RESUMEN
In the present state of overwhelming emergence of drug-unresponsive phenotypes of Leishmania donovani and persistent severe toxicity in conventional anti-leishmanial therapy, in search for novel leads, the aim of this study has been fixed to identify the active extract(s) of Croton caudatus Geisel. var. tomentosus Hook effective against the parasitic protozoans in vitro and in vivo. C. caudatus Geisel. is often used by Chakma and Hmar community, the local tribes of north-east India for medicinal and veterinary purposes. Among the five semi-purified extracts tested, C. caudatus leaves, extracted in hexane and subsequently semi-purified in a column packed with silica gel (70-130 µM; mesh size 60 A°) using ethyl acetate-hexane solvent (9:1), was found to be the most effective growth inhibitor (JDHex) against the Leishmania promastigotes and amastigotes. JDHex significantly altered the biochemical parameters (protein, lipid and carbohydrates) in promastigotes followed by the morphological changes, DNA condensation and subsequent apoptosis in L. donovani. In consequent steps, it has been also proved that JDHex reduced the replication of intracellular amastigotes with concomitant release of nitric oxide and pro-inflammatory cytokines, IL-12 and TNF-α in vitro. Significantly, the 50% inhibitory concentration of JDHex was estimated much lower against the intracellular amastigotes (2.5 µg/mL) in comparison to promastigotes (10 µg/mL). JDHex was also found efficient in reducing parasite burden in spleen and liver when treated in vivo and increased the intracellular IFN-γ and decreased the IL-10 in CD4+ T cells in splenocytes of orally treated animals. The results of this study support the importance in exploration of novel anti-leishmanial leads from C. caudatus Geisel. var. tomentosus Hook. against the L. donovani (MHOM/IN/83/AG83) infection. Partial chemical characterization of JDHex revealed the presence of terpenoids. However, the further chemical investigation of JDHex is warranted.
Asunto(s)
Croton/química , Citocinas/metabolismo , Leishmania donovani/efectos de los fármacos , Leishmaniasis Visceral/prevención & control , Extractos Vegetales/uso terapéutico , Animales , Apoptosis , Citocinas/inmunología , Relación Dosis-Respuesta a Droga , Concentración 50 Inhibidora , Leishmania donovani/crecimiento & desarrollo , Leishmania donovani/ultraestructura , Leishmaniasis Visceral/tratamiento farmacológico , Leishmaniasis Visceral/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Macrófagos/parasitología , Masculino , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica de Rastreo , Óxido Nítrico/metabolismo , Fosfatidilserinas/metabolismo , Extractos Vegetales/farmacología , Hojas de la Planta/química , Bazo/citología , Bazo/efectos de los fármacos , Bazo/inmunologíaRESUMEN
Anticancer and immunostimulatory properties of tryptic digest peptides of Abrus precatorius agglutinin protein (10kDAGP) have already been reported. Here attempt has been made to further validate anticancer properties of 10kDAGP peptides in Ehrlich's ascites carcinoma (EAC) and B16 melanoma (B16M) bearing mice models and to analyze 10kDAGP by anion exchange chromatography and RP-HPLC for obtaining the bioactive fraction from the total peptide pool. 10kDAGP treatment decreased the tumour pack volume by â¼82% for EAC and 58.8% for B16M. It also showed increase in ex vivo proliferation of splenocyte and thymocyte isolated from tumour bearing mice and increase in TNF-α and Interferon-γ in splenocyte culture supernatant. From chromatographic analysis it was found that anionic peptide fraction may be responsible for anti-proliferative activities of 10kDAGP. As most anticancer peptides are cationic in nature, further studies regarding bioactivity of anionic peptide fraction may lead to novel anticancer peptides and pathways of action.
Asunto(s)
Abrus , Antineoplásicos Fitogénicos/uso terapéutico , Carcinoma de Ehrlich/tratamiento farmacológico , Melanoma Experimental/tratamiento farmacológico , Péptidos/uso terapéutico , Aglutininas/química , Animales , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Carcinoma de Ehrlich/patología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Femenino , Células HeLa , Humanos , Melanoma Experimental/patología , Ratones , Ratones Endogámicos C57BL , Péptidos/química , Péptidos/farmacología , Fitoterapia , Bazo/citología , Timo/citología , Carga Tumoral/efectos de los fármacosRESUMEN
A water soluble new arabinoxylan, isolated through hot water extraction from the green leaves of Litsea glutinosa (Lauraceae) was found to contain xylose and arabinose in a molar ratio of nearly 1:3. On the basis of NMR ((1)H, (13)C, DQF-COSY, TOCSY, NOESY, ROESY, HMBC and DEPT-135), GLC and GLC-MS analyses, the backbone was established as (1â4)-α-D-xylopyranosyl residue, substituted at C-2 with one unit of two adjacently linked (1â3)-α-L-arabinofuranosyl residues and the other one was terminated by ß-L-arabinofuranosyl residue. The proposed repeating unit of the molecule was established as: [formula, see text] This molecule showed strong splenocyte, thymocyte, and macrophage activations. The optimum doses of the polysaccharide for splenocyte and thymocyte proliferation were observed at 25 µg/mL and 50 µg/mL, respectively. An enhanced production of NO was observed at 100 µg/mL of the polysaccharide.
Asunto(s)
Litsea/química , Hojas de la Planta/química , Xilanos/química , Xilanos/farmacología , Animales , Secuencia de Carbohidratos , Proliferación Celular/efectos de los fármacos , Isomerismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Datos de Secuencia Molecular , Óxido Nítrico/biosíntesis , Extractos Vegetales/química , Bazo/citología , Bazo/efectos de los fármacos , Timocitos/citología , Timocitos/efectos de los fármacos , Agua/química , Xilanos/aislamiento & purificaciónRESUMEN
An immunostimulating water-insoluble ß-glucan isolated from hot alkaline extract of the fruiting bodies of an edible somatic hybrid mushroom of Pleurotus florida and Calocybe indica var. APK2 showed significant macrophage, splenocyte, and thymocyte activations. On the basis of total hydrolysis, methylation analysis, and NMR experiments ((1)H, (13)C, DQF-COSY, TOCSY, NOESY, DEPT-135, and HSQC), the repeating unit of the polysaccharide is established.
Asunto(s)
Agaricales/química , Agaricales/genética , Macrófagos Peritoneales/efectos de los fármacos , beta-Glucanos/metabolismo , beta-Glucanos/farmacología , Animales , Conformación de Carbohidratos , Metabolismo de los Hidratos de Carbono , Células Cultivadas , Cuerpos Fructíferos de los Hongos/química , Espectroscopía de Resonancia Magnética , Ratones , Bazo/citología , Timocitos/citología , Timocitos/efectos de los fármacos , beta-Glucanos/químicaRESUMEN
A water soluble heteroglycan, isolated from the alkaline extract of the fruit bodies of the somatic hybrid mushroom (PfloVv1aFB), raised through protoplast fusion between the strains of Pleorutus florida and Volverilla volvacea, was found to consist of (1â3)-, (1â6)-, (1â3,4)-linked, and terminal ß-D-Glcp along with (1â2,6)-α-D-Galp and terminal α-D-Manp in a relative proportion of approximately 1:1:1:1:1:1. This polysaccharide exhibited strong immunostimulating activity of macrophages as well as splenocytes and thymocytes. Structural investigation was carried out using sugar analysis, methylation analysis; periodate oxidation study, and NMR experiments ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC). On the basis of the above mentioned experiments, the structure of the repeating unit of the polysaccharide was established as: [formula see text].
Asunto(s)
Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/farmacología , Pleurotus/química , Polisacáridos/química , Polisacáridos/inmunología , Volvariella/química , Adyuvantes Inmunológicos/aislamiento & purificación , Álcalis/química , Animales , Conformación de Carbohidratos , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Frutas/química , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Pleurotus/inmunología , Polisacáridos/aislamiento & purificación , Especies Reactivas de Oxígeno/inmunología , Bazo/citología , Bazo/efectos de los fármacos , Bazo/inmunología , Relación Estructura-Actividad , Timocitos/efectos de los fármacos , Timocitos/inmunología , Volvariella/inmunologíaRESUMEN
Two different glucans (PS-I and PS-II) were isolated from the alkaline extract of the fruiting bodies of an edible mushroom, Pleurotus florida blue variant and the PS-I showed macrophage, splenocyte and thymocyte activations. On the basis of sugar analysis, methylation analysis, periodate oxidation, and NMR studies ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC and HMBC), the structure of the repeating unit of these polysaccharides were established.
Asunto(s)
Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/farmacología , Glucanos/química , Glucanos/farmacología , Pigmentación , Pleurotus/química , Adyuvantes Inmunológicos/aislamiento & purificación , Secuencia de Carbohidratos , Glucanos/aislamiento & purificación , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Datos de Secuencia Molecular , Solubilidad , Bazo/efectos de los fármacos , Bazo/inmunología , Timocitos/efectos de los fármacos , Timocitos/inmunología , Agua/químicaRESUMEN
Collagens presently used in tissue engineering are primarily of bovine or porcine origin. However, a risk of a spongiform encephalopathy epidemic has limited the use of collagen from these sources. Keeping the aforementioned perspective in mind, we explored the possibility of using domestic goat available in the subcontinent as a potential source of collagen for tissue-engineering application. This article delineates the isolation, physico-chemical characterization, biocompatibility study and wound healing application of acid soluble caprine (goat) tendon collagen (GTC). Physico-chemical characterization of 1% acetic acid extracted GTC was done by SDS-PAGE, amino-acid composition analysis, FT-IR and CD spectroscopy. Results revealed that GTC was comprised of type-I collagen. Biocompatibility study showed that GTC augmented cell adhesion, cell cycle progression and proliferation. Immuno-cytochemical analysis in conjugation with traction force microscopy further confirmed a superior focal adhesion complex mediated cell-substrate interaction in GTC. Finally, in vivo study in mice model revealed that GTC has low immunogenicity and it augments healing process significantly. Throughout the study, calf skin collagen (CSC) was used as standard for comparative evaluation. In conclusion, it can be said that GTC may find its application as biomaterial in skin tissue engineering.
Asunto(s)
Materiales Biocompatibles/farmacología , Colágeno/farmacología , Cabras , Piel/citología , Piel/efectos de los fármacos , Ingeniería de Tejidos/métodos , Andamios del Tejido , Actinas/metabolismo , Animales , Materiales Biocompatibles/química , Bovinos , Adhesión Celular/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colágeno/química , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Concentración de Iones de Hidrógeno , Masculino , Ratones , Fosfoproteínas/metabolismo , Piel/inmunología , Solubilidad , Andamios del Tejido/química , Cicatrización de Heridas/efectos de los fármacosRESUMEN
Two different glucans (water-soluble PS-I, water-insoluble PS-II) were isolated from the alkaline extract of the fruit bodies of hybrid mushroom. PS-I was found to consist of only (1â6)-linked ß-D-glucopyranose. PS-II was composed of terminal, (1â3,4)-linked, and (1â3)-linked ß-D-glucopyranosyl moieties in a molar ratio of nearly 1:1:1. PS-I showed macrophages, splenocytes, and thymocytes activation as well as antioxidant property. On the basis of sugar analysis, methylation analysis, and NMR studies ((1)H, (13)C, DEPT-135, TOCSY, DQF-COSY, NOESY, ROESY, HMQC, and HMBC), the structure of the repeating unit of these glucans were established as:
Asunto(s)
Glucanos/química , Glucanos/farmacología , Hibridación Genética , Pleurotus/química , Pleurotus/genética , Volvariella/química , Volvariella/genética , Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/aislamiento & purificación , Adyuvantes Inmunológicos/farmacología , Animales , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Compuestos de Bifenilo/química , Secuencia de Carbohidratos , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Glucanos/aislamiento & purificación , Concentración de Iones de Hidrógeno , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/inmunología , Ratones , Datos de Secuencia Molecular , Picratos/química , Solubilidad , Bazo/citología , Timocitos/citología , Timocitos/efectos de los fármacos , Agua/químicaRESUMEN
A water-soluble polysaccharide of an edible mushroom Calocybe indica var. APK2 showed immunoenhancing (macrophage, splenocyte, thymocyte, and bone marrow activation) and cytotoxic activity toward HeLa cell lines and found to consist of D-glucose, D-galactose, and L-fucose in a molar ratio of nearly 3:1:1. On the basis of acid hydrolysis, methylation analysis, and NMR studies ((1)H, (13)C, DEPT-135, TOCSY, DQF-COSY, NOESY, ROESY, HMQC, and HMBC), the structure of the repeating unit of the fuco-galacto-glucan was established as: [see formula in text].
Asunto(s)
Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/farmacología , Agaricales/química , Antineoplásicos/química , Antineoplásicos/farmacología , Polisacáridos/química , Polisacáridos/farmacología , Adyuvantes Inmunológicos/aislamiento & purificación , Animales , Antineoplásicos/aislamiento & purificación , Secuencia de Carbohidratos , Proliferación Celular/efectos de los fármacos , Células HeLa , Humanos , Espectroscopía de Resonancia Magnética , Ratones , Datos de Secuencia Molecular , Células 3T3 NIH , Polisacáridos/aislamiento & purificaciónRESUMEN
A water-soluble glucan, isolated from the alkaline extract of the fruit bodies of an edible mushroom, Lentinus squarrosulus (Mont.) Singer was found to consist of (1â3,6)-linked, (1â3)-linked, (1â6)-linked, and terminal ß-d-glucopyranosyl moieties in a relative proportion of approximately 1:2:1:1. This polysaccharide showed optimum activation of macrophages as well as splenocytes and thymocytes at 10µg/mL. Structural investigation was carried out using sugar analysis, methylation analysis, periodate oxidation study, and NMR experiments ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC). On the basis of above-mentioned experiments, the structure of the repeating unit of the polysaccharide was established as:
Asunto(s)
Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/aislamiento & purificación , Agaricales/química , Glucanos/química , Glucanos/aislamiento & purificación , Lentinula/química , Adyuvantes Inmunológicos/farmacología , Animales , Secuencia de Carbohidratos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Glucanos/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Ratones , Datos de Secuencia Molecular , Bazo/citología , Timocitos/citología , Timocitos/efectos de los fármacosRESUMEN
A water-soluble glucan isolated from an alkaline extract of fruit bodies of a somatic hybrid mushroom PCH9FB of Pleurotus florida and Calocybe indica var. APK2 strains showed antioxidant properties with immune activation of macrophage, splenocyte, and thymocyte. On the basis of acid hydrolysis, methylation, periodate oxidation and NMR studies ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC) the structure of the repeating unit of the glucan was established as: [structure: see text].
Asunto(s)
Álcalis/química , Antioxidantes/farmacología , Técnicas de Química Analítica/métodos , Glucanos/inmunología , Glucanos/aislamiento & purificación , Hibridación Genética , Pleurotus/química , Animales , Proliferación Celular/efectos de los fármacos , Glucanos/síntesis química , Glucanos/farmacología , Activación de Macrófagos/efectos de los fármacos , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Ratones , Óxido Nítrico/farmacología , Bazo/citología , Timocitos/citología , Timocitos/efectos de los fármacosRESUMEN
Mushroom-derived polysaccharides like ß-glucan are being investigated for therapeutic properties for a long time, but their mode of action of immunomodulatory properties is not well established. In the present study, a heteroglucan from Astraeus hygrometricus designated as AE2 is investigated for its macrophage stimulatory properties using RAW 264.7 cell line. An augmentation of nitric oxide production is observed in the presence of AE2 in a dose-dependent manner due to up-regulation of iNOS (inducible NO synthase) expression; hence NF κB (nuclear factor κB) pathway is investigated. RAW 264.7 cells endured phosphorylation of Ikk (IκB kinase) and subsequently NF κB is translocated to the nucleus. Further, the PKC (protein kinase C) level of the cells enhanced significantly. We also found that AE2 could induce the phosphorylation of p38 MAPK (mitogen-activated protein kinase), ERK1/2 (extracellular-signal-regulated kinase 1/2), MEK (MAPK/ERK kinase) and JNK (c-Jun N-terminal kinase), whereas it failed to induce phosphorylation of JAK2 (Janus kinase 2) and STAT1. These results indicated that the macrophage activation by AE2 might be exerted, at least in part, via MAPKs (mitogen-activated protein kinases) pathway of signal transduction.
Asunto(s)
Basidiomycota/metabolismo , Glucanos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Animales , Línea Celular Tumoral , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos/genética , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Janus Quinasa 2/genética , Janus Quinasa 2/metabolismo , Ratones , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosforilación , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT1/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
A water soluble polysaccharide isolated from the hot aqueous extract of Pleurotusostreatus cultivar was found to contain d-glucose and d-galactose in a molar ratio of nearly 7:1. Structural investigation was carried out using acid hydrolysis, methylation analysis, periodate oxidation, Smith degradation, and NMR studies ((1)H, (13)C, DEPT-135, TOCSY, DQF-COSY, NOESY, ROESY, HMQC, and HMBC). On the basis of the above mentioned experiments the structure of the repeating unit of the polysaccharide was established as: This heteroglycan stimulates macrophages, splenocytes, and thymocytes.
Asunto(s)
Pleurotus/química , Polisacáridos/química , Secuencia de Carbohidratos , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Polisacáridos/aislamiento & purificación , Agua/químicaRESUMEN
Specific targeting of tumor cells to achieve higher drug levels in tumor tissue and to overcome the side effects is the major goal in cancer therapy. Nanoparticles encapsulating a hydrophobic core in their nanoreservoir structure were developed as a carrier for a water-insoluble drug, paclitaxel. In the present study, target-oriented nanoparticles based on biodegradable O-carboxymethyl chitosan modified with stearic acid. The surface of the nanoparticles was modified by covalent attachment of folic acid (FA) by simple carbodimide reaction to achieve tumor cell targeting property. Nanoparticles were prepared by the sonication method without involving any surfactants/emulsifiers. The nanoparticles were characterized by various state-of-the-art techniques, including laser light scattering for particles size distribution, field emission scanning electron microscopy and transmission electron microscope for surface morphology. The drug release property and the cytotoxicity of the drug loaded nanoparticles to both cancerous and noncancerous cells were evaluated in cell culture system. To our knowledge, this is the first study demonstrating a FA modified hydrophobically chitosan with paclitaxel-loaded nanoparticles targeting of folate receptor overexpressing cancer cells.
Asunto(s)
Antineoplásicos Fitogénicos/administración & dosificación , Quitosano/análogos & derivados , Sistemas de Liberación de Medicamentos , Paclitaxel/administración & dosificación , Animales , Antineoplásicos Fitogénicos/farmacología , Quitosano/química , Portadores de Fármacos/química , Ácido Fólico/química , Células HeLa , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Ratones , Células 3T3 NIH , Nanopartículas , Paclitaxel/farmacología , Tamaño de la Partícula , Ácidos Esteáricos/química , Propiedades de SuperficieRESUMEN
Amphiphilic biodegradable succinylchitosan nanoparticles modified with folic acid are described that act as an emulsifier to form nanoparticles. Their molecular structures and physicochemical as well as self-assembly properties are characterized by means of FT-IR, ¹H NMR, FESEM, DLS, and TEM. The nanoparticles are 60-80 nm in size and are not toxic in vitro. They are immobilized with the cytostatic drug doxorubicin. Specific transport of doxorubicin by the nanoparticles into the folate-receptor-overexpressing cancer cells and its biological activity as well as in vitro release are demonstrated. It is shown that under acidic condition more drug is released. The nanoparticles can thus not only specifically deliver doxorubicin to its target, but also release the drug depending on the pH.
Asunto(s)
Quitosano/química , Doxorrubicina/farmacología , Sistemas de Liberación de Medicamentos/métodos , Ácido Fólico/química , Nanopartículas/química , Animales , Ciclo Celular/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Doxorrubicina/química , Citometría de Flujo , Células HeLa , Humanos , Concentración de Iones de Hidrógeno/efectos de los fármacos , Indoles/metabolismo , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Luz , Ratones , Células 3T3 NIH , Nanopartículas/ultraestructura , Tamaño de la Partícula , Dispersión de Radiación , Coloración y Etiquetado , Propiedades de Superficie/efectos de los fármacosRESUMEN
A water-soluble polysaccharide was isolated from the aqueous extract of the fruit bodies of somatic hybrid PCH9FB, obtained through intergeneric protoplast fusion between the strains Pleurotus florida and Calocybe indica var. On the basis of total acid hydrolysis, the polysaccharide was found to contain galactose, fucose, and glucose in a molar ratio of nearly 2:1:2. Methylation analysis and NMR experiments ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC) showed that the structure of the repeating unit present in the polysaccharide was This molecule showed macrophage, splenocyte, thymocyte activation as well as antioxidant property.
Asunto(s)
Agaricales/química , Antioxidantes/metabolismo , Hibridación Genética , Pleurotus/química , Polisacáridos/química , Polisacáridos/inmunología , Agua/química , Animales , Proliferación Celular , Activación de Macrófagos , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/metabolismo , Espectroscopía de Resonancia Magnética , Ratones , Óxido Nítrico/metabolismo , Polisacáridos/aislamiento & purificación , Bazo/citología , Timo/citologíaRESUMEN
Previously, we reported the in vitro anticancer and immunomodulatory effect of a protein fraction designated as Cibacron blue affinity purified protein (CBAEP) obtained from the culinary-medicinal oyster mushroom, Pleurotus ostreatus. In the present study, we investigated the in vivo antitumor potential of CBAEP in different tumor-bearing mice models and studied the detailed mechanism of tumor regression in Dalton lymphoma (DL)-bearing mice. The lethal dose (LD50) of CBAEP was found to be 55 mg/kg body weight and sublethal doses (5 mg/kg and 10 mg/kg body weight) showed a prolonged tumor survival time in DL, Sarcoma-180, and B16F0 melanoma tumor-bearing mice. Further, CBAEP reduced about 35.68 and 51.43% DL cell growth in 5 and 10 mg/kg body weight, respectively. The in vivo CBAEP treatment showed an apoptotic feature as demonstrated in morphological study and sub-G0/G1 population in cell cycle and Western blot of DL cells. CBAEP also activated immunosuppression condition in DL tumor-bearing host. It also stimulated immune cells in the presence of nonspecific immunostunulator (LPS and ConA) ex vivo as well as enhanced Th1 response with production of TNF-α, IFN-γ, and IL-2. Moreover, it activated tumor-associated macrophages and NK cells. The present findings revealed the potent antitumor property of CBAEP, which might help in developing a new anticancer drug.
Asunto(s)
Antineoplásicos/farmacología , Proteínas Fúngicas/uso terapéutico , Linfoma/tratamiento farmacológico , Pleurotus/química , Sarcoma 180/tratamiento farmacológico , Animales , Antineoplásicos/química , Ratones , Ratones Endogámicos C57BLRESUMEN
An immunoenhancing polysaccharide isolated from the aqueous extract of the fruit bodies of the mushroom, Pleurotus florida blue variant, was found to consist of d-glucose and d-galactose in a molar ratio of nearly 5:1. On the basis of sugar analysis, methylation analysis, periodate oxidation, Smith degradation, and NMR studies ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC), the structure of the repeating unit of the polysaccharide was established as: â6)-ß-D-Glcp-(1â3)-α-D-glcp-(1â[6)-ß-D-Glcp-(1]3â α-D=Glcp. The molecule activated macrophages, splenocytes, and thymocytes.
Asunto(s)
Pleurotus/química , Polisacáridos/química , Polisacáridos/farmacología , Agua/química , Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/farmacología , Animales , Secuencia de Carbohidratos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Macrófagos/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Ratones , Datos de Secuencia Molecular , Óxidos de Nitrógeno/metabolismo , SolubilidadRESUMEN
A water-soluble polysaccharide, isolated from the hot water extract of the fruiting bodies of an edible mushroom, Lentinus squarrosulus (Mont.) Singer was found to contain d-galactose, l-fucose, and d-glucose in a molar ratio of nearly 1:1:5. Structural investigation of the polysaccharide was carried out using acid hydrolysis, methylation analysis, periodate oxidation study, and NMR experiments ((1)H, (13)C, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC). On the basis of above-mentioned experiments, the structure of the repeating unit of the polysaccharide was established as: This polysaccharide showed macrophage as well as splenocyte and thymocyte activation.
Asunto(s)
Factores Inmunológicos/química , Factores Inmunológicos/inmunología , Lentinula/química , Polisacáridos/química , Polisacáridos/inmunología , Agua/química , Animales , Secuencia de Carbohidratos , Factores Inmunológicos/aislamiento & purificación , Macrófagos/inmunología , Espectroscopía de Resonancia Magnética , Ratones , Datos de Secuencia Molecular , Monosacáridos/análisis , Polisacáridos/aislamiento & purificación , Bazo/citología , Bazo/inmunología , Timo/citología , Timo/inmunologíaRESUMEN
Mushrooms have been used as food and for therapeutic purpose for decades, and various compounds derived from these have potential biological activities. Application of such agents of natural origin is vital with relevance to an increase in diseases involving immune dysfunction, cancer, and autoimmune conditions in recent years. These compounds belong mainly to polysaccharides, especially beta-D-glucan, glycopeptide/protein complexes, proteoglycans, proteins, and triterpenoids. Among these compounds, polysaccharides and their peptide/protein derivates have extremely important roles in immunomodulating activities. The present study aims to explore the immunomodulatory properties of polysaccharide isolated from the mushroom Astraeus hygrometricus. The polysaccharide isolated by hot alkaline extraction and chromatography was designated as AE2 and studied for its immunostimulatory potential in vivo in a murine model. Macrophages from treated mice showed higher production of nitric oxide and interleukin-1 and also exhibited an increased phagocytic potential. It also enhanced natural killer cell activation and proliferation of splenocytes with an increase in the T-helper (Th) 1 cytokine level of splenocyte culture supernatant. The cytokine level in serum also indicated a Th1 skew. The observed activities were associated with no general toxicity to the organism. The findings will be helpful in considering A. hygrometricus as a potential source of an immunomodulator and in designing further studies to understand its mode of action on immune system.