RESUMO
Brazil-grown outdoor-cultivated Agaricus brasiliensis KA21 fruiting body (KA21) significantly increases the production of serum anti-beta-glucan antibody. Therefore, KA21 ingestion may be useful for the prevention and alleviation of fungal infections. This study aimed to determine the effects of KA21 in fungal infections in animals. KA21 was administered to nine dogs infected with Malassezia. Notably, the anti-beta-glucan antibody titer remained unchanged or tended to decrease in the oral steroid arm, whereas in the non-steroid arm, antibody titer increased in almost all animals after KA21 ingestion. Dogs showing improved clinical symptoms exhibited increased anti-beta-glucan antibody titers. The results of this study suggest that KA21 ingestion may alleviate the symptoms of Malassezia and other fungal infections and that continuous ingestion may help prolong recurrence-free intervals. Additionally, the ingestion of KA21 during oral steroid dosage reduction or discontinuation may enable smoother steroid withdrawal.
Assuntos
Agaricus , Doenças do Cão , Carpóforos , Malassezia , Animais , Cães , Agaricus/química , Carpóforos/química , Malassezia/efeitos dos fármacos , Doenças do Cão/microbiologia , Doenças do Cão/tratamento farmacológico , Dermatomicoses/veterinária , Dermatomicoses/prevenção & controle , Dermatomicoses/tratamento farmacológico , Dermatomicoses/microbiologia , beta-Glucanas/administração & dosagem , beta-Glucanas/farmacologia , Masculino , Brasil , Dermatite/tratamento farmacológico , Dermatite/veterinária , Dermatite/microbiologia , Dermatite/prevenção & controle , Feminino , Anticorpos Antifúngicos/sangueRESUMO
The edible mushroom Agaricus brasiliensis contains a large amount ß-glucan, which is mainly composed of a ß-1,6-glucan structure. In this study, we investigated the effect of A. brasiliensis strain KA21 on the anti-ß-glucan antibody titer in healthy humans and the role of antibodies as an immunomodulator. Twenty-two healthy volunteers were fed the dried fruiting body of A. brasiliensis (900 or 1500 mg/day) for 12 weeks. The anti-ß-glucan antibody titer in the serum was determined by enzyme-linked immunosorbent assay. Immunoglobulin G (IgG) against ß-glucan was significantly upregulated after intake of A. brasiliensis. Murine experiments demonstrated improvement of anti-ß-glucan antibody production after intraperitoneal injection of Agaricus-derived ß-glucan. To understand the role of antibody against ß-glucan in exclusion of pathogenic fungi, we examined the interaction between HL-60 cells and antibody-treated heat-killed Candida albicans. Flow cytometry analysis indicated the upregulation of Candida-positive HL-60 cells after treatment with human IgG, whereas the competitive assay demonstrated that the main epitope of Candida-reacted IgG was the ß-1,6-glucan structure. Binding between HL-60 and IgG-opsonized C. albicans was suppressed by anti-Fcγ receptor 1 (FcγRI) neutralizing antibody. Finally, using FcγRI-expressed cells with the nuclear factor of activated T-cell reporter assay, we demonstrated that higher titers of anti-ß-glucan IgG can induce stronger Fc receptor-mediated cell activation through the formation of an antibody-ß-glucan complex. In conclusion, oral ingestion of A. brasiliensis KA21 promotes anti-ß-glucan antibody production and may contribute to preventing fungal infection through the activation of immune cells by forming antibody-ß-glucan complexes via an FcγR-dependent pathway.
Assuntos
Agaricus , Animais , Formação de Anticorpos , Glucanos , Voluntários Saudáveis , Humanos , Camundongos , beta-GlucanasRESUMO
We investigated whether outdoor-cultivated Agaricus brasiliensis (KA21) could reduce the side effects caused by the anticancer medicine 5-fluorouracil (5-FU). The adverse effects of 5-FU were analyzed in mice by orally administering the drug every day for 5 days. Leukopenia, diarrhea, body weight loss, anorexia, kidney injury, gastrointestinal tract injury, and hair loss were evaluated as side effects. We determined whether these side effects were reduced by the intake of outdoor-cultivated A. brasiliensis. The side effects were reduced in mice receiving the outdoor-cultivated A. brasiliensis but not in those receiving the indoor-cultivated A. brasiliensis. These results suggest that outdoor-cultivated A. brasiliensis is beneficial in reducing the side effects of the anticancer medicine and might, therefore, be useful in improving the quality of life of patients with cancer. Furthermore, because antioxidants have been reported to contribute to the suppression of the side effects of anticancer medicines, the antioxidant activities of different mushrooms were determined. The antioxidant activity of the outdoor-cultivated A. brasiliensis was the highest among all the tested mushrooms. These findings make it clear that the cultivation conditions of A. brasiliensis are important in suppressing the side effects of anticancer medicines. Strong antioxidant activity might be one of the mechanisms through which this pharmacological activity is mediated.
Assuntos
Agaricus/química , Antineoplásicos/efeitos adversos , Antioxidantes/química , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/prevenção & controle , Agricultura , Animais , Feminino , Fluoruracila/efeitos adversos , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The royal sun medicinal mushroom, Agaricus brasiliensis, is a health food material that helps to improve quality of life. A. brasiliensis has long been used as a tea by extraction with cold and hot water. Our group has been investigating the immunopharmacological activities of the A. brasiliensis KA21 strain, which is cultivated outdoors. We prepared cold water (AgCWE) and hot water (AgHWE) extracts of this strain. AgCWE contained a larger proportion of proteins, including enzymes, and showed a brownish color during the extraction process. By contrast, chemical and immunochemical analyses revealed that AgHWE contained large amounts of ß-1,3-/1,6-glucans. In an attempt to elucidate the immunochemical characteristics of AgCWE, reactivities to immunoglobulin (Ig) preparations for intravenous injection were analyzed and compared with standard materials. To characterize brownish high-molecular weight components, standard phenol compounds such as caffeic acid (CA), trans-ferulic acid (FA), and coumaric acid (CouA) were polymerized to brownish polymerized polyphenols (PPPs) (i.e., polymerized CA, polymerized FA, and polymerized CouA) by laccase or peroxidase. The results obtained revealed that intravenous Ig reacted with all PPPs and PPPs cross-reacted with AgCWE and AgHWE. The isotype of the anti-PPP antibody was found to be IgG1, in contrast to that of the ß-glucan antibody, which was mainly IgG2. These results strongly suggest that A. brasiliensis extracts contain immunoreactive components against various classes of Igs.
Assuntos
Agaricus/imunologia , Anticorpos Antifúngicos/imunologia , Misturas Complexas/imunologia , Alimentos , Humanos , Injeções Intravenosas , Lignanas/imunologia , Água , beta-Glucanas/imunologiaRESUMO
Laccases are enzymes that oxidize various aromatic compounds, and therefore they have attracted much attention from the standpoints of medical and industrial applications. We previously isolated the cDNA that codes for a laccase isozyme (Lac2a) from the medicinal mushroom Agaricus brasiliensis (Matsumoto-Akanuma et al., Int. J. Med. Mushrooms, 16, 375-393, 2014). In this study, we first attempted heterologous expression of the wild-type laccase using a Pichia pastoris secretory expression system. However, the trial was unsuccessful most likely because the enzyme was too unstable and degraded immediately after production. Therefore, we improved the stability of the laccase by using a phylogeny-based design method. We created a mutant laccase in which sixteen original residues were replaced with those found in the phylogenetically inferred ancestral sequence. The resulting mutant protein was successfully produced using the P. pastoris secretory expression system and then purified. The designed laccase showed catalytic properties similar to those of other fungal laccases. Moreover, the laccase is highly thermally stable at acidic and neutral pH and is also stable at alkaline pH at moderate temperatures. We expect that the laccase will serve as a useful tool for enzymatic polymerization of di-phenolic compounds.
Assuntos
Agaricus/enzimologia , Lacase/química , Lacase/metabolismo , Proteínas Mutantes/química , Proteínas Mutantes/metabolismo , Mutação , Filogenia , Agaricus/genética , Biocatálise , DNA Complementar/genética , Estabilidade Enzimática/genética , Concentração de Íons de Hidrogênio , Lacase/genética , Lacase/isolamento & purificação , Proteínas Mutantes/genética , Proteínas Mutantes/isolamento & purificação , Oxirredução , Pichia/genética , Pichia/metabolismo , Engenharia de Proteínas , TemperaturaRESUMO
We conducted an open-label study in which food containing Agaricus brasiliensis KA21 was consumed continuously for 12 weeks. A questionnaire for subjective evaluation of the efficacy of this food (hereafter, subjective evaluation questionnaire) revealed significant improvements compared with before its intake; there were improvements in the scores of the amounts of hair loss and gray hair, fatigue and general malaise, eye strain, shoulder stiffness, coldness of extremities, difficulty staying awake during the day, and ease of getting out of bed. These findings suggest that intake of food containing A. brasiliensis KA21 results in the above-mentioned subjectively evaluated improvements, and the possibility that A. brasiliensis KA21 improves the body's immunity. Moreover, no issues regarding the safety of the test food were found.
Assuntos
Agaricus , Dieta , Qualidade de Vida , Atividades Cotidianas , Adulto , Olho , Fadiga/dietoterapia , Feminino , Cabelo , Voluntários Saudáveis , Humanos , Imunidade , Masculino , Pessoa de Meia-Idade , Amplitude de Movimento Articular , Autorrelato , Ombro , Sono , Inquéritos e Questionários , VerdurasRESUMO
BACKGROUND: Agaricus brasiliensis (A. brasiliensis) is a medicinal mushroom that exerts various pharmacological actions. We previously demonstrated that different cultivation conditions altered the activity of the polyphenol-related enzymes from this mushroom. However, the influence of cultivation conditions on the antioxidant activity of the fruiting bodies remains unclear. Therefore, in this study we compared the antioxidative effects of fruiting bodies of A. brasiliensis cultivated outdoors and indoors. In addition, we assessed whether different cultivation methods affected the hepatoprotective effects against CCl4-induced liver injury. METHODS: We assessed the antioxidative effects of mushrooms cultivated in open-air or indoors using the DPPH radical-scavenging assay. Furthermore, we prepared experimental feeds containing outdoor- or indoor-cultivated A. brasiliensis. Acute liver injury was induced by CCl4 injection in mice that consumed feed containing outdoor- or indoor-cultivated A. brasiliensis. The hepatoprotective effects of these mushrooms were then evaluated by monitoring the reduction in the circulating levels of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase. The significance of the differences between the means was assessed using Student's t-test. Finally, histopathological analysis of liver was performed. RESULTS: In the DPPH assay, the antioxidant activity of outdoor-cultivated A. brasiliensis was higher than that of indoor-cultivated mushroom. Moreover, in the mouse model of CCl4-induced hepatitis, the oral administration of outdoor-cultivated A. brasiliensis reduced liver damage significantly, but indoor-cultivated mushrooms failed to inhibit hepatitis. The hepatoprotective effects of outdoor-cultivated A. brasiliensis were observed even when ingestion commenced only 1 day before CCl4 injection, and these effects were not affected by excessive heat treatment. CONCLUSIONS: Outdoor cultivation significantly enhanced the antioxidative activity of A. brasiliensis fruiting bodies. In addition, outdoor-cultivated A. brasiliensis was more effective at protecting against CCl4-induced liver injury in mice than mushrooms grown in a greenhouse.
Assuntos
Agaricus/química , Agricultura/métodos , Antioxidantes/uso terapêutico , Produtos Biológicos/uso terapêutico , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Fígado/efeitos dos fármacos , Alanina Transaminase/sangue , Animais , Antioxidantes/farmacologia , Aspartato Aminotransferases/sangue , Produtos Biológicos/farmacologia , Compostos de Bifenilo/metabolismo , Tetracloreto de Carbono , Doença Hepática Induzida por Substâncias e Drogas/sangue , Carpóforos , Hepatite/sangue , Hepatite/tratamento farmacológico , L-Lactato Desidrogenase/sangue , Fígado/enzimologia , Masculino , Camundongos Endogâmicos ICR , Oxirredução , Picratos/metabolismo , Polifenóis/farmacologia , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêuticoRESUMO
Laccase isozymes have been identified in several fungi. We report the cloning of 4 laccase genes from the medicinal mushroom Agaricus brasiliensis. The lac1 gene contained a 1560-base pair (bp) open reading frame (ORF) encoding 520 amino acids that was interrupted with 14 introns in genomic DNA. The deduced amino acid sequence indicated a multicopper oxidase signature 1 and 2 multicopper oxidase signature 2. The lac2 gene contained a 1566-bp ORF encoding 522 amino acids that was interrupted with 13 introns in genomic DNA. A number of different nucleotides were observed in whole regions containing the substitution of amino acid residues (lac2a and lac2b). The partial DNA fragments of lac3 and lac4 genes were subcloned using the semi-random two-step polymerase chain reaction method. The lac3 and lac4 genes contained coding sequences with a 1575-bp ORF encoding 525 amino acids and a 1584-bp ORF encoding 528 amino acids, respectively. However, the whole complementary DNA fragment of both laccases could not be amplified with polymerase chain reaction against the complementary DNA library; therefore, introns were deduced based on the GT-AG rule and multiple alignment of laccases from other fungi, which showed high identity. All laccases from A. brasiliensis conserved the fungal laccase signature sequence and suggest 2 subfamilies according to the location of introns and phylogenetic analysis. The genes lac2 and lac4 had a high degree of identity, and the lac2a gene was located upstream of the lac4 gene.
Assuntos
Agaricus/enzimologia , Agaricus/genética , Lacase/genética , Sequência de Aminoácidos , Sequência de Bases , Basidiomycota , Clonagem Molecular , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , Fungos , Íntrons , Lacase/química , Modelos Moleculares , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Reação em Cadeia da Polimerase , Conformação Proteica , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de SequênciaRESUMO
The Royal Sun medicinal mushroom, Agaricus brasiliensis, is used as a natural health product. In Japan, however, the quality control of some of these mushroom products has been viewed as a safety problem. Focusing on the quality control of A. brasiliensis KA21, we have performed several safety studies. To date, we have established evidence that this mushroom can be used safely as an immunostimulant and to mediate biochemical parameters associated with obesity or diabetes. Furthermore, to improve the manufacturing process of this mushroom, we have studied the relationship between its pharmaceutical actions and the conditions of its cultivation and thermal management. The purpose of this review is to report the findings of basic and clinical studies of the fruit body of A. brasiliensis KA21.
Assuntos
Agaricus/química , Alimento Funcional , HumanosRESUMO
Agaricus brasiliensis is a well-known medicinal mushroom. We have previously demonstrated that Agaricus-derived polysaccharides exhibit potent antitumor effects; however, the underlying mechanism(s) have not been elucidated yet. In this study, we examined the immunoenhancing activities of Agaricus extracts. Agaricus-derived polysaccharides were characterized as 1,6-ß-glucan with a small amount of 1,3-ß-glucan using anti-ß-glucan antibody and nuclear magnetic resonance analysis. These polysaccharides strongly induced the production of various cytokines from both murine splenocytes and bone marrow-derived dendritic cells in the presence of exogenous granulocyte-macrophage colony-stimulating factor. Polysaccharide-induced cytokine production was significantly reduced in bone marrow-derived dendritic cells derived from dectin-1-deficient mice. Furthermore, a binding assay revealed that the Agaricus-derived polysaccharides can be recognized by dectin-1, a pivotal receptor for 1,3-ß-glucan. Taken together, our results clearly indicate that the immunostimulation induced by Agaricus-derived polysaccharides is exerted, at least in part, via dectin-1 in combination with granulocyte-macrophage colony-stimulating factor.