RESUMEN
We used water-soluble Chitosan obtained by Maillard reaction with glucosamine to microencapsulate soy genistein (Ge) and preserve its biological activity for oral administration. Release of Ge was pH dependent with a super Case II mechanism at pH 1.2 and an anomalous transport with non-Fickian kinetics at pH 6.8. Microencapsulated Ge retained its antioxidant properties in vitro and its daily administration to mice attenuated clinical signs of acute colitis, limited inflammatory reaction and reduced oxidative stress and tissue injury as well. Remarkably, after feeding microencapsulated Ge the production of IL-10 in colonic tissue was restored to levels of untreated controls. According to statistical multivariate analysis, this cytokine was the parameter with the highest influence on the inflammatory/oxidative status. Microencapsulation of Ge with derivatized Chitosan becomes an interesting alternative to develop therapeutic approaches for oxidative inflammatory diseases; our findings suggest that the soy isoflavone could be incorporated into any functional food for application in intestinal inflammation.
Asunto(s)
Antioxidantes/administración & dosificación , Colitis/dietoterapia , Genisteína/administración & dosificación , Administración Oral , Animales , Antioxidantes/química , Antioxidantes/farmacología , Quitosano/química , Colitis/inducido químicamente , Colitis/metabolismo , Citocinas/metabolismo , Suplementos Dietéticos , Modelos Animales de Enfermedad , Composición de Medicamentos/métodos , Femenino , Genisteína/química , Genisteína/farmacología , Interleucina-10/metabolismo , Ratones Endogámicos C57BL , Estrés Oxidativo/efectos de los fármacos , Solubilidad , Glycine max/químicaRESUMEN
Bovine mastitis affects the health of dairy cows and the profitability of herds worldwide. Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens in bovine intramammary infection. Based on the wide range of antimicrobial, mucoadhesive and immunostimulant properties demonstrated by chitosan, we have evaluated therapy efficiency of chitosan incorporation to cloxacillin antibiotic as well as its effect against different bacterial lifestyles of seven CNS isolates from chronic intramammary infections. The therapeutic effects of combinations were evaluated on planktonic cultures, bacterial biofilms and intracellular growth in mammary epithelial cells. We found that biofilms and intracellular growth forms offered a strong protection against antibiotic therapy. On the other hand, we found that chitosan addition to cloxacillin efficiently reduced the antibiotic concentration necessary for bacterial killing in different lifestyle. Remarkably, the combined treatment was not only able to inhibit bacterial biofilm establishment and increase preformed biofilm eradication, but it also reduced intracellular bacterial viability while it increased IL-6 secretion by infected epithelial cells. These findings provide a new approach to prophylactic drying therapy that could help to improve conventional antimicrobial treatment against different forms of bacterial growth in an efficient, safer and greener manner reducing multiresistant bacteria generation and spread.
Asunto(s)
Antibacterianos/uso terapéutico , Quitosano/uso terapéutico , Cloxacilina/uso terapéutico , Mastitis Bovina/tratamiento farmacológico , Infecciones Estafilocócicas/veterinaria , Staphylococcus/efectos de los fármacos , Animales , Antibacterianos/administración & dosificación , Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Bovinos , Quitosano/administración & dosificación , Quitosano/farmacología , Cloxacilina/administración & dosificación , Cloxacilina/farmacología , Femenino , Mastitis Bovina/microbiología , Pruebas de Sensibilidad Microbiana , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/microbiología , Staphylococcus/aislamiento & purificación , Staphylococcus/fisiologíaRESUMEN
Lamina propria dendritic cells (DCs) have a permanent turnover with constitutive migration to mesenteric lymph nodes and replenishment by progenitors. Luminal bacteria and dietary constituents provide key signals that endow DCs their unique properties in vivo. Taking into account that the intestinal immune system is greatly influenced by retinoids, we evaluated in B6 mice 3, 8, 16 and 24 h after feeding a single dose of vitamin A phenotype and function of cells present in mesenteric afferent lymph nodes as well as signals involved in migration. We studied the frequency of CD11c+MHC-II+CD103+CD86+ and RALDH+ DCs by flow cytometry, we determined CCL-21 and D6 levels in tissue homogenates by Western blot, and we co-cultured cells isolated from afferent lymphatics with sorted CD4+ lymphocytes to assess Foxp-3 induction and homing receptor expression. Sixteen hours after vitamin A administration, DCs isolated from afferent lymphatics were able to induce homing receptors and Foxp3 expression in CD4+ lymphocytes. Our results show that a single dose of vitamin A generated a stream of signals and amplified the tolerogenic activity of DCs migrating to lymphoid tissue.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Células Dendríticas/metabolismo , Suplementos Dietéticos , Factores de Transcripción Forkhead/agonistas , Regulación de la Expresión Génica , Receptores Mensajeros de Linfocitos/agonistas , Vitamina A/administración & dosificación , Animales , Antígenos CD/metabolismo , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Movimiento Celular , Células Cultivadas , Técnicas de Cocultivo , Células Dendríticas/citología , Células Dendríticas/inmunología , Femenino , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Tolerancia Inmunológica , Linfa/citología , Linfa/inmunología , Linfa/metabolismo , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Mesenterio , Ratones Endogámicos C57BL , Ratones Transgénicos , Receptores Mensajeros de Linfocitos/genética , Receptores Mensajeros de Linfocitos/metabolismo , Organismos Libres de Patógenos EspecíficosRESUMEN
Larrea divaricata Cav. (jarilla) is a plant with well-documented applications in Argentinean folk medicine. In order to determine if the treatment with a purified fraction named F1 was capable to maintain a state of priming of macrophages after 15 days of mice infection with Candida albicans. Infected and uninfected mice were used. The effect of F1 on: cytosolic protein levels, apoptosis, phagocytosis, reactive oxygen species production, nitric oxide (NO), cell activity, lysosomal activity and the tissue fungal burden were studied. The results showed that F1 increased macrophages yeast phagocytosis and reactive oxygen species and NO production. All these effects were related to a decrease of cell activity and possible apoptosis. In conclusion, it was observed that F1 could induce a state of long-term activation of macrophages, since we observed increased activity of macrophages 15 days after infection, and it could be related to the elimination of C. albicans. These data may suggest that F1 fraction could be useful against disseminated candidiasis in patients and further studies on this field are desirable.
Asunto(s)
Candida albicans , Candidiasis/tratamiento farmacológico , Larrea/química , Macrófagos Peritoneales/metabolismo , Extractos Vegetales/farmacología , Animales , Apoptosis/efectos de los fármacos , Candidiasis/metabolismo , Candidiasis/patología , Células Cultivadas , Femenino , Macrófagos Peritoneales/microbiología , Macrófagos Peritoneales/patología , Ratones , Óxido Nítrico/biosíntesis , Fagocitosis/efectos de los fármacos , Extractos Vegetales/química , Factores de TiempoRESUMEN
Larrea divaricata Cav. (jarilla) is a plant with well-documented applications in folk medicine in Argentina. In this study, we aimed to evaluate functional parameters of peritoneal macrophages isolated from mice injected with three fractions (F1, F2 and F3) of L. divaricata. The response of macrophages against Candida albicans was evaluated. Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, apoptosis was evaluated using Giemsa, acridine orange/ethidium bromide and ladder assay, oxidative burst was assayed using nitroblue tetrazolium test and nitrite production using Griess assay. Cell stimulation and their ability to kill C. albicans in vitro were measured. The number and cell viability were similar to controls. However, we found that F1 induces pre-activation of macrophages, and this pre-activation is enhanced by C. albicans. The effects exerted by F1 make it more important than F2 and F3 for the treatment of disseminated candidiasis in patients with immunodeficiency diseases such as AIDS and chronic granulomatous disease, among others.
Asunto(s)
Candida albicans/inmunología , Factores Inmunológicos/farmacología , Larrea/química , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/inmunología , Extractos Vegetales/farmacología , Animales , Apoptosis , Supervivencia Celular , Células Cultivadas , Fragmentación del ADN , Etidio/metabolismo , Femenino , Factores Inmunológicos/aislamiento & purificación , Masculino , Ratones , Nitritos/metabolismo , Nitroazul de Tetrazolio/metabolismo , Extractos Vegetales/aislamiento & purificación , Estallido Respiratorio , Coloración y Etiquetado/métodos , Sales de Tetrazolio/metabolismo , Tiazoles/metabolismoRESUMEN
Chitosan is a copolymer of N-acetylglucosamine and glucosamine derived from chitin with several applications in pharmaceutical and medical fields. This polysaccharide exhibits adjuvant properties in mucosal immune responses of humans, rats and mice. Characterization of signals elicited by chitosan at the intestinal epithelium could explain its immunomodulatory activity and biocompatibility. We fed normal rats with single doses of chitosan and 16h later, we purified intestinal epithelial cells (IECs) to assess immune and biochemical parameters. Following chitosan administration, mRNA expression and release of several cytokines and chemokines increased, injury markers maintained constitutive levels and MHC type II molecule expression was augmented. IEC supernatants showed higher levels of IL-10, IL-6 and TGF-beta. Arginase activity of IECs increased upon chitosan interaction in vivo and in vitro. Together, after chitosan feeding, mild activation of IECs occurs in vivo, with production of regulatory factors that could be relevant for its biocompatibility and immunomodulatory effects.
Asunto(s)
Quitosano/inmunología , Inmunidad Mucosa , Inmunomodulación , Mucosa Intestinal/inmunología , Administración Oral , Animales , Arginasa/metabolismo , Células Cultivadas , Quimiocinas/inmunología , Células Epiteliales/inmunología , Femenino , Interleucina-10/inmunología , Interleucina-6/inmunología , Mucosa Intestinal/citología , Ratas , Ratas Wistar , Factor de Crecimiento Transformador beta/inmunologíaRESUMEN
BACKGROUND AND AIM: Larrea divaricata Cav. (Zygophyllaceae) is a plant widely used in Argentina. MATERIAL AND METHODS: We isolated different fractions of L. divaricata aqueous extract containing minor amounts of NDGA, and we analyzed these fractions on mouse macrophages. RESULTS: We showed that a fraction without NDGA was capableof activating macrophages, principally through the production of mitochondrial anion superoxide and H(2)O(2). This could be important in the defense of infections. Moreover, this fraction decreased NO level suggesting an anti-inflammatory action. CONCLUSION: These results indicate that NDGA was not the compound responsible for the immunomodulatory action exerted by the aqueous extract from L. divaricata.
Asunto(s)
Factores Inmunológicos/farmacología , Larrea , Macrófagos Peritoneales/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Apoptosis/efectos de los fármacos , Cromatografía en Capa Delgada , Femenino , Larrea/química , Receptores de Lipopolisacáridos/análisis , Macrófagos Peritoneales/metabolismo , Masculino , Masoprocol/farmacología , Ratones , Óxido Nítrico/biosíntesis , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Larrea divaricata is a plant widely used in folk medicine in Argentina. This work aimed to study the mechanisms of decoction activity on the release of oxygen reactive species. Decoction increased the binding of zymosan-FITC and superoxide production. Cadmium decreased the superoxide production as well as malonate and barbital. Decoction decreased the release of hydrogen peroxide. Decoction increased the reduction of MTT but not when malonate and barbital were included. Together, decoction increased the expression of dectin-1 leading to increased superoxide production. It is possible that decoction increases the activity of peroxidase, and decreases the Cu, Zn-superoxide dismutase.
Asunto(s)
Peróxido de Hidrógeno/metabolismo , Larrea , Macrófagos Peritoneales/efectos de los fármacos , Extractos Vegetales/farmacología , Receptores de Complemento/efectos de los fármacos , Superóxidos/metabolismo , Animales , Barbital/farmacología , Cloruro de Cadmio/farmacología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Larrea/química , Lectinas Tipo C , Macrófagos Peritoneales/metabolismo , Masculino , Malonatos/farmacología , Proteínas de la Membrana/metabolismo , Ratones , Proteínas del Tejido Nervioso/metabolismo , Componentes Aéreos de las Plantas , Receptores de Complemento/metabolismo , Zimosan/metabolismoRESUMEN
Several medicinal plants are considered immunomodulatory as they display a variety of anti-inflammatory, antimicrobial and antitumoral effects. Larrea divaricata Cav. (jarilla) (Zygophyllaceae) is a plant widely used in popular medicine to treat tumors, infections, and inflammatory diseases. So far, the immunostimulating activities of Larrea divaricata have not been studied in vivo. In this work, we used healthy mice to assess the immunomodulatory potential of aqueous extracts of Larrea divaricata Cav. We found that Decoction (D) and Infusion (I) from Larrea divaricata Cav showed any acute hepatotoxic activity. Only D at 0.5 mg/kg increased the carrageenan-induced inflammation. Macrophages harvested from treated mice showed no signs of apoptosis. These cells showed a significant increase in NO and TNF-alpha release and exhibited the strongest expression of iNOS. Decoction also increased the phagocytosis of zymosan and the binding of LPS-FITC. The expression of CD14, TLR4 and CR3 was lower in macrophages of mice treated than in controls. Thus, Larrea divaricata was able to prime Mphi in vivo and to induce full activation in vitro. Our finding contribute to characterize the biological activity of Larrea divaricata and to understand the ability of these extracts to enhance immune responses.
Asunto(s)
Factores Inmunológicos/farmacología , Larrea/química , Alanina Transaminasa/sangre , Animales , Antiinflamatorios no Esteroideos , Apoptosis/efectos de los fármacos , Western Blotting , Carragenina , Edema/inducido químicamente , Edema/tratamiento farmacológico , Femenino , Citometría de Flujo , Factores Inmunológicos/uso terapéutico , Técnicas In Vitro , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Masculino , Ratones , Óxido Nítrico/metabolismo , Nitritos/metabolismo , Extractos Vegetales/farmacología , Hojas de la Planta/química , Receptores Depuradores/efectos de los fármacos , Estallido Respiratorio/efectos de los fármacos , Estimulación Química , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Decoction and infusion of Larrea divaricata were tested at apoptotic concentrations (1 and 4 mg/ml) on peritoneal murine macrophages. Consistent changes were observed after incubation with 4 mg/ml decoction. Phagocytosis of zymosan, lysosomal enzyme activity, nitric oxide production, TNF-alpha release, and expression of CD14, TLR4, and CR3 increased significantly. Decoction at 1 and 4 mg/ml increased the binding of LPS-FITC. Apoptosis triggered by L. divaricata decoction is consequence of cell activation. The effects are independent of nordihydroguaiaretic acid. This "activation and death" could be the mechanism of L. divaricata to exert the antituberculosis effect known in folk medicine.
Asunto(s)
Apoptosis/efectos de los fármacos , Larrea/química , Macrófagos/efectos de los fármacos , Fosfatasa Ácida/metabolismo , Animales , Antioxidantes/farmacología , Femenino , Citometría de Flujo , Técnicas In Vitro , Prueba de Limulus , Receptores de Lipopolisacáridos/efectos de los fármacos , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/toxicidad , Masculino , Masoprocol/farmacología , Ratones , Óxido Nítrico/análisis , Óxido Nítrico/biosíntesis , Fagocitosis/efectos de los fármacos , Extractos Vegetales/farmacología , Hojas de la Planta/química , Estallido Respiratorio/efectos de los fármacos , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
We investigated the association with Yersinia infection in patients with arthropathies in our region. To assess the reactivity to articular antigens, the correlation of anti-Yersinia with anti-type I and type II collagen antibodies was studied. Sera from 124 patients with musculoskeletal symptoms, and 47 synovial fluids (SF) from patients with rheumatoid arthritis (RA), spondyloarthopathies (SpA) or osteoarthritis (OA) were examined. Immunoglobulins against Yersinia enterocolitica, type I and type II collagens were determined by enzyme-linked immunosorbent assay. Immunoglobulin (Ig) A to Yersinia lipopolysaccharide (LPS) was present in 13/124 sera (10%) and 3/47 SF (6%). By Western blot, IgA to Yersinia outer proteins (Yops) was found in 14/124 sera (11%) and 2/47 SF (4%). Yersinia DNA from SF was not amplified by polymerase chain reaction. We found a significant correlation with anti-collagen type I but not type II antibodies. These results suggest different reactivity to articular collagen in patients with Yersinia antibodies.