RESUMO
MTX is an effective therapy for autoimmune-inflammatory diseases. The mechanisms that mediate these actions are not completely clear. It is accepted that many of these effects are mediated through the release of adenosine with the activation of the adenosine receptor A2. MTX is used as a steroid sparing agent. An improved in vitro GC cell sensitivity in GC insensitive asthma patients has been demonstrated after MTX treatment. Most GC actions are mediated by the GCR. The effect of MTX on GCRs expression has not been previously evaluated. Therefore, we evaluate if MTX regulates the expression of glucocorticoid receptors, increasing the expression of the active receptor (GCR alpha) and/or decreasing the expression of the dominant negative receptor (GCR beta). We show that MTX increases the mRNA and protein levels of GCR alpha and decreases or leaves unchanged the protein expression of the GCR beta in CEM cells in culture. This effect was also observed in other lymphocytes (Jurkat and Raji) and in PBMNC from healthy volunteers. We also show that upon MTX treatment PBMC from normal volunteers exhibit a higher sensitivity to DEX inhibition on LPS-induced TNF alpha release. To explore if these actions are mediated by adenosine through the adenosine receptor A2 we evaluate the effect of adenosine on the GCRs expression and the effect of an A2 receptor blocker (DMPX) on MTX effects on GCRs expression. Our results show that adenosine does not mimic and DMPX can enhance MTX effects on these receptors. We conclude that MTX increases the GCR alpha/GCR beta ratio of expression in lymphocytes which could mediate its previously reported effects in improving cell glucocorticoid sensitivity. These actions are not mediated by the adenosine receptor A2.
Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Imunossupressores/farmacologia , Linfócitos/metabolismo , Metotrexato/farmacologia , Receptores de Glucocorticoides/biossíntese , Adenosina/farmacologia , Antagonistas do Receptor A2 de Adenosina , Asma/tratamento farmacológico , Asma/imunologia , Asma/metabolismo , Doenças Autoimunes/tratamento farmacológico , Doenças Autoimunes/imunologia , Doenças Autoimunes/metabolismo , Sinergismo Farmacológico , Humanos , Imunossupressores/uso terapêutico , Inflamação/tratamento farmacológico , Inflamação/imunologia , Inflamação/metabolismo , Células Jurkat , Lipopolissacarídeos/farmacologia , Linfócitos/imunologia , Metotrexato/agonistas , Metotrexato/uso terapêutico , Isoformas de Proteínas/biossíntese , Isoformas de Proteínas/imunologia , Receptores de Glucocorticoides/imunologia , Teobromina/agonistas , Teobromina/análogos & derivados , Teobromina/farmacologia , Vasodilatadores/farmacologiaRESUMO
In the present study, we show that adenosine (Ado) affects vaccinia virus (VV) replication in BSC-40 cells. In order to identify its effects on VV replicative cycle we analyzed the synthesis of virus macromolecules in cells incubated with 0.5 mM Ado. A 50% reduction in the steady-state level of virus DNA synthesis was observed. Consequently, virus post-replicative gene expression was also affected. A prolonged synthesis of the F11L early virus protein was also observed and it is likely related to a slow decline in the steady-state level of early mRNAs, as suggested by northern blot analysis of the VGF early transcript. The involvement of cAMP-signaling pathway as mediator of Ado response was also evaluated. Ado stimulated a three-fold increase in cAMP levels in BSC-40 cells and cAMP-mimetic agents reduced virus yield in a dose-dependent manner. Co-incubation of infected cells with H-89 reduced the inhibitory effects of 8-Br-cAMP and Ado on VV yields suggesting PKA involvement. A(2) receptor-mediated activation of PKA was indicated by antagonism of Ado response by theophylline and DMPX. Together, these results indicate that virus DNA replication is the main target of Ado. The mechanism involved is not related to reduction of the pyrimidine nucleotide synthesis. Furthermore, Ado-induced PKA activation modulates negatively an unidentified step of the virus replicative cycle.
Assuntos
Adenosina/farmacologia , Antivirais/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Receptores Purinérgicos P1/metabolismo , Teobromina/análogos & derivados , Vaccinia virus/efeitos dos fármacos , Adenosina/antagonistas & inibidores , Animais , Proteínas de Transporte/farmacologia , Linhagem Celular , Técnicas de Cocultura , AMP Cíclico/análise , AMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/virologia , Inibidores de Fosfodiesterase/farmacologia , Inibidores da Síntese de Proteínas/farmacologia , Antagonistas de Receptores Purinérgicos P1 , Transdução de Sinais , Teobromina/farmacologia , Teofilina/farmacologia , Vaccinia virus/fisiologia , Replicação Viral/efeitos dos fármacosRESUMO
1. The contribution of sensory neurons and mast cells to the oedema evoked by adenosine A1 (N(6)-cyclopentyladenosine, CPA, 3 - 30 nmol site(-1)), A2 (5'N-ethylcarboxamidoadenosine, NECA, 1 - 10 nmol site(-1)) and A3 receptor agonists (N6-[3-iodobenzyl]-N-methyl-5'-carboxiamidoadenosine, IB-MECA, 0.01 - 3 nmol site(-1)) was investigated in the rat skin microvasculature, by the extravascular accumulation of intravenously-injected (i.v.) 125I-albumin. 2. Intradermal (i.d.) injection of adenosine and analogues induced increased microvascular permeability in a dose-dependent manner (IB-MECA > NECA > CPA > adenosine). The non-selective adenosine receptor antagonist theophylline (5 - 50 nmol site(-1)) markedly inhibited adenosine, CPA or NECA but not IB-MECA-induced plasma extravasation. The A1 receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, 0.3 - 3 micromol kg(-1), i.v.) significantly reduced CPA-induced plasma extravasation whereas responses to adenosine, NECA or IB-MECA were unchanged. The A2 receptor antagonist 3,7-dymethyl-1-proprargylxanthine (DMPX, 0.5 - 50 nmol site(-1)) significantly reduced NECA-induced plasma extravasation without affecting responses to adenosine, CPA and IB-MECA. 3. The tachykinin NK1 receptor antagonist (S)-1-[2-[3-(3,4-dichlorphenyl)-1 (3-isopropoxyphenylacetyl) piperidin-3-yl] ethyl]-4-phenyl-1 azaniabicyclo [2.2.2]octane chloride (SR140333), but not the NK2 receptor antagonist (S)-N-methyl-N[4-acetylamino-4-phenyl piperidino)-2-(3,4-dichlorophenyl)butyl]-benzamide (SR48968), significantly inhibited the plasma extravasation evoked by higher doses of adenosine (100 nmol site(-1)), CPA (100 nmol site(-1)), NECA (1 nmol site(-1)) and IB-MECA (0.1 - 1 nmol site(-1)). In rats treated with capsaicin to destroy sensory neurons, the response to higher doses of adenosine, CPA and NECA, but not IB-MECA, was significantly inhibited. 4. The effects of adenosine and analogues were largely inhibited by histamine and 5-hydroxytryptamine (5-HT) antagonists and by compound 48/80 pretreatment. 5. In conclusion, our results provide evidence that adenosine A1 and A2, but not A3, receptor agonists may function as cutaneous neurogenic pro-inflammatory mediators; acting via microvascular permeability-increasing mechanisms that can, depending on dose of agonist and purine receptor under study, involve the tachykinin NK1 receptor and mast cell amines.
Assuntos
Adenosina/análogos & derivados , Adenosina/farmacologia , Proteínas Sanguíneas/efeitos dos fármacos , Permeabilidade Capilar/efeitos dos fármacos , Pele/efeitos dos fármacos , Adenosina-5'-(N-etilcarboxamida)/farmacologia , Animais , Proteínas Sanguíneas/metabolismo , Capsaicina/farmacologia , Relação Dose-Resposta a Droga , Feminino , Injeções Intradérmicas , Soluções Isotônicas/farmacologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/fisiologia , Antagonistas dos Receptores de Neurocinina-1 , Neurônios Aferentes/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Piperidinas/farmacologia , Agonistas do Receptor Purinérgico P1 , Antagonistas de Receptores Purinérgicos P1 , Quinuclidinas/farmacologia , Ratos , Ratos Wistar , Receptores da Neurocinina-2/antagonistas & inibidores , Pele/irrigação sanguínea , Pele/metabolismo , Substância P/análogos & derivados , Substância P/farmacologia , Teobromina/análogos & derivados , Teobromina/farmacologia , p-Metoxi-N-metilfenetilamina/farmacologiaRESUMO
The role of adenosine on regulation of the (Na(+)+K(+))ATPase activity present in the Malpighian tubules isolated from Rhodnius prolixus was investigated. Adenosine decreases the (Na(+)+K(+)) ATPase specific activity by 88%, in a dose-dependent manner, with maximal effect at a concentration of 10(-9) M. This effect was mimicked by N(6)-cyclohexyladenosine (CHA) at 10(-8) M, an agonist for A(1) adenosine receptor, and was reversed by 10(-9) M 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), an antagonist for A(1) adenosine receptor. On the other hand, 5'-N-ethyl-carboxamide adenosine (NECA), an agonist for A(2) adenosine receptor, used in the range of 10(-9)-10(-5) M, did not change the (Na(+)+K(+))ATPase specific activity. In the same way, 10(-8) M 3, 7-dimethyl-1-propargylxanthine (DMPX), an antagonist for A(2) adenosine receptor, did not modify the inhibitory effect of adenosine. These data suggest that the inhibitory effect of adenosine on the (Na(+)+K(+))ATPase specific activity present in Malpighian tubules from Rhodnius prolixus is mediated by A(1) adenosine receptor activation. Arch.
Assuntos
Adenosina/farmacologia , Túbulos de Malpighi/enzimologia , Rhodnius/enzimologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Adenosina-5'-(N-etilcarboxamida)/farmacologia , Animais , Receptores Purinérgicos P1/metabolismo , Rhodnius/efeitos dos fármacos , Teobromina/análogos & derivados , Teobromina/farmacologia , Xantinas/farmacologiaRESUMO
The effects of pentoxifylline on skin flap survival were studied in rabbits. A total of 40 rabbits had caudally based single-pedicle flaps measuring 4 x 14 cm raised on the mid dorsum of each animal. Twenty of these rabbits were given intraperitoneal injections of pentoxifylline in doses of 24 mg/kg per day beginning 48 hours prior to flap construction and continued daily for 7 days postoperatively. The remaining 20 control rabbits received intraperitoneal injections of saline in equal volumes as the experimental groups. At the end of 7 days, viable flap length was visually inspected and measured in all 40 rabbits. There was no significant difference in skin flap viability in rabbits treated with pentoxifylline compared to the control group.
Assuntos
Sobrevivência de Enxerto/efeitos dos fármacos , Pentoxifilina/farmacologia , Retalhos Cirúrgicos , Teobromina/análogos & derivados , Animais , Masculino , Coelhos , Fatores de TempoRESUMO
Out of 235 patients with recent cerebral transient ischaemic attacks, 208 subjects were available for final evaluation after 6 months' randomised treatment with either pentoxifylline (PTX 1200 mg/day) or a combination (ASAD) of acetylsalicylic acid (ASA, 1050 mg/day) and dipyridamole (D, 150 mg/day). Prevention of TIA, stroke or death attributable to previous events were endpoint criteria. The pentoxifylline group (n = 100) exhibited no recurrent episodes in 86 patients (86%). TIA occurred in 9 patients, stroke in 5 patients and there was 1 death. In the ASAD group (n = 108) no recurrence of ischaemic episodes was recorded in 82 cases (75.9%). TIA occurred in 20 patients, stroke in 6 patients and there were 3 deaths of vascular origin. Side effects were recorded in 4 ASAD and 1 PTX patients. The total rate of recurrence was 14% with PTX as compared to 24.1% with ASAD treatment.
Assuntos
Aspirina/uso terapêutico , Dipiridamol/uso terapêutico , Ataque Isquêmico Transitório/tratamento farmacológico , Pentoxifilina/uso terapêutico , Inibidores da Agregação Plaquetária/uso terapêutico , Teobromina/análogos & derivados , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Seventy-six patients with labyrinthine diseases of vascular origin were treated in a 6-week double-blind comparative study with either 400 mg pentoxifylline ('Trental') or 75 mg cinnarizine 3-times daily. Clinical evaluations, supported by audiological tests and vectornystagmography, were carried out before and after treatment. Statistical analysis of the results showed pentoxifylline to be globally superior to cinnarizine and especially to have a more intense antivertiginous effect. No significant differences were observed between the two drugs in respect of tinnitus and hearing loss therapy. Side-effects were occasional, mild and well tolerated in the pentoxifylline group, and more pronounced and frequent with cinnarizine.
Assuntos
Cinarizina/uso terapêutico , Doenças do Labirinto/tratamento farmacológico , Pentoxifilina/uso terapêutico , Teobromina/análogos & derivados , Idoso , Audiometria , Cinarizina/efeitos adversos , Ensaios Clínicos como Assunto , Método Duplo-Cego , Eletronistagmografia , Feminino , Humanos , Doenças do Labirinto/fisiopatologia , Masculino , Pessoa de Meia-Idade , Pentoxifilina/efeitos adversos , Distribuição AleatóriaRESUMO
Sixty patients (37 males, 23 females, age range 7 to 34 years) suffering from sickle cell disease were treated with pentoxifylline (1200 mg/day per os, Trental 400 t.i.d.) or placebo in a double-blind randomized study of six week duration. Observation of pain frequency, intensity and duration of pain events as well as determination of various laboratory and hematologic parameters were maintained on weekly or biweekly basis, respectively, throughout the study. At the end of the trial the number of patients (14) with pain periods and the number of pain events (57) and the mean duration of pain events (4.1 days) in the drug group were significantly better than in the placebo group with 25 patients with pain, 219 pain events and 8.8 days mean pain duration. Hematocrit, red cell count, hemoglobin and platelet aggregation showed slight but significant positive changes with no corresponding alteration with placebo. The over all assessment of the therapeutic response showed 21 clearly improved patients with pentoxifylline compared to 10 such patients with placebo (p less than 0.05). Two patients on pentoxifylline and four on placebo experienced transient side effects of nausea and gastric intolerance.
Assuntos
Anemia Falciforme/tratamento farmacológico , Pentoxifilina/uso terapêutico , Teobromina/análogos & derivados , Adolescente , Anemia Falciforme/sangue , Anemia Falciforme/fisiopatologia , Ensaios Clínicos como Assunto , Método Duplo-Cego , Contagem de Eritrócitos , Feminino , Hematócrito , Humanos , Masculino , Dor/fisiopatologia , Placebos , Agregação Plaquetária , Distribuição AleatóriaRESUMO
The subcutaneous injection of the methylxanthine derivative pentoxifylline (3,7, dimethyl-1-(5-oxo-hexyl)-xanthine) was able to induce, 3 h later, a significant reduction of benzodiazepine binding sites in rat cerebral cortex. When tested in vitro, pentoxifylline displaced 3H-flunitrazepam from specific binding sites in a competitive manner. For this effect pentoxifylline was about 10 times more potent than caffeine. When given in two daily injections for 5 days, pentoxifylline brought about a significant elevation in the number of cortical benzodiazepine binding sites. Neither acute nor chronic pentoxifylline treatment modified cortical muscarinic cholinergic binding sites. Pentoxifylline has negligible affinity for cortical muscarinic receptors in vitro. These results suggest that pentoxifylline is able to affect the cortical benzodiazepine receptors differentially, depending on time of drug administration.