RESUMEN
A major drawback of radiotherapy is the accelerated growth of the surviving tumor cells. Radiotherapy generates a variety of lipids that bind to the receptor for platelet-activating factor, expressed by cells in the tumor microenvironment. In the present study, using the TC-1 tumor cell line, we found that irradiation induced a twofold increase in receptor expression and generated agonists of receptor. Irradiated cells induced a 20-fold increase in live TC-1 proliferation in vitro. Furthermore, subcutaneous co-injection of irradiated TC-1 cells with TC-1 expressing luciferase (TC-1 fluc+) markedly increased TC-1 fluc+ proliferation in a receptor-dependent way. Moreover we used a human carcinoma cell line not expressing the PAF receptor (KBM) and the same cell transfected with the receptor gene (KBP). Following co-injection of live KBP cells with irradiated KBM in RAG mice, the tumor growth was significantly increased compared with tumor formed following co-injection of live KBM with irradiated KBM. This tumor cell repopulation correlated with increased infiltration of tumor-promoting macrophages (CD206+). We propose that receptor represents a possible target for improving the efficacy of radiotherapy through inhibition of tumor repopulation.
RESUMEN
Macrophage interaction with oxidized low-density lipoprotein (oxLDL) leads to its differentiation into foam cells and cytokine production, contributing to atherosclerosis development. In a previous study, we showed that CD36 and the receptor for platelet-activating factor (PAFR) are required for oxLDL to activate gene transcription for cytokines and CD36. Here, we investigated the localization and physical interaction of CD36 and PAFR in macrophages stimulated with oxLDL. We found that blocking CD36 or PAFR decreases oxLDL uptake and IL-10 production. OxLDL induces IL-10 mRNA expression only in HEK293T expressing both receptors (PAFR and CD36). OxLDL does not induce IL-12 production. The lipid rafts disruption by treatment with ßCD reduces the oxLDL uptake and IL-10 production. OxLDL induces co-immunoprecipitation of PAFR and CD36 with the constitutive raft protein flotillin-1, and colocalization with the lipid raft-marker GM1-ganglioside. Finally, we found colocalization of PAFR and CD36 in macrophages from human atherosclerotic plaques. Our results show that oxLDL induces the recruitment of PAFR and CD36 into the same lipid rafts, which is important for oxLDL uptake and IL-10 production. This study provided new insights into how oxLDL interact with macrophages and contributing to atherosclerosis development.
Asunto(s)
Antígenos CD36/metabolismo , Interleucina-10/biosíntesis , Lipoproteínas LDL/metabolismo , Macrófagos/citología , Macrófagos/metabolismo , Microdominios de Membrana/metabolismo , Glicoproteínas de Membrana Plaquetaria/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animales , Gangliósido G(M1)/metabolismo , Células HEK293 , Humanos , Masculino , Proteínas de la Membrana/metabolismo , Ratones Endogámicos C57BL , Placa Aterosclerótica/metabolismo , Placa Aterosclerótica/patología , Transporte de ProteínasRESUMEN
OxLDL is recognized by macrophage scavenger receptors, including CD36; we have recently found that Platelet-Activating Factor Receptor (PAFR) is also involved. Since PAFR in macrophages is associated with suppressor function, we examined the effect of oxLDL on macrophage phenotype. It was found that the presence of oxLDL during macrophage differentiation induced high mRNA levels to IL-10, mannose receptor, PPAR γ and arginase-1 and low levels of IL-12 and iNOS. When human THP-1 macrophages were pre-treated with oxLDL then stimulated with LPS, the production of IL-10 and TGF- ß significantly increased, whereas that of IL-6 and IL-8 decreased. In murine TG-elicited macrophages, this protocol significantly reduced NO, iNOS and COX2 expression. Thus, oxLDL induced macrophage differentiation and activation towards the alternatively activated M2-phenotype. In murine macrophages, oxLDL induced TGF- ß , arginase-1 and IL-10 mRNA expression, which were significantly reduced by pre-treatment with PAFR antagonists (WEB and CV) or with antibodies to CD36. The mRNA expression of IL-12, RANTES and CXCL2 were not affected. We showed that this profile of macrophage activation is dependent on the engagement of both CD36 and PAFR. We conclude that oxLDL induces alternative macrophage activation by mechanisms involving CD36 and PAFR.
Asunto(s)
Antígenos CD36/metabolismo , Regulación de la Expresión Génica , Lipoproteínas LDL/química , Macrófagos/citología , Glicoproteínas de Membrana Plaquetaria/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animales , Arginasa/metabolismo , Línea Celular , Ciclooxigenasa 2/metabolismo , Humanos , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Lectinas Tipo C/metabolismo , Activación de Linfocitos , Macrófagos/metabolismo , Masculino , Receptor de Manosa , Lectinas de Unión a Manosa/metabolismo , Ratones , Ratones Endogámicos C57BL , Monocitos/citología , Óxido Nítrico/química , Óxido Nítrico Sintasa de Tipo II/metabolismo , PPAR gamma/metabolismo , Fenotipo , Receptores de Superficie Celular/metabolismoRESUMEN
INTRODUCTION: Ischemia and reperfusion injury (IRI) are mainly caused by leukocyte activation, endothelial dysfunction and production of reactive oxygen species. Moreover, IRI can lead to a systemic response affecting distant organs, such as the lungs. AIM: The objective was to study the pulmonary inflammatory systemic response after renal IRI. METHODS: Male C57Bl/6 mice were subjected to 45 min of bilateral renal ischemia, followed by 4, 6, 12, 24 and 48 h of reperfusion. Blood was collected to measure serum creatinine and cytokine concentrations. Bronchoalveolar lavage fluid (BALF) was collected to determine the number of cells and PGE(2) concentration. Expressions of iNOS and COX-2 in lung were determined by Western blot. Gene analyses were quantified by real time PCR. RESULTS: Serum creatinine increased in the IRI group compared to sham mainly at 24 h after IRI (2.57 +/- 0.16 vs. 0.43 +/- 0.07, p < 0.01). The total number of cells in BAL fluid was higher in the IRI group in comparison with sham, 12 h (100 x 10(4) +/- 15.63 vs. 18.1 x 10(4) +/- 10.5, p < 0.05) 24 h (124 x 10(4) +/- 8.94 vs. 23.2 x 10(4) +/- 3.5, p < 0.05) and 48 h (79 x 10(4) +/- 15.72 vs. 22.2 x 10(4) +/- 4.2, p < 0.05), mainly by mononuclear cells and neutrophils. Pulmonary COX-2 and iNOS were up-regulated in the IRI group. TNF-alpha, IL-1beta, MCP-1, KC and IL-6 mRNA expression were up-regulated in kidney and lungs 24 h after renal IRI. ICAM-1 mRNA was up-regulated in lungs 24 h after renal IRI. Serum TNF-alpha, IL-1beta and MCP-1 and BALF PGE(2) concentrations were increased 24 h after renal IRI. CONCLUSION: Renal IRI induces an increase of cellular infiltration, up-regulation of COX-2, iNOS and ICAM-1, enhanced chemokine expression and a Th1 cytokine profile in lung demonstrating that the inflammatory response is indeed systemic, possibly leading to an amplification of renal injury.
Asunto(s)
Riñón/fisiopatología , Neumonía , Daño por Reperfusión , Síndrome de Respuesta Inflamatoria Sistémica , Animales , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Quimiocinas/sangre , Quimiocinas/inmunología , Ciclooxigenasa 2/inmunología , Citocinas/sangre , Citocinas/inmunología , Molécula 1 de Adhesión Intercelular/inmunología , Riñón/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Óxido Nítrico Sintasa de Tipo II/inmunología , Neumonía/etiología , Neumonía/inmunología , Neumonía/fisiopatología , Daño por Reperfusión/complicaciones , Daño por Reperfusión/inmunología , Daño por Reperfusión/fisiopatología , Síndrome de Respuesta Inflamatoria Sistémica/inmunología , Síndrome de Respuesta Inflamatoria Sistémica/fisiopatologíaRESUMEN
In alveolar macrophages, leukotriene (LT) B(4) and cysteinyl LTs (LTC(4), LTD(4) and LTE(4)) both enhance Fc gamma receptor (Fc gammaR)-mediated phagocytosis. In the present study we investigated the role of specific PKC isoforms (PKC-alpha and -delta), the MAP kinases p38 and ERK 1/2, and PI3K in mediating the potentiation of Fc gammaR-mediated phagocytosis induced by addition of leukotrienes to the AMs. It was found that exogenously added LTB(4) and LTD(4) both enhanced PKC-delta and -alpha phosphorylation during Fc gammaR engagement. Studies with isoform-selective inhibitors indicated that exogenous LTB(4) effects were dependent on both PKC-alpha and -delta, while LTD(4) effects were exclusively due to PKC-delta activation. Although both exogenous LTB(4) and LTD(4) enhanced p38 and ERK 1/2 activation, LTB(4) required only ERK 1/2, while LTD(4) required only p38 activation. Activation by both LTs was dependent on PI3K activation. Effects of endogenous LTs on kinase activation were also investigated using selective LT receptor antagonists. Endogenous LTB(4) contributed to Fc gammaR-mediated activation of PKC-alpha, ERK 1/2 and PI3K, while endogenous cysLTs contributes to activation of PKC-delta, p38 and PI3K. Taken together, our data show that the capacities of LTB(4) and LTD(4) to enhance Fc gammaR-mediated phagocytosis reflect their differential activation of specific kinase programs.
Asunto(s)
Leucotrieno B4/fisiología , Leucotrieno D4/fisiología , Macrófagos Alveolares/inmunología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteína Quinasa C/metabolismo , Receptores de IgG/fisiología , Animales , Células Cultivadas , Femenino , Leucotrieno B4/farmacología , Leucotrieno D4/farmacología , Macrófagos Alveolares/efectos de los fármacos , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Fagocitosis , Inhibidores de las Quinasa Fosfoinosítidos-3 , Isoformas de Proteínas/antagonistas & inhibidores , Isoformas de Proteínas/metabolismo , Proteína Quinasa C/antagonistas & inhibidores , Ratas , Ratas Wistar , Receptores de IgG/inmunologíaRESUMEN
Neutrophils are important effector cells of tissue injury in several pathological conditions, among them, immune complexes (IC)-induced inflammation and tissue injury. There is evidence that endothelins modulate IC-induced tissue injury in experimental models in vivo. In the present study we investigated the effect of endothelins on neutrophil activation by IC in vitro. To this purpose, pre-formed insoluble immune complexes were used to stimulate human neutrophils and production of leukotriene B(4) (LTB(4)) and hydrogen peroxyde (H(2)O(2)) were measured as indicative of phospholipase A(2) and oxidative burst activation and myeloperoxidase (MPO) release as indicative of cell degranulation. The effect of endothelins (ETs) in these events induced by IC was then examined. We found that IC stimulated all three events in human neutrophils. Addition of ET-1 but not ET-2 or ET-3 to the IC-stimulated neutrophils potentiated LTB(4) but not H(2)O(2) production. The endothelins added to resting neutrophils did not induce LTB(4) production but they were effective to stimulate H(2)O(2) production. The increased MPO activity induced by IC was not affected by endothelins nor did they stimulate the release of this enzyme in resting cells. These results show that endothelins are able to activate some neutrophil functions and to upregulate the IC-induced production of the pro-inflammatory molecule LTB(4). These data indicate that products of endothelial cells, such as endothelins, can be involved in the potentiation of neutrophil-dependent tissue injury.
Asunto(s)
Complejo Antígeno-Anticuerpo/farmacología , Endotelina-1/farmacología , Endotelina-2/farmacología , Endotelina-3/farmacología , Neutrófilos/efectos de los fármacos , Anticuerpos/inmunología , Degranulación de la Célula/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/farmacología , Leucotrieno B4/biosíntesis , Activación Neutrófila , Neutrófilos/metabolismo , Neutrófilos/fisiología , Peroxidasa/metabolismo , Estallido Respiratorio/efectos de los fármacos , Albúmina Sérica Bovina/inmunologíaRESUMEN
Alterations in arachidonic acid (AA) metabolism have been reported to occur in diabetes mellitus. The present study was carried out to verify if these alterations are due to the relative lack of insulin or to high levels of blood glucose. Male Wistar rats were rendered diabetic by alloxan injection (42 mg/kg, i.v.), 10 or 30 days before the experiments. Some diabetic rats received a single dose (4 IU, s.c.) of NPH insulin 2 h before an intratracheal instillation of lipopolysaccharide (LPS, 750 microg) or saline. Six hours after LPS challenge, the following parameters were analysed: blood glucose levels, total and differential leukocyte counts in bronchoalveolar lavage (BAL) fluid; linoleic acid and AA content in blood neutrophils (HPLC), and levels of prostaglandin (PG)E(2) in BAL (ELISA). Relative to controls, a reduced number of neutrophils (18%) and decreased amounts of PGE(2) (40%) were observed in the BAL fluid of diabetic rats in response to LPS. A single dose of insulin was not able to reduce blood sugar levels to normal values, but instead resulted in the normalization of both leukocyte migration to the lungs and levels of PGE(2). Accordingly, these abnormalities might be primarily linked to a continuing insulin deficiency rather than to secondary hyperglycaemia occurring in the diabetic rat. In conclusion, data presented suggest that insulin might regulate neutrophil migration and generation of PGE(2) during the course of acute lung injury induced by LPS.
Asunto(s)
Dinoprostona/biosíntesis , Insulina/fisiología , Lipopolisacáridos/farmacología , Neumonía/metabolismo , Aloxano , Animales , Ácido Araquidónico/análisis , Ácido Araquidónico/metabolismo , Líquido del Lavado Bronquioalveolar/citología , Ciclooxigenasa 2/biosíntesis , Diabetes Mellitus Experimental/metabolismo , Ácido Linoleico/análisis , Masculino , Neutrófilos/fisiología , Ratas , Ratas WistarRESUMEN
OBJECTIVE: The effect of bradykinin (B(1) or B(2)) receptor antagonists was studied in allergic and immune-complex-induced lung inflammation. METHODS: Lungs of BALB/c mice were examined 24 h after induction of lung inflammation, either allergic (ovalbumin-sensitized submitted to two aerosol of antigen, one week apart) or immune-complex induced (intratracheal instillation of IgG antibodies followed by intravenous antigen). The bradykinin B(2) receptor antagonist, HOE-140 or bradykinin B(1) receptor antagonist, R-954 were given intraperitoneally (100 microg/kg), 30 min before induction. RESULTS: In allergic inflammation, pre-treatment with R-954 reduced eosinophil infiltration into the lungs, mucus secretion and the airway hyperreactivity to methacholine. Pre-treatment with HOE-140 increased eosinophil infiltration but did not affect the other parameters. In immune-complex inflammation, HOE-140 increased neutrophil infiltration but not their activation nor the hemorrhagic lesions. R-594 pre-treatment did not change the parameters examined. CONCLUSION: These results show important modulatory effects of bradykinin B(1) and B(2) receptor antagonists in both models of lung inflammation.
Asunto(s)
Antagonistas de los Receptores de Bradiquinina , Bradiquinina/análogos & derivados , Hipersensibilidad , Enfermedades del Complejo Inmune , Neumonía/inmunología , Neumonía/prevención & control , Animales , Bradiquinina/uso terapéutico , Antagonistas del Receptor de Bradiquinina B1 , Antagonistas del Receptor de Bradiquinina B2 , Broncoconstricción/efectos de los fármacos , Eosinófilos/patología , Masculino , Cloruro de Metacolina/administración & dosificación , Ratones , Ratones Endogámicos BALB C , Moco/metabolismo , Ovalbúmina/inmunología , Neumonía/patologíaRESUMEN
The participation of endothelins (ETs) in a model of neutrophil-dependent lung injury induced by intrabronchial instillation of rabbit antibodies to ovalbumin followed by i.v. injection of the antigens (Arthus reaction) was investigated. Hemorrhagic lesions were evaluated by measuring the extravasations of hemoglobin in lung parenchyma. From 5 min to 24 h after the Arthus reaction (AR), endothelin (ir-ET) levels in bronchoalveolar lavage fluid (BALF) and in plasma were measured by radioimmunoassay. BALF levels of ir-ET were not different between control and AR animals for the first 90 min after the antigen challenge but increased from 2 to 24 h after induction of AR. ET levels in the plasma did not change from the respective controls over the same 24 h period. Increased ir-ET in BALF was not affected by pretreatment with L-NAME (30 mg/kg, i.v.). A PAF antagonist (BN52021; 5 and 10 mg/kg, i.v.) increased ET content in BALF and decreased the intensity of the AR. Thiorphan (2 mg/kg, i.v.) inhibited the AR-induced hemorrhagic lesions in lungs. An ET(A) receptor antagonist, BQ-123 (1 mg/kg, i.v.) potentiated, whereas the ET(B) antagonist, BQ-788 (1 mg/kg, i.v.) inhibited the lung hemorrhage. It is concluded that ETs are released during and play a role in the lung AR.
Asunto(s)
Reacción de Arthus/inmunología , Endotelinas/metabolismo , Hemorragia/inmunología , Enfermedades Pulmonares/inmunología , Neumonía/inmunología , Animales , Complejo Antígeno-Anticuerpo , Reacción de Arthus/sangre , Reacción de Arthus/etiología , Líquido del Lavado Bronquioalveolar/química , Diterpenos/farmacología , Antagonistas de los Receptores de la Endotelina A , Antagonistas de los Receptores de la Endotelina B , Endotelinas/análisis , Endotelinas/sangre , Fibrinolíticos/farmacología , Ginkgólidos , Hemoglobinas/análisis , Hemorragia/etiología , Hemorragia/metabolismo , Lactonas/farmacología , Enfermedades Pulmonares/etiología , Enfermedades Pulmonares/metabolismo , Masculino , Neutrófilos/inmunología , Oligopéptidos/farmacología , Ovalbúmina , Péptidos Cíclicos/farmacología , Piperidinas/farmacología , Neumonía/patología , Ratas , Ratas WistarRESUMEN
OBJECTIVE: The involvement of arachidonic acid (AA) and PGE2 during the E. coli lipopolysaccharide (LPS)-induced acute lung injury was investigated. MATERIAL: Adult male Wistar rats were used. For in vitro studies, rat neutrophils, bronchoalveolar lavage (BAL) fluid, and lug vascular endothelium were used, as described below. TREATMENT: Rats were given an intratracheal injection of LPS (750 microg). METHODS: Total and differential cell counts in BAL fluid; enzyme-linked immunoassay (ELISA) analyses of TNF-alpha, IL-1beta, LTB4 and PGE2 in BAL, and immunohistochemical detection of ICAM-1 on lung vascular endothelium were performed six h after LPS challenge. Fatty acid composition of blood neutrophils and plasma was analyzed by HPLC. RESULTS: Rats instilled with LPS presented a sixty three-fold increase in the number of neutrophils in BAL (from 0.5 x 10(6) to 31.5 x 10(6) cells), accompanied by increased levels of TNF-alpha and IL-1beta (p < 0.001), and a three-fold increase in ICAM-1 expression on vascular endothelium. The content of AA in blood neutrophils was reduced by 50%, whereas the level of PGE2 in BAL was increased by 3.5 fold, without changes in the levels of LTB4. CONCLUSIONS: These findings suggest that AA and PGE2 are associated with LPS challenge.
Asunto(s)
Ácido Araquidónico/metabolismo , Dinoprostona/metabolismo , Escherichia coli , Lipopolisacáridos/farmacología , Neutrófilos/metabolismo , Neumonía/inducido químicamente , Neumonía/metabolismo , Enfermedad Aguda , Animales , Líquido del Lavado Bronquioalveolar/citología , Inmunohistoquímica , Molécula 1 de Adhesión Intercelular/metabolismo , Interleucina-1/metabolismo , Recuento de Leucocitos , Leucotrieno B4/metabolismo , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Pulmón/patología , Masculino , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE AND DESIGN: Oxidized low-density lipoproteins (oxLDL) and protein fractions obtained by size exclusion chromatography of oxLDL were tested for vascular permeability effects on topical application to the hamster cheek pouch. MATERIALS: The hamster cheek pouch was prepared for intravital microscopy observations of macromolecular leakage at post capillary venules (=leaks) with FITC-dextran as tracer. TREATMENT: OxLDL (0.1 mg/ml), PAF (platelet activation factor, 50-100 nM) and protein fractions of oxLDL (10 microg/ml) were applied topically to hamster cheek pouches. RESULTS: Application of oxLDL and PAF resulted in reversible increases in the number of leaks. The PAF-antagonist WEB 2170, L-NAME and a beta(2)-adrenoceptor agonist inhibited (P<0.01) almost completely the macromolecular leakage induced with oxLDL or PAF. Protein fractions were found to be more effective than unfractionated oxLDL in inducing plasma leakage as calculated on mg/ml-basis. CONCLUSION: Hamster oxLDL is a potent inducer of macromolecular leakage increase in the hamster cheek pouch microcirculation. The principal effect is mediated by PAF-like structures produced by the oxidation of the LDL-particle but oxLDL also contains low molecular weight proteins that could contribute to the overall vascular permeability increasing effect of ox LDL.
Asunto(s)
Apolipoproteínas B/farmacología , Permeabilidad Capilar/efectos de los fármacos , Lipoproteínas LDL/farmacología , Fragmentos de Péptidos/farmacología , Agonistas de Receptores Adrenérgicos beta 2 , Agonistas Adrenérgicos beta/farmacología , Animales , Mejilla/irrigación sanguínea , Mejilla/fisiología , Cromatografía en Gel , Cricetinae , Inhibidores Enzimáticos/farmacología , Lipoproteínas LDL/química , Masculino , Mesocricetus , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo III , Fragmentos de Péptidos/química , Glicoproteínas de Membrana Plaquetaria/agonistas , Receptores Acoplados a Proteínas G/agonistas , Terbutalina/farmacologíaRESUMEN
The present study was undertaken to investigate the effects of parenteral lipid emulsions (LE) enriched with n-3 fatty acids (n-3 FA) in experimental acute colitis. Seventy-four adult male Wistar rats were randomized into six groups, five of which had acetic acid-induced colitis. The animals received a fat-free diet and water ad libitum in individual metabolic cages. By a central venous catheter, saline was infused (0.5 ml/h) into the control groups CS (without colitis) and CC (with colitis), while the test groups received specific LE for 7 days. The n-3/n-6 FA ratio and the lipidic compositions regarding long chain (LCT) and medium chain (MCT) triglycerides were: group L--1:7.7 (LCT, n = 12), M--1:7.0 (MCT and LCT, n = 12), LW-3--1:4.5 (LCT plus n-3 FA, n = 12) and MW-3--1:3.0 (MCT and LCT plus n-3 FA, n = 13). The frequency of diarrhea, oral intake/body weight ratio, intestinal alterations, macrophage cellularity were evaluated and colonic concentrations of leukotrienes (LTB4, LTC4), prostaglandins (PGE2) and thromboxanes (TXB2) were measured. Groups M, MW-3 and LW-3 had less diarrhea than the CC group (P<0.05). Average oral intake/body weight ratio in MW-3 animals was comparable to the CS and better than the CC group. n-3 FA treated rats (LW-3 and MW-3) presented less intestinal inflammatory alterations than CC rats. Mucosal ulcer formation in MW-3 group did not differ from CS rats. M and MW-3 rats had less macrophages in the colon than the CC group. Compared with CC group, lower concentrations of LTB4 in the CS, LW-3 and MW-3 groups; of PGE2 in the CS, M and MW-3 groups; and of TXB2 in the CS and MW-3 groups were found. Mean concentrations of LTC4 did not differ among the groups. Thus, a LCT-containing LE with a low n-3-n-6 ratio does not modify inflammatory colitis manifestations; LE with a high n-3-n-6 ratio reduces diarrhea, preserves oral intake-weight ratio, attenuates morphological consequences and decreases colonic concentrations of inflammatory mediators; MCT/LCT-containing LE with 1:3 n-3-n-6 ratio exerts the most profound beneficial impact on the inflammatory response.
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Colitis Ulcerosa/terapia , Emulsiones Grasas Intravenosas/uso terapéutico , Ácidos Grasos Omega-3/uso terapéutico , Nutrición Parenteral/métodos , Enfermedad Aguda , Animales , Colitis Ulcerosa/metabolismo , Colitis Ulcerosa/patología , Colon/patología , Eicosanoides/metabolismo , Ácidos Grasos Omega-3/administración & dosificación , Macrófagos/patología , Masculino , Ratas , Ratas WistarRESUMEN
The effect of a receptor binding protein for tumor necrosis factor (TNFrbp) on cell infiltration, bronchial hyperreactivity, and release of inflammatory mediators were studied following antigen challenge in sensitized rats. A 3-fold increase in total cell number, mainly neutrophils and eosinophils, was noted in bronchoalveolar lavage (BAL) fluid 8 hours after antigen challenge. Antigen challenge also induced a significant hyperreactivity of the lower bronchus to carbachol and serotonin, but did not affect the reactivity of the trachea and upper bronchus. This increased responsiveness of the lower bronchus was transient, being detected 8 hours but not 24 hours after antigen challenge. Thromboxane B2 (TxB2), prostaglandin E2 (PGF2), and nitric oxide (NO) levels increased in the BAL fluid of sensitized rats 8 hours after antigen challenge by 197%, 172%, and 173%, respectively. TNFrbp treatment reduced by 83% the antigen-induced cell infiltration, with neutrophils being the cells most affected. The bronchial hyperreactivity induced by antigen challenge was also significantly inhibited by TNFrbp, whereas TxB2, PGE2, and NO levels in the BAL fluid were not affected. In our animal model, the cell infiltration and bronchial hyperreactivity appear to be mediated to some extent by TNF, but not by prostanoids nor NO.
Asunto(s)
Hiperreactividad Bronquial/inmunología , Bronconeumonía/inmunología , Proteínas Portadoras/farmacología , Receptores del Factor de Necrosis Tumoral , Animales , Hiperreactividad Bronquial/patología , Hiperreactividad Bronquial/prevención & control , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Bronconeumonía/patología , Bronconeumonía/prevención & control , Carbacol/farmacología , Proteínas Portadoras/uso terapéutico , Recuento de Células , Movimiento Celular/efectos de los fármacos , Dinoprostona/análisis , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Eosinófilos/efectos de los fármacos , Técnicas In Vitro , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Músculo Liso/fisiología , Neutrófilos/efectos de los fármacos , Óxido Nítrico/análisis , Ratas , Ratas Wistar , Receptores Tipo I de Factores de Necrosis Tumoral , Serotonina/farmacología , Tromboxano B2/análisis , Receptores Señuelo del Factor de Necrosis TumoralRESUMEN
Bacterial translocation is an important source of pancreas infection in acute pancreatitis. The effect of platelet-activating factor (PAF) in the pathogenesis of acute pancreatitis has been proved in various studies. The aim of this study was to determine whether potent PAF antagonists influence bacterial translocation in acute pancreatitis. Acute pancreatitis was induced in 62 Wistar rats by injection of 2.5% sodium taurocholate into the biliopancreatic duct. The rats treated with PAF factor antagonists received intravenous injection of WEB-2170 (10 mg/kg), lexipafant (5 mg/kg), and BN-52021 (5 mg/kg) 30 minutes before induction of acute pancreatitis. Six hours after induction of acute pancreatitis, bacteriologic cultures and histologic scoring of tissues were performed. There was a statistically significant reduction in bacterial translocation to the mesenteric lymph nodes and liver but not to the pancreas of the rats treated with PAF antagonists. No significant increase in the intestinal bacterial population of any group was found. There were no statistical differences between the pancreatic histologic scores of the groups. PAF antagonists reduced bacterial translocation to distant sites other than the pancreas, preventing the bacterial dissemination that occurs in the early phase of acute pancreatitis and may have beneficial effects on the evolution of this disease.
Asunto(s)
Traslocación Bacteriana/efectos de los fármacos , Diterpenos , Leucina/análogos & derivados , Pancreatitis/fisiopatología , Factor de Activación Plaquetaria/antagonistas & inhibidores , Enfermedad Aguda , Animales , Azepinas/farmacología , Ginkgólidos , Imidazoles/farmacología , Lactonas/farmacología , Leucina/farmacología , Masculino , Extractos Vegetales/farmacología , Premedicación , Ratas , Ratas Wistar , Triazoles/farmacologíaRESUMEN
In the present study we investigated the role of platelet-activating factor (PAF) and prostaglandins in experimental Leishmania (Leishmania) amazonensis infection and the relationship between these mediators and nitric oxide (NO) production. Mouse peritoneal macrophages elicited with thioglicolate were infected with leishmania amastigotes, and the infection index determined 48 h later. The course of infection was monitored for 5 weeks in mice infected in the footpad with promastigotes by measuring the footpad swelling and parasite load in regional lymph nodes and spleen. The addition of PAF to C57BL/6 mouse macrophages significantly inhibited parasite growth and induced NO production. Treatment of macrophages with a selective PAF antagonist, WEB2086, increased the infection, indicating that endogenously produced PAF regulates macrophage ability to control leishmania infection. This effect of PAF was abolished by addition of the inhibitor of NO synthesis, L-NAME, to the cultures. The addition of prostaglandin E(2) significantly increased the infection and NO production. Treatment with cyclo-oxygenase inhibitor, indomethacin, reduced the infection and PAF-induced release of NO. Thus, the increased NO production induced by PAF seems to be mediated by prostaglandins. The more-selective inhibitors of cyclo-oxygenase 2, nimesulide and NS-398, had no significant effect. Thus, antileishmanial activity correlates better with the presence of PAF or absence of prostaglandins than with NO production. In vivo treatment with PAF antagonists significantly increased leishmania lesions, as well as the parasite load, in regional lymph nodes and spleens. These findings indicate that PAF is essential for the control of leishmania infection.
Asunto(s)
Leishmaniasis/etiología , Factor de Activación Plaquetaria/fisiología , Animales , Ciclooxigenasa 2 , Dinoprostona/farmacología , Isoenzimas/fisiología , Leishmaniasis/inmunología , Ganglios Linfáticos/parasitología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Óxido Nítrico/biosíntesis , Factor de Activación Plaquetaria/antagonistas & inhibidores , Prostaglandina-Endoperóxido Sintasas/fisiología , Bazo/parasitología , Tioglicolatos/farmacologíaRESUMEN
In the present study we propose a mathematical approach to improve the analysis of NK and LAK activities measured by MTT assay adapted for murine cells. We found that to calculate NK activity, high E:T ratios should be used (up to 50:1) and the phenomenon fits to a linear least-squares analysis. However, 5-fold less effector cells (10:1, E:T) should be used to detect LAK activity and the phenomenon has a nonlinear exponential behavior. Using this approach, we showed that EDTA inhibits LAK but not NK activity whereas PGE(2) inhibits NK but not LAK activity. In conclusion, this analytical approach allowed the discrimination between NK and LAK activities and exposed differences between these two cytotoxic activities.
Asunto(s)
Pruebas Inmunológicas de Citotoxicidad/métodos , Células Asesinas Activadas por Linfocinas/inmunología , Células Asesinas Naturales/inmunología , Bazo/inmunología , Sales de Tetrazolio , Tiazoles , Animales , Citotoxicidad Inmunológica/efectos de los fármacos , Dinoprostona/farmacología , Ácido Edético/farmacología , Células Asesinas Activadas por Linfocinas/citología , Células Asesinas Naturales/citología , Masculino , Ratones , Ratones Endogámicos C57BL , Bazo/citologíaRESUMEN
The effect of fat-rich diets on the acute inflammatory response was examined. Male Wistar rats aged 21 days were fed, for 6 weeks, with a control diet (4% fat content), or a control diet supplemented with coconut or soybean oils (15% fat content). Carrageenan-induced paw oedema and pleurisy were evaluated. Prostaglandin (PG) E2 and leukotriene (LT) C4/D4 concentrations were determined in the pleural exudate (ELISA). Pleural samples were tested for their effect on cutaneous vascular permeability of control rats and the effect of a LTD4 receptor antagonist (L660-711; 10 mg/kg; i.v.) examined. Relative to controls, rats fed both fat-rich diets presented a significant reduction in protein leakage and oedema formation without affecting the number of migrating leukocytes. Production of LTC4/D4 in pleural exudate was significantly increased from 1.8 +/- 0.2 ng/ml in controls to 2.8 +/- 0.2 and 3.0 +/- 0.3 ng/ml in animals fed coconut and soybean oil enriched diets, respectively, without changes in PGE2 production. The activity of these samples on cutaneous vascular permeability was 50% reduced, returning to control values after treatment of testing animals with a LTD4 receptor antagonist. Rats fed fat-rich diets presented a reduced inflammatory response due, at least in part, to the LTC4/D4 mediated vasoconstrictor effect.
Asunto(s)
Reacción de Fase Aguda/dietoterapia , Grasas de la Dieta/administración & dosificación , Leucotrieno C4/metabolismo , Leucotrieno D4/metabolismo , Reacción de Fase Aguda/inducido químicamente , Reacción de Fase Aguda/metabolismo , Animales , Permeabilidad Capilar/efectos de los fármacos , Carragenina , Aceite de Coco , Dinoprostona/metabolismo , Edema/inducido químicamente , Edema/metabolismo , Ensayo de Inmunoadsorción Enzimática , Exudados y Transudados/metabolismo , Miembro Posterior/efectos de los fármacos , Antagonistas de Leucotrieno/farmacología , Masculino , Aceites de Plantas/administración & dosificación , Derrame Pleural/metabolismo , Pleuresia/inducido químicamente , Pleuresia/metabolismo , Propionatos/farmacología , Quinolinas/farmacología , Ratas , Ratas Wistar , Piel/irrigación sanguínea , Piel/efectos de los fármacos , Aceite de Soja/administración & dosificaciónRESUMEN
The aim of this study was to examine antigen-induced lung cell migration, eosinophil activation, and pulmonary reactivity of Wistar rats exposed to a new sensitization technique. The animals were sensitized with a single subcutaneous implant of a fragment of heat coagulated hen egg white and challenged 21 days later with an intratracheal injection of heat-aggregated ovalbumin (EWI). For comparison, another group of rats were sensitized by an intraperitoneal injection of ovalbumin in alum as adjuvant, with one booster on day 14 and challenge on day 21 post immunization (OVA/AL). Twenty-four hours after antigen challenge, the EWI group presented a higher number of eosinophils in the bronchoalveolar lavage (BAL) (4.85 +/- 1.43 x 10(6)) than the OVA/AL group (0.2 +/- 0.06 x 10(6)) or the control group, where the level of eosinophils were essentially undetectable. Levels of eosinophil peroxidase activity were increased in the cell-free BAL and homogenates of lung tissue in the EWI group (12.10 +/- 2.97 mg/mL and 36.14 +/- 7.21 ng/mg, respectively), but not in the OVA/AL group (4.83 +/- 1.4 ng/mL and 11.95 +/- 2.54 ng/mg, respectively), as compared with controls (5.16 +/- 1.65 ng/mL and 12.13 +/- 1.74 ng/mg, respectively). Thromboxane B2 levels were also increased in the BAL of EWI group (2.89 +/- 0.54 ng/mL) but not the OVA/AL group (1.13 +/- 0.23 ng/mL) as compared with controls (1.14 +/- 0.19 ng/mL). In contrast, the levels of prostaglandin E2 in the BAL were increased in both groups (456.4 +/- 11.8 pg/mL in the EWI group and 303.5 +/- 31.7 pg/mL in the OVA/AL group) as compared with controls (205.7 +/- 29.7 ng/mL). Moreover, only the EWI group developed increased pulmonary reactivity to serotonin (around two-fold), 24 hours after antigen challenge. The extent of lung eosinophil migration and activation and the pulmonary hyperreactivity induced by this novel sensitization procedure without adjuvants represents a significant improvement over existing experimental models of asthma.