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1.
Life Sci ; 238: 116922, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31634463

RESUMO

AIMS: Nitric oxide (NO) has a critical, but not well understood, influence in the physiology of the lower urinary tract. We evaluated the effect of NO/phosphodiesterase (PDE)5 signaling in voiding dysfunction in the sickle cell disease (SCD) mouse, characterized by low NO bioavailability. MAIN METHODS: Adult SCD (Sickle) and wild-type (WT) male mice were treated daily with sodium nitrate (10 mM) or vehicle. After 18 days, blood was obtained for nitrite measurement, urethra was collected for organ bath study, and bladder and urethra were collected for Western blot analysis of PDE5 phosphorylation (Ser-92) (activated form). Non-anesthetized mice underwent evaluation of urine volume by void spot assay. eNOS phosphorylation (Ser-1177) and nNOS phosphorylation (Ser-1412) (positive regulatory sites) were evaluated in the bladder and urethra of untreated mice. KEY FINDINGS: Sickle mice exhibited decreased eNOS, nNOS, and PDE5 phosphorylation in the bladder and urethra, decreased plasma nitrite levels, increased relaxation of phenylephrine-contracted urethral tissue to an NO donor sodium nitroprusside, and increased total urine volume, compared with WT mice. Nitrate treatment normalized plasma nitrite levels, relaxation of urethra to sodium nitroprusside, PDE5 phosphorylation in the urethra and bladder, and urine volume in Sickle mice. SIGNIFICANCE: Derangement in PDE5 activity associated with basally low NO bioavailability in the bladder and urethra contributes to the molecular basis for voiding abnormalities in Sickle mice. Inorganic nitrate supplementation normalized voiding in Sickle mice through mechanisms likely involving upregulation of PDE5 activity. These findings suggest that interventions targeting dysregulatory NO/PDE5 signaling may ameliorate overactive bladder in SCD.


Assuntos
Anemia Falciforme/fisiopatologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Nitratos/administração & dosagem , Óxido Nítrico/metabolismo , Uretra/efeitos dos fármacos , Bexiga Urinária/efeitos dos fármacos , Administração Oral , Animais , Masculino , Camundongos , Óxido Nítrico Sintase Tipo I/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Fosforilação , Uretra/metabolismo , Uretra/patologia , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia
2.
Molecules ; 24(18)2019 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-31546796

RESUMO

Polymethylene-interrupted polyunsaturated fatty acids (PMI-PUFAs) are emerging functional lipids with proven antioxidant and anti-inflammatory effects. In this study, a typical PMI-PUFA, sciadonic acid (C20:3, 5c 11c 14c), was enriched in the kernel oil of Torreya fargesii (T. fargesii) by fractionation. Fractionated kernel oil of T. fargesii (containing 25% sciadonic acid) showed equal stability and similar radical scavenging ability compared with the non-fractionated oil. In anti-inflammatory tests, fractionated kernel oil was shown to inhibit the activity of phosphodiesterase (PDE-5, efficiency 80% at 133.7 µg/mL) and lipoxygenase-5 (LOX-5, efficiency 65% at 66.7 µg/mL) more effectively than the non-fractionated oil. This shows that increasing the amount of sciadonic acid can enhance the anti-inflammatory effect of the kernel oil. This research also indicates that fractionation is a feasible way to obtain sciadonic acid-rich functional oil with potential pharmacological effects.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Óleos Vegetais/química , Sementes/química , Taxaceae/química , Animais , Araquidonato 5-Lipoxigenase/metabolismo , Compostos de Bifenilo/química , Fracionamento Químico , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Orelha/patologia , Edema/patologia , Ácidos Graxos/análise , Depuradores de Radicais Livres/análise , Camundongos , Picratos/química
3.
Eur J Pharmacol ; 861: 172600, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31401155

RESUMO

This study was performed to examine the protective effects of icariin (ICA) on ischemic random skin flaps. A rat random-pattern skin flap model was established, and animals in the low-dose and high-dose experimental groups were administered ICA intraperitoneally at doses of 40 and 80 mg/kg, respectively, once daily for 7 days after the initial surgery. Control rats received vehicle according to the same schedule. Survival rates were observed and recorded using transparent graph paper, and flaps were obtained and stained with hematoxylin and eosin (H&E). The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the flap tissue were assessed. The blood flow volume was determined by the laser Doppler method, and tissue expression levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), interleukin-1ß, and phosphodiesterase 5 (PDE5) were scored immunohistochemically. The levels of proinflammatory cytokines, including tumor necrosis factor-α and IL-6, were determined by enzyme-linked immunosorbent assay. The main flap survival area was significantly larger in rats treated with ICA than in vehicle-treated controls. H&E staining showed an inhibitory effect of ICA on inflammation, especially at the high dose. In addition, ICA treatment was associated with decreases in the tissue MDA level, proinflammatory cytokine production, and the level of PDE5, but increases in SOD activity, blood flow volume, and the level of VEGF expression. The findings of the present study suggest that ICA is a potential therapeutic agent for random-pattern skin flap necrosis in the clinical setting.


Assuntos
Flavonoides/farmacologia , Pele/patologia , Retalhos Cirúrgicos , Sobrevivência de Tecidos/efeitos dos fármacos , Animais , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Isquemia/metabolismo , Isquemia/patologia , Isquemia/fisiopatologia , Masculino , Malondialdeído/metabolismo , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacos , Pele/irrigação sanguínea , Superóxido Dismutase/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
4.
Andrologia ; 51(9): e13371, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31347717

RESUMO

In comparison to other antidepressant drugs, erectile dysfunction (ED) is more pronounced in paroxetine use. On the other hand, orange (Citrus sinensis) peels commonly consumed in various forms are used in folkloric medicine for ED management. Thus, this study evaluated the effect of orange peels infusion on sexual behaviour, nitric oxide (NO) level and some enzymes (arginase, phosphodiesterase-5 [PDE-5], acetylcholinesterase [AChE] and adenosine deaminase [ADA]) in paroxetine-treated rats. Erectile dysfunction was induced with paroxetine (10 mg/kg body weight). The animals were grouped into five (n = 6): normal rats; paroxetine-induced rats; paroxetine-induced rats treated with sildenafil citrate (5 mg/kg); paroxetine-induced rats treated with orange peels infusion (50 mg/kg); Paroxetine induced rats treated with orange peel infusions (100 mg/kg). The results revealed a significant decrease in sexual behaviour, NO level and the activities of antioxidant enzymes, while there was a significant increase in arginase, PDE-5, AChE and ADA activities in paroxetine-induced rats. However, orange peel infusions ameliorated erectile dysfunction in paroxetine-treated rats. This study showed some possible biochemical basis underlying the use of orange peels infusion in erectile dysfunction management.


Assuntos
Antidepressivos de Segunda Geração/toxicidade , Antioxidantes/administração & dosagem , Citrus sinensis/química , Disfunção Erétil/tratamento farmacológico , Paroxetina/toxicidade , Extratos Vegetais/administração & dosagem , Acetilcolinesterase/metabolismo , Adenosina Desaminase/metabolismo , Animais , Arginase/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Modelos Animais de Doenças , Disfunção Erétil/induzido quimicamente , Disfunção Erétil/patologia , Feminino , Proteínas Ligadas por GPI/metabolismo , Humanos , Masculino , Proteínas de Membrana/metabolismo , Óxido Nítrico/metabolismo , Ereção Peniana/efeitos dos fármacos , Pênis/efeitos dos fármacos , Pênis/patologia , Ratos , Comportamento Sexual/efeitos dos fármacos , Citrato de Sildenafila/administração & dosagem , Resultado do Tratamento
5.
Artigo em Inglês | MEDLINE | ID: mdl-31028932

RESUMO

In this study the nitric oxide (NO)-soluble guanylate cyclase (sGC) and phosphodiesterase-5 (PDE-5) pathway was characterized in tortoise Chelonoidis carbonaria aorta. Concentration response curves (CCR) to ATP, ADP, AMP, adenosine and histamine were performed in the presence and absence of L-NAME in aorta pre-contracted with ACh (3 µM). CCR to SNP, BAY 41-2272 (sGC stimulator), BAY 60-2770 (sGC activator) and tadalafil (PDE-5 inhibitor) were constructed in the presence and absence of ODQ (10 µM). ATP (pEC50 6.1 ±â€¯0.1), ADP (pEC50 6.0 ±â€¯0.2), AMP (pEC50 6.8 ±â€¯0.1) and histamine (pEC50 6.8 ±â€¯0.12) relaxed Chelonoidis aorta and the addition of L-NAME reduced their efficacy (p < .05). Adenosine effects (pEC50 6.6 ±â€¯0.1) were not changed in the presence of L-NAME. SNP (pEC50 7.5 ±â€¯0.7; Emax 102.2 ±â€¯2.5%), BAY 41-2272 (pEC50 7.3 ±â€¯0.2; Emax 130.3 ±â€¯10.2%), BAY 60-2770 (pEC50 11.4 ±â€¯0.1; Emax 130.3 ±â€¯6.5%) and tadalafil (pEC50 6.7 ±â€¯0.3; Emax 121.3 ±â€¯15.3%) relaxed Chelonoidis aorta. The addition of ODQ reduced the SNP and tadalafil maximum response (p < .05) and promoted 63 fold right shift on BAY 41-2272 curve. In contrast, no alteration was observed on BAY 60-2770 response. Transcriptomic analysis for eNOS and sGC were found in aorta and brain libraries with high homology when compared with human transcripts. The NO-sGC-PDE-5 is functionally present in Chelonoidis aorta with a functional and genomic similarity to mammalian vessels. Unlike most of mammalian vessels, ACh did not cause endothelium-dependent relaxation in Chelonoidis carbonaria aortic rings.


Assuntos
Aorta/efeitos dos fármacos , Aorta/metabolismo , Óxido Nítrico/metabolismo , Transcriptoma/efeitos dos fármacos , Tartarugas , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Histamina/farmacologia , Masculino , Agonistas Purinérgicos/farmacologia , Guanilil Ciclase Solúvel/genética , Guanilil Ciclase Solúvel/metabolismo , Vasoconstrição/efeitos dos fármacos
6.
Prostate ; 79(8): 909-919, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30958912

RESUMO

BACKGROUND: Benign prostatic hyperplasia (BPH) is the most common urological disease in elderly men, but the underlying pathophysiological mechanisms are complex and not fully understood. Phosphodiesterase type 5 inhibitors (PDE5-Is) used to treat BPH could upregulate the cyclic guanosine monophosphate (cGMP)-dependent protein kinase G (PKG) signaling, which was shown to blunt inflammation in the prostate. Our previous findings indicate that CD8+ T cells promote the proliferation of BPH epithelial cells (BECs) in low androgen conditions through secretion of CCL5; however, the role of the cGMP/PKG pathway in the process is unclear. METHODS: Paraffin-embedded tissues were used for expression quantity of CD8+ T cells, CCL5, cyclin D1, and PDE5 protein by immunohistology in prostate specimens which were/were not treated with finasteride 5 mg daily for at least 6 months before surgery. BPH-1 cells were cocultured with or without CD8 + T cells or PDE5-Is in low androgen conditions for 4 days. The conditioned media, BPH-1 cells, and CD8 + T cells were harvested for the subsequent experiments. The quantitative polymerase chain reaction was used for assaying the level of messenger RNA expression of CCL5. CCL5 in the conditioned media was detected by the enzyme-linked immunosorbent assay. The effect of PDE5-Is on cocultured BPH-1/CD8 + T-cell proliferation was detected by the cell counting kit-8. A high-fat diet (HFD)-induced prostatic hyperplasia rat model was used to investigate the effect of cGMP/PKG activation in CD8 + T cells in vivo. RESULTS: CD8+ T-cell infiltration into human BPH tissues was positively correlated with the expression of CCL5, cyclin D1, and PDE5, whereas in an HFD-induced prostatic hyperplasia rat model, the activation of the cGMP/PKG signaling by a PDE5-I could suppress the CD8 + T-cell infiltration and the CCL5 and cyclin D1 expression. Furthermore, the activation of the cGMP/PKG pathway inhibited CCL5 secretion by CD8 + T cells by downregulating nuclear factor-κB p65 phosphorylation, which reduced the growth of BPH-1 through CCL5/STAT5/CCND1 signaling. CONCLUSIONS: Our results indicate that the upregulation of the cGMP/PKG/p65 signaling reduces CCL5 secretion in CD8 + T cells, which in turn decreases the proliferation of BECs in low androgen conditions, suggesting that the combination of 5α reductase inhibitors lowering androgen levels and PDE5-Is may be a novel, more effective treatment for BPH patients.


Assuntos
Linfócitos T CD8-Positivos/metabolismo , Quimiocina CCL5/metabolismo , Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Fator de Transcrição RelA/metabolismo , Animais , Linfócitos T CD8-Positivos/patologia , Linhagem Celular , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Ciclina D1/metabolismo , Regulação para Baixo , Humanos , Masculino , Hiperplasia Prostática/metabolismo , Hiperplasia Prostática/patologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais
8.
Am J Physiol Regul Integr Comp Physiol ; 316(6): R704-R715, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-30892912

RESUMO

Nitric oxide (NO) is a potent vasodilator, which improves perfusion and oxygen delivery during tissue hypoxia in terrestrial animals. The vertebrate dive response involves vasoconstriction in select tissues, which persists despite profound hypoxia. Using tissues collected from Weddell seals at necropsy, we investigated whether vasoconstriction is aided by downregulation of local hypoxia signaling mechanisms. We focused on NO-soluble guanylyl cyclase (GC)-cGMP signaling, a well-known vasodilatory transduction pathway. Seals have a lower GC protein abundance, activity, and capacity to respond to NO stimulation than do terrestrial mammals. In seal lung homogenates, GC produced less cGMP (20.1 ± 3.7 pmol·mg protein-1·min-1) than the lungs of dogs (-80 ± 144 pmol·mg protein-1·min-1 less than seals), sheep (-472 ± 96), rats (-664 ± 104) or mice (-1,160 ± 104, P < 0.0001). Amino acid sequences of the GC enzyme α-subunits differed between seals and terrestrial mammals, potentially affecting their structure and function. Vasoconstriction in diving Weddell seals is not consistent across tissues; perfusion is maintained in the brain and heart but decreased in other organs such as the kidney. A NO donor increased median GC activity 49.5-fold in the seal brain but only 27.4-fold in the kidney, consistent with the priority of cerebral perfusion during diving. Nos3 expression was high in the seal brain, which could improve NO production and vasodilatory potential. Conversely, Pde5a expression was high in the seal renal artery, which may increase cGMP breakdown and vasoconstriction in the kidney. Taken together, the results of this study suggest that alterations in the NO-cGMP pathway facilitate the diving response.


Assuntos
Encéfalo/irrigação sanguínea , Caniformia/metabolismo , Circulação Cerebrovascular , Mergulho , Guanilato Ciclase/metabolismo , Rim/irrigação sanguínea , Circulação Renal , Vasoconstrição , Animais , Caniformia/genética , GMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Regulação Enzimológica da Expressão Gênica , Guanilato Ciclase/genética , Homeostase , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Sistemas do Segundo Mensageiro , Especificidade da Espécie
9.
Biomed Pharmacother ; 111: 1029-1035, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30841416

RESUMO

Sexual dysfunction is a side effect of the antidepressant drug paroxetine. Anogeissus leiocarpus is a medicinal plant with a wide range of biological activities which include antioxidant and antiulcer properties. With these in mind, we investigated the effect of Anogeissus leiocarpus stem bark extract on paroxetine-induced sexual dysfunction in male Wistar rats. Forty-two adult male Wistar rats were divided into seven experimental groups: normal control, PAR (10 mg/kg), PAR + sildenafil (5 mg/kg), ALE (50 and 100 mg/kg) and PAR + ALE (50 and 100 mg/kg). The experiment lasted for 21 days, after which the rats were subjected to sexual behavioral test. Various biochemical assays (phosphodiesterase-5, arginase, acetylcholinesterase, nitric oxide and MDA) were carried out on the penile tissue homogenate. From our findings, paroxetine significantly altered sexual behavior in male rats and increased phosphodiesterase-5, arginase and acetylcholinesterase activities with a concomitant decrease in nitric oxide level. Furthermore, paroxetine altered antioxidant status which revealed by increased MDA level and reduced thiol level. However, treatment with Anogeissus leiocarpus stem bark extract reversed the altered sexual behavior in male rats and boosted antioxidant status. In addition, administration of Anogeissus leiocarpus stem bark extract resulted in a significant attenuation of phosphodiesterase-5, arginase and acetylcholinesterase activities in paroxetine-induced rats. In view of the aforementioned findings, Anogeissus leiocarpus could be considered a promising natural agent in erectile dysfunction management.


Assuntos
Antioxidantes/metabolismo , Óxido Nítrico/metabolismo , Paroxetina/farmacologia , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Comportamento Sexual/efeitos dos fármacos , Disfunções Sexuais Fisiológicas/tratamento farmacológico , Animais , Arginase/metabolismo , Combretaceae/química , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Disfunção Erétil/tratamento farmacológico , Disfunção Erétil/metabolismo , Masculino , Malondialdeído/metabolismo , Pênis/efeitos dos fármacos , Pênis/metabolismo , Ratos , Ratos Wistar , Disfunções Sexuais Fisiológicas/metabolismo , Citrato de Sildenafila/farmacologia
10.
Curr Comput Aided Drug Des ; 15(4): 318-333, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30767749

RESUMO

OBJECTIVE: To screen the phytochemicals for phosphodiesterase 5A (PDE5A) inhibitory potential and identify lead scaffolds of antihypertensive phytochemicals using in silico docking studies. METHODS: In this perspective, reported 269 antihypertensive phytochemicals were selected. Sildenafil, a PDE5A inhibitor was used as the standard. In silico docking study was carried out to screen and identify the inhibiting potential of the selected phytochemicals against PDE5A enzyme using vLife MDS 4.4 software. RESULTS: Based on docking score, π-stacking, H-bond and ionic interactions, 237 out of 269 molecules were selected which have shown one or more interactions. Protein residue Gln817A was involved in H-boding whereas Val782A, Phe820A and Leu804A were involved in π-stacking interaction with ligand. The selected 237 phytochemicals were structurally diverse, therefore 82 out of 237 molecules with one or more tricycles were filtered out for further analysis. Amongst tricyclic molecules, 14 molecules containing nitrogen heteroatom were selected for lead scaffold identification which finally resulted in three different basic chemical backbones like pyridoindole, tetrahydro-pyridonaphthyridine and dihydro-pyridoquinazoline as lead scaffolds. CONCLUSION: In silico docking studies revealed that nitrogen-containing tetrahydro-pyridonaphthyridine and dihydro-pyridoquinazoline tricyclic lead scaffolds have emerged as novel PDE5A inhibitors for antihypertensive activity. The identified lead scaffolds may provide antihypertensive lead molecules after its optimization.


Assuntos
Anti-Hipertensivos/química , Anti-Hipertensivos/farmacologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Inibidores da Fosfodiesterase 5/química , Inibidores da Fosfodiesterase 5/farmacologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/química , Desenho de Drogas , Descoberta de Drogas , Humanos , Ligantes , Simulação de Acoplamento Molecular , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Software
11.
Curr Top Med Chem ; 19(4): 305-315, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30747070

RESUMO

BACKGROUND: PDE5A is a phosphodiesterase which specifically hydrolyzes the cGMP to GMP. It takes part in several physiological and pathological pathways and is considered an important drug target. Currently, PDE5 inhibitors (ex; Sildenafil, Tadalafil) available in the market are not only being used for the treatment of erectile dysfunction but at the same time, they are also in clinical trials being investigated as anticancer agents. MATERIALS & METHODS: In this work, we have examined pyrazolo [4,3-c]quinolin-3-ones as PDE5A inhibitors. Pyrazolo [4,3-c]quinolin-3-ones are the class of tricyclic heterocyclic derivatives having a variety of therapeutically interesting drug candidates known for their anti-inflammatory, anti-viral, anti-anxiety and anti-cancer activity. Therefore, synthetic methods providing access to pyrazolo [4, 3-c] quinolin-3-ones are immensely valuable. Here, we are reporting a simple but efficient route for the synthesis of novel 8-morpholino-2-aryl - 2, 5-dihydro-3H-pyrazolo [4, 3-c] quinolin-3-one derivatives. RESULTS: Further, molecular docking studies of synthesized compounds with human PDE5A protein showed that all the compounds exhibited good docking score in comparison with known inhibitors. In addition, all the synthesized molecules were evaluated against HCT116 cell lines for their antitumor activity. CONCLUSION: Among all the synthesized compounds, compound 5a, 5d, and 6e showed better cytotoxicity. Thus, these derivatives can be studied as potential inhibitors of PDE5A.


Assuntos
Antineoplásicos/farmacologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Inibidores da Fosfodiesterase 5/química , Inibidores da Fosfodiesterase 5/farmacologia , Pirazóis/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Células HCT116 , Humanos , Modelos Moleculares , Estrutura Molecular , Inibidores da Fosfodiesterase 5/síntese química , Pirazóis/síntese química , Pirazóis/química , Quinolonas/síntese química , Quinolonas/química , Quinolonas/farmacologia , Relação Estrutura-Atividade
12.
Res Vet Sci ; 125: 397-400, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29174607

RESUMO

Pulmonary hypertension (PH), remains a challenging disease with a large impact on both humans and meat-type chickens. PH is characterized by the onset of idiopathic pulmonary arterial hypertension leading to right ventricular failure. In this experiment relative gene expression of adenosine A1 receptor (ADORA1), serotonin transporter (SERT), phosphodiesterase 5A (PDE5) and troponin T2 were compared in hearts from broilers with acute right ventricular failure and from healthy birds. There were major increases in adenosine A1 receptor (177%) and serotonin transporter (475%), and more modest but significant increases in PDE5 (146%) and troponin T2 (140%) gene expressions in broilers with right ventricular failure compared to healthy birds (P<0.01). This novel report shows that pulmonary hypertension related gene expression in broilers is similar to that in humans. This molecular similarity between PH in broilers and human patients suggests, first, that they will make a suitable animal model for study PH in humans, but also that the literature on PH in humans may be profitably applied to the study of PH in broilers.


Assuntos
Galinhas/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Receptor A1 de Adenosina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Troponina T/metabolismo , Regulação para Cima , Animais , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Regulação da Expressão Gênica , Insuficiência Cardíaca/metabolismo , Insuficiência Cardíaca/veterinária , Hipertensão Pulmonar/veterinária , Masculino , Miocárdio/metabolismo , Doenças das Aves Domésticas/metabolismo , Receptor A1 de Adenosina/genética , Troponina T/genética
13.
J Ethnopharmacol ; 229: 167-179, 2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-30339977

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Selected Peruvian Amazon plants are macerated into sugar cane distillates to prepare alcoholic beverages used to improve male sexual performance. The tree bark from Campsiandra angustifolia Spruce ex Benth (Fabaceae), Swartzia polyphylla DC (Fabaceae), Minquartia guianensis Aubl. (Olacaceae) and Thynantus panurensis (Bureau) Sandwith (Bignoniaceae) usually are used as crude drugs in mixtures of several ingredients. AIM OF STUDY: Describe the chemical composition of the most traded traditional male enhancer beverages, namely "Levántate Lazaro" and "Siete veces sin sacarla", and their single crude drug constituents, as well as their inhibitory activity towards the enzyme phosphodiesterase-5. The presence of pro-sexual drugs such as Sildenafil® and derivatives was assessed in the samples. MATERIALS AND METHODS: Single plant constituents and the preparation mixtures were purchased in the Mercado Belen (Iquitos, Peru). Chemical profiling was carried out by HPLC-DAD-ESI-MS/MS. The extracts were assessed for phosphodiesterase-5 inhibition. The occurrence of pro-sexual drugs was determined by HPLC-DAD-ESI-MS/MS. RESULTS: Chemical profiling allowed the identification of condensed tannins as the main constituents of C. angustifolia and S. polyphylla, hydrolysable tannins for M. guianensis, and C-glycosides for T. panurensis. The traditional preparations showed similar composition compared to the crude drugs. At 200 µg/mL, the traditional preparation "Levántate Lázaro" and "Siete veces sin sacarla" inhibited the phosphodiesterase-5 by 49.88% and 27.90%, respectively. No adulterations with pro-sexual drugs were found in the samples. From the crude drugs, low effect was found for the extracts of S. polyphylla and T. panurensis and high activity for C. angustifolia which inhibited the enzyme by 89.37% and 81.32% at 200 and 100 µg/mL, respectively. CONCLUSION: The traditional preparations used to improve sexual performance in the Peruvian Amazon showed activity as phosphodiesterase-5 inhibitors. The most active ingredient of the traditional preparations was C. angustifolia, with some contribution from T. panurensis. These results encourage additional studies, including animal models to confirm the male enhancer effect of the preparations.


Assuntos
Afrodisíacos/farmacologia , Magnoliopsida , Inibidores da Fosfodiesterase 5/farmacologia , Preparações de Plantas/farmacologia , Afrodisíacos/química , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Etanol/farmacologia , Humanos , Masculino , Peru , Inibidores da Fosfodiesterase 5/química , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/farmacologia , Casca de Planta , Preparações de Plantas/química
14.
Chem Biol Drug Des ; 93(4): 419-429, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30381900

RESUMO

Phosphodiesterase type 5 (PDE-5) is an important enzyme involved in the hydrolysis of cyclic guanosine monophosphate (cGMP) to guanosine monophosphate (GMP). The inhibition of this protein leads to the accumulation of cGMP in cells with various biological and therapeutic effects. Several PDE-5 inhibitors exist, with Tadalafil being one of the most commonly studied and used in clinical therapy. In this study, we applied Molecular Dynamics simulations coupled to the ABF (Adaptive Biasing Force) method to study the effect of the mutation on the Gln817 residue (Q817G). The results of the free energy profiles made clear that the affinity of the inhibitor for PDE-5 is dependent on the amino acid residue Gln817. The hydrogen bond made between the side chain of glutamine and the indole ring of Tadalafil results in the stabilization of the ligand in the catalytic site. Despite the prominent role of this interaction, it is important to highlight the contribution of other residues of the catalytic domain for the stabilization of the compound, due to the set of polar, hydrophobic and electrostatic interactions performed by specific amino acid residues.


Assuntos
Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Inibidores da Fosfodiesterase 5/química , Tadalafila/química , Sítios de Ligação , Domínio Catalítico , GMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/química , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Humanos , Ligações de Hidrogênio , Ligantes , Simulação de Dinâmica Molecular , Mutagênese Sítio-Dirigida , Inibidores da Fosfodiesterase 5/metabolismo , Tadalafila/metabolismo , Termodinâmica
15.
Biochemistry ; 58(6): 799-808, 2019 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-30532959

RESUMO

Forster resonance energy transfer (FRET)-based biosensors have been recently applied to the study of biological pathways. In this study, a new biosensor was validated for the first time in live HEK293 and steroidogenic MLTC-1 cell lines for studying the effect of the PDE5 inhibitor on the hCG/LH-induced steroidogenic pathway. The sensor improves FRET between a donor (D), the fluorescein-like diarsenical probe that can covalently bind a tetracysteine motif fused to the PDE5 catalytic domain, and an acceptor (A), the rhodamine probe conjugated to the pseudosubstrate cGMPS. Affinity constant ( Kd) values of 5.6 ± 3.2 and 13.7 ± 0.8 µM were obtained with HEK293 and MLTC-1 cells, respectively. The detection was based on the competitive displacement of the cGMPS-rhodamine conjugate by sildenafil; the Ki values were 3.6 ± 0.3 nM (IC50 = 2.3 nM) in HEK293 cells and 10 ± 1.0 nM (IC50 = 3.9 nM) in MLTC-1 cells. The monitoring of both cAMP and cGMP by bioluminescence resonance energy transfer allowed the exploitation of the effects of PDE5i on steroidogenesis, indicating that sildenafil enhanced the gonadotropin-induced progesterone-to-testosterone conversion in a cAMP-independent manner.


Assuntos
Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Inibidores da Fosfodiesterase 5/metabolismo , Progesterona/biossíntese , Citrato de Sildenafila/metabolismo , Testosterona/biossíntese , Animais , Arsenicais/química , Técnicas Biossensoriais/métodos , Domínio Catalítico , Linhagem Celular Tumoral , Gonadotropina Coriônica/farmacologia , AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/química , Cisteína/química , Transferência Ressonante de Energia de Fluorescência/métodos , Corantes Fluorescentes/química , Células HEK293 , Humanos , Hormônio Luteinizante/farmacologia , Camundongos , Inibidores da Fosfodiesterase 5/farmacologia , Progesterona/metabolismo , Ligação Proteica , Rodaminas/química , Citrato de Sildenafila/farmacologia , Testosterona/metabolismo
16.
Iran Biomed J ; 23(4): 280-6, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30388886

RESUMO

Background: This study aimed to investigate Levisticum officinale hydroalcoholic extract (LOHE) effect on both cGMP signaling pathway and phosphodiesterase 5 (PDE5) gene expression pattern and to examine the role of LOHE in apoptosis induction of MCF-7 and MDA-MB-468 cell lines. Methods: The half maximal inhibitory concentration (IC50) of LOHE was examined in both cell lines using the MTT assay. Using IC50 values of LOHE on both cells, the type of cell death was detected by flowcytometric analysis. The values of PDE5 and cGMP were evaluated by real-time PCR and ELISA methods, respectively. Results: The IC50 values were measured as 150 µg/ml for MDA-MB-468 and 200 µg/ml for MCF-7. At 12 hour of treatment, a significant decrease in the PDE5 expression and maximum increase in the amount of intracellular cGMP were observed (p < 0.05). However, these effects were more noticeable in MDA-MB-468 triple-negative cells. Conclusion: Our data suggest that LOHE extract could be a potential source for new strategies towards targeting both PDE5 and cGMP signaling pathways.


Assuntos
Apoptose/efeitos dos fármacos , GMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Regulação para Baixo/efeitos dos fármacos , Levisticum/química , Extratos Vegetais/farmacologia , Transdução de Sinais , Água/química , Álcoois/química , Proliferação de Células/efeitos dos fármacos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Células MCF-7 , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos
17.
J Neuropathol Exp Neurol ; 78(2): 191-194, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30590671

RESUMO

Cerebral small vessel disease in deep penetrating arteries is a major cause of lacunar infarcts, white matter lesions and vascular cognitive impairment. Local cerebral blood flow in these small vessels is controlled by endothelial-derived nitric oxide, which exerts a primary vasodilator stimulus on vascular myocytes, via cytoplasmic cyclic GMP. Here, we investigated whether the cGMP-degrading enzyme phosphodiesterase-5 (PDE5) is present in small penetrating arteries in the deep subcortical white matter of older people. Frontal cortical tissue blocks were examined from donated brains of older people (n = 42, 24 male: 18 female, median age 81, range: 59-100 years). PDE5, detected by immunohistochemical labeling, was graded as absent, sparse, or abundant in vascular cells within small arteries in subcortical white matter (vessel outer diameter: 20-100 µm). PDE5 labeling within arterial myocytes was detected in all cases. Degree of PDE5 expression (absent, sparse, or abundant) was not associated with age or with neuropathological diagnosis of small vessel disease. In conclusion, PDE5 is present in vascular myocytes within small penetrating arteries in older people. This is a potential molecular target for pharmacological interventions.


Assuntos
Encéfalo/enzimologia , Artérias Cerebrais/enzimologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Músculo Liso Vascular/enzimologia , Substância Branca/enzimologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Células Musculares/enzimologia
18.
Bioorg Med Chem Lett ; 29(2): 267-270, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30509781

RESUMO

We describe the design, synthesis and evaluation of a series of N2,N4-diaminoquinazoline analogs as PDE5 inhibitors. Twenty compounds were prepared and these were assessed in terms of their PDE5 and PDE6 activity, ex-vivo vasodilation response, mammalian cytotoxicity and aqueous solubility. Molecular docking was used to determine the binding mode of the series and this was demonstrated to be consistent with the observed SAR. Compound 15 was the most active PDE5 inhibitor (IC50 = 0.072 ±â€¯0.008 µM) and exhibited 4.6-fold selectivity over PDE6. Ex-vivo assessment of 15 and 22 in a rat pulmonary artery vasodilation model demonstrated EC50s of 1.63 ±â€¯0.72 µM and 2.28 ±â€¯0.74 µM respectively.


Assuntos
Antineoplásicos/farmacologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Desenho de Drogas , Inibidores da Fosfodiesterase 5/farmacologia , Quinazolinas/farmacologia , Células A549 , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Inibidores da Fosfodiesterase 5/síntese química , Inibidores da Fosfodiesterase 5/química , Quinazolinas/síntese química , Quinazolinas/química , Ratos , Relação Estrutura-Atividade
19.
Biomed Res Int ; 2018: 6452965, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30498760

RESUMO

In this study, we aimed to assess the effect and possible underlying mechanism of Anacardium occidentale leaves extract on male sexual behaviors in stress-exposed rats. Male Wistar rats were orally given A. occidentale extract at doses of 25, 100, and 200 mg/kg BW before 12-hour-immobilization exposure for 14 days. Sexual behaviors, serum testosterone and corticosterone levels, TH-positive cells density in nucleus accumbens (NAc) and ventral tegmental area (VTA), MAO-B activity in NAc and medial preoptic area (MPOA), testis histology together with phosphodiesterase type-5 ( PDE-5) activity, and endothelial nitric oxide synthase (eNOS) expression in penis were evaluated after treatment. All doses of extract improved male sexual behaviors, suppressed MAO-B in NAc, enhanced TH-positive cells density in NAc, suppressed PDE-5 in penis, and enhanced interstitial cell of Leydig. The increase of serum testosterone, TH-positive cells density in VTA, eNOS expression in penis, and the decreased serum corticosterone were observed at some doses. Therefore, the sexual enhancing effect of extract occurred mainly via the improved dopaminergic and testicular functions. PDE-5 suppression in penis also played the role especially in the increased intromission behavior. Therefore, A. occidentale leaves extract is the potential protective agent against sexual dysfunction. However, further researches are necessary.


Assuntos
Anacardium/química , Dopamina/metabolismo , Extratos Vegetais/farmacologia , Comportamento Sexual Animal/efeitos dos fármacos , Estresse Psicológico/fisiopatologia , Testículo/fisiopatologia , Animais , Corticosterona/sangue , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/enzimologia , Células Endoteliais/patologia , Masculino , Monoaminoxidase/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Pênis/efeitos dos fármacos , Pênis/enzimologia , Folhas de Planta/química , Ratos Wistar , Estresse Psicológico/sangue , Testículo/efeitos dos fármacos , Testosterona/sangue
20.
Biochem Biophys Res Commun ; 505(3): 685-691, 2018 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-30292404

RESUMO

BACKGROUND: Sildenafil, a phosphodiesterase 5 (PDE5) inhibitor, is known to increase the intracellular level of cyclic guanosine monophosphate (cGMP), which causes vasodilation. However, the effect of sildenafil on human hair follicles (hHFs) is unknown. OBJECTIVE: The purpose of this study was to determine the role of sildenafil in hair growth. METHODS: We investigated the expression of PDE5 in human dermal papilla cells (hDPCs) and hHFs. The effects of sildenafil on hDPC proliferation were evaluated using BrdU assays. The mRNA expression of growth factors and extracellular signal-regulated kinase (ERK) phosphorylation were investigated using real-time PCR and western blotting, respectively. Additionally, anagen induction and perifollicular vessel formation were evaluated using an in vivo mice model. RESULTS: We confirmed high expression of PDE5 in hDPCs and hHFs. Sildenafil enhances proliferation of hDPCs and up-regulates the mRNA expression of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), which are responsible for hair growth. Additionally, sildenafil up-regulates the levels of phosphorylated ERK and accelerates anagen induction by stimulating perifollicular vessel formation after topical application in mice. CONCLUSION: Our study demonstrates for the first time, the significant therapeutic potential of sildenafil on hair growth and its potential use in treatment of alopecia.


Assuntos
Derme/efeitos dos fármacos , Folículo Piloso/efeitos dos fármacos , Inibidores da Fosfodiesterase 5/farmacologia , Citrato de Sildenafila/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Derme/citologia , Derme/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Cabelo/efeitos dos fármacos , Cabelo/crescimento & desenvolvimento , Folículo Piloso/citologia , Folículo Piloso/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Camundongos Endogâmicos C57BL , Fosforilação/efeitos dos fármacos
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