ABSTRACT
The aim of current study was to determinate ex vivo and chromatographic fingerprint by HPLC of four extracts of Euphorbia furcillata K. Ethyl acetate extract of Euphorbia furcillata (EaEEf) was the most effective and potent extract (Emax=98.69±1.24%) and its effect was partially endothelium-dependent. Functional vasorelaxant mechanism of action of EaEEf was determinate, EaEEf showed efficient relaxation of KCl [80 mM]-induced contraction and norepinephrine and CaCl2 contraction curves showed diminution of maximal contraction in the presence of EAEEf and EaEEf-relaxation curve was shifted to the right in the presence of L-NAME (nitric oxide synthase inhibitor) and ODQ (guanylate cyclase inhibitor). Chromatographic fingerprints analysis suggests presence of diterpenoid such as abietane, tigliane, and ingenane skeletons. Our experiments suggest the EaEEf vasorelaxant activity could be attributed to diterpenoid molecules whose mechanism involves nitric oxide production and calcium channel blockade.
Se determinoÌ el efecto vasorrelajante ex vivo y los perfiles cromatograÌficos mediante HPLC de cuatro extractos de Euphorbia furcillata K.. El extracto de acetato de etilo de E. furcillata (EaEEf) fue el maÌs eficaz y potente en la contraccioÌn inducida por norepinefrina (Emax=98.69±1.24%) y el efecto fue parcialmente dependiente del endotelio vascular. Se determinoÌ el mecanismo de accioÌn vasorrelajante para EaEEf, este mostroÌ ser eficaz sobre la contraccioÌn inducida por KCl [80 mM] y la curva de contraccioÌn en respuesta a norepinefrina y CaCl2 en presencia de EaEEf mostroÌ disminucioÌn en la contraccioÌn maÌxima, mientras que la curva de relajacioÌn de EaEEf en presencia de L-NAME (inhibidor de oÌxido niÌtrico sintasa) y ODQ (inhibidor de guanilato ciclasa) se desplazoÌ hacia la derecha. El anaÌlisis cromatograÌfico de EaEEf sugiere la presencia de moleÌculas diterpenoides como abietano, tigliano y esqueletos de ingenano. Nuestros resultados sugieren que el efecto vasorrelajante de EaEEf podriÌa atribuirse a moleÌculas diterpenoides, cuyo mecanismo de accioÌn involucra la produccioÌn de oÌxido niÌtrico y bloqueo de canales de calcio.
Subject(s)
Animals , Male , Rats , Vasodilator Agents/pharmacology , Plant Extracts/pharmacology , Euphorbia/chemistry , Calcium Channel Blockers/metabolism , Chromatography, High Pressure Liquid , Rats, Wistar , Cyclic GMP/metabolism , Nitric Oxide/metabolismABSTRACT
The aim of this study was to investigate the effect and action mechanism of quercetin on penile corpus cavernosum smooth muscle (PCCSM). PCCSM precontracted with phenylephrine (Phe) was treated with four different concentrations of quercetin (10−7, 10−6, 10−5 and 10−4 M). PCCSM were preincubated with N-Nitro-L-arginine methyl ester hydrochloride (L-NAME) and 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) to block nitric oxide synthase and guanylate cyclase, respectively. The changes in PCCSM tension were recorded, and cyclic nucleotides in the perfusate were measured by radioimmunoassay. The interactions of quercetin with phosphodiesterase type 5 inhibitors (PDE5-Is) such as sildenafil, udenafil and mirodenafil, were also evaluated. PCCSM relaxation induced by quercetin occurred in a concentrationdependent manner. The application of quercetin to PCCSM pre-treated with L-NAME and ODQ significantly inhibited the relaxation. Quercetin significantly increased cGMP in the perfusate. Furthermore, quercetin enhanced PDE5-Is-induced relaxation of PCCSM. Quercetin relaxed the PCCSM by activating the NO-cGMP signaling pathway, and it may be a therapeutic candidate or an alternative treatment for patients with erectile dysfunction who do not completely respond to PDE5-Is.
Subject(s)
Humans , Male , Erectile Dysfunction , Guanylate Cyclase , Muscle, Smooth , NG-Nitroarginine Methyl Ester , Nitric Oxide Synthase , Nucleotides, Cyclic , Phenylephrine , Phosphodiesterase 5 Inhibitors , Quercetin , Radioimmunoassay , Relaxation , Sildenafil CitrateABSTRACT
Estudos publicados nas duas últimas décadas sugerem um aumento do risco de doença cardiovascular (DCV) em pacientes com periodontite, mas os mecanismos fisiopatológicos dessa associação ainda não estão completamente esclarecidos. Uma vez que foi demonstrado aumento da ativação plaquetária e do estresse oxidativo na periodontite, o objetivo do presente estudo foi investigar a via L-arginina-óxido nítrico (NO)- guanosina monofosfato cíclica (GMPc) e parâmetros de estresse oxidativo em plaquetas de pacientes com periodontite, bem como avaliar o efeito do tratamento periodontal não-cirúrgico nessas variáveis. Um total de 10 pacientes sem periodontite (periodontalmente saudáveis ou com gengivite) e 10 pacientes com periodontite participaram do estudo. A avaliação clínica, laboratorial e experimental foi realizada no início do estudo e 90 dias após realização da terapia periodontal básica (grupo periodontite). A avaliação clínica periodontal incluiu registros de: profundidade de bolsa à sondagem (PBS), nível de inserção (NIC), percentual de placa e percentual de sangramento à sondagem. Os seguintes experimentos foram realizados: influxo de L-arginina; atividade e expressão das enzimas óxido nítrico sintase e da arginase; expressão das enzimas guanilato ciclase solúvel e fosfodiesterase 5; determinação dos níveis intraplaquetários de GMPc; agregação plaquetária; avaliação do estresse oxidativo (atividade oxidante total, atividade das enzimas antioxidantes catalase e da superóxido dismutase - SOD); medição dos níveis de proteína C reativa (CRP) e de fibrinogênio. Os resultados obtidos no início do estudo demonstraram ativação do influxo de L-arginina em plaquetas via sistema y+L nos pacientes com periodontite, bem como concentrações intraplaquetárias de GMPc diminuídas e aumento sistêmico da CRP. Após o tratamento periodontal, observou-se redução do percentual de sítios com PBS ≥ 6 mm, NIC 4-5 mm e NIC ≥ 6 mm, aumento nos níveis de GMPc, para níveis ...
Studies published over the last two decades have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients, but the physiopathological mechanisms involved in this association are not yet clear. Since it has been demonstrated an enhancement on both platelet activation and oxidative stress on periodontitis patients, the aim of this study was to investigate the L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway on platelets from periodontitis patients, and the effect of non-surgical periodontal treatment in these variables. A total of 10 patients without periodontitis (periodontal healthy controls or gingivitis patients) and 10 periodontitis patients were included in this study. The clinical, laboratorial, and experimental evaluations were performed at the beginning of the study and 90 days after the basic periodontal therapy (periodontitis group). The clinical periodontal evaluation included the measurements of probing pocket depth (PPD), clinical attachment level (CAL), plaque percentage, and percentage of bleeding on probing. The following experiments were performed: L-arginine influx; nitric oxide synthase and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; measurement of intraplatelet cGMP levels; platelet aggregation; oxidative stress evaluation (total oxidant activity and activity of both antioxidant enzymes catalase and superoxide dismutase SOD); measurement of C reactive protein (CRP) and fibrinogen. The initial results demonstrated an activation of L-arginine influx in platelets from periodontitis patients via y+L system, reduced intraplatelet cGMP levels and increased CRP. After periodontal treatment, it was observed reduction on percentage of sites with PPD ≥ 6 mm, CAL 4-5 mm and CAL ≥ 6 mm, enhancement on cGMP levels, to levels comparables to patients without periodontitis, accompanied by a higher activity of both antioxidant ...
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Arginine/metabolism , Cyclic GMP/metabolism , Oxidative Stress , Nitric Oxide/metabolism , Periodontitis/therapy , Blood Platelets/enzymology , Blood Platelets/metabolism , C-Reactive Protein , Cardiovascular Diseases/etiology , Fibrinogen , Platelet Activation , Platelet AggregationABSTRACT
Estudos publicados nas duas últimas décadas sugerem um aumento do risco de doença cardiovascular (DCV) em pacientes com periodontite, mas os mecanismos fisiopatológicos dessa associação ainda não estão completamente esclarecidos. Uma vez que foi demonstrado aumento da ativação plaquetária e do estresse oxidativo na periodontite, o objetivo do presente estudo foi investigar a via L-arginina-óxido nítrico (NO)- guanosina monofosfato cíclica (GMPc) e parâmetros de estresse oxidativo em plaquetas de pacientes com periodontite, bem como avaliar o efeito do tratamento periodontal não-cirúrgico nessas variáveis. Um total de 10 pacientes sem periodontite (periodontalmente saudáveis ou com gengivite) e 10 pacientes com periodontite participaram do estudo. A avaliação clínica, laboratorial e experimental foi realizada no início do estudo e 90 dias após realização da terapia periodontal básica (grupo periodontite). A avaliação clínica periodontal incluiu registros de: profundidade de bolsa à sondagem (PBS), nível de inserção (NIC), percentual de placa e percentual de sangramento à sondagem. Os seguintes experimentos foram realizados: influxo de L-arginina; atividade e expressão das enzimas óxido nítrico sintase e da arginase; expressão das enzimas guanilato ciclase solúvel e fosfodiesterase 5; determinação dos níveis intraplaquetários de GMPc; agregação plaquetária; avaliação do estresse oxidativo (atividade oxidante total, atividade das enzimas antioxidantes catalase e da superóxido dismutase - SOD); medição dos níveis de proteína C reativa (CRP) e de fibrinogênio. Os resultados obtidos no início do estudo demonstraram ativação do influxo de L-arginina em plaquetas via sistema y+L nos pacientes com periodontite, bem como concentrações intraplaquetárias de GMPc diminuídas e aumento sistêmico da CRP. Após o tratamento periodontal, observou-se redução do percentual de sítios com PBS ≥ 6 mm, NIC 4-5 mm e NIC ≥ 6 mm, aumento nos níveis de GMPc, para níveis ...
Studies published over the last two decades have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients, but the physiopathological mechanisms involved in this association are not yet clear. Since it has been demonstrated an enhancement on both platelet activation and oxidative stress on periodontitis patients, the aim of this study was to investigate the L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway on platelets from periodontitis patients, and the effect of non-surgical periodontal treatment in these variables. A total of 10 patients without periodontitis (periodontal healthy controls or gingivitis patients) and 10 periodontitis patients were included in this study. The clinical, laboratorial, and experimental evaluations were performed at the beginning of the study and 90 days after the basic periodontal therapy (periodontitis group). The clinical periodontal evaluation included the measurements of probing pocket depth (PPD), clinical attachment level (CAL), plaque percentage, and percentage of bleeding on probing. The following experiments were performed: L-arginine influx; nitric oxide synthase and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; measurement of intraplatelet cGMP levels; platelet aggregation; oxidative stress evaluation (total oxidant activity and activity of both antioxidant enzymes catalase and superoxide dismutase SOD); measurement of C reactive protein (CRP) and fibrinogen. The initial results demonstrated an activation of L-arginine influx in platelets from periodontitis patients via y+L system, reduced intraplatelet cGMP levels and increased CRP. After periodontal treatment, it was observed reduction on percentage of sites with PPD ≥ 6 mm, CAL 4-5 mm and CAL ≥ 6 mm, enhancement on cGMP levels, to levels comparables to patients without periodontitis, accompanied by a higher activity of both antioxidant ...(AU)
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Arginine/metabolism , Cyclic GMP/metabolism , Oxidative Stress , Nitric Oxide/metabolism , Periodontitis/therapy , Blood Platelets/enzymology , Blood Platelets/metabolism , C-Reactive Protein , Cardiovascular Diseases/etiology , Fibrinogen , Platelet Activation , Platelet AggregationABSTRACT
This study was undertaken to investigate an involvement of nitroxergic innervation in gastric smooth muscle of rat. Isometric tension study, the measurement of single cell length, NADPH diaphorase stain of smooth muscle layers and neuronal nitric oxide synthase (nNOS) western blotting were performed. Sodium nitroprusside (SNP), a nitric oxide donor, relaxed the muscle strips precontracted by acetylcholine (ACh) in a concentration-dependent manner. Pretreatment of L-arginine decreased the contraction induced by electric field stimulation (EFS). Pretreatment of NG-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, increased the EFS-induced contractions. LY 83583, a guanylate cyclase (GC) inhibitor, reversed the inhibitory actions of L-arginine on the muscle contractions. The effects of L-Arginine, L-NAME and LY 83583 on ACh-induced contractions were not significant. L-arginine reduced the EFS-induced contraction in circular muscle, whereas L-NAME enhanced the EFS-induced contraction in longitudinal strips. By EFS, the phasic contractions appeared approximately 20~25 seconds later. L-NAME significantly shortened the delay time to about 2~3 seconds. In single cell study, ACh contracted gastric smooth muscle cells, SNP relaxed the cells, and the latter also inhibited the ACh-induced contraction. LY 83583 enhanced the ACh-induced contraction and antagonized SNP-induced relaxation. NADPH diaphorase activity was assessed by a histochemistry, nitroblue tetrazolium (NTB) staining. Positive staining was observed in both circular and longitudinal muscle layers. L-arginine increased the staining, while L-NAME decreased the staining. Western blotting for nNOS proved the presence of nNOS in rat gastric smooth muscle. EFS and additional Ca2+ increased nNOS protein expression. These results suggest that in rat stomach, both circular and longitudinal muscle layers are innervated with nitroxergic nerves which relax the gastric smooth muscle via NO-cGMP pathway.