RESUMEN
PURPOSE:To study the anti-inflammatory actions of electroacupuncture (EAc) on an experimental colitis model in mice.METHODS:Thirty-eight male Swiss mice, divided in five groups, were subjected to induction of colitis by TNBS in 50% ethanol. Saline (SAL) and ethanol (ETNL) groups served as controls. TNBS+EAc and TNBS+ dexamethasone subgroups were treated with EAc 100Hz and dexamethasone (DEXA) 1 mg/Kg/day, respectively. After three days, a colon segment was obtained for quantification of myeloperoxidase (MPO) activity, immunohistochemistry for iNOS, malondialdehyde (MDA) and cytokines (IL-1β and IL-10).RESULTS:Neutrophilic activity, assayed as MPO activity, was significantly higher in the TNBS colitis group than that in the saline control group. TNBS+EAc group showed suppression of IL-10 in the colon. EAc treatment significantly reduced the concentration of MDA and the expression of iNOS, as compared to the other groups. CONCLUSION: Electroacupuncture 100Hz applied to acupoint ST-36 promotes an anti-inflammatory action on the TNBS-induced colitis, mediated by increase of IL-10 and decrease of iNOS expression.(AU)
Asunto(s)
Animales , Ratones , Electroacupuntura/veterinaria , Interleucina-10 , Trinitrobencenos , Colitis/inducido químicamente , Óxido Nítrico SintasaRESUMEN
PURPOSE:To study the anti-inflammatory actions of electroacupuncture (EAc) on an experimental colitis model in mice.METHODS:Thirty-eight male Swiss mice, divided in five groups, were subjected to induction of colitis by TNBS in 50% ethanol. Saline (SAL) and ethanol (ETNL) groups served as controls. TNBS+EAc and TNBS+ dexamethasone subgroups were treated with EAc 100Hz and dexamethasone (DEXA) 1 mg/Kg/day, respectively. After three days, a colon segment was obtained for quantification of myeloperoxidase (MPO) activity, immunohistochemistry for iNOS, malondialdehyde (MDA) and cytokines (IL-1β and IL-10).RESULTS:Neutrophilic activity, assayed as MPO activity, was significantly higher in the TNBS colitis group than that in the saline control group. TNBS+EAc group showed suppression of IL-10 in the colon. EAc treatment significantly reduced the concentration of MDA and the expression of iNOS, as compared to the other groups. CONCLUSION: Electroacupuncture 100Hz applied to acupoint ST-36 promotes an anti-inflammatory action on the TNBS-induced colitis, mediated by increase of IL-10 and decrease of iNOS expression.
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Animales , Ratones , Colitis/inducido químicamente , Electroacupuntura/veterinaria , Trinitrobencenos , Óxido Nítrico SintasaRESUMEN
Resolvins of the D series are generated from docosahexaenoic acid, which are enriched in fish oils and are believed to exert beneficial roles on diverse inflammatory disorders, including inflammatory bowel disease (IBD). In this study, we investigated the anti-inflammatory effects of the aspirin-triggered resolvin D1 (AT-RvD1), its precursor (17(R)-hydroxy docosahexaenoic acid [17R-HDHA]) and resolvin D2 (RvD2) in dextran sulfate sodium (DSS)- or 2,4,6-trinitrobenzene sulfonic acid-induced colitis. Our results showed that the systemic treatment with AT-RvD1, RvD2, or 17R-HDHA in a nanogram range greatly improved disease activity index, body weight loss, colonic damage, and polymorphonuclear infiltration in both colitis experimental models. Moreover, these treatments reduced colonic cytokine levels for TNF-α, IL-1ß, MIP-2, and CXCL1/KC, as well as mRNA expression of NF-κB and the adhesion molecules VCAM-1, ICAM-1, and LFA-1. Furthermore, AT-RvD1, but not RvD2 or 17R-HDHA, depended on lipoxin A4 receptor (ALX) activation to inhibit IL-6, MCP-1, IFN-γ, and TNF-α levels in bone marrow-derived macrophages stimulated with LPS. Similarly, ALX blockade reversed the beneficial effects of AT-RvD1 in DSS-induced colitis. To our knowledge, our findings showed for the first time the anti-inflammatory effects of resolvins of the D series and precursor 17R-HDHA in preventing experimental colitis. We also demonstrated the relevant role exerted by ALX activation on proresolving action of AT-RvD1. Moreover, AT-RvD1 showed a higher potency than 17R-HDHA and RvD2 in preventing DSS-induced colitis. The results suggest that these lipid mediators possess a greater efficacy when compared with other currently used IBD therapies, such as monoclonal anti-TNF, and have the potential to be used for treating IBD.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Aspirina/farmacología , Colitis/tratamiento farmacológico , Ácidos Docosahexaenoicos/farmacología , Animales , Anticuerpos Monoclonales/farmacología , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/metabolismo , Moléculas de Adhesión Celular/biosíntesis , Moléculas de Adhesión Celular/inmunología , Colitis/inducido químicamente , Colitis/inmunología , Colitis/metabolismo , Citocinas/biosíntesis , Citocinas/inmunología , Sulfato de Dextran/toxicidad , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , ARN Mensajero/biosíntesis , ARN Mensajero/inmunología , Receptores de Formil Péptido/antagonistas & inhibidores , Receptores de Formil Péptido/inmunología , Receptores de Formil Péptido/metabolismo , Trinitrobencenos/efectos adversos , Trinitrobencenos/farmacología , Contaminantes Químicos del Agua/efectos adversos , Contaminantes Químicos del Agua/farmacologíaRESUMEN
The aqueous fraction of the ethanolic extract of the plant CISSAMPELOS SYMPODIALIS (Menispermaceae) was previously described to inhibit B cell function. The alkaloid warifteine is the major component of this extract. In the present study we investigated the effect of warifteine on B lymphocyte function and characterized its mechanism of action. Purified splenic murine B lymphocytes were stimulated with either Toll-like receptor (TLR) ligands (LPS, Pam (3)Cys and CpG oligodeoxynucleotides) or anti-IgM antibody and the effect of warifteine on B cell response was investigated. Warifteine inhibited both the proliferative response and immunoglobulin (Ig) secretion induced by these stimuli. Kinetics studies demonstrated that warifteine blocked B cell function even when added after 24 h of a 72 h culture. The inhibitory effect of warifteine was also detected in cultures activated by phorbol myristate acetate and calcium ionophore. We investigated the signal transduction pathways blocked by warifteine. It did not modify the total protein phosphorylation pattern in LPS and anti-IgM-stimulated B cell cultures. It did, however, decrease the rise in intracellular calcium levels, the phosphorylation of the mitogen activated protein kinase (MAPK) ERK and the intranuclear levels of the transcription factor NFkappaB. Warifteine also induced an increase in cAMP and its effect on LPS-induced proliferation was mimicked by the control adenyl cyclase activator forskolin. IN VIVO Ig production induced by the TI-2 antigen TNP-ficoll was also inhibited by warifteine. Taking together, our data suggest that warifteine is a potent inhibitor of B cell response both IN VITRO and IN VIVO and that this effect may be due to the induction of increased intracellular cAMP levels, suggesting that this substance may be useful as a modulator of B cell function.
Asunto(s)
Alcaloides/farmacología , Linfocitos B/efectos de los fármacos , Cissampelos/química , Factores Inmunológicos/farmacología , Extractos Vegetales/farmacología , Adenilil Ciclasas/metabolismo , Alcaloides/aislamiento & purificación , Animales , Anticuerpos Antiidiotipos , Linfocitos B/metabolismo , Calcimicina/farmacología , Técnicas de Cultivo de Célula , Proliferación Celular/efectos de los fármacos , Colforsina/farmacología , AMP Cíclico/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Ficoll/análogos & derivados , Inmunoglobulinas/metabolismo , Factores Inmunológicos/aislamiento & purificación , Ionóforos/farmacología , Ligandos , Lipopolisacáridos , Ratones , Ratones Endogámicos BALB C , FN-kappa B/metabolismo , Fosforilación , Extractos Vegetales/química , Hojas de la Planta , Transducción de Señal/efectos de los fármacos , Bazo/inmunología , Acetato de Tetradecanoilforbol , Receptores Toll-Like , TrinitrobencenosRESUMEN
We have evaluated the adjuvant action of jacalin, a lectin obtained from seeds of Artocarpus integrifolia, on humoral immune response against the trinitrophenyl (TNP) hapten when conjugated to it and to Trypanosoma cruzi. The protective effect of parasite-specific antibodies generated in mice immunized with epimastigote forms of T. cruzi plus jacalin was also evaluated by determining the parasitemia levels of animals after infection with 1000 trypomastigote forms. Immunization of mice with trinitrophenylated jacalin (TNP-JAC) in saline resulted in an antibody response to the TNP hapten that was eight and 16 times higher than that found in mice immunized with TNP-human gamma globulin (TNP-HGG) or TNP-bovine serum albumin (TNP-BSA), respectively. In addition, immunization with either a lysate or viable epimastigote forms of T. cruzi in the presence of jacalin resulted in a marked increase in the levels of anti-T. cruzi antibodies. The protective action of antibodies against acute infection by T. cruzi was evident when mice were immunized with 1.0 x 10(5) epimastigotes plus jacalin. These animals had a significantly lower parasitemia than those immunized with epimastigotes alone. In contrast, mice immunized with 1.0 x 10(6) epimastigotes developed very low levels of parasitemia, regardless of the presence of jacalin. These data suggest that jacalin is a potent adjuvant in the humoral response to TNP and T. cruzi, and that the protective action of the T. cruzi-specific antibodies depends on the number of parasites used in the immunization protocol.
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Anticuerpos Antiprotozoarios/sangre , Enfermedad de Chagas/prevención & control , Lectinas/inmunología , Lectinas de Plantas , Trinitrobencenos/inmunología , Trypanosoma cruzi/inmunología , Adyuvantes Inmunológicos , Animales , Femenino , Haptenos/inmunología , Ratones , Ratones Endogámicos BALB C , VacunaciónRESUMEN
Immunological memory is embodied in the rapid and enhanced immune responsiveness to previously encountered antigens. Classically, memory would depend on the presence of small resting long-lived specific lymphocytes which, through clonal expansion after priming with antigen, would be present at higher frequencies than in naive animals. Here we report that T-cell-reconstituted athymic mice, which lack recent thymic emigrants, mount a primary response to a T-cell-dependent antigen, but do not develop memory or the capacity to produce specific anti-TNP IgG1 antibodies during the secondary immune response. On the other hand, if thymocytes are continuously provided during the secondary response, a typical secondary immune response is achieved with high levels of specific IgG1. These results lead us to propose that the development of humoral immunological memory cannot be explained solely by the long life span of primed T lymphocytes, but is rather a dynamic state dependent on the continuous presence of recent thymic emigrants and qualitative functional differences in responder T cells.