RESUMO
Using intravital microscopy, we examined the role played by B(1) receptors in leukocyte trafficking across mouse mesenteric postcapillary venules in vivo. B(1) receptor blockade attenuated interleukin (IL)-1beta-induced (5 ng intraperitoneally, 2 h) leukocyte-endothelial cell interactions and leukocyte emigration ( approximately 50% reduction). The B(1) receptor agonist des-Arg(9)bradykinin (DABK), although inactive in saline- or IL-8-treated mice, caused marked neutrophil rolling, adhesion, and emigration 24 h after challenge with IL-1beta (when the cellular response to IL-1beta had subsided). Reverse transcriptase polymerase chain reaction and Western blot revealed a temporal association between the DABK-induced response and upregulation of mesenteric B(1) receptor mRNA and de novo protein expression after IL-1beta treatment. DABK-induced leukocyte trafficking was antagonized by the B(1) receptor antagonist des-arg(10)HOE 140 but not by the B(2) receptor antagonist HOE 140. Similarly, DABK effects were maintained in B(2) receptor knockout mice. The DABK-induced responses involved the release of neuropeptides from C fibers, as capsaicin treatment inhibited the responses. Treatment with the neurokinin (NK)(1) and NK(3) receptor antagonists attenuated the responses, whereas NK(2), calcitonin gene-related peptide, or platelet-activating factor receptor antagonists had no effect. Substance P caused leukocyte recruitment that, similar to DABK, was inhibited by NK(1) and NK(3) receptor blockade. Mast cell depletion using compound 48/80 reduced DABK-induced leukocyte trafficking, and DABK treatment was shown histologically to induce mast cell degranulation. DABK-induced trafficking was inhibited by histamine H(1) receptor blockade. Our findings provide clear evidence that B(1) receptors play an important role in the mediation of leukocyte-endothelial cell interactions in postcapillary venules, leading to leukocyte recruitment during an inflammatory response. This involves activation of C fibers and mast cells, release of substance P and histamine, and stimulation of NK(1), NK(3), and H(1) receptors.
Assuntos
Leucócitos/imunologia , Receptores da Bradicinina/imunologia , Vênulas/imunologia , Animais , Bradicinina/análogos & derivados , Bradicinina/farmacologia , Capilares/imunologia , Movimento Celular , Endotélio Vascular/citologia , Endotélio Vascular/imunologia , Expressão Gênica , Interleucina-1/imunologia , Interleucina-1/farmacologia , Leucócitos/citologia , Leucócitos/efeitos dos fármacos , Masculino , Veias Mesentéricas/imunologia , Mesentério/irrigação sanguínea , Camundongos , Camundongos Knockout , Receptor B1 da Bradicinina , Receptor B2 da Bradicinina , Receptores da Bradicinina/biossíntese , Receptores da Bradicinina/genética , Substância P/imunologiaRESUMO
Crohn's disease and ulcerative colitis are inflammatory bowel diseases (IBD) characterized by chronic relapsing mucosal inflammation. Tumour necrosis factor (TNF)-α, a known agonist of the mitogen-activated protein kinase (MAPK) pathway, is a key cytokine in this process. We aimed first to determine whether p38 MAPK is activated in IBD inflamed mucosa, and then studied the effect of four different p38α inhibitory compounds on MAPK phosphorylation and secretion of proinflammatory cytokines by IBD lamina propria mononuclear cells (LPMCs) and organ culture biopsies. In vivo phospho-p38α and p38α expression was evaluated by immunoblotting on intestinal biopsies from inflamed areas of patients affected by Crohn's disease and ulcerative colitis, and from normal mucosa of sex- and age-matched control subjects. Both mucosal biopsies and isolated LPMCs were incubated with four different p38α selective inhibitory drugs. TNF-α, interleukin (IL)-1ß and IL-6 were measured in the organ and cell culture supernatants by enzyme-linked immunosorbent assay. We found higher levels of phospho-p38α in the inflamed mucosa of IBD patients in comparison to controls. All the p38α inhibitory drugs inhibited p38α phosphorylation and secretion of TNF-α, IL-1ß and IL-6 from IBD LPMCs and biopsies. Activated p38α MAPK is up-regulated in the inflamed mucosa of patients with IBD. Additionally, all the p38α selective inhibitory drugs significantly down-regulated the activation of the MAPK pathway and the secretion of proinflammatory cytokines.
Assuntos
Citocinas/metabolismo , Inibidores Enzimáticos/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Adolescente , Adulto , Western Blotting , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Feminino , Humanos , Imidazóis/farmacologia , Mediadores da Inflamação/metabolismo , Doenças Inflamatórias Intestinais/enzimologia , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Masculino , Pessoa de Meia-Idade , Niacinamida/farmacologia , Técnicas de Cultura de Órgãos , Fosforilação/efeitos dos fármacos , Piridinas/farmacologia , Pirimidinas/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Adulto Jovem , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidoresRESUMO
Kinins are important peptide mediators of a diverse range of physiological and pathological functions of the cardiovascular system. The kinin peptides exert their effects by selective activation of two distinct G-protein coupled receptors termed B(1) and B(2). The principal kinin peptides involved in the acute regulation of cardiovascular function during normal physiology are bradykinin (BK) and Lys-BK which produce their effects via activation of B(2) receptors. The B(1) receptor is activated by the des-Arg(9)kinin metabolites namely des-Arg(9)BK and Lys-des-Arg(9)BK, the synthesis of which are increased during inflammation. The B(1) receptor, which is not constitutively expressed, is induced in various pathologies relating to inflammation. Recent investigations into the molecular mechanisms of B(1) receptor induction and their distribution and function in the cardiovascular system have shown that following an inflammatory stimulus the B(1) receptor is induced and may play an important role in modulation of cardiovascular function. This review summarises recent studies on B(1) receptor expression and function in the cardiovascular system and discusses the role of these receptors in regulation of circulatory homeostasis and their potential as therapeutic targets.
Assuntos
Bradicinina/análogos & derivados , Endotélio Vascular/metabolismo , Sistema Calicreína-Cinina/fisiologia , Músculo Liso Vascular/metabolismo , Receptores da Bradicinina/metabolismo , Choque Séptico/metabolismo , Animais , Arteriosclerose/metabolismo , Bradicinina/metabolismo , Humanos , Inflamação , Precondicionamento Isquêmico Miocárdico , Isquemia Miocárdica/metabolismo , Receptor B1 da Bradicinina , Receptor B2 da BradicininaRESUMO
5-HT stimulated cyclic AMP generation in human colonic circular smooth muscle in a concentration-dependent fashion (EC50 = 229.1 nM). DAU 6236 also increased cyclic AMP formation and was a partial agonist relative to 5-HT. GR 113808 inhibited the cyclic AMP formation induced by 5-HT with a -log Ki value of 9.1 and an apparent pA2 value of 9.2. Ondansetron and methysergide failed to inhibit cyclic AMP formation induced by 5-HT. These results indicate that the 5-HT4 receptors of human colonic circular muscle mediate relaxation and inhibition of spontaneous contractions via formation of cyclic AMP.
Assuntos
Colo/metabolismo , AMP Cíclico/biossíntese , Músculo Liso/metabolismo , Receptores de Serotonina/metabolismo , Colo/efeitos dos fármacos , Humanos , Técnicas In Vitro , Músculo Liso/efeitos dos fármacos , Serotonina/farmacologia , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologiaRESUMO
1. The nature of the receptor coupling mechanism of the 5-hydroxytryptamine4 (5-HT4) receptor in the circular smooth muscle of the human colon has been further investigated. 2. 5-HT stimulated cyclic AMP generation and caused a relaxation in a concentration-dependent fashion, with EC50 values of 175.5 and 274.9 nM respectively. DAU 6236 increased cyclic AMP formation and caused a relaxant effect but was a partial agonist relative to 5-HT. 3. The 5-HT4 receptor antagonist, GR 113808, inhibited cyclic AMP formation and relaxation induced by 5-HT with -log Ki values of 9.1 (cyclic AMP) and 8.9 (relaxation) and apparent pA2 values of 9.2 (cyclic AMP) and 9.5 (relaxation). 4. Ondansetron and methysergide failed to inhibit cyclic AMP formation or the relaxation induced by 5-HT. 5. The phosphodiesterase inhibitor, IBMX, produced a concentration-dependent relaxation (EC50 = 30 microM) and at 1 microM it enhanced the 5-HT-induced relaxation producing a leftward shift of the 5-HT concentration-effect curve with a concentration-ratio of 4.1. Rolipram caused a concentration-dependent relaxation (EC50 = 564.8 nM) and at 200 nm caused a leftward shift of the concentration-effect curve to 5-HT with a concentration-ratio of 5.5. 6. Application of the adenylyl cyclase inhibitor, SQ 22536 (0.1 mM), and the protein kinase inhibitors, H7 (100 nM) and H89 (200 nM), inhibited the relaxant effect of 5-HT inducing a rightward shift of the concentration-effect curve with concentration-ratios of 10.1, 2.7 and 4.2 respectively. 7. Forskolin stimulated cyclic AMP production and caused a relaxation. The maximum relaxant effect of forskolin (6 microM, 13.8 +/- 1.9 cm.s) was not significantly different from the maximum relaxant effect of 5-HT (10 microM, 12.7 +/- 4.9 cm.s). However, the cyclic AMP levels stimulated by forskolin (6 microM, 49.3 +/- 6.6 pmol mg-1) were markedly greater than those stimulated by 5-HT (10 microM, 7.6 +/- 2.0 pmol mg-1). 8. In conclusion, these results indicate that the 5-HT4 receptors of the circular smooth muscle of human colon mediate relaxation and inhibition of spontaneous contractions via activation of adenylyl cyclase, formation of cyclic AMP and activation of protein kinase A.
Assuntos
Benzimidazóis/farmacologia , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , AMP Cíclico/biossíntese , Indóis/farmacologia , Relaxamento Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Receptores de Serotonina/fisiologia , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologia , Serotonina/farmacologia , Transdução de Sinais , Sulfonamidas/farmacologia , 1-Metil-3-Isobutilxantina/farmacologia , Análise de Variância , Colo/efeitos dos fármacos , AMP Cíclico/antagonistas & inibidores , Relação Dose-Resposta a Droga , Humanos , Ondansetron/farmacologia , Inibidores de Fosfodiesterase/farmacologia , Receptores de Serotonina/efeitos dos fármacos , Receptores 5-HT4 de Serotonina , Transdução de Sinais/efeitos dos fármacosRESUMO
1. The pharmacological properties of 5-hydroxytryptamine (5-HT), the 5-HT4 receptor agonists, DAU 6236 and SC 53116 and the 5-HT4 receptor antagonist, GR 1130808, were studied in the rat oesophagus, rat ileum and human colon. 2. 5-HT relaxed the longitudinal muscle of the rat oesophagus and rat ileum and the circular muscle of the human colon. Absolute values of relaxation were measured and showed the order of the maximum responses, rat oesophagus >> human colon > rat ileum with EC50 values of 189 +/- 15 nM, 157 +/- 4 nM, 306 +/- 72 nM, respectively. 5-HT also inhibited the spontaneous contractions of the human colon with an EC50 value of 119 +/- 1 nM. The effect of 5-HT on the human colon was not affected by methysergide (10 microM) or ondansetron (1 microM). 3. The use of the uptake and metabolism inhibitors, cocaine (30 microM) and pargyline (100 microM), did not increase the potency of 5-HT in the rat oesophagus or human colon. In the rat oesophagus, cocaine (30 microM) produced a reduction in carbachol-induced tone of 22.2 +/- 0.6% and reduced the 5-HT maximum effect by 52.0 +/- 0.4%. 4. The compounds, DAU 6236 and SC 53116, showed a different pattern of potencies and efficacies in the rat oesophagus, rat ileum and human colon compared to 5-HT. DAU 6236 relaxed the human colonic circular muscle with an EC50 value of 129 +/- 16 nM but its efficacy was less than that of 5-HT. DAU 6236 (1 microM) also antagonized the 5-HT-induced relaxation of the human colon with a dose-ratio of 9.9. In the rat oesophagus and rat ileum, DAU 6236 was inactive in the majority of tissues. In the minority of oesophagus tissues that did respond the EC50 value was 1.2 +/- 0.7 microM. DAU 6236 also antagonized the effect of 5-HT in the rat oesophagus in a non-surmountable fashion. SC 53116 relaxed the rat oesophagus with an EC50 value of 91 +/- 4 nM, with an efficacy less than that observed to 5-HT; however, at 200 nM it did not antagonize the 5-HT-induced relaxation of the rat oesophagus. SC 53116 showed no agonist activity in the rat ileum and human colon, but at 1 microM it did antagonize the effect of 5-HT in the human colon with a dose-ratio of 11.3 +/- 0.3. 5. GR 113808 competitively antagonized the 5-HT4 receptor-mediated relaxation of the rat oesophagus with a pA2 value of 8.59 (8.18-9.00) against 5-HT and 9.05 (8.79-9.31) against SC 53116. GR 113808(0.01 microM) also antagonized the 5-HT-induced relaxation of human colonic circular muscle with an apparent pA2 value of 9.02 +/- 0.12. However at 1 microM the apparent pA2 value was significantly lower than that measured at 0.01 and 0.1 microM. GR 113808 (0.01 microM) antagonized the 5-HT4 receptor-mediated relaxation of the rat ileum with an apparent pA2 value of 9.30 +/- 0.21.6. In conclusion, these studies have shown that the human colon, rat oesophagus and rat ileum contain functional 5-HT4 receptors. However, the 5-HT4 receptor agonists displayed differences in these tissues making it necessary to be cautious when extrapolating from animal to human tissue. This emphasizes the importance of the use of human tissue in the development of therapeutic drugs.
Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes , Colo/fisiologia , Esôfago/fisiologia , Íleo/fisiologia , Receptores de Serotonina/fisiologia , Animais , Benzamidas/farmacologia , Benzimidazóis/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Feminino , Humanos , Indóis/farmacologia , Masculino , Modelos Biológicos , Músculo Liso/fisiologia , Pirróis/farmacologia , Ratos , Ratos Wistar , Receptores de Serotonina/efeitos dos fármacos , Serotonina/fisiologia , Agonistas do Receptor de Serotonina/metabolismo , Sulfonamidas/farmacologiaRESUMO
1 We have investigated the role of kinin B1 receptor induction in the endotoxemic rat heart and elucidated the mechanisms underlying B1 receptor-mediated coronary vasodilation. We also investigated the role of these receptors in endotoxin-induced hypotension. 2 Endotoxin treatment induced cardiac B1 receptor mRNA expression and promoted a coronary vasodilation response to des-Arg9bradykinin (DABK; ED50 = 149.4 pmol, n = 9) ex vivo peaking at 6 h. The B1 receptor antagonist des-Arg9-[Leu8]-BK (DALBK, 30 nM) significantly (P<0.05) inhibited the DABK-induced response (pA2 = 8.4, n = 5) whilst HOE140 (B2 receptor antagonist, 10 nM) was inactive (n = 4). 3 Removal of the endothelium or infusion with indomethacin (5 microM), but not L-NAME (300 microM) or ODQ (1 microM), inhibited (>85%, P<0.05, n = 5) the DABK-induced response. DABK caused a dose-dependent release of the prostacyclin metabolite, 6-keto-PGF1a (Emax = 0.3 ng ml-1, n = 6). 4 In vitro perfusion of hearts with endotoxin (1 microg ml-1, n = 6) or interleukin-1beta (5 ng ml-1, n = 6) induced B1 receptor mRNA expression and promoted a time-dependent vasodilation response to DABK. 5 Endotoxin treatment (6 h) in vivo promoted a hypotensive response to DABK (ED50 = 29.7 nmol kg-1, n = 10) which was antagonised by DALBK (3-6 nmol kg-1 min-1, P<0.05, n = 7). DALBK (3 nmol kg-1 min-1) and des-Arg10HOE140 (B1 receptor antagonist, 30 nmol kg-1 min-1) produced a 5.3% (n = 6, P<0.05) and 8.8% (n = 5, P<0.05) reversal, respectively, of endotoxin-induced hypotension. 6 In summary, we have shown that in endotoxemia activation of B1 receptors causes coronary vasodilation via endothelial prostacyclin release. Additionally, B1 receptor antagonists partially reversed endotoxin-induced hypotension. Therefore activation of B1 receptors may have a role to play in the vascular changes associated with endotoxemia.
Assuntos
Bradicinina/análogos & derivados , Vasos Coronários/efeitos dos fármacos , Endotoxemia/metabolismo , Miocárdio/metabolismo , Receptores da Bradicinina/biossíntese , Vasodilatação/efeitos dos fármacos , Animais , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Bradicinina/farmacologia , Antagonistas dos Receptores da Bradicinina , Circulação Coronária/efeitos dos fármacos , Circulação Coronária/fisiologia , Endotélio Vascular/fisiologia , Endotoxemia/fisiopatologia , Hipotensão/etiologia , Hipotensão/fisiopatologia , Masculino , Tono Muscular/fisiologia , Óxido Nítrico/biossíntese , RNA Mensageiro/biossíntese , Ratos , Ratos Wistar , Receptor B1 da Bradicinina , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
1. The effects of endotoxaemia on coronary vasodilator responses to bradykinin (BK), sodium nitroprusside (SNP) and nicardipine were investigated in the rat isolated heart perfused at constant flow ex vivo. 2. Dose-dependent reductions in coronary perfusion pressure reaching a maximum of 56+/-3 and 57+/-5 mmHg were observed for BK and SNP respectively. The BK response was biphasic, consisting of a rapid dilator response that was insensitive to N(G)nitro-L-arginine methyl ester (L-NAME, 0.1 mM) and a second slower component whose duration was attenuated by L-NAME. 3. Hearts obtained from rats treated with endotoxin (2.5 mg kg(-1), i.p.) for 2 or 6 h had increased basal coronary perfusion pressure and reduced vasodilator responses to BK or SNP. Dilator responses to nicardipine were not affected by endotoxin treatment. In vitro perfusion of hearts from endotoxin-treated rats with L-NAME (0.1 mM) restored SNP responses to control values. 4. Treatment with dexamethasone (1 mg kg(-1)), 1 h before endotoxin did not alter the endotoxin-induced impairment of dilator responses to BK or SNP. 5. These results show that coronary microvascular responses are altered following endotoxin exposure. Endotoxin results in increased coronary microvascular tone despite induction of NO synthase and inhibits the dilator response to BK and SNP, vasodilators that act via the release of NO. Responses to SNP in endotoxin-treated hearts were restored to control values in the presence of L-NAME suggesting that enhanced endogenous NO synthesis might saturate guanylate cyclase resulting in reduced response to NO donors. The reduced response to vasodilators and increased coronary resistance might be important in determining the response of the coronary circulation to systemic inflammation and infection.
Assuntos
Circulação Coronária/efeitos dos fármacos , Endotoxinas/farmacologia , Miocárdio/metabolismo , Óxido Nítrico/metabolismo , Animais , Bradicinina/farmacologia , Vasos Coronários/efeitos dos fármacos , Relação Dose-Resposta a Droga , Endotoxemia/induzido quimicamente , Escherichia coli , Lipopolissacarídeos/farmacologia , Microcirculação/efeitos dos fármacos , Nicardipino/farmacologia , Nitroprussiato/farmacologia , Ratos , Ratos Wistar , Resistência Vascular/efeitos dos fármacos , Resistência Vascular/fisiologia , Vasodilatadores/farmacologiaRESUMO
Distension of the rat intestine causes a capsaicin-sensitive, pressure-dependent depressor response which is indicative of nociception. A hypersensitivity of jejunal distension which possibly involves tachykinin NK2 receptors and is restricted to areas with mast cell hyperplagia is observed in rats infected 30 days previously with Nippostrongylus brasiliensis. This study aimed to further investigate the role of mast cells, tachykinins and kinins in this intestinal hypersensitivity. The activity of a mast cell stabilizer (doxantrazole), kinin antagonists (des-Arg 10-[Leu9]-kallidin, B1, HOE 140, B2) and tachykinin antagonists (CP 99, 994, NK1, SR 142801, NK3) were tested against the distension-induced depressor responses in control and post-infected rats. The 30-day post-infection-induced hypersensitivity was significantly reduced by the mast cell stabilizer doxantrazole. The hypersensitivity had resolved in 90-day post-infected rats when mast cells levels had normalized. Des-Arg 10-[Leu9]-kallidin and HOE 140 did not inhibit the depressor responses in controls but produced a significant inhibition in 30-day post-infected rats. CP 99,994 inhibited the depressor responses in post-infected rats with an equal potency to that in control rats. SR 142801 was inactive in both groups. In conclusion, mast cells and kinin-mediated nociception appear to be involved in post-infection intestinal hypersensitivity whereas tachykinin NK1 and NK3 receptors do not.
Assuntos
Enteropatias/fisiopatologia , Enteropatias/parasitologia , Jejuno/fisiopatologia , Mastócitos/fisiologia , Receptores da Bradicinina/fisiologia , Infecções por Strongylida/fisiopatologia , Animais , Cateterismo , Masculino , Nippostrongylus , Ratos , Ratos Wistar , Receptor B1 da Bradicinina , Receptor B2 da Bradicinina , Receptores de Taquicininas/fisiologia , Infecções por Strongylida/complicações , Fatores de TempoRESUMO
The 5-HT (5-hydroxytryptamine)-induced contractile biphasic concentration-effect curve in rat isolated jejunum was investigated. The pEC50 values for the first and second phases were 8.0 and 6.1, respectively. The responses were insensitive to atropine (0.1 microM), ketanserin (2 microM), (-)-pindolol (5 microM), yohimbine (0.1 microM) and GR 113808 ({1-[2-(methyl-sulphonylamino)ethyl]-4-piperidinyl}methyl 1-methyl-1 H-indole-3-carboxylate, 1 microM) but susceptible to cocaine (10 microM). The low affinity phase was blocked by tetrodotoxin (1 microM), ondansetron (1 microM) and SR48968 (S)-N-methyl-N-[4-(4-acetylamino-4-phenyl piperidino)-2-(3,4-dichlorophenyl)butyl]benzamide, 0.1 microM). The high affinity phase was antagonised non-surmountably by fluoxetine (1 microM) methysergide (0.1 microM), spiperone (0.1 microM) and methiothepin (0.1 microM). Ritanserin (0.01-0.1 microM) and mesulergine (0.01-0.1 microM) acted as surmountable, competitive antagonists with pA2 values of 8.0 and 8.1, respectively. Clozapine (0.1 microM) was a surmountable antagonist with an apparent pA2 value of 8.0. The rank potency order of the 5-HT receptor agonists was 5-CT (5-carboxyamidotryptamine) > or = 5-HT = 5-methoxytryptamine > or = alpha-methyl-5-HT > > 8-OH-DPAT ((+/-)-2-dipropyl-amino-8-hydroxy-1,2,3, 4-tetrahydronaphthalene) > dipropyl-5-CT > renzapride = sumatriptan. The responses to 5-HT and 5-CT were not potentiated by pargyline (10 and 100 microM). It is suggested that rat jejunum contains a neuronal 5-HT3 receptor facilitating neurokinin release and a contractile smooth muscle 5-HT receptor with a pharmacological operational profile similar to the cloned 5-ht7 receptor.
Assuntos
Jejuno/metabolismo , Receptores de Serotonina/metabolismo , Serotonina/metabolismo , Animais , Atropina/farmacologia , Benzamidas/farmacologia , Relação Dose-Resposta a Droga , Técnicas In Vitro , Masculino , Metisergida/farmacologia , Inibidores da Monoaminoxidase/farmacologia , Contração Muscular/efeitos dos fármacos , Ondansetron/farmacologia , Piperidinas/farmacologia , Ratos , Ratos Wistar , Antagonistas da Serotonina/farmacologia , Tetrodotoxina/farmacologiaRESUMO
Distension of the rat intestine causes a cardiovascular response which is indicative of nociception. Since tachykinins are involved in nociception, we tested the effect of neurokinin receptor antagonists against the distension-induced response. The jejunal distension-induced depressor responses were inhibited in a dose-dependent fashion by CP 99,994 (+)-(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine, tachykinin NK1 receptor antagonist, ED50 = 0.8 mg/kg) and SR 48968 (S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4-dichloropheny l)butyl]benzamide, tachykinin NK2 receptor antagonist, ED50 = 0.7 mg/kg). SR 142801 (S)-(N)-(1-(3-(1-benzoyl-3-(3,4-dichlorophenyl)piperidin-3-yl)prop yl)-4-phenylpiperidin-4-yl)-N-methylacetamide, tachykinin NK3 receptor antagonist, 0.3-10 mg/kg) did not significantly affect the depressor responses to jejunal distension. In addition, CP 99,994 (3 mg/kg) and SR 48968 (3 and 10 mg/kg) reduced sensitivity to distension as revealed by a 2.7-fold (CP 99.994, 3 mg/kg), 2.6-fold (SR 48968, 3 mg/kg) and 4.7-fold (SR 48968, 10 mg/kg) increase in the threshold pressure. Intestinal compliance was not affected by the antagonists. In conclusion, these results suggest that tachykinin NK1 and NK2 but not NK3 receptors are possibly involved in the rat jejunal distension pain response.
Assuntos
Analgésicos não Narcóticos/farmacologia , Jejuno/fisiopatologia , Dor/tratamento farmacológico , Receptores de Taquicininas/antagonistas & inibidores , Animais , Benzamidas/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Masculino , Antagonistas dos Receptores de Neurocinina-1 , Piperidinas/farmacologia , Pressão , Ratos , Ratos Wistar , Receptores da Neurocinina-2/antagonistas & inibidores , Receptores da Neurocinina-3/antagonistas & inibidores , Análise de RegressãoRESUMO
Distension of the rat intestine causes a depressor response which is predictive of nociception. This study investigated the effects of previous infection with Nippostrongylus (N.) brasiliensis on the sensitivity to intestinal distension and the role of tachykinin NK2 receptors. The tachykinin NK2 receptor antagonist, SR48968 (S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4-dichloropheny l)butyl]benzamide) inhibited the nociceptive response (ED50 = 0.7 mg/kg) in control rats. In post-N. brasiliensis-infected rats sensitivity to intestinal distension was increased which was accompanied by an increase in the apparent potency value of SR48968 (ED50 = 0.1 mg/kg). The hypersensitivity was limited to areas of hypermastocytosis. It is concluded that the post-inflammatory changes that occur in post-infected rats increase visceral sensitivity and the apparent potency of tachykinin NK2 receptor antagonists.
Assuntos
Doenças do Colo/fisiopatologia , Enteropatias Parasitárias/fisiopatologia , Doenças do Jejuno/fisiopatologia , Nippostrongylus , Receptores da Neurocinina-2/fisiologia , Infecções por Strongylida/fisiopatologia , Animais , Benzamidas/farmacologia , Colo/enzimologia , Colo/patologia , Colo/fisiopatologia , Doenças do Colo/enzimologia , Doenças do Colo/patologia , Dilatação Patológica , Enteropatias Parasitárias/enzimologia , Enteropatias Parasitárias/patologia , Doenças do Jejuno/enzimologia , Doenças do Jejuno/patologia , Jejuno/enzimologia , Jejuno/patologia , Jejuno/fisiopatologia , Masculino , Mastócitos/patologia , Medição da Dor , Peroxidase/metabolismo , Piperidinas/farmacologia , Ratos , Ratos Wistar , Receptores da Neurocinina-2/antagonistas & inibidoresRESUMO
Functional 5-HT4 receptors have been reported to be present in numerous isolated tissue preparations including the rat oesophagus, guinea-pig ileum, and human colon. The pharmacological properties of the novel, potent and selective 5-HT4 receptor antagonists SB203186 (1-piperidinyl)ethyl 1H-indole 3-carboxylate), SB205008 (1-butyl-1-methyl-4-piperidinylmethyl)-8-amino-7-chloro-1,4-ben zodioxan-5- carboxylate iodide), and SB207710 (1-butyl-4-piperidinylmethyl)-8-amino-7-iodo-1,4 benzo-dioxan-5-carboxylate) were studied in these tissues. The nature of antagonism of the 5-HT-induced effects was investigated on the above isolated tissue preparations. 5-HT produced its effect with the following EC50 values: 400 +/- 0.4 nM (rat oesophagus, n = 20), 154 +/- 14 nM (guinea-pig ileum, n = 9) and 144 +/- 0.1 nM (human colon, n = 9). SB207710 (0.03-1 nM), SB205008 (1.0-10 nM), and SB203186 (10-100 nM) antagonised the 5-HT4 receptor-mediated relaxations of the carbachol-contracted rat isolated oesophagus against 5-HT with pKB values of 10.9 +/- 0.1, 9.5 +/- 0.1, and 9.0 +/- 0.1 respectively without effecting the maximum response. On the guinea-pig ileum peristaltic reflex preparation, SB207710 (0.01-1 nM) did not modify the reflex but it behaved as an antagonist of the 5-HT-induced facilitation with a pA2 value of 9.9 +/- 0.2.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Dioxanos/farmacologia , Indóis/farmacologia , Peristaltismo/efeitos dos fármacos , Piperidinas/farmacologia , Receptores de Serotonina/metabolismo , Antagonistas da Serotonina/farmacologia , Animais , Carbacol/farmacologia , Colo/fisiologia , Esôfago/fisiologia , Feminino , Cobaias , Humanos , Íleo/fisiologia , Técnicas In Vitro , Masculino , Músculo Liso/efeitos dos fármacos , Ratos , Ratos Wistar , Receptores de Serotonina/efeitos dos fármacosRESUMO
The effects of chronic infusion with 5-hydroxytryptamine (5-HT, 75 micrograms/kg per hour) for 5 or 10 days in vivo on the responses of rat oesophagus, fundus and jejunum to 5-HT and partial 5-HT receptor agonists in vitro were investigated. In the rat oesophagus, chronic treatment produced rightward shifts of the 5-HT4 receptor-mediated concentration-effect curves to 5-HT (dose-ratio = 3.8, day 5 and 2.8, day 10) and SC 53116 (1-S,8-S)-4-amino-5-chloro-N-[(hexahydro-1H-pyrrolizin-1-yl) methyl]-2-methoxy-benzamide hydrochloride, dose-ratio = 7.1, day 5 and 8.9, day 10) as compared to control tissues. The maximum effect of 5-HT and SC 53116 in rat oesophagus was reduced following the 10 day treatment. The 5-HT2B receptor-mediated contractile effect of 5-HT on rat fundus from treated animals responded with a significantly reduced potency (dose-ratio = 4.1, 5-day; 4.2, 10-day) compared to control tissues. The maximum response to 5-HT was reduced in tissues from animals treated for 10 days. The concentration-effect curve to the partial agonist 1-(3-chlorophenyl)-piperazine dihydrochloride (mCPP) was shifted to the right in fundic tissue from treated animals (dose-ratio = 2.5, 5-day; 2.8, 10-day) compared to control tissues. The maximum response to mCPP was also reduced in tissues from 5-HT treated animals. It has recently been shown in a preliminary characterisation that in rat jejunum, 5-HT produces a biphasic concentration-effect curve which is mediated by a putative 5-ht7 (first phase) and 5-HT3 (second phase) receptor mechanism. In the present study 5-HT produced a control biphasic concentration-effect curve in rat jejunum with a pEC50-1 value of 8.5 +/- 3.5 and a pEC50-2 value of 5.9 +/- 0.3 and maximum response values of 43.8 (32.5-55.0)% (Emax1) and 65.3 (41.7-88.9)% (Emax 2) of the response to acetylcholine. In jejunal tissues from treated animals, 5-HT produced its contractile effect in vitro with a 3.7 and 2.8 fold (5-day) and a 1.3 and 1.4 fold (10-day) rightward shift of the first and second phase respectively of the control concentration-effect curve to 5-HT. The maximum response produced by 5-HT in the first phase in jejunal tissues from animals treated for 10 days was significantly reduced by 49.1%. These findings represent the first report that chronic infusion of 5-HT produces a residual desensitisation of the 5-HT4, 5-HT2B, and putative 5-ht7/5-HT3 receptor-mediated responses in rat oesophagus, fundus and jejunum respectively when measured in vitro.
Assuntos
Esôfago/efeitos dos fármacos , Jejuno/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Serotonina/farmacologia , Estômago/efeitos dos fármacos , Animais , Benzamidas/farmacologia , Esôfago/metabolismo , Mucosa Gástrica/metabolismo , Infusões Intravenosas , Jejuno/metabolismo , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Liso/metabolismo , Piperazinas/farmacologia , Pirróis/farmacologia , Ratos , Serotonina/administração & dosagem , Agonistas do Receptor de Serotonina/farmacologiaRESUMO
BACKGROUND: The probiotic Bifidobacterium longum NCC3001 normalizes anxiety-like behavior and hippocampal brain derived neurotrophic factor (BDNF) in mice with infectious colitis. Using a model of chemical colitis we test whether the anxiolytic effect of B. longum involves vagal integrity, and changes in neural cell function. Methods Mice received dextran sodium sulfate (DSS, 3%) in drinking water during three 1-week cycles. Bifidobacterium longum or placebo were gavaged daily during the last cycle. Some mice underwent subdiaphragmatic vagotomy. Behavior was assessed by step-down test, inflammation by myeloperoxidase (MPO) activity and histology. BDNF mRNA was measured in neuroblastoma SH-SY5Y cells after incubation with sera from B. longum- or placebo-treated mice. The effect of B. longum on myenteric neuron excitability was measured using intracellular microelectrodes. KEY RESULTS: Chronic colitis was associated with anxiety-like behavior, which was absent in previously vagotomized mice. B. longum normalized behavior but had no effect on MPO activity or histological scores. Its anxiolytic effect was absent in mice with established anxiety that were vagotomized before the third DSS cycle. B. longum metabolites did not affect BDNF mRNA expression in SH-SY5Y cells but decreased excitability of enteric neurons. CONCLUSIONS & INFERENCES: In this colitis model, anxiety-like behavior is vagally mediated. The anxiolytic effect of B. longum requires vagal integrity but does not involve gut immuno-modulation or production of BDNF by neuronal cells. As B. longum decreases excitability of enteric neurons, it may signal to the central nervous system by activating vagal pathways at the level of the enteric nervous system.
Assuntos
Ansiolíticos/uso terapêutico , Ansiedade/tratamento farmacológico , Bifidobacterium/metabolismo , Colite , Trato Gastrointestinal , Probióticos , Nervo Vago , Animais , Ansiedade/fisiopatologia , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/genética , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Linhagem Celular , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colite/fisiopatologia , Sulfato de Dextrana/farmacologia , Sistema Nervoso Entérico/citologia , Sistema Nervoso Entérico/efeitos dos fármacos , Sistema Nervoso Entérico/fisiologia , Fezes/microbiologia , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/inervação , Trato Gastrointestinal/microbiologia , Humanos , Masculino , Camundongos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Placebos , Probióticos/farmacologia , Probióticos/uso terapêutico , Vagotomia , Nervo Vago/anatomia & histologia , Nervo Vago/fisiologiaAssuntos
Saúde , Psicanálise , Atitude Frente a Saúde , Caráter , Ética , Humanos , Estilo de Vida , Saúde Mental , Princípios Morais , Teoria Psicanalítica , Autoimagem , Comportamento SocialRESUMO
Leucocyte-endothelial cell interactions are prerequisite to leucocyte infiltration and intestinal inflammation. GI270384X is a novel inhibitor of ICAM-1 and E-selectin expression and inhibits leucocyte adhesion and improves experimental colitis. We hypothesized that GI270384X maybe effective in treatment of visceral hyperalgaesia. Visceromotor behavioural responses to colorectal distension (CRD) were obtained in naïve rats or rats treated with zymosan (3 h) or 2,4,6-trinitrobenzene sulphonic acid (TNBS) (4 and 30 days) or rats exposed to acute restraint stress. Studies were also performed in a high-anxiety genetic model of colonic hyperalgaesia using Wistar-Kyoto (WKY) rats. Rats were treated orally with GI270384X or vehicle either prior to or after the administration of sensitizing stimulus. The visceromotor response to CRD was significantly enhanced in all models. GI270384X attenuated the enhanced responses to distension induced by inflammatory stimuli (TNBS and zymosan) and in the high-anxiety WKY rats; however, the drug did not inhibit the hypersensitivity induced by acute restraint stress. GI270384X was most potent in the models of acute inflammatory hyperalgaesia with a minimum efficacious dose (MED) of 0.3 and 1 mg kg(-1) observed in the TNBS and zymosan models respectively. The compound was less potent in the chronic and postinflammatory models with an MED of 10 and 30 mg kg(-1) observed in the WKY and 30-day TNBS models respectively. These findings show for the first time that inhibition of leucocyte-endothelial cell interactions can have a beneficial effect on visceral hyperalgaesia associated with inflammatory and chronic anxiety states, but is less effective against stress-associated visceral hyperalgaesia.
Assuntos
Moléculas de Adesão Celular/metabolismo , Colo/efeitos dos fármacos , Colo/fisiologia , Células Endoteliais/metabolismo , Motilidade Gastrointestinal/efeitos dos fármacos , Hiperalgesia/tratamento farmacológico , Polietilenoglicóis , Purinas , Animais , Adesão Celular/efeitos dos fármacos , Moléculas de Adesão Celular/genética , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colo/anatomia & histologia , Colo/patologia , Dilatação Patológica , Selectina E/genética , Selectina E/metabolismo , Células Endoteliais/citologia , Humanos , Hiperalgesia/fisiopatologia , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Leucócitos/citologia , Leucócitos/metabolismo , Masculino , Polietilenoglicóis/farmacologia , Polietilenoglicóis/uso terapêutico , Purinas/farmacologia , Purinas/uso terapêutico , Ratos , Ratos Endogâmicos WKY , Ratos Sprague-Dawley , Restrição Física , Estresse Psicológico , Ácido Trinitrobenzenossulfônico/farmacologia , Zimosan/farmacologiaRESUMO
1. 5-Hydroxytryptamine (5-HT) has been shown to cause a consistent secretory effect in the rat small intestine only when administered luminally or by close intraarterial infusion. Intraluminal 5-HT-induced secretion is possibly mediated by 5-HT4 receptors. Therefore, it was decided to investigate the effect of 5-HT and selective 5-HT4 receptor agonists (SC 53116 and DAU 6236) on intestinal fluid transport in rat jejunum and ileum. The study also investigated the effect of a selective 5-HT4 receptor antagonist (GR 113808) against the intraluminally administered 5-HT. 2. 5-HT receptor agonists and antagonists were administered intraluminally in pentobarbitone-anesthetized rats. Changes in intestinal fluid transport across the intestinal wall were measured by a single pass technique. 3. Intraluminal 5-HT produced significant antiabsorptive effects is both the jejunum and ileum. The 5-HT-induced responses were blocked by intraluminal administration of the 5-HT4 receptor antagonist GR 113808. The 5-HT4 agonist SC 53116 induced antiabsorptive effects in both regions of the small intestine, but DAU 6236 did not affect the rates of fluid transport. 4. The results indicate that a 5-HT4 receptor has a role in the luminal 5-HT-induced antiabsorptive effect on intestinal fluid transport in the rat.
Assuntos
Absorção Intestinal/efeitos dos fármacos , Intestino Delgado/efeitos dos fármacos , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologia , Serotonina/farmacologia , Animais , Benzamidas/farmacologia , Benzimidazóis/farmacologia , Líquidos Corporais , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Indóis/farmacologia , Intestino Delgado/fisiologia , Masculino , Pirróis/farmacologia , Ratos , Ratos Wistar , Sulfonamidas/farmacologiaRESUMO
We assessed here the effect of the glucocorticoid-regulated protein lipocortin 1 (LC1) in a model of rat myocardial ischemia reperfusion. Treatment of animals with human recombinant LC1 at the end of a 25-min ischemic period significantly reduced the extent of infarct size in the area at risk as measured 2 h later, with approximately 50% inhibition at the highest dose tested of 50 microg per rat (equivalent to 5.4 nmol/kg). The protective effect of LC1 was abolished by protein denaturation and not mimicked by the structurally related protein annexin V. A combination of electron and light microscopy techniques demonstrated the occurrence of the myocardial damage at the end of the reperfusion period, with loss of fiber organization. LC1 provided a partial and visible protection. The dose-dependent protection afforded by LC1 was paralleled by lower values of myeloperoxidase activity, tumor necrosis factor a, and macrophage inflammatory protein-1a. The functional link between migrated leukocytes and the myocardial damage was confirmed by electron and light microscopy, and a significantly lower number of extravasated leukocytes was counted in the group of rats treated with LC1 (50 microg). In conclusion, we demonstrate for the first time that LC1 reduces the leukocyte-dependent myocardial damage associated with an ischemia-reperfusion procedure.