Avaliação do extrato do fungo caripia montagnei e de agonistas de PPAR - α no processo inflamatório

The mushrooms have been object of intense research in view of its potential raising of application in different sectors of the pharmacology and alimentary industry. Among diverse bioactive composites of polyssacharides nature that exist in the fungus the glucans are much searched. These are polymers of glucose and classified as the type of glicosidic linking [α, β]. Peroxisome proliferator-activated receptors (PPARs), ranscription factors belonging to the family of nuclear receptors that bind themselves o specific agonists, have shown their importance in controlling the inflammatory process. The aim of this study was to perform a chemical characterization of extract rom the mushroom Caripia montagnei, assess its antiinflammatory and antibacterial effect and determine if this effect occurs via PPAR. This mushroom is composed of carbohydrates (63.3±4.1%), lipids (21.4l±0.9%) and proteins (2.2± 0.3%). The aqueous solution resulting from the fractionation contained carbohydrates (98.7±3.3%) and protein (1.3±0.25%). Analyses of infrared spectrophotometry and of nuclear magnetic esonance demonstrated that the extract of mushroom C. montagnei is rich in β-glucans. In hioglycolate-induced peritonitis, the C. montagnei glucans (50 mg/kg) educed the inflammatory process in 65.5±5.2% and agonists, pharmacological igands, for PPAR: Wy-14643 (49.3±6.1%), PFOA (48.9±3.8%) and clofibrate in 45.2±3.2%. Sodium diclofenac showed a reduction of 81.65±0.6%. In the plantar edema, the glucans from C. montagnei (50 mg/kg) and L-NAME reduced the edema to a similar degree 91.4±0.3% and 92.8±0,5 %, respectively. In all the groups tested, nitric oxide (NO), an inflammation mediator, showed a significant reduction in the nitrate/nitrite levels when compared to the positive control (P<0.001). The C. montagnei glucans did not show cytotoxicity in the concentrations tested (2.5, 5.0, 10.0, 20.0 and 40.0 µg/100 µL). Antibacterial activity demonstrated that, unlike total extract, there was no inhibition of bacterial growth. The C. montagnei glucans show great potential for antiinflammatory applications. This effect suggests that it is mediated by PPAR activation and by COX and iNOS inhibition

Effect of myrrh and thyme on Trichinella spiralis enteral and parenteral phases with inducible nitric oxide expression in mice

Trichinellosis is a serious disease with no satisfactory treatment. We aimed to assess the effect of myrrh ( Commiphora molmol ) and, for the first time, thyme ( Thymus vulgaris L.) against enteral and encysted (parenteral) phases of Trichinella spiralis in mice compared with albendazole, and detect their effect on inducible nitric oxide synthase (iNOS) expression. Oral administration of 500 mg/kg of myrrh and thyme led to adult reduction (90.9%, 79.4%), while 1,000 mg/kg led to larvae reduction (79.6%, 71.3%), respectively. Administration of 50 mg/kg of albendazole resulted in adult and larvae reduction (94.2%, 90.9%). Positive immunostaining of inflammatory cells infiltrating intestinal mucosa and submucosa of all treated groups was detected. Myrrh-treated mice showed the highest iNOS expression followed by albendazole, then thyme. On the other hand, both myrrh and thyme-treated groups showed stronger iNOS expression of inflammatory cells infiltrating and surrounding encapsulated T. spiralis larvae than albendazole treated group. In conclusion, myrrh and thyme extracts are highly effective against both phases of T. spiralis and showed strong iNOS expressions, especially myrrh which could be a promising alternative drug. This experiment provides a basis for further exploration of this plant by isolation and retesting the active principles of both extracts against different stages of T. spiralis.

Identificación de subpoblaciones de macrófagos y linfocitos T en pulmones de pacientes fallecidos por influenza A H1N1

La infección por el virus de la influenza es un problema de salud pública por su rápida diseminación y alta morbilidad. Los casos graves de infección por éste virus presentan hipercitocinemia, la cual se ha asociado a una respuesta inmune adquirida desfavorable y un mal pronóstico para el paciente. Se han realizado hasta ahora experimentos en suero de pacientes o en tejido pulmonar en modelos de animales, sin embargo, no se tienen reportes del microambiente en el pulmón de pacientes fallecidos por el virus de la influenza. Objetivo: Determinar las subpoblaciones de macrófagos y linfocitos T y su correlación con el daño tisular en pulmón de pacientes fallecidos por influenza A H1N1 y otras enfermedades respiratorias. Material y Métodos: Se identificó y cuantificó el virus de influenza A H1N1 (pdm)09 e influenza A estacional por qRT-PCR, así como se determinó los niveles de citocinas y marcadores de macrófagos por qRT-PCR (IL-2, IL-4,IL-6, IL- 10, IL-12, IL-17, IL-23, IFN-, TGFβ, TNFα, Arg1, Retnlb e iNOS). Se realizaron tinciones de HyE para la determinación del daño tisular. Por otra parte, se realizaron tinciones de inmunohistoquímica para analizar las poblaciones celulares CD14+, CD206+, CD4+, CD8+, FOXP3+ y citocinas (IL-4, IL-10, IL-17 e IFN-) in situ. Resultados: Se determinó la presencia del virus de la influenza A en 10 muestras, de las cuales 4 fueron H1N1 (pdm)09. Además, se obtuvieron 5 muestras de neumonía por Coccidioides spp., y 6 de neumonías de origen bacteriano. No existe diferencia en el daño causado por el virus de la influenza A H1N1 (pdm)09 e influenza A estacional a nivel histopatológico. Las células CD14+ y CD4+ se encontraron aumentadas para todos los grupos de neumonías sin importar el agente etiológico, excepto CD4 para las neumonías bacterianas. Las células que expresaron Foxp3 solo se encontraron aumentadas en el grupo de coccidioidomicosis y neumonías bacterianas. No se encontró diferencia significativa entre los grupos de estudio para las células CD8+, excepto para las neumonías bacterianas. La expresión génica relativa de IL-6 se encontró aumentada 3000 veces la expresión génica en el grupo de influenza pandémica A H1N1 (pdm)09, mientras en influenza estacional se encontró una disminución de 0.08 veces de su expresión. INOS se encontró con expresión disminuida 0.5 veces para los grupos de influenza pandémica, influenza estacional y coccidioidomicosis. La expresión génica de IL-10 se encontró solamente para el grupo de influenza A H1N1 (pdm)09 (P=0.01). Resistin Like Beta se encontró con expresión disminuida solo para el grupo de influenza A H1N1 (pdm)09. Existe un aumento de células positivas para IL4 en todos los grupos, excepto neumonías bacterianas. No se encontró diferencia significativa de células positivas para IL-10 en los grupos de estudio. Existe un aumento de células positivas para IL-17 en influenza A H1N1p/2009, influenza A y neumonías bacterianas. IFN se encontró aumentado en los grupos de neumonía comparados con el control. Conclusiones: Ambos grupos de neumonías por influenza A se caracterizan por daño tisular y un exacerbado ambiente inflamatorio; solamente CD206 es capaz de diferenciar entre influenza A H1N1(pdm)09 e influenza estacional

Anti-oxidative and anti-neuroinflammatory effects of ethyl acetate extract fractions of Suaeda asparagoides MIq

Purpose: To evaluate the in vitro antioxidant and anti-neuroinflammatory effects of Suaeda asparagoides ethylacetate extract (SAE) in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Methods: The antioxidative activity of SAE was evaluated by measuring 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity spectrometrometrically. Cell viability was evaluated by 3-(4, 5dimethylthiazol-2-yl)-2, 5- diphenyl-tetrazolium bromide (MTT) assay, while LPS-stimulated BV-2 microglia were used to study the expression and production of inflammatory mediators, including nitric oxide (NO), inducible NO synthase (iNOS) and tumor necrosis alpha (TNF-α). Results: Pretreatment with SAE prior to LPS treatment significantly inhibited excessive production of NO (p < 0.001 at 20, 40, 80 and 100 μg/mL) in a dose-dependent manner, and was associated with down-regulation of expression of inducible nitric oxide synthase (iNOS). SAE also suppressed the LPSinduced increase in TNF-α level (p < 0.01at concentrations of 40 and 80 μg/mL) in BV-2 cells. Furthermore, DPPH-generated free radicals were inhibited by SAE in a concentration-dependent manner. Conclusion: These results indicate that SAE possesses strong anti-oxidant properties, and inhibits excessive production of pro-inflammatory mediators, including NO, iNOS and TNF-α, in LPS-stimulated BV-2 cells

Antiviral activity and mechanism of action of arbidol against Hantaan virus infection

Purpose: To investigate the activity and mechanism of action of arbidol against Hantaan virus (HTNV) activity by modulating inflammation via TLR-4 pathway. Methods: HUVEC cells infected with HTNV 76-118 were treated with serially diluted arbidol solutions at -2h (2 h before viral infection, pre-treatment mode), 0 h (at the same time as viral infection, simultaneous treatment mode) or 2 h (2 h after viral infection, post-treatment mode). The transcript levels of TLR4 were detected by semi-quantitative reverse transcription-PCR (RT-PCR) at 6, 12, 18, and 24 h later. The levels of iNOS and TNF-α were examined using enzyme-linked immunosorbent assay (ELISA). Results: Pre-treatment with arbidol, rather than simultaneous treatment or post-treatment, effectively inhibited up-regulation of cellular TLR4 expression (up to 40 ± 6.1 % inhibition) and activity of supernatant iNOS induced by HTNV infection(up to 44.1 ± 9.4 % inhibition), as well as in a LPSstimulated inflammatory endothelial cell. Arbidol decreased the elevated TNF-α levels induced by LPS stimulation. Conclusion: These results are the first evidence that arbidol modulates viral PRRs signaling and its consequential inflammatory cytokine/chemokine response during hantavirus infection.

Avaliação do extrato do fungo caripia montagnei e de agonistas de PPAR - α no processo inflamatório

The mushrooms have been object of intense research in view of its potential raising of application in different sectors of the pharmacology and alimentary industry. Among diverse bioactive composites of polyssacharides nature that exist in the fungus the glucans are much searched. These are polymers of glucose and classified as the type of glicosidic linking [α, β]. Peroxisome proliferator-activated receptors (PPARs), ranscription factors belonging to the family of nuclear receptors that bind themselves o specific agonists, have shown their importance in controlling the inflammatory process. The aim of this study was to perform a chemical characterization of extract rom the mushroom Caripia montagnei, assess its antiinflammatory and antibacterial effect and determine if this effect occurs via PPAR. This mushroom is composed of carbohydrates (63.3±4.1%), lipids (21.4l±0.9%) and proteins (2.2± 0.3%). The aqueous solution resulting from the fractionation contained carbohydrates (98.7±3.3%) and protein (1.3±0.25%). Analyses of infrared spectrophotometry and of nuclear magnetic esonance demonstrated that the extract of mushroom C. montagnei is rich in β-glucans. In hioglycolate-induced peritonitis, the C. montagnei glucans (50 mg/kg) educed the inflammatory process in 65.5±5.2% and agonists, pharmacological igands, for PPAR: Wy-14643 (49.3±6.1%), PFOA (48.9±3.8%) and clofibrate in 45.2±3.2%. Sodium diclofenac showed a reduction of 81.65±0.6%. In the plantar edema, the glucans from C. montagnei (50 mg/kg) and L-NAME reduced the edema to a similar degree 91.4±0.3% and 92.8±0,5 %, respectively. In all the groups tested, nitric oxide (NO), an inflammation mediator, showed a significant reduction in the nitrate/nitrite levels when compared to the positive control (P<0.001). The C. montagnei glucans did not show cytotoxicity in the concentrations tested (2.5, 5.0, 10.0, 20.0 and 40.0 µg/100 µL). Antibacterial activity demonstrated that, unlike total extract, there was no inhibition of bacterial growth. The C. montagnei glucans show great potential for antiinflammatory applications. This effect suggests that it is mediated by PPAR activation and by COX and iNOS inhibition