RESUMEN
In addition to their pigment properties, the potential health benefits of anthocyanins have made them a subject of interest in recent years. This study aimed to obtain purified anthocyanin fractions from native Mexican black bean cultivars using Amberlite XAD-7 resin column and HPCCC and evaluate their anti-inflammatory properties using RAW 264.7 cells. The major anthocyanins in the purified anthocyanin fractions were delphinidin 3-glucoside (61.8%), petunidin 3-glucoside (25.2%), and malvidin 3-glucoside (12.2%). Purified anthocyanin fractions at 12.5 µg/mL effectively prevented LPS-induced ERK1/ERK2 phosphorylation and reduced the protein expression of COX-2 and mRNA expression of iNOS. Results showed that purified anthocyanin fractions have the potential to modulate the inflammatory response by inhibiting the production of pro-inflammatory mediators through the ERK1/ERK2 and NF-κB pathways. This study suggests that anthocyanins from black beans could be used as a natural strategy to help modulate inflammation-associated diseases.
Asunto(s)
Antocianinas , Antiinflamatorios , FN-kappa B , Extractos Vegetales , Antocianinas/farmacología , Antocianinas/química , Antocianinas/aislamiento & purificación , Ratones , Células RAW 264.7 , Animales , Extractos Vegetales/química , Extractos Vegetales/farmacología , Extractos Vegetales/aislamiento & purificación , Antiinflamatorios/farmacología , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , FN-kappa B/genética , FN-kappa B/metabolismo , FN-kappa B/inmunología , Distribución en Contracorriente , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Inflamación/metabolismo , Inflamación/tratamiento farmacológico , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Ciclooxigenasa 2/inmunología , Cromatografía Líquida de Alta Presión , México , Phaseolus/química , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismoRESUMEN
BACKGROUND: Brazilian berry is a fruit popularly known as "Jaboticaba," rich in bioactive compounds with antioxidant and anti-inflammatory properties. Senescence and overweight are increasing worldwide and are considered risk factors to prostatic pathogenesis mainly due to oxidative and inflammatory processes induction. Thus, this study aimed to evaluate the effect of two increasing doses of the patented jaboticaba peel extract (PJE) on oxidative-stress and inflammation in the prostate of aging or high-fat-fed aging mice. METHODS: PJE and/or high-fat diet (HFD) treatments started with 11-month-old mice and lasted 60 days. The levels or the immunoexpression of different inflammatory (nuclear factor κB [NFκB], CD3+, cyclooxygenase 2 [COX-2], toll-like receptor 4 [TLR4], phosphorylated signal transducers and activators of transcription 3 [pSTAT-3], tumor necrosis factor α [TNF-α], interleukin 6 [IL-6], and IL-1ß) and oxidative-stress (catalase, superoxide dismutase 2 [SOD2], glutathione reductase [GSR], reduced glutathione, and glutathione peroxidase 3 [GPx3]) related molecules were analyzed by western-blotting, immunohistochemistry, and enzyme-linked immunosorbent assays. RESULTS: Both PJE doses reduced the levels of oxidative-stress-related molecules (GPx3, GSR, catalase), lipid peroxidation (4-hydroxynonenal), inflammatory mediators (COX-2, TNF-α, and pSTAT-3) and CD3+ T cells number, which were associated with the maintenance of the glandular morphological integrity in aging and HFD-fed-aging mice. Nevertheless, only the high PJE dose reduced the NFκB and TLR4 levels in aging mice; and SOD2, IL-6, and IL-1ß levels in HFD-aging mice. Aging itself promoted an oxidative inflammation in the prostate, interfering in the levels of the different oxidative-stress, lipid peroxidation, and inflammatory mediators evaluated, in association with high incidence of prostate epithelial and stromal damages. The HFD intake intensified aging alterations, showing an unfavorable prostatic microenvironment prone to oxidative and inflammatory damages. CONCLUSIONS: PJE exerted a dose-dependent effect controlling inflammation and oxidative-stress in aging and HFD-fed aging mice prostate. This fact contributed to prostate microenvironment balance recovery, preserving the tissue architecture of this gland. Thus, the PJE emerges as a potential therapy to prevent inflammation and oxidative stress in the prostate.
Asunto(s)
Frutas/química , Myrtaceae/química , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/farmacología , Prostatitis/tratamiento farmacológico , Factores de Edad , Animales , Antiinflamatorios/química , Antiinflamatorios/farmacología , Antioxidantes/química , Antioxidantes/farmacología , Ciclooxigenasa 2/biosíntesis , Ciclooxigenasa 2/inmunología , Dieta Alta en Grasa , Relación Dosis-Respuesta a Droga , Interleucina-1beta/sangre , Interleucina-6/sangre , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Extractos Vegetales/química , Prostatitis/inmunología , Prostatitis/metabolismo , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunologíaRESUMEN
Wound healing involves the interaction of blood cells, proteins, proteases, growth factors, and extracellular matrix components. Inflammation is one of the first events occurring during this process. Previously, we showed that the N-Methyl-(2S,4R)-trans-4-Hydroxy-L-Proline (NMP) from Sideroxylon obtusifolium leaves (a Brazilian medicinal species) presents an anti-inflammatory action. Considering inflammation as an important event in the wound healing process, the objectives were to investigate the topical effects of the NMP gel on a mice wound-induced model. Male Swiss mice were divided into 4 groups: Sham (surgical procedure only), Control (gel-base treated), and 3% or 10% NMP gel-treated groups. Measurements of wound areas and microscopic analyses (HE [hematoxylin-eosin] and PSR [picrosirius red] stainings) were carried out, at the 7th and 12th, days after the wound induction. Furthermore, immunohistochemical assays for iNOS (inducible nitric oxide synthase) and COX-2 (cyclooxygenase-2) and biochemical measurements for TBARS (thiobarbituric acid reactive substances), GSH (glutathione), and myeloperoxidase (MPO) were also performed, at the second day after the wound induction. The work showed that NMP decreases the wound areas, after topical application, relatively to the Sham and Control groups. In addition, microscopic alterations were reduced and collagen deposition was increased, at the 7th and 12th days, in the 10% NMP group. While iNOS and COX-2 immunostainings and GSH contents increased, in relation to the Sham and Control groups, TBARS and MPO decreased. Altogether, the results showed NMP to improve the wound healing process, by upregulating iNOS and COX-2 activities, reducing lipid peroxidation and MPO activity, and increasing GSH contents. In addition, NMP certainly contributes to the increased collagen deposition. These data may stimulate translational studies dealing with the possible use of NMP from Sideroxylon obtusifolium or from other sources for the management of wound healing.
Asunto(s)
Antiinflamatorios/administración & dosificación , Antioxidantes/administración & dosificación , Extractos Vegetales/administración & dosificación , Prolina/administración & dosificación , Sapotaceae/química , Cicatrización de Heridas/efectos de los fármacos , Heridas y Lesiones/tratamiento farmacológico , Animales , Antiinflamatorios/química , Antioxidantes/química , Colágeno/genética , Colágeno/inmunología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Glutatión/inmunología , Humanos , Masculino , Ratones , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/inmunología , Peroxidasa/genética , Peroxidasa/inmunología , Extractos Vegetales/química , Prolina/análogos & derivados , Heridas y Lesiones/genética , Heridas y Lesiones/inmunología , Heridas y Lesiones/fisiopatologíaRESUMEN
This study investigated the efficacy of the vaccine in liver of mice infected with the Trypanosoma cruzi (T. cruzi) and immunized with AdASP-2. For this purpose, histopathological analysis and gene expression of COX-2, TNF-alpha, TNFR, iNOS, cytochrome C, caspase-3, TLR4, IL-6 and IL10 were evaluated. The following groups were used in this study: Group 1 - Control Group (CTRL) animals received AdßGal vehicle; Group 2 - Infected Group (TC) animals were infected with T. cruzi; Group 3 - Immunized Group (AdASP-2): animals were immunized by AdASP-2 vaccine; Group 4 - Immunized and Infected Group (AdASP-2+TC) animals were infected with T. cruzi and immunized by AdSP-2 vaccine. A significant decrease of amastigote nests was noticed in the group of animals that were immunized with AdASP-2 and infected on the same day. COX-2 and TNF-alpha gene expressions increased in TC group, whereas TNF-alpha decreased in the TC+AdASP-2 group. TNFR expression was high in AdASP-2+TC group. iNOS expression was high for all experimental groups whereas cytochrome C decreased for all experimental groups. Caspase 3 increased in TC and TC+AdASP-2 groups. The gene expression of TLR4 and IL-10 showed an increase in AdASP-2+TC group. Finally, hepatic fibrosis was noticed to TC and AdASP-2â¯+â¯TC groups. Taken together, our results demonstrated that vaccination with AdASP-2 was effective against the acute phase of experimental Chagas disease as a result of a more powerful and rapid immune response closely related to expression of some inflammatory genes, such as iNOS, TNF-alpha, TLR 4, and IL-10.
Asunto(s)
Cardiomiopatía Chagásica/inmunología , Cirrosis Hepática/inmunología , Hígado/inmunología , Neuraminidasa/inmunología , Vacunas Antiprotozoos/inmunología , Trypanosoma cruzi/inmunología , Adenoviridae , Animales , Caspasa 3/inmunología , Cardiomiopatía Chagásica/parasitología , Cardiomiopatía Chagásica/patología , Cardiomiopatía Chagásica/prevención & control , Ciclooxigenasa 2/inmunología , Citocromos c/inmunología , Citocinas/inmunología , Femenino , Hígado/parasitología , Hígado/patología , Cirrosis Hepática/parasitología , Cirrosis Hepática/patología , Cirrosis Hepática/prevención & control , Ratones , Óxido Nítrico Sintasa de Tipo II/inmunología , Receptor Toll-Like 4/inmunologíaRESUMEN
Chronic obstructive pulmonary disease (COPD) is related to inflammatory process caused by smoking habit. In this scenario, the anti-inflammatory protein Annexin A1 (AnxA1) may represent a therapeutic alternative. We performed experiments to evaluate the effects of the AnxA1 mimetic peptide Ac2-26 in an initial COPD model by physiological, histopathological, biochemical and immunohistochemical analyses. Weight loss, increased blood pressure, reductions in the pulmonary frequency and ventilation, loss of tracheal cilia, enlargement of the pulmonary intra-alveolar spaces and lymphoid tissue found in untreated smoke-exposed group were attenuated by AnxA1 peptide treatment. The Ac2-26 administration also protected against leukocytes influx in bronchoalveolar lavage (BAL), lung and trachea, and it also led to decreased hemoglobin, glucose, cholesterol, gamma glutamyl transferase and aspartato aminotransferase levels. Similarly, reduction of proinflammatory mediators and higher concentration of anti-inflammatory cytokine were found in macerated lung supernatant, blood plasma and BAL in the treated animals. Besides Ac2-26 group showed reduced tissue expressions of AnxA1, cyclooxygenase-2 and metalloproteinase-9, but formylated peptide receptor 2 (FPR2) overexpression. Our results all together highlighted the protective role of the Ac2-26 mimetic peptide in COPD with promising perspectives.
Asunto(s)
Anexina A1/uso terapéutico , Antiinflamatorios/uso terapéutico , Péptidos/uso terapéutico , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Animales , Anexina A1/farmacología , Antiinflamatorios/farmacología , Líquido del Lavado Bronquioalveolar/citología , Ciclooxigenasa 2/inmunología , Citocinas/inmunología , Femenino , Macrófagos/inmunología , Mastocitos/inmunología , Péptidos/farmacología , Enfermedad Pulmonar Obstructiva Crónica/inducido químicamente , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Enfermedad Pulmonar Obstructiva Crónica/patología , Ratas Wistar , Humo , Productos de TabacoRESUMEN
ABSTRACT Objectives: Determine immunohistochemical expression of Phosphatase and tensin homolog (PTEN), Phosphatidylinositol 3 kinase (PI3K), Cycloxygenase-2 (COX2) and one proliferation marker (Ki67) in colorectal polyps and correlate with clinical and pathological data in search of carcinogenic pathways. Methods: The reports of 297 polyps diagnosed through endoscopy were reviewed for parameters including age, gender, prior colorectal cancer, the presence of multiple polyps, and polyps' location, appearance and size. Was conducted a microscopic morphometric computerized analysis of immunohistochemical expression using, the selected antibodies and correlated with clinical and pathological variables. Results: The tissue immunohistochemical expression was higher in right colon polyps for the proliferation marker and Phosphatidylinositol 3 kinase (p ≤ 0.0001 and 0.057 respectively). Cycloxygenase-2 and Phosphatase and tensin homolog demonstrated higher tissue immunoexpression in pedunculated polyps (p = 0.009 and 0.002 respectively). Cycloxygenase-2 exhibited higher immunoexpression in larger polyps (p = 0.005). Phosphatidylinositol 3 kinase, Cycloxygenase-2, Phosphatase and tensin homolog and the proliferation marker exhibited higher immunoexpression in high-grade dysplastic polyps (p = 0.031, 0.013, 0.044 and <0.001 respectively). Phosphatase and tensin homolog labeling was higher in polyps with high-grade dysplasia and lower in some of serrated lesions (p = 0.044). Conclusions: The greater expression of the proliferation marker and Phosphatidylinositol 3 kinase in the right colon may be related to right-sided colorectal carcinogenesis. The proliferation marker, Cycloxygenase-2 and Phosphatidylinositol 3 kinase results can be associated with progression of polyps to colorectal cancer. The higher Phosphatase and tensin homolog expression suggests its attempt to control the cell cycle.
RESUMO Objetivos: Determinar a expressão imuno-histoquímica de Fosfatase homóloga a tensina (PTEN), Fosfatidilinositol-3-cinase (PI3K), Ciclooxigenase-2 (COX2) e um marcador de proliferação (Ki67) em pólipos colorretais e correlacionar com dados clínicos e patológicos buscando sua correspondência na carcinogênese. Métodos: Revisados 297 pólipos diagnosticados através de endoscopia quanto a idade, gênero, história de câncer colorretal, número, localização, aparência e tamanho dos pólipos. Realizadas as avaliações morfométricas computadorizadas das expressões imuno-histoquímicas dos marcadores selecionados, que foram correlacionadas com variáveis clínicas e patológicas. Resultados: A expressão do marcador de proliferação e da Fosfatidilinositol-3-cinase foi maior nos pólipos do cólon direito (p = <0,0001 e 0.057 respectivamente). Ciclooxigenase-2 e Fosfatase homóloga a tensina demonstraram maior imunoexpressão em pólipos pediculados (p = 0,009 e 0,002, respectivamente). Ciclooxigenase-2 expressou mais em pólipos maiores (p = 0,005). Fosfatidilinositol-3-cinase, Ciclooxigenase-2, Fosfatase homóloga a tensina e o marcador de proliferação expressaram mais em pólipos com displasia de alto grau (p = 0,031, 0,013, 0,044 e <0,001, respectivamente). Fosfatase homóloga a tensina marcou mais pólipos com displasia de alto grau que lesões serrilhadas (p = 0,044). Conclusões: A maior expressão do marcador de proliferação e Fosfatidilinositol-3-cinase à direita pode estar relacionada à carcinogênese do lado direito do cólon. Os resultados do marcador de proliferação, Ciclooxigenase-2 e Fosfatidilinositol-3-cinase podem ser associados à progressão dos pólipos para câncer. A expressão aumentada de Fosfatase homóloga a tensina sugere tentativa de controle do ciclo celular.
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Neoplasias Colorrectales/diagnóstico , Pólipos del Colon/patología , Antígeno Ki-67/inmunología , Fosfohidrolasa PTEN/inmunología , Ciclooxigenasa 2/inmunología , Fosfatidilinositol 3-Quinasa/inmunologíaRESUMEN
Riparins, natural alkaloids of the alkamide group, can be synthesized by simple methods, enhancing their potential application in pharmaceutical development. Here, the pharmacological properties of riparins were investigated in in vitro and in vivo assays of pain and inflammation in Swiss mice. Inflammatory mediators were measured by radioimmunoassay and Real-Time PCR. Riparins I, II, III and IV (1.56-100 mg/kg; ip) produced dose-related antinociceptive effects in the formalin test, exhibiting ED50 values of 22.93, 114.2, 31.05 and 6.63 mg/kg, respectively. Taking the greater potency as steering parameter, riparin IV was further investigated. Riparin IV did not produce antinociceptive effect on the tail flick, suggesting that its antinociception is not a centrally-mediated action. In fact, riparin IV (1.56-25 mg/kg) produced dose-related antinociceptive and antiedematogenic effects on the complete Freund's adjuvant (CFA)-induced paw inflammation in mice. During CFA-induced inflammation, riparin IV did not modulate either the production of cytokines, TNF-α and IL-10, or COX-2 mRNA expression. On the other hand, riparin IV decreased the PGE2 levels in the inflamed paw. In in vitro assays, riparin IV did not exhibit suppressive activities in activated macrophages. These results indicate, for the first time, that riparin IV induces antinociceptive and anti-inflammatory effects, possibly through the inhibition of prostanoid production.
Asunto(s)
Alcaloides/farmacología , Analgésicos/farmacología , Antiinflamatorios/farmacología , Benzamidas/farmacología , Edema/tratamiento farmacológico , Dolor/tratamiento farmacológico , Animales , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Dinoprostona/antagonistas & inhibidores , Dinoprostona/biosíntesis , Dinoprostona/inmunología , Edema/inducido químicamente , Edema/inmunología , Edema/patología , Adyuvante de Freund/efectos adversos , Expresión Génica , Inflamación , Interleucina-10/genética , Interleucina-10/inmunología , Masculino , Ratones , Dolor/inducido químicamente , Dolor/inmunología , Dolor/patología , Manejo del Dolor , Dimensión del Dolor , Percepción del Dolor/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
AIM: In the present study, we examine the effects of copper oxide nanoparticles (CuNP) on macrophage immune response and the signaling pathways involved. MATERIALS & METHODS: A peritonitis model was used to determine in vivo immune cells recruitment, while primary macrophages were used as an in vitro model for the cellular and molecular analysis. RESULTS: In vivo, CuNP induce significant macrophages recruitment to the site of injection. In vitro, in LPS-stimulated primary macrophages, the co-treatment with CuNP inhibited the production of NO in a dose-dependent manner. The mechanism underlying NO and proinflammatory cytokines inhibition was associated with an increased arginase activity. Macrophage stimulation with CuNP did not provoke any cytokine secretion; however, arginase inhibition promoted TNFα and MIP-1ß production. In addition, CuNP induced the expression of COX-2 and the production of PGE2 through arginase activation. CONCLUSION: Our results demonstrate that CuNP activate arginase and suppress macrophage innate immune response.
Asunto(s)
Arginasa/inmunología , Cobre/inmunología , Citocinas/inmunología , Dinoprostona/inmunología , Macrófagos/efectos de los fármacos , Nanopartículas , Óxido Nítrico/inmunología , Animales , Arginasa/química , Células Cultivadas , Cobre/química , Ciclooxigenasa 2/inmunología , Activación Enzimática/efectos de los fármacos , Macrófagos/enzimología , Macrófagos/inmunología , Ratones Endogámicos C57BL , Nanopartículas/química , Transducción de Señal/efectos de los fármacosRESUMEN
PURPOSE: Galectin-1 (Gal-1) is a ß-galactoside-binding protein with diverse biological activities in the pathogenesis of inflammation but has been poorly investigated in terms of ocular inflammation. In the present study, we monitored the anti-inflammatory effects of Gal-1 using the in vivo rodent model of endotoxin-induced uveitis (EIU) and in vitro assays with human RPE (ARPE-19) cells. METHODS: For this purpose, EIU was induced by subcutaneous sterile saline injection of 0.1 ml of lipopolysaccharide (LPS, 1 mg/Kg) in the rat paw, which was maintained under these conditions for 24 h. The therapeutic efficacy of recombinant Gal-1 (rGal-1) was tested in the EIU animals by intraperitoneal inoculation (3 µg/100 µl per animal) 15 min after the LPS injection. In vitro studies were performed using LPS-stimulated ARPE-19 cells (10 µg/ml) for 2, 8, 24 and 48 h, treated or not with rGal-1 (4 µg/ml) or dexamethasone (Dex, 1.0 µM). RESULTS: Gal-1 treatment attenuated the histopathological manifestation of EIU via the inhibition of polymorphonuclear cells (PMN) infiltration in the eye and by causing an imbalance in adhesion molecule expression and suppressing interleukin (IL)-1ß, IL-6, and monocyte chemotactic protein-1 (MCP-1) productions. Immunohistochemical and western blotting analyses revealed significant upregulation of Gal-1 in the eyes induced by EIU after 24 h. In the retina, there was no difference in the Gal-1 expression, which was high in all groups, demonstrating its structural role in this region. To better understand the effects of Gal-1 in the retina, in vitro studies were performed using ARPE-19 cells. Ultrastructural immunocytochemical analyses showed decreased levels of endogenous Gal-1 in LPS-stimulated cells (24 h), while Dex treatment upregulated this protein. The protective effects of rGal-1 on LPS-stimulated cells were associated with the significant reduction of the release of cytokines (IL-8 and IL-6), similar to Dex treatment. Furthermore, rGal-1 and Dex inhibited cyclooxygenase-2 (COX-2) expression in LPS-stimulated cells, as shown by immunofluorescence. CONCLUSIONS: Overall, this study identified potential roles for Gal-1 in ocular inflammation, especially uveitis, and may lead to future therapeutic approaches.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Células Epiteliales/efectos de los fármacos , Galectina 1/farmacología , Epitelio Pigmentado de la Retina/efectos de los fármacos , Uveítis/tratamiento farmacológico , Uveítis/genética , Animales , Antiinflamatorios no Esteroideos/inmunología , Antiinflamatorios no Esteroideos/metabolismo , Línea Celular , Quimiocina CCL2/antagonistas & inhibidores , Quimiocina CCL2/genética , Quimiocina CCL2/inmunología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Dexametasona/farmacología , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Células Epiteliales/patología , Galectina 1/genética , Galectina 1/inmunología , Regulación de la Expresión Génica , Humanos , Inyecciones Subcutáneas , Interleucina-1beta/antagonistas & inhibidores , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Interleucina-6/antagonistas & inhibidores , Interleucina-6/genética , Interleucina-6/inmunología , Lipopolisacáridos , Infiltración Neutrófila/efectos de los fármacos , Ratas , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/farmacología , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/patología , Uveítis/inducido químicamente , Uveítis/inmunologíaRESUMEN
Leishmanioses are chronic parasitic diseases and host responses are associated with pro- or anti-inflammatory cytokines involved, respectively, in the control or exacerbation of infection. The relevance of other inflammatory mediators and stress markers has not been widely studied and there is a need to search for biomarkers to leishmaniasis. In this work, the stress and inflammatory molecules p38 mitogen-activated protein kinase, cyclooxygenase-2, migration inhibitory factor, macrophage inflammatory protein 2, heat shock protein 70 kDa, vascular endothelial factor (VEGF), hypoxia-inducible factors (HIF-1α and HIF-2α), heme oxygenase and galectin-3 expression were assessed immunohistochemically in self-controlled lesions in C57BL/6 mice and severe lesions in Balb/c mice infected with Leishmania amazonensis. The results indicated that the majority of molecules were expressed in the cutaneous lesions of both C57BL/6 and Balb/c mice during various phases of infection, suggesting no obvious correlation between the stress and inflammatory molecule expression and the control/exacerbation of leishmanial lesions. However, the cytokine VEGF was only detected in C57BL/6 footpad lesions and small lesions in Balb/c mice treated with antimonial pentavalent. These findings suggest that VEGF expression could be a predictive factor for murine leishmanial control, a hypothesis that should be tested in human leishmaniosis.
Asunto(s)
Ciclooxigenasa 2/inmunología , Citocinas/inmunología , Inflamación/inmunología , Leishmaniasis Cutánea/inmunología , Factor A de Crecimiento Endotelial Vascular/inmunología , Animales , Biomarcadores/análisis , Ciclooxigenasa 2/biosíntesis , Modelos Animales de Enfermedad , Inmunohistoquímica , Leishmania/inmunología , Leishmaniasis Cutánea/parasitología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Óxido Nítrico/biosíntesis , Nitroimidazoles/uso terapéutico , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Proteínas Quinasas p38 Activadas por Mitógenos/biosíntesis , Proteínas Quinasas p38 Activadas por Mitógenos/inmunologíaRESUMEN
Inflammatory bowel disease (IBD) is a common chronic gastrointestinal disorder characterized by alternating periods of remission and active intestinal inflammation. Flavonoids exert several biological activities, which are mainly related to their ability to inhibit inflammatory process and/or to their antioxidant properties, and are able to regulate the immune response. The aim of this study was to evaluate whether phenolic compounds present in grape juice could reduce the inflammatory effects induced by experimental colitis. A total of 41 male Wistar rats were randomized into seven groups, as follows: G1--Sham group: sham induced-colitis rats; G2--(2,4,6-rinitrobenzenesulfonic acid) TNBS group: nontreated induced-colitis; G3--2% grape juice control group; G4--1% grape juice 24h after TNBS colitis induction; G5--1% grape juice on day 7 after colitis induction; G6--2% grape juice 24h after colitis induction; G7--2% grape juice on day 7 after colitis induction. Genotoxicity was evaluated by comet assay. Immunohistochemistry was determined using the streptavidin-biotin-peroxidase method being analyzed in control (normal tissue) and "hot spot" areas i.e., presenting inflammatory process being graded as 1 (weak), 2 (moderate), or 3 (strong). Both parameters were evaluated in the cytoplasm of epithelial or inflammatory cells. TNF-immunoexpression and iNOS were reduced after drinking grape juice 24 h or after 7 days for all doses tested. COX-2 was reduced in the groups exposed to 1% grape juice 24 h or 7 days of exposure. The grape juice at 1% dose in the last 7 days of treatment as well as grape juice at 2% dose decreased the peripheral blood genotoxicity. Taken together, the grape juice mainly at 1% dose exerts anti-inflammatory effects in chronic colitis caused by TNBS as a result of down regulation in the expression of pro-inflammatory cytokines and reduction of genotoxicity in peripheral blood cells.
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Antiinflamatorios/uso terapéutico , Bebidas , Colitis/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Vitis , Animales , Antiinflamatorios/farmacología , Colitis/inducido químicamente , Colitis/inmunología , Colitis/patología , Colon/efectos de los fármacos , Colon/inmunología , Colon/patología , Ensayo Cometa , Ciclooxigenasa 2/inmunología , Daño del ADN/efectos de los fármacos , Frutas , Masculino , Óxido Nítrico Sintasa de Tipo II/inmunología , Fitoterapia , Extractos Vegetales/farmacología , Ratas Wistar , Ácido Trinitrobencenosulfónico , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Activation of the platelet-activating factor receptor (PAFR) in macrophages is associated with suppressor phenotype. Here, we investigated the PAFR in murine dendritic cells (DC). Bone marrow-derived dendritic cells (BALB/c) were cultured with GM-CSF and maturation was induced by LPS. The PAFR antagonists (WEB2086, WEB2170, PCA4248) and the prostaglandin (PG) synthesis inhibitors (indomethacin, nimesulide and NS-398) were added before LPS. Mature and immature DCs expressed PAFR. LPS increased MHCII, CD40, CD80, CD86, CCR7 and induced IL-10, IL-12, COX-2 and PGE2 expression. IL-10, COX-2 and PGE2 levels were reduced by PAFR antagonists and increased by cPAF. The IL-10 production was independent of PGs. Mature DCs induced antigen-specific lymphocyte proliferation. PAFR antagonists or PG-synthesis inhibitors significantly increased lymphocyte proliferation. It is proposed that PAF has a central role in regulatory DC differentiation through potentiation of IL-10 and PGE2 production.
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Células de la Médula Ósea/metabolismo , Células Dendríticas/metabolismo , Dinoprostona/metabolismo , Glicoproteínas de Membrana Plaquetaria/genética , Receptores Acoplados a Proteínas G/genética , Linfocitos T/metabolismo , Animales , Presentación de Antígeno , Antígenos CD/genética , Antígenos CD/inmunología , Células de la Médula Ósea/citología , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/inmunología , Diferenciación Celular , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Inhibidores de la Ciclooxigenasa/farmacología , Células Dendríticas/citología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Regulación de la Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Antígenos de Histocompatibilidad Clase II/genética , Antígenos de Histocompatibilidad Clase II/inmunología , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-12/genética , Interleucina-12/inmunología , Lipopolisacáridos/farmacología , Ratones , Ratones Endogámicos BALB C , Inhibidores de Agregación Plaquetaria/farmacología , Glicoproteínas de Membrana Plaquetaria/agonistas , Glicoproteínas de Membrana Plaquetaria/inmunología , Cultivo Primario de Células , Receptores CCR7/genética , Receptores CCR7/inmunología , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/inmunología , Transducción de Señal , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunologíaRESUMEN
Curcumin is the active compound in the extract of Curcuma longa rhizomes with anti-inflammatory properties mediated by inhibition of intracellular signalling. SOCS and MAPKinases are involved in the signalling events controlling the expression of IL-6, TNF-α and PGE2, which have important roles on chronic inflammatory diseases. The aim was to assess if these pathways are involved in curcumin-mediated effects on LPS-induced expression of these cytokines in macrophages. RAW 264.7 murine macrophages were stimulated with Escherichia coli LPS in the presence and absence of non-cytotoxic concentrations of curcumin. Curcumin potently inhibited LPS-induced expression of IL-6, TNF-α and COX-2 mRNA and prevented LPS-induced inhibition of SOCS-1 and -3 expression and the inhibition of the activation of p38 MAPKinase by modulation of its nuclear translocation. In conclusion, curcumin potently inhibits expression of LPS-induced inflammatory cytokines in macrophages via mechanisms that involve modulation of expression and activity of SOCS-1 and SOCS-3 and of p38 MAPK.
Asunto(s)
Curcumina/farmacología , Inmunidad Innata , Macrófagos/inmunología , Proteínas Supresoras de la Señalización de Citocinas/inmunología , Proteínas Quinasas p38 Activadas por Mitógenos/inmunología , Animales , Ciclooxigenasa 2/inmunología , Citocinas/inmunología , Interleucina-6/inmunología , Lipopolisacáridos/farmacología , Ratones , ARN Mensajero/inmunología , Transducción de Señal/inmunología , Proteína 1 Supresora de la Señalización de Citocinas , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The in vivo modified forms of low-density lipoprotein (LDL) are important for the formation of foam cells and as mediators of the immuno-inflammatory process involved in the progression of atherosclerosis. Electronegative LDL, LDL(-), is a LDL subfraction with pro-inflammatory properties that is present in human blood. To investigate possible atheroprotective effects, an anti-LDL(-) single-chain variable fragment (scFv) was expressed in the methylotrophic yeast Pichia pastoris and its activity was evaluated in vitro against macrophages and in experimental atherosclerosis in Ldlr(-/-) mice. The recombinant 2C7 scFv was produced in a yield of 9.5 mg of protein/L. The specificity and affinity of purified 2C7 scFv against LDL(-) was confirmed by ELISA. To assess the activity of 2C7 scFv on foam cell formation, RAW 264.7 macrophages were exposed to LDL(-) in the presence or absence of 2C7 scFv. The 2C7 scFv inhibited the uptake of LDL(-) by macrophages in a dose-dependent manner, and internalization of LDL(-) by these cells was found to be mediated by the CD36 and CD14 receptor. In addition, compared with untreated cells, lipid accumulation in macrophages was decreased, and the expression of Cd36, Tlr-4 and Cox-2 was downregulated in macrophages treated with 2C7 scFv. Importantly, compared with untreated mice, the treatment of Ldlr(-/-) mice with 2C7 scFv decreased the atherosclerotic lesion area at the aortic sinus. In conclusion, our data show that 2C7 scFv inhibits foam cell formation and atherosclerotic plaque development by modulating the expression of genes relevant to atherogenesis. These results encourage further use of this antibody fragment in the development of new therapeutic strategies that neutralize the pro-atherogenic effects of LDL(-).
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Aterosclerosis/prevención & control , Lipoproteínas LDL/inmunología , Proteínas Recombinantes/farmacología , Anticuerpos de Cadena Única/farmacología , Animales , Afinidad de Anticuerpos/inmunología , Especificidad de Anticuerpos/inmunología , Aterosclerosis/genética , Aterosclerosis/inmunología , Antígenos CD36/genética , Antígenos CD36/inmunología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/inmunología , Clonación Molecular , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Ensayo de Inmunoadsorción Enzimática , Células Espumosas/efectos de los fármacos , Células Espumosas/inmunología , Células Espumosas/metabolismo , Expresión Génica/efectos de los fármacos , Expresión Génica/inmunología , Lípidos/análisis , Lipoproteínas LDL/sangre , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Pichia/genética , Receptores de LDL/genética , Receptores de LDL/inmunología , Receptores de LDL/metabolismo , Proteínas Recombinantes/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Anticuerpos de Cadena Única/genética , Anticuerpos de Cadena Única/inmunología , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/inmunologíaRESUMEN
Nymphaea mexicana Zucc. is an aquatic plant species which belongs to the family Nymphaea and is commonly known as the yellow water lily. The aim of this work was to study the in vitro antiinflammatory effects of phenolic compounds isolated from the flowers of Nymphaea mexicana Zucc. Seven phenolic compounds including vanillic acid, 4-methoxy-3,5-dihydroxybenzoic acid, (2R,3R)-3,7-dihydroxyflavanone, naringenin (4), kaempferol 3-O-(3-O-acetyl-a-L-rhamnopyranoside), kaempferol 3-O-(2-O-acetyl-a-L-rhamnopyranoside), and quercetin 3-(30 0-acetylrhamnoside) (7) were isolated from the flowers of Nymphaea mexicana Zucc. These results revealed that compound 4 has the most prominent inhibitory effect on the LPS-stimulated nitric oxide (NO), monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-a) production in RAW 264.7 macrophages. In addition, compound 4 also inhibited LPS-mediated induction of protein expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), and phospho-ERK in RAW 264.7 macrophages. Thus, compound 4 from the flowers of Nymphaea mexicana Zucc. may provide a potential therapeutic approach for inflammation-associated disorders.
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Antiinflamatorios/farmacología , Flores/química , Nymphaea/química , Fenoles/farmacología , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Quimiocina CCL2/genética , Quimiocina CCL2/inmunología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Óxido Nítrico/inmunología , Fenoles/química , Fenoles/aislamiento & purificación , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Pharmacological therapy is widely used in the treatment of muscle injuries. On the other hand, low-level laser therapy (LLLT) arises as a promising nonpharmacological treatment. The aim of this study was to analyze the effects of sodium diclofenac (topical application) and LLLT on morphological aspects and gene expression of biochemical inflammatory markers. We performed a single trauma in tibialis anterior muscle of rats. After 1 h, animals were treated with sodium diclofenac (11.6 mg g(-1) of solution) or LLLT (810 nm; continuous mode; 100 mW; 3.57 W cm(-2) ; 1, 3 or 9 J; 10, 30 or 90 s). Histological analysis and quantification of gene expression (real-time polymerase chain reaction-RT-PCR) of cyclooxygenase 1 and 2 (COX-1 and COX-2) and tumor necrosis factor-alpha (TNF-α) were performed at 6, 12 and 24 h after trauma. LLLT with all doses improved morphological aspects of muscle tissue, showing better results than injury and diclofenac groups. All LLLT doses also decreased (P < 0.05) COX-2 compared to injury group at all time points, and to diclofenac group at 24 h after trauma. In addition, LLLT decreased (P < 0.05) TNF-α compared both to injury and diclofenac groups at all time points. LLLT mainly with dose of 9 J is better than topical application of diclofenac in acute inflammation after muscle trauma.
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Antiinflamatorios no Esteroideos/farmacología , Diclofenaco/farmacología , Terapia por Luz de Baja Intensidad , Músculo Esquelético/efectos de la radiación , Traumatismos de los Tejidos Blandos/radioterapia , Animales , Biomarcadores/análisis , Ciclooxigenasa 1/genética , Ciclooxigenasa 1/inmunología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Expresión Génica/efectos de la radiación , Inflamación/prevención & control , Masculino , Proteínas de la Membrana/genética , Proteínas de la Membrana/inmunología , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/lesiones , Músculo Esquelético/metabolismo , Ratas , Ratas Wistar , Traumatismos de los Tejidos Blandos/tratamiento farmacológico , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Muscle injuries represent ca 30% of sports injuries and excessive stretching of muscle causes more than 90% of injuries. Currently the most used treatments are nonsteroidal anti-inflammatory drugs (NSAIDs), however, in last years, low-level laser therapy (LLLT) is becoming an interesting therapeutic modality. The aim of this study was to evaluate the effect of single and combined therapies (LLLT, topical application of diclofenac and intramuscular diclofenac) on functional and biochemical aspects in an experimental model of controlled muscle strain in rats. Muscle strain was induced by overloading tibialis anterior muscle of rats. Injured groups received either no treatment, or a single treatment with topical or intramuscular diclofenac (TD and ID), or LLLT (3 J, 810 nm, 100 mW) 1 h after injury. Walking track analysis was the functional outcome and biochemical analyses included mRNA expression of COX-1 and COX-2 and blood levels of prostaglandin E2 (PGE2 ). All treatments significantly decreased COX-1 and COX-2 gene expression compared with injury group (P < 0.05). However, LLLT showed better effects than TD and ID regarding PGE2 levels and walking track analysis (P < 0.05). We can conclude that LLLT has more efficacy than topical and intramuscular diclofenac in treatment of muscle strain injury in acute stage.
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Antiinflamatorios no Esteroideos/farmacología , Diclofenaco/farmacología , Terapia por Luz de Baja Intensidad , Músculo Esquelético/efectos de la radiación , Traumatismos de los Tejidos Blandos/radioterapia , Esguinces y Distensiones/radioterapia , Animales , Biomarcadores/análisis , Terapia Combinada , Ciclooxigenasa 1/genética , Ciclooxigenasa 1/inmunología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Dinoprostona/sangre , Expresión Génica/efectos de la radiación , Inyecciones Intramusculares , Masculino , Proteínas de la Membrana/genética , Proteínas de la Membrana/inmunología , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/lesiones , Músculo Esquelético/metabolismo , Ratas , Ratas Wistar , Traumatismos de los Tejidos Blandos/tratamiento farmacológico , Esguinces y Distensiones/tratamiento farmacológicoRESUMEN
Oral mucositis is an important dose-limiting and costly side effect of cancer chemotherapy. Soluble proteins obtained of the latex of Calotropis procera have been extensively characterized as anti-inflammatory in different experimentally induced inflammatory conditions, including arthritis and sepsis. In this study, the phytomodulatory laticifer proteins (LP) were challenged to regress the inflammatory events associated with 5-fluorouracil-induced oral mucositis. We also evaluated the expression of pro-inflammatory cytokines and inducible enzymes, such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). Oral mucositis was induced in hamsters by two injections of 5-fluorouracil (5-FU; 60 and 40 mg/kg, i.p., on experimental days 1 and 2, respectively). LP (5 mg/kg, i.p.) was injected 24 h before and 24 h after mechanical trauma of the cheek pouches. A normal control group received only saline. On day 10, the animals were sacrificed, and the cheek pouches were excised for macroscopic and histopathological analysis, myeloperoxidase activity measurement, and immunohistochemical assessment of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), iNOS, and COX-2. LP significantly inhibited macroscopic histopathological scores and myeloperoxidase activity compared with the 5-FU control group. 5-Fluorouracil also induced marked immunostaining of TNF-α, IL-1ß, iNOS, and COX-2 on inflamed conjunctive and epithelial tissue compared with the normal control group. Such damage was significantly inhibited (p < 0.05) by LP treatment compared with the 5-FU group. These findings demonstrate an anti-inflammatory effect of LP on 5-FU-induced oral mucositis. The protective mechanism appears to involve inhibition of the expression of iNOS, COX-2, TNF-α, and IL-1ß.
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Antimetabolitos Antineoplásicos/efectos adversos , Calotropis/química , Fluorouracilo/efectos adversos , Factores Inmunológicos/inmunología , Látex/química , Proteínas de Plantas/uso terapéutico , Estomatitis/prevención & control , Animales , Cricetinae , Ciclooxigenasa 2/biosíntesis , Ciclooxigenasa 2/inmunología , Citocinas/biosíntesis , Citocinas/inmunología , Modelos Animales de Enfermedad , Regulación hacia Abajo , Inmunohistoquímica , Factores Inmunológicos/biosíntesis , Masculino , Mesocricetus , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Óxido Nítrico Sintasa de Tipo II/inmunología , Peroxidasa/metabolismo , Proteínas de Plantas/administración & dosificación , Proteínas de Plantas/aislamiento & purificación , Estomatitis/inducido químicamente , Estomatitis/inmunología , Estomatitis/patologíaRESUMEN
OBJECTIVE: The aim was to investigate the relationship between patient clinical background, histological features, and immunoexpression of COX-2 and IL-10 in oral pyogenic granuloma (PG). DESIGN: Paraffin-embedded samples of oral PG (n=57) were prepared for histological and immunohistochemical assessment. Based on the histological features, the samples were categorised into lobular capillary hemangioma (LCH) and non-LCH subtypes. The epithelial lining, angiogenic index, inflammatory infiltrate density, and interstitial fibrosis, were assessed in haematoxylin-eosin stained sections. In addition, the marker expression estimation (stained cells/total cell number) was used to assess immunoreactivity for each sample. RESULTS: Although there were no significant differences between histological subtypes regarding demographic and clinical parameters, mean values of microvessel count and inflammatory infiltrate density were significantly greater in the non-LCH PG subtype. Also, whilst cellular immunolocalisation patterns of COX-2 and IL-10 were similar, mean values of expression estimation of each immunomarker were significantly higher in non-LCH PGs in comparison with LCH subtypes. Furthermore, significant variations for immunohistochemical parameters were evident regarding to angiogenic index and inflammatory infiltrate density, but not concerning demographic and clinical data. Finally, linear regression analysis showed a significant positive correlation between the expression estimation of the two immunomarkers. CONCLUSION: These findings suggest a role for COX-2 and IL-10 in the etiopathogenesis of oral PG and indicate that LCH and non-LCH histological subtypes represent different stages in the evolution of a single lesion with varying degrees of proliferative, angiogenic, and inflammatory activity.
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Ciclooxigenasa 2/metabolismo , Granuloma Piogénico/metabolismo , Interleucina-10/metabolismo , Adulto , Análisis de Varianza , Biomarcadores/metabolismo , Biopsia , Distribución de Chi-Cuadrado , Ciclooxigenasa 2/inmunología , Femenino , Granuloma Piogénico/inmunología , Humanos , Inmunohistoquímica , Interleucina-10/inmunología , Modelos Lineales , Masculino , Distribución Aleatoria , Reproducibilidad de los Resultados , Coloración y Etiquetado , Estadísticas no ParamétricasRESUMEN
The effects of Leishmania mexicana metacyclic promastigotes upon MAP kinase signalling in mouse bone marrow macrophages and subsequent expression of the disease regulatory proteins iNOS and COX-2 were studied. At a ratio of 5:1, promastigotes caused a marked increase in phosphorylation of the three major MAP kinases, ERK, p38 and JNK. MAP kinase signalling was substantially reduced in TLR-4(-/-) but not TLR-2(-/-) deficient macrophages and completely abolished in double TLR-2/4(-/-) macrophages. A similar outcome was observed using cysteine peptidase B deficient amastigotes. Furthermore, whilst promastigotes had no independent effect on iNOS or COX-2 expression, they prolonged the induction of these proteins stimulated by LPS and enhanced PGE(2) and NO production. Induction of COX-2 and iNOS was also TLR-4 dependent. Blockade of either PGE(2) or NO production with indomethacin or l-NAME reversed promastigote inhibition of LPS induced IL-12 production. Promastigotes also increased macrophage arginase-1 expression and enhanced arginase activity, both of which were substantially reduced in TLR-4 but not TLR-2 deficient macrophages. Surprisingly, arginase inhibition by Nor-NOHA also caused a reversal of promastigote mediated inhibition of macrophage IL-12 production. These data demonstrate for the first time the role of TLR-4 in mediating the effects of L. mexicana promastigotes on MAP kinase activation, up-regulation of COX-2, iNOS as well as arginase-1 expression in macrophages and further shows that PGE(2), NO and arginase activity all contribute substantially to the inhibition of host cell IL-12 production.