 100 nM, 6-cyanodopamine also significantly potentiated the RIEVD contractions induced by adrenaline and dopamine. The potentiation of both noradrenaline and adrenaline contractions by 6-cyanodopamine was present in tetrodotoxin-pretreated tissues. Co-incubation of 6-cyanodopamine (100 pM) with 6-nitrodopamine (10 pM), caused significant leftward shifts and increased maximal responses of the concentration-response curves to noradrenaline, even in the presence of tetrodotoxin. Immunohistochemistry on RIEVD revealed the presence of tyrosine hydroxylase in both the cytoplasm of epithelial cells of the mucosae and in the nerve fibers. Conclusions The identification of the basal release of epithelium-derived 6-cyanodopaine from rat isolated vas deferens and its remarkable ability to potentiate noradrenaline- and EFS induced contractions of this tissue, demonstrate that epithelial cells modulate smooth muscle contractility. Disclosure No.

(314) 6-NITRODOPAMINE IS A MAJOR MEDIATION THAT POTENTIATES CONTRACTIONS OF HUMAN ISOLATED EPIDIDYMAL VAS DEFERENS INDUCED BY NORADRENALINE AND ELECTRIC FIELD STIMULATION

Abstract Introduction Contractions of the vas deferens have an important role in the propulsion of the spermatozoa and occur due to excitatory inputs from a network of sympathetic nerves. When stimulated, these nerves co-release noradrenaline and ATP as the primary chemical messengers. However, 6-nitrodopamine (6-ND) was recently identified as a novel mediator of rat and human epididymal vas deferens (HEVD) contractility providing a new insight on the physiological process of the male ejaculation. Objective We sought to investigate the interaction of 6-ND in contractions induced by noradrenaline and electric-field stimulation on the HEVD. Methods Human epididymal vas deferens was obtained from 35 patients aged 28-59 years old who had undergone vasectomy surgery at Hospital e Maternidade Salvalus, São Paulo, BR. The HEVD strips (1.5 cm) were mounted in a 10-mL glass chamber filled with a warm (37°C) and oxygenated (95%O2: 5%CO2) Krebs–Henseleit solution. The tissues were allowed to stabilize for 45 minutes under a resting tension of 10 mN, and the isometric tension was registered using a PowerLab system (ADInstruments). The HEVD strips viability was tested with potassium chloride (80 mM). HEVD strips were pretreated with 6-ND (0.1, 1, 10 and 100 nM for 30 min) and a cumulative concentration-response curve to noradrenaline was performed. Also, the strips pretreated with 6-ND and its respective controls were submitted to electric-field stimulation (EFS) (60 V for 20 sec, at 2–32 Hz in square-wave pulses, 0.3 ms pulse width, and 0.1 ms delay), using a Grass S88 stimulator. Results Pre-incubation with 6-ND 0.1 nM had no effect on the noradrenaline-induced contraction curve. However, at 1, 10 and 100 nM, 6-ND causes significantly leftward shift on the noradrenaline concentration-response curve (6-ND 100 nM; pEC50: 4.9 ± 0.1 vs 4.5 ± 0.2; Emax: 54.8 ± 3.2 vs 31.2 ± 3.3; 6-ND vs control, respectively; p < 0.05. Similar results were found on contractions induced by EFS. Preincubation with 10 and 100 nM of 6-ND, but not 1 nM, significantly increased the contractions induced by EFS at the frequencies of 4 to 32 Hz. Conclusions The findings establish the 6-nitrodopamine as an important mediator of HEVD contractions by causing remarkable potentiation of noradrenaline and EFS induced- contraction. Disclosure No.

(292) THE ROLE OF PRE-SYNAPTIC β1-ADRENOCEPTORS IN RAT ISOLATED EPIDIDYMAL VAS DEFERENS CONTRACTILITY

Abstract Introduction The absence of a low-affinity site for propranolol after reserpinisation or rauwolscine treatment suggests a dependence on intact catecholamine stores and α2-adrenoceptors, reflecting β-blockers’ action. In guinea-pig vas deferens, β-adrenoceptor activation increases noradrenaline release while decrease ATP release. Isoprenaline reduces nerve-induced responses in rat vas deferens. Objective To investigate the effect of presynaptic beta 1 adrenergic agonists in rat vas deferens. Methods Adult male Wistar rats were euthanized with isoflurane, then epididymal vas deferens were isolated, placed in Krebs–Henseleit’s solution, and mounted in tissue baths for isometric tension recording using a PowerLab system. The effect of β-adrenoceptor agonist RO363 (100 nM), and antagonists betaxolol (1 μM), metoprolol (1 μM), and atenolol (1 μM) on adrenaline (ADR), noradrenaline (NA) and dopamine (DA) induced contractile responses was assessed. Electric-field stimulation (EFS) and the effect of the sodium channel antagonist tetrodotoxin (TTX 1 μM) were also evaluated. All the procedures were approved by an Ethic Committee. Data were analyzed using nonlinear regression to determine pEC50 values and expressed as mean ± SEM. Results The data showed that RO363 had no effect on DA but caused a concentration-dependent leftward shift on the NA and ADR curves, abolished by TTX pretreatment. RO363 enhanced tissue contraction in response to EFS across all frequencies. Atenolol, betaxolol and metoprolol abolished the effect of RO363 on NA and ADR curves. Conclusions Findings reveal that the β1-adrenoceptor agonist RO363 selectively modulates contractile responses to NA and ADR, indicating a specific interaction with these neurotransmitters sensitive to sodium channel blockade by TTX. The abolition of RO363’s effect by β1-adrenoceptor antagonists further underscores the specificity of β1-adrenoceptor involvement. Results suggest that β1-adrenoceptors are crucial in regulating neurotransmitter-induced contractility in the vas deferens, potentially through mechanisms dependent on intact catecholamine stores and α2-adrenoceptors. This study enhances understanding of adrenergic modulation in male reproductive physiology and may inform future research on pharmacological manipulation of vas deferens contractility. Disclosure No.

(294) 6-NITRODOPAMINE IS AN ENDOGENOUS MEDIATOR OF RAT SEMINAL VESICLES CONTRACTILITY

Abstract Introduction 6-Nitrodopamine (6-ND) is released from rat and human isolated vas deferens and acts as an endogenous modulator of vas deferens contractility. Objective To investigate whether rat isolated seminal vesicles (RISV) present basal release of 6-ND, the mechanisms involved in the release, and the role of 6-ND on tissue contractility. Methods The local ethics committee approved the experimental protocols. Adult male Wistar rats (280 to 320 g) were used to perform the experiments. After isoflurane (5% for 2 min) induced euthanasia, the tissues were removed and one pair placed in 3 mL of Krebs–Henseleit’s solution at 37°C for 30 min, then an aliquot was used to analyze the concentrations of 6-ND, dopamine, noradrenaline, and adrenaline by Liquid Chromatography with tandem mass spectrometry (LC–MS/MS). The effects of the mechanical removal of the epithelium, the chronic treatment with L-NAME (20 mg/rat/day) in the drinking water and the pre-incubation of the RISV with L-NAME (100 μM), tetrodotoxin (1 μM), GKT137831 (1 μM) and H2O2 (100 μM) were also evaluated by LC–MS/MS. For functional studies, a RISV strip was mounted vertically in a glass chamber tissue bath containing 10-mL KHS with a fine sewing thread, one end attached through a loop to a metal hook and the other end tied to an isometric force transducer under a resting tension of 10mN and the isometric tension was registered using a PowerLab system (ADInstruments, Sdney, Australia). After tissue viability verification with noradrenaline (10 μM), concentration-response curves with 6-ND and classical catecholamines were performed, in addition interaction protocols among those drugs, including pre-incubation with tetrodotoxin, were assessed. Lastly, the expression of tyrosine hydroxylase was investigated by both immunohistochemistry and fluorescence in-situ hybridization. Results 6-ND was the major released catecholamine compared with noradrenaline, adrenaline, and dopamine (3.2 ± 0.7, 0.6 ± 0.4, 0.2 ± 0.1 and 0.3 ± 0.1 ng/mL, respectively). Mechanical removal of the epithelium caused significant reduction in 6-ND basal release, this depletion was also observed with the chronic treatment and the pre-incubation with L-NAME. In contrast, pre-incubation with either GKT137831 or H2O2 caused selective increases in the 6-ND release. Pre-incubation with tetrodotoxin had no effect on the release of catecholamines. All catecholamines produced concentration-dependent RISV contractions, but 6-ND was 30x less potent than the others. However, at low concentration (0.1 nM), 6-ND significantly potentiated noradrenaline-, adrenaline- and dopamine-induced contractions, which were not observed in presence of tetrodotoxin. Immunohistochemistry and fluorescence in-situ hybridization revealed the presence and expression of tyrosine hydroxylase in RISV epithelial cells. Conclusions This is the first demonstration of 6-ND release from epithelium and the results indicate that epithelial-derived 6-ND modulates rat seminal vesicle contractility. Disclosure No.

The pivotal role of neuronal nitric oxide synthase in the release of 6-nitrodopamine from mouse isolated vas deferens

6-Nitrodopamine (6-ND) is released from rat and human vas deferens and is considered a major mediator of both tissues contractility. The contractions induced by 6-ND are selectively blocked by both tricyclic antidepressants and α1-adrenoceptor antagonists. Endothelial nitric oxide synthase (eNOS) is the major isoform responsible for 6-ND release in mouse isolated heart, however the origin of 6-ND in the vas deferens is unknown. Here it was investigated by LC-MS/MS the basal release of 6-ND from isolated vas deferens obtained from control, eNOS−/−, nNOS−/−, and iNOS−/− mice. In addition, it was evaluated in vitro vas deferens contractility following electric field stimulation (EFS).Basal release of 6-ND was significantly reduced in nNOS−/− mice compared to control mice, but not decreased when the vas deferens were obtained from either eNOS−/− or iNOS−/− mice. Pre-incubation of the vas deferens with tetrodotoxin (1 μM) significantly reduced the basal release of 6-ND from control, eNOS−/−, and iNOS−/− mice but had no effect on the basal release of 6-ND from nNOS−/− mice. EFS-induced frequency-dependent contractions of the vas deferens, which were significantly reduced when the tissues obtained from control, eNOS−/− and iNOS−/− mice, were pre-incubated with l-NAME, but unaltered when the vas deferens was obtained from nNOS−/− mice. In addition, the EFS-induced contractions were significantly smaller when the vas deferens were obtained from nNOS−/− mice.The results clearly demonstrate that nNOS is the main NO isoform responsible for 6-ND release in mouse vas deferens and reinforces the concept of 6-ND as a major modulator of vas deferens contractility.

Interaction of endocannabinoid system and cyclooxygenase metabolites with fatty acid amide hydrolase and cyclooxygenase enzyme activities on contractile responses in rat vas deferens tissue.

The endocannabinoid system and prostaglandins are important modulators in the genitourinary system. This study aimed to investigate the possible interactions between the endocannabinoid system and the cyclooxygenase (COX) pathway on rat vas deferens. For this purpose, the concentration responses of the endocannabinoid anandamide, prostaglandin F2α analog latanoprost, and prostaglandin E1 analog misoprostol on the electrical field stimulation (EFS)-induced contractile responses were obtained. The concentration responses to anandamide were obtained again in the presence of nonselective COX inhibitor flurbiprofen and prostaglandin analogs, while the concentration responses of latanoprost and misoprostol were obtained in the presence of cannabinoid receptor antagonists and fatty acid amide hydrolase (FAAH) enzyme inhibitor URB597. FAAH, COX-1, and COX-2 enzyme levels in vas deferens tissue samples were also determined. The cumulative addition of anandamide was not different from the vehicle; however, the EFS-induced contractile responses were significantly increased with the incubation of latanoprost or flurbiprofen in the prostatic portion. Flurbiprofen and misoprostol decreased FAAH enzyme levels in both portions of the vas deferens, while latanoprost induced the inhibition in the prostatic portion. The cumulative administration of latanoprost and misoprostol significantly enhanced the contractile responses in the prostatic portion. This effect of latanoprost was significantly antagonized by URB597 and AM251. The enhancing effect of misoprostol was antagonized by anandamide, URB597, AM251, and AM630. Anandamide, AM251, AM630, and URB597 decreased enzyme levels of COX-1 and COX-2 in both portions of the vas deferens. These results demonstrate an intricate crosstalk between endocannabinoids and prostaglandins in modulation of the vas deferens contractility.

6-nitrodopamine is a major endogenous modulator of human vas deferens contractility.

Rat isolated vas deferens releases 6-nitrodopamine (6-ND), and the spasmogenic activity of this novel catecholamine is significantly reduced by tricyclic compounds such as amitriptyline, desipramine, and carbamazepine and by antagonists of the α1 -adrenergic receptors such as doxazosin, tamsulosin, and prazosin. To investigate the liberation of 6-ND by human epididymal vas deferens (HEVDs) and its pharmacological actions. The in vitro liberation of 6-ND, dopamine, noradrenaline, and adrenaline from human vas deferens was evaluated by LC-MS/MS. The contractile effect of the catecholamines in HEVDs was investigated in vitro. The action of tricyclic antidepressants was evaluated on the spasmogenic activity ellicited by the catecholamines and by the electric-field stimulation (EFS). The tissue was also incubated with the inhibitor of nitric oxide (NO) synthase L-NAME and the release of catecholamines and the contractile response to EFS were assessed. 6-ND is the major catecholamine released from human vas deferens and its synthesis/release is inhibited by NO inhibition. The spasmogenic activity elicited by EFS in the human vas deferens was blocked by tricyclic antidepressants only at concentrations that selectively antagonize 6-ND induced contractions of the human vas deferens, without affecting the spasmogenic activity induced by dopamine, noradrenaline, and adrenaline in this tissue. Incubation of the vas deferens with L-NAME reduced both the 6-ND release and the contractions induced by EFS. 6-ND should be considered a major endogenous modulator of human vas deferens contractility and possibly plays a pivotal role in the emission process of ejaculation. It offers a novel and shared mechanism of action for tricyclic antidepressants and α1 -adrenergic receptor antagonists.

Effects and safety of combined hypericum perforatum and quercetine in premature ejaculation: A pilot study

ABSTRACT Introduction Premature ejaculation (PE) is a common, and under treated medical condition that affects men and their partners. Advances in clinical research in premature ejaculation (PE) over the last years have led to the development of a variety of new treatment options, including dietary supplements. Hypericum perforatum (HP), a natural supplement has demonstrated pharmacologic action to inhibit serotonin reuptake and that inhibits rat and human vas deferens contractility. These results might explain delayed ejaculation described in patients receiving HP. Quercetin is a naturally occurring bioflavonoid found in high concentrations in red wine, onions, and green tea. Therapy with the bioflavonoid quercetin provides significant symptomatic improvement in most men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). Subjects with PE often show symptoms and signs compatible with CP/CPPS, as confirmed by many different studies, CP/CP are an organic risk factor for PE. Objective The purpose of the present investigation was to assess the safety and preliminary evidence of the effectiveness of a supplement that combined HP and quercetin for the treatment of PE. Methods A one-group pretest-posttest design was used in the current research. Main outcome measures were assessed using the mean stopwatch-measured Intravaginal Ejaculatory Latency Time (IELT) at baseline and after 12 weeks. The collected data were tabulated and analyzed using the Statistical Package for the Social Sciences version 22 (SPSS). Quantitative data are expressed as means ± standard deviations (SDs). For the comparison of paired variables the Wilcoxon test was used. Premature Ejaculation Diagnostic Tool (PEDT) was used for the diagnosis and as a secondary measure. The study inclusion criteria to be selected were as follows: Being over 18 years old, being in a heterosexual relationship for at least the last 6 months, having a score ≥ 11 in the PEDT and a mean self-reported IELT ≤ 2 minutes. Exclusion criteria included a history of alcohol abuse or dependence, having received medication or psychological treatment for PE in the last 6 months, suffering from diabetes or habitual use of recreational drugs. A total of 12 subjects, aged between 27 and 58 years of age (mean = 40.50 years, standard deviation (SD) = 8.31) were treated with 1 daily tablet of 650 mg of Myhixel Max®. Each tablet contained Hypereric dry extract 433.64mg, Sophora japonica L. (95% quercetin 72.27mg), 68.65mg quercetin and anti-caking agents (magnesium stearate and silico dioxide). Results In the present study, there was a significant improvement in mean IELT (mean pre = 69.6 seg (SD) = 37.8 mean post = 146.4 seg (SD) = 67.2, Wilcoxon test pre vs post p-value = 0.008). Conclusions Oral therapy with HP and quercetin was associated with a significant increase in mean IELT from baseline, with no adverse effects related to the treatment. We can conclude that this supplement in daily use has the potential to become an effective and safe treatment for patients suffering from PE, long-term durability of the response is yet to be determined. Future studies with, larger group of patients; larger follow-up and group control are needed. Disclosure Work supported by industry: yes, by New Wellness Concept S.L. A consultant, employee (part time or full time) or shareholder is among the authors (New Wellness Concept. S.L).

Chronic Effects of Loxapine, Iloperidone, Paliperidone on Mice Isolated Vas Deferens Contractility

Objective: Erectile dysfunction is a usual side effect of antipsychotic medications; this causes patients to avoid using drugs. The aim of this study to investigate the effects of iloperidone, paliperidone and loxapine on serotonin (5-HT), noradrenaline (NA), adenosine triphosphate (ATP) and potassium chloride (KCl)-induced contractions of the vas deferens in mice.Materials and Methods: The mice were randomly divided into experimental groups and treated by ip injection of drugs for 21 days. After the treatment, the effects of drugs were investigated on 5-HT, ATP, NA and KCl-induced contractile responses in the epididymal and prostatic portions of mice vas deferens strips. Results: 5-HT-induced contractile responses were significantly increased while ATP-induced contractile responses were significantly decreased in the both portions of the vas deferens obtained from the iloperidone, paliperidone and loxapine-treated groups. However, these drugs had no significant effect on NA- and KCl-induced contractions of mice vas deferens. Conclusion: These results showed that serotonin and ATP-induced contractions of vas deferens were affected by the chronic treatments of iloperidone, paliperidone, and loxapine. In mice chronically treated with these drugs, serotonergic and purinergic receptors may contribute to changes in vas deferens contractions that cause erectile dysfunction.

Chronic Effects of Atypic Antipsychotics on Mice Isolated Vas Deferens Contractility: An In vitro Study

Aims: Antipsychotic drugs are known to be commonly associated with sexual dysfunction. The aim of this study to investigate the effects of sertindole, asenapine and ziprasidone on serotonin (5-HT), noradrenaline (NA), adenosine triphosphate (ATP) and potassium chloride (KCl)-induced contractions of the isolated vas deferens in mice.
 Study design: All the drugs were administered intraperitoneally (i.p.) in a volume of 0.1 ml per 10 g body weight of mice.
 Place and Duration of Study: Department of Pharmacology, Kocaeli University, Animal Research Center, between May 2018 and August 2019.
 Methodology: The mice were randomly divided into groups (n=7) as follows: saline; sertindole 1.3 mg/kg; sertindole 2.5 mg/kg; asenapine 0.05 mg/kg; asenapine 0.075 mg/kg; ziprasidone 1 mg/kg; ziprasidone 2 mg/kg once daily for 21 days. Mice receiving only the vehicle (0.9% saline, i.p.) served as control group. After 21 days of treatment, the effects of drugs were investigated on 5-HT, NA, ATP and KCl-induced contractions of isolated vas deferens.
 Results: The results showed that serotonin-induced contractions of vas deferens were affected by the chronic treatments of sertindole, asenapine and ziprasidone, however these drugs had no significant effect on NA, ATP, and KCl-induced contractions of mice vas deferens.
 Conclusion: Serotonergic receptors may contribute to changes in vas deferens contractions in mice with chronic treatment of sertindole, asenapine and ziprasidone. Thus, our results may explain one of the causes of sexual dysfunction of sertindole, asenapine and ziprasidone.

Resveratrol and quercetin attenuate depressive-like behavior and restore impaired contractility of vas deferens in chronic stress-exposed rats: involvement of oxidative stress and inflammation.

Chronic stress is associated with male sexual problems including ejaculatory dysfunctions. The aim of this study was to determine whether resveratrol (RS) or quercetin (QE) has protective effects on vas deferens (VD) contractility in the unpredictable chronic mild stress (UCMS) rat model of depression. Animals were separated into six groups: control, control + RS and control + QE, stress, stress + RS, and stress + QE. Stress groups were subjected to UCMS procedure for 5weeks. Animals in treatment groups were injected intraperitoneally with RS (20mg/kg) or QE (30mg/kg) for 5weeks during UCMS period. UCMS caused depressive-like behaviors and enhanced systemic levels of corticosterone. The nerve-evoked contractile responses of VD significantly impaired and, noradrenaline- and ATP-induced contractile responses of VD significantly increased in stressed rats. UCMS exposure also markedly enhanced oxidative stress and inflammation in VD tissues. Treatment with RS or QE significantly ameliorated all the aforementioned parameters. The current study demonstrated that RS or QE protected against chronic stress-induced VD dysfunction by their antioxidant and anti-inflammatory effects on VD, suggesting that oxidative stress and inflammation may be synergistic parts in the development of VD dysfunction associated with chronic stress-induced depression.

<i>Lannea acida</i> improves the emission phase of ejaculation of mature male rat: effects on vas deferens and seminal vesicles

Objective: The barks of Lannea acida (L. acida) are used in traditional medicine as aphrodisiac. This work was undertaken to evaluate the effects of L. acida on vas deferens contractility (in vitro test) and seminal vesicles pressure (ex-copula test) of mature rat.Methods: For the in vitro test, rats were sacrificed by cervical dislocation and the vas deferens removed and mounted in an isolated organ bath containing aerated physiological Krebs solution for the recording of the effects of aqueous extract of L. acida (0.91-14.54 mg/ml) and KCl (13.25-106 mg/ml). For the ex-copula test, the seminal vesicles pressure of urethane-anesthetized rats was measured after urethral and tactile stimulations, intravenous administration of NaCl 0.9% (1 ml/kg), dopamine (5 mg/kg) or plant extract (5-40 mg/kg).Results: Similar to KCl (13.25-106 mg/ml), the aqueous extract of L. acida concentration-dependently contracted the vas deferens. Furthermore, urethral and tactile stimulations as well as intravenous administration of dopamine or plant extract induced contractions of the seminal vesicles characterized by an increase of the pressure. This increase induced by the plant extract was dose-dependent with significant effects obtained at doses 20 mg/kg, 30 mg/kg and 40 mg/kg.Conclusion: These results show that L. acida improves the emissive phase of ejaculation through contraction of vas deferens and seminal vesicle smooth muscles.Keywords: Lannea acida, emission phase of ejaculation, vas deferens, seminal vesicles, rat

Effect of a Fructose Rich Diet in Vas Deferens Contractility in Rats

Objective: We aimed to investigate the effects of a Carbohydrate-rich diet on vas deferens contractility and histology. Materials and Methods: Twenty mature male rats were subjected into two groups. The control group received regular food and water and the study group received regular food and a carbohydrate-rich liquid diet mixture of 37–40% fructose, 30–36% sucrose and 27–30% glucose instead of water and prepared stock diet was diluted with 50% water. Animals were weighted and sacrificed after six weeks and the vas deferens has been evaluated with in vitro studies. Five rats from both the control and study groups, were separated into subgroups for dapoxetine administration 4 hours before they were sacrificed. Results: At the beginning of this study, the mean body weights were 228.6±6.7 g (range: 205-237) and 231.4±9.6 g (range: 202-243) in control and study groups, respectively. The mean body weights were 247.1±4.8 g (236-252) in the control group and 318.8±9.3 g (314-326) in the study group ( p 0.05). Responses to Adenosine Three Phosphate (ATP) were also not significant ( p >0.05). The contractile responses at the same frequencies of electrical field stimulation (EFS) were similar in both groups ( p >0.05). Histological examinations showed no abnormalities in either group. Conclusion: Carbohydrate-rich diet caused no increase in fibrotic activity, and pharmacological and histological properties remained stable, but caused significant weight gain over a short time period. Furthermore, dapoxetine had no effect on the contractility of the vas deferens.

Effect of noni (Morinda citrifolia Linn.) fruit and its bioactive principles scopoletin and rutin on rat vas deferens contractility: an ex vivo study.

This study examined the effect of methanolic extract of Morinda citrifolia Linn. (MMC) and its bioactive principles, scopoletin and rutin, on dopamine- and noradrenaline-evoked contractility in isolated rat vas deferens preparations. MMC (1–40 mg/mL), scopoletin (1–200 μg/mL), and rutin hydrate (0.6–312.6 μg/mL) dose-dependently inhibited the contractility evoked by submaximal concentrations of both dopamine and noradrenaline, respectively. Haloperidol and prazosin, reference dopamine D2, and α 1-adrenoceptors antagonists significantly reversed the dopamine- and noradrenaline-induced contractions, respectively, in a dose-dependent manner. Interestingly, MMC per se at higher doses (60–100 mg/mL) showed dose-dependent contractile response in rat vas deferens which was partially inhibited by high doses of haloperidol but not by prazosin. These results demonstrated the biphasic effects of MMC on dopaminergic system; that is, antidopaminergic effect at lower concentrations (<40 mg/mL) and dopaminergic agonistic effect at higher concentrations (>60 mg/mL). However, similar contractile response at high doses of scopoletin (0.5–5 mg/mL) and rutin hydrate (0.5–5 mg/mL) per se was not observed. Therefore, it can be concluded that the bioactive principles of MMC, scopoletin, and rutin might be responsible for the antidopaminergic and antiadrenergic activities of MMC.

Purinergic contraction of the rat vas deferens in L-NAME-induced hypertension: effect of sildenafil

Hypertension (HTN) is a risk factor for erectile dysfunction, but its effect on vas deferens (VD) contractility and the ejaculatory response has not been delineated. NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, was used for induction of nitric oxide (NO)-deficient HTN. Our aim was to evaluate the effects of L-NAME-induced HTN on rat VD contractility and to determine whether sildenafil affects VD contractility. A total of 36 male rats were divided into (1) control, (2) L-NAME-HTN, (3) sildenafil treated L-NAME-HTN groups. Group 2 was treated with L-NAME (40 mg kg(-1) per day) in drinking water for 4 weeks. Group 3 received sildenafil (1.5 mg kg(-1) per day, by oral gavage) concomitantly with L-NAME. The prostatic portion of the VD was subjected to electrical field stimulation (EFS, 1-20 Hz), and the P2X(1) agonist alpha,beta-methylene ATP (alpha,beta-meATP, 100 micromol L(-1)-1 micromol L(-1)) and the alpha1-adrenoceptor agonist phenylephrine (Phe, 100 micromol L(-1)-1 mmol L(-1)) were used to construct concentration-response curves. These experiments were repeated in the presence of P2X receptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS, 30 micromol L(-1)). VD contractions in response to EFS, alpha,beta-meATP and Phe were significantly enhanced by L-NAME. Sildenafil treatment in the L-NAME group improved the contractile response of VD to EFS (20 Hz). In the presence of PPADS, the enhanced contractile response of VD to EFS and alpha,beta-meATP in hypertensive rats was reversed. In the rat model of chronic NO depletion, the purinergic and adrenergic components and EFS affect VD contractility. The VD contractile response may be mediated more by the purinergic system than the adrenergic system, and sildenafil may alter the ejaculatory response in men with PE.

Comparative study of α 2-adrenoceptors in Fischer 344 and Lewis rats. Evidence for clonidine-induced place aversion

Fischer 344 (F344) and Lewis rat strains have been shown to exhibit different vulnerability to development or maintenance of opioid seeking behaviours probably due to differences in the endogenous opioid system. Since opioid and α 2-adrenergic mechanisms closely interact in nociception and substance abuse, strain differences may be expected to affect α 2-adrenoceptor-mediated events. The sensitivity of these two strains to α 2-adrenoceptor-mediated antinociception has been reported to be markedly different. In this work we have further studied the function of α 2-adrenoceptors in F344 and Lewis rats by means of several in vivo and in vitro procedures. Comparative studies of [ 3H]RX821002 and [ 35S]GTPγS binding revealed that α 2-adrenoceptors could be slightly more responsive to agonist stimulation in the brain cortex of F344 rats, which is in agreement with previous antinociception studies. However, these differences were modest, not observed in the spinal cord and did not translate into functional differences concerning the effects of clonidine on vas deferens contractility and body temperature. Conditioning experiments showed that a moderate dose of clonidine, which is relevant in antinociceptive and opioid antiwithdrawal studies, induces a robust place aversion which is also equivalent in F344 and Lewis rats. This finding underlies the consistency of the effect and its independency of genetic differences between both rat strains. It seems therefore that the pharmacological properties of α 2-adrenoceptors are similar in F344 and Lewis rats, and thus the previously reported differences in clonidine-induced antinociception could be attributed to other factors such as dissimilar endogenous function of specific noradrenergic pathways.