RESUMEN

Mushroom ß-D-glucans can be isolated from several species, including the widely consumed Agaricus bisporus. Besides immunomodulatory responses, some ß-D-glucans may exhibit direct antitumoral effects. It was previously observed that a ß-(1→6)-D-glucan (BDG16) has indirect cytotoxicity on triple-negative breast cancer cells. In this study, the cytotoxicity of this same glucan was observed on estrogen receptor-positive (ER+) breast cancer cells (MCF-7). Cell viability was determined by multiple methods to assess metabolic activity, lysosomal membrane integrity, and adhesion capacity. Assays to evaluate cell respiration, cell cycle, apoptosis, necroptosis, and oxidative stress were performed to determine the action of BDG16 on MCF-7 cells. A gradual and significant cell viability reduction was observed when the cells were treated with BDG16 (10-1000 µg/mL). This result could be associated with the inhibition of the basal state respiration after incubation with the ß-D-glucan. The cells showed a significant arrest in G1 phase population at 1000 µg/mL, with no induction of apoptosis. However, an increase in necrosis and necroptosis at the same concentration was observed. No difference in oxidative stress-related molecules was observed. Altogether, our findings demonstrate that BDG16 directly induces toxicity in MCF-7 cells, primarily by impairing mitochondrial respiration and promoting necroptosis. The specific mechanisms that mediate this action are being investigated.

Asunto(s)

Agaricus , Antineoplásicos , Apoptosis , Neoplasias de la Mama , Supervivencia Celular , Estrés Oxidativo , Receptores de Estrógenos , Agaricus/química , Humanos , Células MCF-7 , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Femenino , Supervivencia Celular/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Receptores de Estrógenos/metabolismo , Antineoplásicos/farmacología , Antineoplásicos/química , beta-Glucanos/farmacología , beta-Glucanos/química

RESUMEN

C. brachyspora, a widespread dematiaceous fungus, was evaluated in this study to optimize the production of exopolysaccharides (CB-EPS). Optimization was performed using response surface methodology, and the best production yielded 75.05% of total sugar at pH 7.4, with 0.1% urea, after 197 h. The obtained CB-EPS showed typical signals of polysaccharides, which was confirmed by FT-IR and NMR. The HPSEC analysis indicated a polydisperse polymer, showing a non-uniform peak, with an average molar mass (Mw) of 24,470 g/mol. The major monosaccharide was glucose (63.9 Mol%), followed by mannose (19.7 Mol%), and galactose (16.4 Mol%). Methylation analysis encountered derivatives that indicated the presence of a ß-d-glucan and a highly branched glucogalactomannan. CB-EPS was tested on murine macrophages to verify its immunoactivity, and the treated cells were able to produce TNF-α, IL-6, and IL-10. However, the cells did not produce superoxide anions or nitric oxide nor stimulated phagocytosis. The results demonstrated an indirect antimicrobial activity of macrophages by stimulating cytokines, showing another biotech applicability for the exopolysaccharides produced by C. brachyspora.

Asunto(s)

Macrófagos , Polisacáridos , Animales , Ratones , Espectroscopía Infrarroja por Transformada de Fourier , Polisacáridos/farmacología , Biotecnología

RESUMEN

Liquidambar styraciflua L. is an aromatic species, popularly used in traditional Chinese medicine to treat diarrhea, dysentery, coughs, and skin sores. The present study was designed to investigate the chemical composition and biological potential of extracts obtained from the fruits of this plant. For the chemical evaluation, it was used mainly liquid and gas chromatography, plus NMR, and colorimetric methods. The aqueous extract (EA) originated two other fractions: an aqueous (P-EA) and an ethanolic (S-EA). The three extracts were composed of proteins, phenolic compounds, and carbohydrates in different proportions. The analyses showed that the polysaccharide extract (P-EA) contained pectic polysaccharides, such as acetylated and methyl esterified homogalacturonans together with arabinogalactan, while the fraction S-EA presented phenolic acids and terpenes such as gallic acid, protocathecuic acid, liquidambaric acid, combretastatin, and atractyloside A. EA, P-EA, and S-EA showed antioxidant activity, with IC50 values of 4.64 µg/mL, 16.45 µg/mL, and 3.67 µg/mL, respectively. The cytotoxicity followed the sequence S-EA > EA > P-EA, demonstrating that the toxic compounds were separated from the non-toxic ones by ethanol precipitation. While the fraction S-EA is very toxic to any cell line, the fraction P-EA is a promising candidate for studies against cancer due to its high toxicity to tumoral cells and low toxicity to normal cells.

Asunto(s)

Antineoplásicos , Liquidambar , Frutas , Extractos Vegetales/farmacología , Extractos Vegetales/química , Liquidambar/química , Antioxidantes/farmacología , Antioxidantes/química , Antineoplásicos/farmacología

RESUMEN

The plant Miconia albicans (Sw.) Triana has been popularly used in Brazil to treat chronic inflammatory disturbances, such as osteoarthritis. This disease affects 250 million people worldwide, and is associated with intense pain and loss of articular function. There is a lack of information about the phytochemistry and bioactivity of M. albicans. Therefore, this study determined the chemical composition of some extracts and evaluated their cytotoxicity, along with their antioxidant and anti-inflammatory, activities using in vitro models. Aqueous and ethanolic extracts were prepared. Afterwards, a liquid-liquid partition was developed using chloroform, ethyl acetate, and n-butanol. The extracts were characterized by LC-MS, and their biological activities were evaluated on epithelial cells (Vero), tumoral hepatic cells (Hep-G2), and THP-1 macrophages. LC-MS analyses identified several flavonoids in all fractions, such as quercetin, myricetin, and their glycosides. The crude extracts and n-butanol fractions did not present cytotoxicity to the cells. The non-toxic fractions presented significant antioxidant activity when evaluated in terms of DPPH scavenging activity, lipid peroxidation, and ROS inhibition. THP-1 macrophages treated with the n-butanol fraction (250 µg/mL) released fewer pro-inflammatory cytokines, even in the presence of LPS. In the future, it will be necessary to identify the phytochemicals that are responsible for anti-inflammatory effects for the discovery of new drugs. In vivo studies on M. albicans extracts are still required to confirm their possible mechanisms of action.

Asunto(s)

Melastomataceae , 1-Butanol , Antiinflamatorios/química , Antioxidantes/química , Cloroformo , Citocinas , Flavonoides/farmacología , Glicósidos , Humanos , Lipopolisacáridos , Fitoquímicos/farmacología , Extractos Vegetales/química , Quercetina/farmacología , Especies Reactivas de Oxígeno

RESUMEN

A fucoxylomannan (FXM) was isolated from the mushroom Ganoderma lucidum through alkaline extraction followed by dialysis, freeze-thawing, and fractionation by Fehling's solution. The main chain of FXM presented α-d-Manp-(1→4)-linked units, and some of them were branched at O-6 position by α-l-Fucp-(1→2)-ß-d-Xylp groups. Its Mw was 35.9 kDa. FXM was tested on melanoma B16-F10 cells and it showed cell viability and cell density reduction, as well as antiproliferative effect, through cell cycle arrest. Additionally, the anchorage-independent clonogenic capacity of such cells was significantly reduced by FXM, decreasing the number of cells by colony and the colonies area. No effect on viability neither in proliferation of non-tumoral Balb c/3T3 fibroblasts was observed. These results indicate that FXM is a promising anti-proliferative compound impairing pivotal tumorigenic mechanisms, eliciting this polysaccharide to be further explored as an antimelanoma drug.

Asunto(s)

Agaricales , Ganoderma , Reishi , Cuerpos Fructíferos de los Hongos , Polisacáridos/farmacología , Diálisis Renal

RESUMEN

Mushroom ß-d-glucans have demonstrated immunomodulatory activity, which is initiated by their recognition by specific receptors on immune system cells surfaces. Studies indicated that ß-d-glucans may present a synergistic effect with chemotherapy drugs. In this study, a linear ß-(1 â†’ 6)-d-glucan (B16), isolated from A. bisporus and previously characterized (Mw: 8.26 × 104 g/mol), was evaluated about its capacity to modulate THP-1 macrophages towards an M1 phenotype and induce an antitumoral activity. This was evidenced by the production of pro-inflammatory markers upon B16 treatment (30; 100 µg/mL). The breast tumor cells (MDA-MB-231) viability was not affected by treatment with B16, however, their viability markedly decreased upon treatment with the drug doxorubicin. The results showed a synergic effect of B16 and doxorubicin, which reduced the viability of MDA-MB-231 cells by 31%. Furthermore, B16 treatment provided a sustainable M1 state environment and contributed to increase the sensitivity of breast cancer cells to the doxorubicin treatment.

Asunto(s)

Agaricus/química , Antibióticos Antineoplásicos/farmacología , Doxorrubicina/farmacología , Factores Inmunológicos/farmacología , Macrófagos/efectos de los fármacos , Polisacáridos/farmacología , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Animales , Antibióticos Antineoplásicos/química , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Doxorrubicina/química , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Factores Inmunológicos/química , Macrófagos/inmunología , Ratones , Fenotipo , Polisacáridos/química , Neoplasias de la Mama Triple Negativas/inmunología , Neoplasias de la Mama Triple Negativas/patología

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: The polysaccharides of the millenary mushroom Ganoderma lucidum (GL) have been shown for decades to present anti-tumor activities, but few studies evaluated its importance on cancer stem cells and EMT process. Cancer stem cells (CSC) drive the development of carcinoma and are also involved in cancer treatment failure, being a good target for treatment success. Also, the process of epithelial-mesenchymal transition (EMT) is involved in metastasis and cancer relapse. Besides that, the increasing incidence worldwide of oral squamous cell carcinoma (OSCC) became a public health issue with a high rate of metastasis and poor quality of life for patients during and after treatment. AIM OF THE STUDY: to evaluate G. lucidum polysaccharides (GLPS) in vitro effects on OSCC, focusing on hallmarks associated with tumorigenesis using the SCC-9, a squamous cells carcinoma lineage from the tongue. MATERIALS AND METHODS: SCC-9 cells were treated in vitro for 72h with different GLPS concentrations. The controls cells were maintained with culture media only and cisplatin was used as treatment control. After the treatment period, the cells were evaluated. RESULTS: GLPS treatment changed cell morphology and granularity, delayed migration, decreased colony, and impaired sphere formation, thereby leading to a non-invasive and less proliferative behavior of tumoral cells. Additionally, GLPS downregulated CSC, EMT, and drug sensitivity (ABC) markers. CONCLUSIONS: These results show that the natural product GLPS has the potential to be an important ally for tongue squamous cell carcinoma treatment, bringing the millenary compound to modern therapy, providing a basis for future studies and the improvement of life quality for OSCC patients.

Asunto(s)

Polisacáridos/farmacología , Reishi/química , Carcinoma de Células Escamosas de Cabeza y Cuello/tratamiento farmacológico , Neoplasias de la Lengua/tratamiento farmacológico , Antineoplásicos/administración & dosificación , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Línea Celular Tumoral , Cisplatino/farmacología , Relación Dosis-Respuesta a Droga , Transición Epitelial-Mesenquimal/efectos de los fármacos , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Humanos , Células Madre Neoplásicas/efectos de los fármacos , Polisacáridos/administración & dosificación , Polisacáridos/aislamiento & purificación , Neoplasias de la Lengua/patología

RESUMEN

BACKGROUND & AIMS: Fungal ß-glucans have been considered as biological response modifiers (BRMs) promoting stimulation of immune system according to numerous scientific publications performed in vitro and in vivo. Some clinical trials involving such compounds started to be published since 1980's. This systematic review aimed to compile and compare clinical studies using these ß-glucans as adjuvants on patients undergoing cancer treatment. Healthy subjects and ß-glucans from other sources were excluded. METHODS: It was developed according to PRISMA-P guidelines (PROSPERO registered n. CRD42020151539), using PICO criteria and the following databases: PubMed, Scielo and LILACS. RESULTS: We found 1018 articles and after removing duplicated records, select by title/abstract and full-text, only 9 studies remained and 7 more were manually added, totalizing 16 trials involving 1650 patients, with arm sizes varying from 9 until 200 patients. The selected studies (published since 1992-2018) included subjects with diagnosis of 9 types of cancer. The studies used different sources of ß-glucans, such as yeast (Saccharomyces cerevisiae), mushrooms (Lentinula edodes and Schizophyllum commune) and non-described fungal sources. CONCLUSIONS: It was observed that the administration of ß-glucan is safe and well-tolerated. Most of the trials pointed that concomitant administration of ß-glucan with chemo or radiotherapy reduced the immune depression caused by such treatments and/or accelerated the recovery of white blood cells counts. However, some articles also commented that no statistical difference was encountered between ß-glucan treated vs. control groups, which gives a controversial conclusion about the ß-glucan effects. The great diversity among the methodology studies and insufficient information was an impeditive for achieving profound statistical analysis, therefore a narrative report of the included studies was performed indicating that further evidences are required to determine the efficacy of this adjuvant in the cancer treatment.

Asunto(s)

Hongos/inmunología , Factores Inmunológicos/inmunología , Factores Inmunológicos/uso terapéutico , Neoplasias/tratamiento farmacológico , Neoplasias/inmunología , beta-Glucanos/inmunología , beta-Glucanos/uso terapéutico , Terapia Combinada/métodos , Humanos , Neoplasias/terapia

RESUMEN

A ß-D-glucan was obtained from the edible mushroom Pholiota nameko by hot aqueous extraction and purification. NMR and methylation analyses of the purified fraction (GHW-PN, 1.46% yield) indicated the presence of a (1 → 3)-linked ß-D-glucan, highly substituted (~27%) at O-6 by single units of ß-D-Glcp or by (1 → 6)-ß-D-Glcp fragments. The ß-glucan (at 0.5, 1, and 2%) showed shear thinning behavior and when the concentration of the solution increased, there was an increase in apparent viscosity. The ß-D-glucan presented gel-like behavior and thermal stability under a simulated pasteurization process, suggesting its potential as a thickening and gelling agent in products submitted to temperature variations. The ß-D-glucan at 0.3, 1.0 and 3.0 mg kg-1 significantly inhibited the inflammatory pain in 24.8, 56.9 and 82.3%, respectively, in the formalin-induced nociception in mice. The results pointed out that the ß-D-glucan (GHW-PN) isolated from P. nameko presents potential application for the food industry or for medical purposes.

Asunto(s)

Analgésicos/química , Analgésicos/farmacología , Antiinflamatorios/química , Antiinflamatorios/farmacología , Glucanos/química , Glucanos/farmacología , Pholiota/química , Animales , Formaldehído/farmacología , Geles , Masculino , Ratones , Nocicepción/efectos de los fármacos , Reología

RESUMEN

An aqueous extract containing polysaccharides was obtained from the giant mushroom Macrocybe titans, and it was purified by amylase treatment, freeze-thawing process and dialysis. The purified fraction (ESP) was analyzed by HPSEC and GC-MS which showed a homogenous polysaccharide with Mw 14.2 × 103 g/mol composed by galactose and fucose. NMR and methylation analysis of ESP confirmed the presence of a fucogalactan with a (1 → 6)-linked α-d-Galp main chain partially substituted at O-2 by non reducing end units of α-l-Fucp residues in the side chain. Its biological activity was evaluated against murine melanoma cells B16-F10. The fucogalactan did not alter the viability, proliferative capacity and morphology of cells. However, this polysaccharide was able to reduce the cell migration in vitro at 40% (100 µg/mL) and 33% (250 µg/mL). The results obtained showed that Macrocybe titans fucogalactan is a promising agent capable of altering melanoma cell migration without decrease the cell viability.

Asunto(s)

Agaricales/química , Movimiento Celular/efectos de los fármacos , Galactanos/farmacología , Melanoma Experimental/patología , Melanoma/patología , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Galactanos/química

RESUMEN

A crude fraction (SCW) was extracted with cold water from Pholiota nameko and showed mannose (24.1%), galactose (44.9%) and glucose (31%). Purification procedures resulted in a polysaccharide fraction (bG-PN), that showed only glucose. NMR and methylation analyses of bG-PN indicated a ß-d-glucan-(1→3)-linked, substituted at O-6 by ß-d-Glcp or (1→6)-linked ß-d-Glcp side chains. Rheological studies of crude and purified fractions at the same concentration showed similar shear-thinning behavior and gel-like structure which indicates no need to isolate the polymer to achieve some desirable rheological properties. SCW (at 1% and 2%) and bG-PN (at 2%) presented thermal stability during heating and cooling, suggesting that the physical structure of gels (SCW and bG-PN at 2%) and viscoelastic fluid (SCW at 1%) formed were not altered in the tested temperature range. Our results suggest that P. nameko ß-d-glucans can be applied in different food preparations as thickener or gelling agents modifying their rheological properties.

Asunto(s)

beta-Glucanos/química , Pholiota , Reología , Sustancias Viscoelásticas/química , Sustancias Viscoelásticas/aislamiento & purificación , beta-Glucanos/aislamiento & purificación

RESUMEN

D-Glucans from edible mushrooms present diversified chemical structures. The most common type consists of a backbone of ß-D-glucose (1→3)-linked frequently branched at O-6 by ß-D-glucose residues as side chains. However it is possible to distinguish α-, ß- and mixed D-glucans. Further discrimination could be made on the basis of glycosidic bond position in a pyranoid ring, distribution of specific glycosidic bonds along the chain, branching and molecular weight. The present manuscript reviews the processes of extraction, purification and chemical characterization of D-glucans, such as NMR studies, methylation analysis, Smith degradation, and some other methodologies employed in carbohydrate chemistry characterization. In addition, these polysaccharides are important because they can provide many therapeutic benefits related to their biological activity in animals and humans, either immunostimulatory activity, inhibiting tumor growth, as well as exerting antinociceptive and anti-inflammatory action, among others, which are usually attached to their structure, molecular weight and degree of branching.

Asunto(s)

Agaricales/química , Fraccionamiento Químico/métodos , Glucanos/química , Glucanos/aislamiento & purificación , Animales , Glucanos/farmacología , Humanos

RESUMEN

ß-D-Glucan, a polysaccharide isolated from an edible mushroom Pleurotus pulmonarius (Fr.) Quel., presented antinociceptive activity in mice. This study evaluated the involvement of transient receptor potential (TRP) channels and protein kinase C (PKC) on antinociceptive effect of a (1→3),(1→6)-linked ß-D-glucan (GL) in mice. Intraperitoneal administration of GL potently inhibited nociceptive responses induced by intraplantar injections of capsaicin, cinnamaldehyde, menthol, acidified saline and phorbol myristate acetate (PMA). Moreover, Western blot analysis revealed that GL treatment also prevented PMA-induced PKCɛ activation. Collectively, present results demonstrate that GL could constitute an attractive molecule of interest for the development of new analgesic drugs.

Asunto(s)

Analgésicos/farmacología , Glucanos/farmacología , Pleurotus/química , Proteína Quinasa C/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Canales Iónicos Sensibles al Ácido , Acroleína/análogos & derivados , Acroleína/farmacología , Analgésicos/aislamiento & purificación , Animales , Capsaicina/farmacología , Activación Enzimática/efectos de los fármacos , Glucanos/aislamiento & purificación , Masculino , Mentol/farmacología , Ratones , Proteínas del Tejido Nervioso/metabolismo , Nocicepción/efectos de los fármacos , Proteína Quinasa C/metabolismo , Inhibidores de Proteínas Quinasas/aislamiento & purificación , Canales de Sodio/metabolismo , Canales Catiónicos TRPV/metabolismo , Acetato de Tetradecanoilforbol/farmacología

RESUMEN

UNLABELLED: The present study evaluated the antinociceptive effect of (1→3),(1→6)-linked ß-glucan (GL) isolated from Pleurotus pulmonarius (Fr.) Quel. in mice and its possible mechanism of action. Intraperitoneal administration of GL inhibited glutamate-induced licking with an ID(50) of 0.34 mg/kg and inhibition of 96% ± 3%. The treatment of animals with GL (1 mg/kg i.p.) inhibited nociception induced by intrathecal injection of N-methyl-D-aspartic acid, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, kainate and interleukin -1ß in 67% ± 13%, 89% ± 11%, 74% ± 9%, and 75% ± 7%, respectively, but not the nociceptive response induced by (±)-1-aminocyclopentane-trans-1,3-dicarboxylic acid, substance P, and tumor necrosis factor-α. Moreover, GL (30 mg/kg i.p.) also reduced mechanical allodynia caused by partial sciatic nerve ligation for 2 hours, with inhibition of 47% ± 10% observed 0.5 hours after treatment. When given chronically (twice a day) over 7 days, GL reversed the mechanical allodynia caused by partial sciatic nerve ligation (inhibition of 45% ± 13% to 60% ± 8%). Interestingly, GL did not affect the locomotor activity of mice in an open field test with doses that produce antinociceptive effects. Our findings show that GL inhibits acute and neuropathic pain in mice through mechanisms that involve the inhibition of ionotropic glutamate receptors and the interleukin -1ß pathway. PERSPECTIVE: This article presents the antinociceptive activity of GL in acute and neuropathic pain with participation of ionotropic glutamate receptors and pro-inflammatory cytokines (interleukin-1ß). After further experiments, this compound may represent a new pharmacological agent for the treatment of clinical pain.

Asunto(s)

Analgésicos/farmacología , Glucanos/farmacología , Neuralgia/tratamiento farmacológico , Neuralgia/metabolismo , Pleurotus/química , Receptores de Glutamato/fisiología , Transducción de Señal/efectos de los fármacos , Enfermedad Aguda , Analgésicos/uso terapéutico , Animales , Citocinas/fisiología , Modelos Animales de Enfermedad , Femenino , Glucanos/uso terapéutico , Inyecciones Intraperitoneales , Interleucina-1beta/fisiología , Masculino , Ratones , Neuralgia/fisiopatología , Transducción de Señal/fisiología