RESUMEN
Seven ginsenosides, namely Rg6 (1), F4 (2), Rk3 (3), Rh4 (4), Rs3 (5), Rs4 (6) and Rs5 (7) isolated from processed ginseng were evaluated for their effects on platelet aggregation induced by adenosine diphosphate (ADP), collagen, arachidonic acid (AA) and U46619 (thromboxane A2 mimetic drug). Ginsenosides Rg6, F4 and Rk3 showed inhibitory activity (IC50 = 76 microM, 114 microM and 128 microM, respectively) on AA-induced platelet aggregation. The corresponding IC50 values were comparable to that of acetylsalicylic acid (ASA) (63 microM). Compared to ASA (IC50 = 468 microM) ginsenosides Rg6, F4, Rk3 and Rh4 were found to be more inhibitive (IC50 = 286 microM, 87 microM, 187 microM and 119 microM, respectively) against U46619-induced aggregation. On the other hand, most of the ginsenosides (Rg6, F4, Rh4, Rs3, Rs5) showed negligible effects on ADP and collagen-induced platelet aggregation. The acetylated ginsenosides (Rs3, Rs4 and Rs5) had only mild effects on aggregation induced by four stimulators.
Asunto(s)
Ginsenósidos/farmacología , Panax/química , Inhibidores de Agregación Plaquetaria , Agregación Plaquetaria/efectos de los fármacos , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/antagonistas & inhibidores , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Acetilación , Adenosina Difosfato/antagonistas & inhibidores , Adenosina Difosfato/farmacología , Animales , Cromatografía Líquida de Alta Presión , Colágeno/antagonistas & inhibidores , Colágeno/farmacología , Ginsenósidos/aislamiento & purificación , Técnicas In Vitro , Masculino , Extractos Vegetales/química , Raíces de Plantas/química , Ratas , Ratas Sprague-Dawley , Espectrofotometría Ultravioleta , Vasoconstrictores/antagonistas & inhibidores , Vasoconstrictores/farmacologíaRESUMEN
The roots of two Paeoniaceae family members have long been used as traditional medicines in Korea, China, and Japan. Dry roots of Paeonia lactiflora and dry root bark of P. suffruticosa are used under the traditional names of Paeoniae Radix and Moutan Cortex, respectively. Both Paeoniae Radix and Moutan Cortex have been used as remedies for cardiovascular diseases, for improving blood circulation, or for other uses. It was postulated that both plants may contain common active constituents that contribute to inhibiting blood coagulation and/or platelet aggregation. Eighteen compounds, which have been reported to be present in both plant medicines, were evaluated for their effects on platelet aggregation and blood coagulation. Paeonol (5), paeoniflorin (9), benzoylpaeoniflorin (11), and benzoyloxypaeoniflorin (12) were found to be the major common active constituents and they would collectively contribute to improving blood circulation through their inhibitory effects on both platelet aggregation and blood coagulation. In addition, methylgallate (4), (+)-catechin (7), paeoniflorigenone (8), galloylpaeoniflorin (13), and daucosterol (16) may also take part in improving blood circulation by inhibiting ether platelet aggregation and/or blood coagulation.
Asunto(s)
Anticoagulantes/química , Anticoagulantes/farmacología , Paeonia/química , Inhibidores de Agregación Plaquetaria/química , Inhibidores de Agregación Plaquetaria/farmacología , Animales , Humanos , Técnicas In Vitro , Corea (Geográfico) , Masculino , Extractos Vegetales/química , Extractos Vegetales/farmacología , Agregación Plaquetaria/efectos de los fármacos , Ratas , Ratas Sprague-DawleyRESUMEN
We examined our hypothesis that (S)-1-(alpha-naphthylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (CKD712) inhibits apoptosis in myocardial ischemia and reperfusion (I/R) injury in vivo via activation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway and by reducing inflammation during I/R. To do this, we induced a 30-min period of ischemia by occlusion of the left anterior descending coronary artery of the rat followed by a 2-h (for phosphorylation of Akt), 6-h (for biochemical analysis), or 24-h (for functional analysis) period of reperfusion to determine the effect of CKD712 treatment. Pretreatment with CKD712 significantly improved myocardial function as evidenced by an increase in the +/-dP/dt and a decrease in the infarct size, which were antagonized by a PI3K inhibitor, wortmannin (WT). Interestingly, CKD712 increased the phosphorylation of Akt and cAMP-response element-binding protein and increased the expression of the Bcl-2 gene, but it reduced the expression of the Bax gene. CKD712 decreased not only the expression but also the activity of the caspase-3 protein in the myocardium after reperfusion. Thus, all of the antiapoptotic effects of CKD712 were significantly inhibited by WT. Furthermore, the antiapoptotic effects of CKD712 and its inhibition by WT in myocardium after reperfusion were confirmed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling staining. Finally, CKD712 was found to reduce the serum levels of the high-mobility group box 1 protein, tumor necrosis factor-alpha, and the cardiac troponin I protein in addition to tissue levels of malondialdehyde and myeloperoxidase activity in I/R hearts. Taken together, both the activation of PI3K/Akt and its anti-inflammatory action prevent apoptosis in myocardial I/R injury by CKD712.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Apoptosis/fisiología , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Miocardio/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/fisiología , Tetrahidroisoquinolinas/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Activación Enzimática/fisiología , Depuradores de Radicales Libres/farmacología , Depuradores de Radicales Libres/uso terapéutico , Masculino , Isquemia Miocárdica/tratamiento farmacológico , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/patología , Daño por Reperfusión Miocárdica/enzimología , Daño por Reperfusión Miocárdica/patología , Miocardio/patología , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Tetrahidroisoquinolinas/farmacologíaRESUMEN
We found that CKD712, an S enantiomer of YS49, strongly inhibited inducible nitric oxide synthase (iNOS) and NO induction but showed a weak inhibitory effect on cyclooxygenase-2 (COX-2) and PGE(2) induction in LPS-stimulated RAW 264.7 cells. We, therefore, investigated the molecular mechanism(s) responsible for this by using CKD712 in LPS-activated RAW264.7 cells. Treatment with either SP600125, a specific JNK inhibitor or TPCK, a NF-kappaB inhibitor, but neither ERK inhibitor PD98059 nor p38 inhibitor SB203580, significantly inhibited LPS-mediated iNOS and COX-2 induction. CKD712 inhibited NF-kappaB (p65) activity and translocation but failed to prevent JNK activation. However, AG490, a specific JAK-2/STAT-1 inhibitor, efficiently prevented LPS-mediated iNOS induction but not the induction of COX-2, and CKD712 completely blocked STAT-1 phosphorylation by LPS, suggesting that the NF-kappaB and JAK-2/STAT-1 pathways but not the JNK pathway are important for CKD712 action. Interestingly, CKD712 induced heme oxygenase 1 (HO-1) gene expression in LPS-treated cells. LPS-induced NF-kappaB and STAT-1 activation was partially prevented by HO-1 overexpression. Furthermore, HO-1 siRNA partly reversed not only the LPS-induced NF-kappaB activation and STAT-1 phosphorylation but also inhibition of these actions by CKD 712. Additionally, silencing HO-1 by siRNA prevented CKD712 from inhibiting iNOS expression but not COX-2. When examined plasma NO and PGE(2) levels and iNOS and COX-2 protein levels in lung tissues of mice injected with LPS (10 mg/kg), pretreatment with CKD712 greatly prevented NO and iNOS induction in a dose-dependent manner and slightly affected PGE(2) and COX-2 production as expected. Taken together, we conclude that inhibition of JAK-2/STAT-1 pathways by CKD 712 is critical for the differential inhibition of iNOS and COX-2 by LPS in vitro and in vivo where HO-1 induction also contributes to this by partially modulating JAK-2/STAT-1 pathways.
Asunto(s)
Ciclooxigenasa 2/genética , Hemo-Oxigenasa 1/metabolismo , Janus Quinasa 2/metabolismo , Activación de Macrófagos/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo II/genética , Factor de Transcripción STAT1/metabolismo , Tetrahidroisoquinolinas/farmacología , Alcaloides/farmacología , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Dinoprostona/biosíntesis , Activación Enzimática/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Silenciador del Gen/efectos de los fármacos , Lipopolisacáridos/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/enzimología , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Modelos Biológicos , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Fosforilación/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Tetrahidroisoquinolinas/químicaRESUMEN
Aspirin is known to suppress platelet function markedly. However, aspirin at concentrations higher than 1 mM was observed to augment 1.3 microM U46619 (a stable thromboxane receptor (TP receptor) agonist) induced human platelet aggregation in this study. Moreover, at a concentration as low as 250 microM aspirin increased the aggregation induced by U46619 in 13% of normal and healthy individuals. The degree of platelet aggregation and the amount of ATP release were enhanced in U46619 stimulated platelet rich plasma by the addition of aspirin (>250 microM). U46619 was previously reported to inhibit forskolin-stimulated adenyl cyclase and to reduce the cAMP formation. Both of the augmentation effects of aspirin on U46619-induced aggregation and ATP release were blocked by MeSAMP, a P2Y(12) receptor antagonist. U46619 induced aggregation was suppressed by the addition of ADP scavenger (CP/CPK) with no significant change on ATP measured and the effect of CP/CPK could not be reversed by aspirin. In addition, aspirin augmented the inhibitory effect of U46619 on the cAMP production. Our present results suggested that the potentiation effect of aspirin on U46619 induced aggregation was related with the secreted ADP and the subsequent P2Y(12)/Gi related signaling.
Asunto(s)
Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Aspirina/farmacología , Plaquetas , Inhibidores de Agregación Plaquetaria/farmacología , Agregación Plaquetaria/efectos de los fármacos , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Adenilil Ciclasas/metabolismo , Adulto , Plaquetas/efectos de los fármacos , Plaquetas/fisiología , AMP Cíclico/metabolismo , Inhibidores de la Ciclooxigenasa/farmacología , Femenino , Humanos , Masculino , Óxido Nítrico Sintasa/antagonistas & inhibidores , Antagonistas del Receptor Purinérgico P2 , Receptores Purinérgicos P2Y12 , Transducción de Señal/fisiología , Vasoconstrictores/farmacología , Adulto JovenRESUMEN
Impaired responsiveness to epinephrine and other catecholamines (CA) were previously reported in platelets of 20 approximately 30% healthy Japanese and Koreans. In the present study, the possible mechanisms of different responsiveness to CA in platelets of CA hypo-responders (CA-HY) and CA good-responders (CA-GR) were investigated. Increased platelet-leukocyte conjugate (PLC) formations were observed with whole blood of CA-GR than with that of CA-HY in both non-stimulated [mean fluorescence intensity (MFI) values: 1.33 +/- 0.26 vs. 1.16 +/- 0.19] and ADP (MFI: 5.54 +/- 3.46 vs. 2.15 +/- 1.13) or TRAP (MFI: 5.11 +/- 2.32 vs. 3.38 +/- 1.47) activated states. The platelets of CA-GR, when stimulated with ADP (10 microM), released approximately twice the amount of ATP than those of CA-HY (0.88 +/- 0.65 and 0.45 +/- 0.36 nmole, respectively). Nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) levels were significantly higher in non-stimulated PRP of CA-HY than in that of CA-GR (70.3 +/- 24.1 microM and 14.1 +/- 4.9 nM vs. 41.1 +/- 15.8 microM and 6.7 +/- 2.4 nM, respectively). The platelet-monocyte conjugation induced with either ADP or TRAP was significantly reduced in CA-GR with the addition of linsidomine, a NO donor, (MFI: 2.78 +/- 0.43 vs. 3.73 +/- 0.90, or 4.28 +/- 0.95 vs. 5.76 +/- 1.33, respectively). Moreover, the degree of platelet aggregation and the ATP secretion induced by epinephrine in CA-GR were significantly retarded with the addition of either linsidomine or 8-Bromo-cGMP (a cGMP analog) with more substantial effects on ATP release than aggregation. The results suggested that elevated NO and/or cGMP plasma levels may be responsible for the lower platelet aggregation and PLC formation observed in CA-HY than that in CA-GR.
Asunto(s)
Plaquetas , Catecolaminas/farmacología , GMP Cíclico/sangre , Leucocitos/metabolismo , Óxido Nítrico/sangre , Agregación Plaquetaria , Adenosina Trifosfato/metabolismo , Adulto , Pueblo Asiatico , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , AMP Cíclico/sangre , GMP Cíclico/análogos & derivados , GMP Cíclico/metabolismo , Femenino , Humanos , Antígenos Comunes de Leucocito/metabolismo , Leucocitos/citología , Masculino , Molsidomina/análogos & derivados , Molsidomina/farmacología , Nitratos/metabolismo , Nitritos/metabolismo , Agregación Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/fisiología , Inhibidores de Agregación Plaquetaria/farmacología , Adulto JovenRESUMEN
Four dammarane glycosides, namely ginsenosides Rk1 (1), Rg5 (2), 20(S)-Rg3 (3), and 20(R)-Rg3 (4), isolated from a new processed ginseng, were evaluated for their inhibitory activity against platelet aggregation induced by adenosine diphosphate (ADP), collagen, arachidonic acid (AA) and U46619 (thromboxane A2 mimetic agent). Ginsenoside Rk1 and Rg5 inhibited AA-induced platelet aggregation in a dose dependent manner. Their activity against AA-induced platelet aggregation were found to be 8-22 fold higher than that of a known antiplatelet drug acetylsalicylic acid (ASA). They also inhibited U46619-induced platelet aggregation. Ginsenoside 20(S)-Rg3 and 20(R)-Rg3 showed mild inhibitory activity against AA and U46619-induced aggregation.
Asunto(s)
Ginsenósidos/farmacología , Panax/química , Inhibidores de Agregación Plaquetaria/farmacología , Agregación Plaquetaria/efectos de los fármacos , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Animales , Ácido Araquidónico/farmacología , Colágeno/farmacología , Relación Dosis-Respuesta a Droga , Ginsenósidos/aislamiento & purificación , Técnicas In Vitro , Masculino , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Vasoconstrictores/farmacologíaRESUMEN
Optically active tetrahydroisoquinoline alkaloids, (R)-(+)-higenamine (1R) and (S)-(-)-higenamine (1 S), and their optically active 1-naphthylmethyl analogues (2 and 3), were synthesized by enantioselective hydrogenation of the corresponding dihydroisoquinoline intermediates 7 as a key step. The evaluation of the platelet anti-aggregation effect demonstrated clearly that the (S)-(-)-enantiomers, 1S, 2S, and 3S, had higher inhibitory potency than the corresponding (R)-(+)-antipodes, 1R, 2R, and 3R, respectively, to platelet aggregation induced by epinephrine. 1S enantiomer was superior to the corresponding 1R enantiomer in attenuating all of the disseminated intravascular coagulation (DIC) and multiple organ failure (MOF) parameters tested, while the S enantiomers 2S and 3S ameliorated some of the DIC and MOF parameters more effectively than the corresponding antipodes 2R and 3R.
Asunto(s)
Alcaloides/síntesis química , Coagulación Intravascular Diseminada/tratamiento farmacológico , Agregación Plaquetaria/efectos de los fármacos , Tetrahidroisoquinolinas/síntesis química , Alcaloides/química , Amidas/química , Animales , Catálisis , Química Farmacéutica/métodos , Cromatografía Líquida de Alta Presión , Modelos Animales de Enfermedad , Diseño de Fármacos , Epinefrina/química , Concentración 50 Inhibidora , Modelos Químicos , Ratas , Estereoisomerismo , Tetrahidroisoquinolinas/químicaRESUMEN
Overexpression of the gene for heme oxygenase (HO)-1 leads to a reduction in pressor responsiveness to angiotensin II (Ang II) in experimental animals. Using rat vascular smooth muscle cells (VSMCs), we tested whether YS 49 [1-(alpha-naphtylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline] inhibits Ang II-stimulated proliferation of VSMCs via induction of HO-1. YS 49 induced HO-1 protein production in a dose-and time-dependent manner in VSMCs. Treatment with YS 49 significantly and dose-dependently inhibited Ang II-induced VSMC proliferation, ROS production, and phosphorylation of JNK, but not P38 MAP kinase or ERK1/2. The antiproliferation effect of YS 49 was reversed by pretreatment with the HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX), or with hemoglobin, a carbon monoxide (CO) scavenger. Similarly, VSMC proliferation, ROS production and phosphorylation of JNK by Ang II were significantly inhibited in VSMCs transfected with the HO-1 gene. Thus, HO-1 and the HO-1 product CO play, at least in part, a crucial role in Ang II-stimulated VSMC proliferation through the regulation of ROS production and JNK phosphorylation. Therefore, YS 49 has potential as a therapeutic strategy for the pathogenesis of Ang II-related vascular diseases such as hypertension and atherosclerosis, via the induction of HO-1 gene activity.
Asunto(s)
Angiotensina II/metabolismo , Hemo-Oxigenasa 1/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Músculo Liso Vascular/citología , Miocitos del Músculo Liso/fisiología , Especies Reactivas de Oxígeno/metabolismo , Tetrahidroisoquinolinas/metabolismo , Animales , Proliferación Celular , Células Cultivadas , Inducción Enzimática , Hemo-Oxigenasa 1/genética , Sistema de Señalización de MAP Quinasas/fisiología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Miocitos del Músculo Liso/citología , RatasRESUMEN
A methanol extract of Sophora japonica was subjected to anti-platelet activity guided fractionation affording the isolation of four flavonoids and six flavonoid-glycosides: biochanin A (1), irisolidone (2), genistein (3), sissotrin (4), sophorabioside (5), genistin (6), tectoridin (7), apigenin (8), quercitrin (9), and rutin (10). The structure of each compound was determined by a variety of spectroscopic methods. Among the compounds, 1, 3, and 7 showed approximately 2.5-6.5 fold greater inhibitory effects on arachidonic acid (AA) and U46619 induced platelet aggregation (IC50: 19.9 and 99.8 microM; 20.3 and 53.8 microM; 25.9 and 123.4 microM, respectively) than acetylsalicylic acid (ASA, IC50: 63.0 and 350.0 microM). Compound 2 was an approximately 22-40 fold stronger inhibitor than ASA on AA and U46619 induced aggregation (IC50: 1.6 and 15.6 microM, respectively).
Asunto(s)
Flavonoides/farmacología , Inhibidores de Agregación Plaquetaria , Sophora/química , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/antagonistas & inhibidores , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Adenosina Difosfato/antagonistas & inhibidores , Adenosina Difosfato/farmacología , Animales , Ácido Araquidónico/antagonistas & inhibidores , Ácido Araquidónico/farmacología , Colágeno/antagonistas & inhibidores , Colágeno/farmacología , Flavonoides/aislamiento & purificación , Frutas/química , Glicósidos/aislamiento & purificación , Glicósidos/farmacología , Técnicas In Vitro , Espectroscopía de Resonancia Magnética , Hojas de la Planta/química , Tallos de la Planta/química , Agregación Plaquetaria/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Espectrofotometría Infrarroja , Espectrofotometría Ultravioleta , Tromboxano A2/farmacologíaRESUMEN
Oxidative stress plays an important role in the pathophysiology of several vascular diseases such as atherosclerosis, and great attention has been placed on the protective role of heme oxygenase-1 (HO-1) for vasculature against oxidant-induced injury. We tested whether the protective effects of YS 51, 1-(beta-naphtyl-methyl)-6,7-dihydroxy-1,2,3,4,-tetrahydroisoquinoline, against hydrogen peroxide (H2O2)-induced cell injury is associated with HO-1 activity in bovine aortic endothelial cells (BAEC). YS 51 increased HO-1 expression and activity in concentration-dependent manners (10-100 microM) and time-dependent manners (1, 3, 6, 18 h), which were correlated well with its protective effect against H2O2-induced injury. Zinc protoporphyrin IX (ZnPP IX), a HO inhibitor, significantly inhibited the effect of YS 51 (50 microM). In contrast, [Ru(CO)3(Cl)2]2 (CORM-2, a CO releasing molecule) but not bilirubin protected against H2O2-induced injury. Oxyhemoglobin (HbO2) used as a CO scavenger significantly inhibited the protective effect of both YS 51 and CORM-2. Furthermore, both YS 51 and CORM-2 significantly reduced H2O2-induced intracellular reactive oxygen species (ROS) production; however, this was counteracted by ZnPP IX, HbO2 and deferoxamine. We found evidence for the involvement of PI3/Akt kinase and ERK1/2 pathways in HO-1 induction by YS-51. Taken together, we conclude that CO is, at least, responsible for the YS 51-mediated protective action of endothelial cells against oxidant stress via HO-1 gene induction, involving the activation of the PI3/Akt and ERK1/2 kinase pathways. Thus, YS 51 may be useful in oxidative stress-induced vascular disorders.
Asunto(s)
Antioxidantes/farmacología , Monóxido de Carbono/metabolismo , Células Endoteliales/efectos de los fármacos , Hemo-Oxigenasa 1/biosíntesis , Peróxido de Hidrógeno/toxicidad , Estrés Oxidativo/efectos de los fármacos , Tetrahidroisoquinolinas/farmacología , Animales , Western Blotting , Bovinos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Células Endoteliales/enzimología , Células Endoteliales/metabolismo , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The effects of higenamine and its 1-naphthyl analogs, YS-49 and YS-51, on thromboxane A(2) (TXA(2)) formation from arachidonic acid (AA) and aggregation in platelets, were investigated. YS-49 and YS-51 (IC(50); 32.8 and 39.4 microM respectively) exhibited much stronger inhibitory effects on TXA(2) formation than higenamine (IC(50); 2.99 mM). The higher inhibitory potencies of YS-49 and YS-51 (IC(50): 3.3 and 5.7 microM respectively) than higenamine (IC(50): 140 microM) on AA induced rat platelet aggregation was presumed to be the result of low inhibitory effect of higenamine than YS-49 and YS-51 on TXA(2) production from AA. Among the present three compounds, the more hydrophobic naphthylmethyl groups were supposed to be more favorable than p-hydroxybenzyl moiety, at 1-position of the tetrahydroisoquinoline ring, to display the inhibitory effects on TXA(2) production and AA induced aggregation of platelets. In addition, higenamine, YS-49 and YS-51 were observed directly antagonistic on TXA(2) receptor (TP receptors) by displaying inhibitory effects to U46619 (TXA(2) mimetic) induced platelet aggregation, however all of the three compounds showed similar order of inhibitory potencies. The present results are suggestive that YS-49 and YS-51 exert their inhibitory effects on AA-induced platelet aggregation partly by inhibiting the production of TXA(2) from AA and partly by directly blocking the TP receptor, in addition to the previously reported effects on alpha(2)-adrenergic receptor. On the other hand, higenamine is supposed to antagonize AA-induced platelet aggregation by mostly directly blocking the TP receptor.
Asunto(s)
Alcaloides/farmacología , Plaquetas/efectos de los fármacos , Fibrinolíticos/farmacología , Agregación Plaquetaria/efectos de los fármacos , Tetrahidroisoquinolinas/farmacología , Tromboxano A2/metabolismo , Adulto , Animales , Femenino , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
Activation of the inducible nitric oxide synthase (iNOS) pathway contributes to inflammation-induced osteoporosis by suppressing bone formation and causing osteoblast apoptosis. We investigated the mechanism of action by which YS-51S, a synthetic isoquinoline alkaloid, inhibits iNOS expression and nitric oxide (NO) production in ROS 17/28 osteoblast cells activated with the mixture of TNF-alpha, IFN-gamma and LPS (MIX). YS-51S, concentration- and time-dependently, increased heme oxygenase (HO-1) expression. Treatment with YS-51S 1 h prior to MIX significantly reduced MIX-induced NO production and iNOS expression with the IC50 to NO production of 47+/-3.3 microM. Electrophoretic mobility shift assay (EMSA) and western blot analysis showed that YS-51S inhibited MIX-mediated activation and translocation of NF-kappaB to nucleus by suppressing the degradation of its inhibitory protein IkappaBalpha in cytoplasm. YS-51S also reduced NF-kappaB-luciferase activity. In addition, an HO-1 inhibitor ZnPPIX, antagonized the inhibitory effect of YS-51S on iNOS expression and DNA strand break induced by MIX, indicating prevention of NO production by YS-51S is associated with HO-1 activity. Moreover, YS-51S inhibited the oxidation of cytochrome c(2+) by peroxynitrite (PN). Our results indicated that YS-51S may be beneficial in NO-mediated inflammatory conditions such as rheumatoid arthritis by alleviating iNOS expression and NO-mediated cell death of osteoblast with 1) inducing HO-1 expression, 2) interfering the activation of NF-kappaB and 3) quenching of PN.
Asunto(s)
Núcleo Celular/metabolismo , Hemo Oxigenasa (Desciclizante)/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Tetrahidroisoquinolinas/farmacología , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Antioxidantes/química , Antioxidantes/farmacología , Línea Celular Tumoral , Citocromos c/química , Inhibidores Enzimáticos/farmacología , Hemo Oxigenasa (Desciclizante)/antagonistas & inhibidores , Hemo Oxigenasa (Desciclizante)/genética , Quinasa I-kappa B/metabolismo , Interferón gamma/farmacología , Lipopolisacáridos/farmacología , Luciferasas/genética , Luciferasas/metabolismo , FN-kappa B/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Oxidación-Reducción/efectos de los fármacos , Ácido Peroxinitroso/química , Fosforilación/efectos de los fármacos , Protoporfirinas/farmacología , ARN Interferente Pequeño/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Estereoisomerismo , Tetrahidroisoquinolinas/química , Transfección , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
An aryltetralin lactone, (-)-(1S,2R,3R)-1-(3',4'-methylenedioxyphenyl)-3-(hydroxymethyl)-6,7-dimethoxy-1,2,3,4-tetrahydronaphthalene-2-carboxylic acid lactone, was isolated from the methanol extract of the aerial parts of Bupleurum falcatum and named bupleurumin. The structure of the compound was determined on the basis of chemical and spectroscopic methods, including two-dimensional NMR spectrometry (e.g., 1H- and 13C-NMR, homo- and heteronuclear COSY, HMBC). Bupleurumin showed an 8-fold potent inhibitory effect (IC50: 47.5 uM) compared to that of acetylsalicylic acid (ASA, IC50 : 420 uM) on collagen induced platelet aggregation and comparable effects as ASA on arachidonic acid-induced platelet aggregation.
Asunto(s)
Bupleurum/química , Lactonas/aislamiento & purificación , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Tetrahidronaftalenos/aislamiento & purificación , Animales , Lactonas/química , Lactonas/farmacología , Espectroscopía de Resonancia Magnética , Inhibidores de Agregación Plaquetaria/química , Inhibidores de Agregación Plaquetaria/farmacología , Ratas , Ratas Sprague-Dawley , Tetrahidronaftalenos/química , Tetrahidronaftalenos/farmacologíaRESUMEN
It has been suggested that nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) may act as a mediator of cytokine-induced effects on bone turn-over. NO is also recognized as an important factor in bone remodeling, i.e., participating in osteoblast apoptosis in an arthritic joint. The components of Agastache rugosa are known to have many pharmacological activities. In the present study, we investigated the effects of Agastache rugosa leaf extract (ELAR) on NO production and the iNOS expression in ROS 17/2.8 cells activated by a mixture of inflammatory cytokines including TNF-alpha and IL-1beta. A preincubation with ELAR significantly and concentration-dependently reduced the expression of iNOS protein in ROS 17/2.8 cells activated with the cytokine mixture. Consequently, the NO production was also significantly reduced by ELAR with an IC50 of 0.75 mg/mL. The inhibitory mechanism of iNOS induction by ELAR prevented the activation and translocation of NF-kappaB (p65) to the nucleus from the cytosol fraction. Furthermore, ELAR concentration-dependently reduced the cellular toxicity induced by sodium nitroprusside, an NO-donor. These results suggest that ELAR may be beneficial in NO-mediated inflammatory conditions such as osteoporosis.
Asunto(s)
Agastache/química , Interleucina-1/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Hojas de la Planta/química , Factor de Necrosis Tumoral alfa/farmacología , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Interleucina-1/fisiología , FN-kappa B/metabolismo , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico Sintasa de Tipo II , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Extractos Vegetales/farmacología , Transporte de Proteínas , Ratas , Factor de Necrosis Tumoral alfa/fisiologíaRESUMEN
In the course of our work on anti-platelet constituents from plants, five phenolic compounds, magnolol, honokiol, obovatol, methyl caffeate, and syringin, were isolated from the methanol extracts of the barks and fruits of Magnolia obovata. The compounds were identified based on the spectroscopic data. Methyl caffeate was isolated for the first time from the genus Magnolia and it showed 3 approximately 4-folds higher potency than ASA. The activities of obovatol and honokiol were comparable to ASA. Magnolol and syringin showed only very mild inhibitory effects to all the stimulators.
Asunto(s)
Magnolia/química , Inhibidores de Agregación Plaquetaria/farmacología , Animales , Secuencia de Carbohidratos , Frutas/química , Cromatografía de Gases y Espectrometría de Masas , Técnicas In Vitro , Espectroscopía de Resonancia Magnética , Masculino , Datos de Secuencia Molecular , Corteza de la Planta/química , Extractos Vegetales/farmacología , Agregación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Espectrofotometría Infrarroja , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Nine phenolic (1-9) and two furan type (10, 11) compounds, were isolated from the methanolic extract of the tuber of Gastrodia elata Blume (Orchidaceae) in the course of continuing search for platelet anti-aggregating plant components. Compound 1 was identified as 4,4'-dihydroxybenzyl sulfone, a novel compound for the best of our knowledge. Compound 10, 5-hydroxymethyl-2-furancarboxaldehyde, was isolated for the first time from this plant. Compound 1 (IC50; 83 microM) was about four times more inhibitory to U46619 induced aggregation than ASA (IC50; 340 microM). Compound 9, 4,4'-dihydroxy-dibenzylether, (IC50; 5 microM, 3 microM and 33 microM, respectively) was 10-80 fold more potent than ASA (IC50; 420 microM, 53 microM and 340 microM respectively) to collagen, epinephrine and U46619 induced aggregation, although it is less active than ASA to AA induced aggregation.
Asunto(s)
Furanos/farmacología , Gastrodia , Fenoles/farmacología , Inhibidores de Agregación Plaquetaria/farmacología , Agregación Plaquetaria/efectos de los fármacos , Animales , Furanos/química , Furanos/aislamiento & purificación , Fenoles/química , Fenoles/aislamiento & purificación , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Tubérculos de la Planta , Agregación Plaquetaria/fisiología , Inhibidores de Agregación Plaquetaria/química , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , RatasRESUMEN
Five coumarins, isoimperatorin (1), pabulenol (2), isooxypeucedanin (3), oxypeucedanin hydrate (4) and osthol (5) were isolated from the MeOH extract of Angelica genuflexa in the course of searching for anti-platelet and anti-coagulant components from plants. Pabulenol (2) was isolated from A. genuflexa for the first time. The five compounds isolated from A. genuflexa, together with decursinol angelate (6), decursin (7) and nodakenin (8) from A. gigas were evaluated for their effects on platelet aggregation and blood coagulation. Compounds 2, 5, 6 and 7 were observed to be either equally effective or 2-4 times more inhibitory than ASA in both arachidonic acid and U46619 (TXA2 mimetic) induced platelet aggregations.
Asunto(s)
Angelica/química , Cumarinas/aislamiento & purificación , Cumarinas/farmacología , Raíces de Plantas/química , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Inhibidores de Agregación Plaquetaria/farmacología , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Animales , Ácido Araquidónico/farmacología , Aspirina/farmacología , Benzopiranos/química , Benzopiranos/aislamiento & purificación , Benzopiranos/farmacología , Plaquetas/efectos de los fármacos , Butiratos/química , Butiratos/aislamiento & purificación , Butiratos/farmacología , Colágeno/farmacología , Cumarinas/química , Furocumarinas/química , Furocumarinas/aislamiento & purificación , Furocumarinas/farmacología , Glucósidos/química , Glucósidos/aislamiento & purificación , Glucósidos/farmacología , Medicina de Hierbas , Compuestos Heterocíclicos con 3 Anillos/química , Compuestos Heterocíclicos con 3 Anillos/aislamiento & purificación , Compuestos Heterocíclicos con 3 Anillos/farmacología , Corea (Geográfico) , Masculino , Medicina Tradicional de Asia Oriental , Metanol , Nefelometría y Turbidimetría/métodos , Extractos Vegetales/química , Agregación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/química , Ratas , Ratas Sprague-DawleyRESUMEN
Five pentacyclic triterpenoids, oleanolic acid (1), hederagenin (2), ursolic acid (3), tormentic acid (4) and myrianthic acid (5), were isolated from the methanol extract of the leaves of Campsis grandiflora, and structures of the compounds were established by the spectroscopic methods. Compounds 2, 3, 4, and 5 were isolated for the first time from the genus Campsis. All of the compounds (IC50: 45.3, 32.8, 82.6, 42.9 and 46.2 microM respectively) were as equivalently inhibitive as acetylsalicylic acid (IC50: 57.0 microM) on epinephrine induced platelet aggregation.
Asunto(s)
Bignoniaceae , Inhibidores de Agregación Plaquetaria/farmacología , Agregación Plaquetaria/efectos de los fármacos , Triterpenos/farmacología , Animales , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Hojas de la Planta , Agregación Plaquetaria/fisiología , Inhibidores de Agregación Plaquetaria/química , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Triterpenos/química , Triterpenos/aislamiento & purificaciónRESUMEN
Although it was thought that platelets did not play a significant role in the pathogenesis of venous thromboembolism (VTE), several studies demonstrated that a marked activation of platelets occurs in patients with VTE. We carried out a case-control study to investigate the effect of the T744C P2RY12 polymorphism on the risk of VTE in the Korean population. We enrolled 154 consecutive patients with VTE and 415 healthy controls. Genotype frequencies for patients with TT, TC, and CC were 71.4%, 24.7%, and 3.9% and in the controls, 68.2%, 30.1%, and 1.7%, respectively. T744C P2RY12 polymorphism did not significantly affect the risk of VTE. Our study shows that T744C P2RY12 polymorphism did not significantly affect the risk of VTE in the Korean population.