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1.
Food Funct ; 10(10): 6633-6643, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31555775

RESUMO

In this study, we investigated the potential bioactivities of an ethanol extract of Hericium novae-zealandiae and four of its constituents, namely hericenone C, hericene B, ergosterol and ergosterol peroxide. The proliferation of three prostate cancer cell lines, namely DU145, LNCaP and PC3, was evaluated after treatment with the extract and constituents. It was found that both the ethanol extract and ergosterol peroxide possess anti-proliferative activities to the three prostate cancer cell lines. Ergosterol peroxide was considered likely to be one of the major compounds responsible for the anti-proliferative effect of the ethanol extract. Subsequently, the results of RT-qPCR assay showed two possible mechanisms for these anti-proliferative activities. One is apoptosis, supported by the up-regulation of CASP3, CASP8, CASP9, and an increase in the ratio of Bax/Bcl2. The other is anti-inflammation, indicated by the down-regulation of IL6 and up-regulation of IL24. The ethanol extract also exhibited antioxidant and AChE inhibitory (though weak) activities. However, none of the four compounds were found to account for these latter two activities. This is the first report of the bioactivities, and the corresponding active ingredients of lipophilic constituents from H. novae-zealandiae.


Assuntos
Agaricales/química , Extratos Vegetais/análise , Extratos Vegetais/farmacologia , Apoptose/efeitos dos fármacos , Caspases/genética , Caspases/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ergosterol/análise , Ergosterol/isolamento & purificação , Ergosterol/farmacologia , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucinas/genética , Interleucinas/metabolismo , Nova Zelândia , Fenóis/análise , Fenóis/isolamento & purificação , Fenóis/farmacologia , Extratos Vegetais/isolamento & purificação
2.
Molecules ; 24(18)2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31514398

RESUMO

Inspired by the significant bioactivity of ergosterol peroxide, we designed and synthesized four fluorescent coumarin and ergosterol peroxide conjugates 8a-d through the combination of ergosterol peroxide with 7-N,N-diethylamino coumarins fluorophore. The cytotoxicity of synthesized conjugates against three human cancer cells (HepG2, SK-Hep1, and MCF-7) was evaluated. The results of fluorescent imaging showed that the synthesized conjugates 8a-d localized and enriched mainly in mitochondria, leading to significantly enhanced cytotoxicity over ergosterol peroxide. Furthermore, the results of biological functions of 8d showed that it could suppress cell colony formation, invasion, and migration; induce G2/M phase arrest of HepG2 cells, and increase the intracellular ROS level.


Assuntos
Antineoplásicos/farmacologia , Ergosterol/análogos & derivados , Mitocôndrias/metabolismo , Ciclo Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Desenho de Fármacos , Ergosterol/biossíntese , Ergosterol/química , Ergosterol/farmacologia , Humanos , Mitocôndrias/efeitos dos fármacos , Invasividade Neoplásica , Fenômenos Ópticos , Espécies Reativas de Oxigênio/metabolismo
3.
Biosci Biotechnol Biochem ; 83(12): 2280-2287, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31412751

RESUMO

The increasing number of patients suffering from allergic diseases is a global health problem. Grifola frondosa is an edible mushroom consumed as a health food in Asia, and has recently been reported to have anti-allergic effects. We previously reported that G. frondosa extract (GFE) and its active components, ergosterol and its derivatives, inhibited the antigen-induced activation of RBL-2H3 cells. Here, we demonstrated that GFE and ergosterol also had an inhibitory effect on the degranulation of bone marrow-derived mast cells (BMMCs) and alleviated anaphylactic cutaneous responses in mice. Using an air pouch-type allergic inflammation mouse model, we confirmed that oral administration of GFE and ergosterol suppressed the degranulation of mast cells in vivo. Our findings suggest that G. frondosa, including ergosterol as its active component, reduces type I allergic reactions by suppressing mast cell degranulation in mice, and might be a novel functional food that prevents allergic diseases.


Assuntos
Degranulação Celular/efeitos dos fármacos , Ergosterol/farmacologia , Grifola/química , Hipersensibilidade/prevenção & controle , Mastócitos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Permeabilidade Capilar/efeitos dos fármacos , Linhagem Celular , Modelos Animais de Doenças , Alimento Funcional , Liberação de Histamina/efeitos dos fármacos , Hipersensibilidade/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , beta-N-Acetil-Hexosaminidases/antagonistas & inibidores
4.
Fitoterapia ; 138: 104289, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31386896

RESUMO

Three newly isolated ergosterols, psathergosterols A-C (1-3), together with two known ones (4 and 5), have been isolated from cultures of the basdiomycete Psathyrella candolleana. Their structures with the absolute configuration were elucidated by means of spectroscopic methods and the single crystal X-ray diffraction. Compounds 2-4 exhibited certain cytotoxicities to five human cancer cell lines (HL-60, SMMC-7721, A-549, MCF-7, SW480).


Assuntos
Antineoplásicos/farmacologia , Basidiomycota/química , Ergosterol/farmacologia , Antineoplásicos/isolamento & purificação , Linhagem Celular Tumoral , China , Ensaios de Seleção de Medicamentos Antitumorais , Ergosterol/isolamento & purificação , Humanos , Estrutura Molecular
5.
Biomed Pharmacother ; 118: 109273, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31374354

RESUMO

Omphalia lapidescens Schroet. is a medicinal macrofungus in China that has shown good antitumor activity in recent research. To explore the potential cytotoxic compounds from O. lapidescens, the ethyl acetate soluble fraction of a 95% ethanol extract with cytotoxic activity was phytochemically investigated. A new tetranorlanostane triterpenoid (1) and a new ergosteroid (4), together with two known lanostanes (2 and 3) and six known Δ22-ergosteroids (5-10), were isolated. Notably, compound 1 possesses a new 24,25,26,27-tetranorlanostane carbon skeleton. All isolates, except compound 8, were tested for cytotoxic activities using a human breast cancer cell line (MDA-MB-231) and a human gastric cancer cell line (HGC-27) in vitro. The new nortriterpenoid (1) exhibited no obvious cytotoxicity, while the known triterpenoid (3) showed significant cytotoxicity against MDA-MB-231 and HGC-27 cells. Compound 4 exhibited the highest cytotoxicity against MDA-MB-231 and HGC-27 cells, with IC50 values of 11.33 ±â€¯2.18 and 12.28 ±â€¯3.64 µM, respectively. Furthermore, Hoechst fluorescence 33342 staining and Western blot tests demonstrated that compound 4 induced apoptosis in MDA-MB-231 cells by downregulating procaspase-3 expression and upregulating the expression ratio of Bax/Bcl-2.


Assuntos
Ergosterol/isolamento & purificação , Ergosterol/farmacologia , Fungos/química , Triterpenos/isolamento & purificação , Triterpenos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Ergosterol/química , Humanos , Espectroscopia de Ressonância Magnética , Triterpenos/química
6.
Eur J Pharmacol ; 860: 172543, 2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31323223

RESUMO

Ergosterol peroxide has been shown to exhibit anti-tumor, antioxidant and anti-bacterial properties. However, the effects of ergosterol peroxide isolated from the herbal Baphicacanthus cusia root on influenza virus infection remain poorly understood. In the present study, ergosterol peroxide (compound 22) was obtained from the B. cusia root and subjected to investigation regarding its immunoregulatory effect on influenza A virus (IAV)-induced inflammation in A549 human alveolar epithelial cells. The structure of compound 22 isolated from B. cusia root. was elucidated by NMR analyses. Structure determination showed that the chemical structure of compound 22 closely resembles that of ergosterol peroxide. We observed that ergosterol peroxide treatment significantly suppressed IAV-induced upregulation of RIG-I expression. Additionally, ergosterol peroxide inhibited the activation of RIG-I downstream signaling pathways, including p38 MAP kinase and NF-κB, which ultimately resulted in the reduced production of an array of pro-inflammatory mediators and interferons (IFN-ß and IFN-λ1). Interestingly, inhibitory effects of ergosterol peroxide on the expression of IFNs did not affect the expression of antiviral effectors or enhance viral replication. On the other hand, ergosterol peroxide effectively abolished the amplified production of pro-inflammatory mediators in cells pretreated with IFN-ß (500 ng/ml) prior to IAV infection. Moreover, Annexin V and Hoechst 33258 staining revealed that increased apoptosis of IAV-infected cells was reversed by the presence of ergosterol peroxide. Our findings suggest that ergosterol peroxide from the B. cusia root suppressed IAV-associated inflammation and apoptosis via blocking RIG-I signaling, which may serve as a supplementary approach to the treatment of influenza.


Assuntos
Apoptose/efeitos dos fármacos , Proteína DEAD-box 58/metabolismo , Ergosterol/análogos & derivados , Vírus da Influenza A Subtipo H1N1/fisiologia , Transdução de Sinais/efeitos dos fármacos , Células A549 , Animais , Cães , Ergosterol/química , Ergosterol/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/patologia , Inflamação/virologia , Mediadores da Inflamação/metabolismo , Interferons/biossíntese , Células Madin Darby de Rim Canino
7.
J Steroid Biochem Mol Biol ; 194: 105435, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31352023

RESUMO

Factors that can modify the bioavailability of orally administered vitamin D are not yet widely known. Ergosterol is a common fungal sterol found in food which has a chemical structure comparable to that of vitamin D. This study aimed to investigate the effect of ergosterol on vitamin D metabolism. Therefore, 36 male wild type-mice were randomly subdivided into three groups (n = 12) and received a diet containing 25 µg vitamin D3 and either 0 mg (control), 2 mg or 7 mg ergosterol per kg diet for 6 weeks. To elucidate the impact of ergosterol on hepatic hydroxylation of vitamin D, human hepatoma cells (HepG2) were treated with different concentrations of ergosterol. Concentrations of vitamin D3 and 25-hydroxyvitamin D3 (25(OH)D3) in cells, livers and kidneys of mice and additionally 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) in serum were quantified by LC-MS/MS. The concentration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) in serum was analyzed by commercially-available enzyme immuno assay. The concentrations of cholesterol and triglycerides were analyzed in livers of mice by photometric assays. Analyses revealed that mice receiving 7 mg/kg ergosterol with their diet had 1.3-, 1.7- and 1.5-times higher concentrations of vitamin D3 in serum, liver and kidney, respectively, than control mice (P < 0.05), whereas no significant effects were observed in mice fed 2 mg/kg ergosterol. The hydroxylation of vitamin D remained unaffected by dietary ergosterol, since the concentration of 25-hydroxyvitamin D3 in serum and tissues and the concentrations of 1,25(OH)2D3 and 24,25(OH)2D3 in serum were not different between the three groups of mice. The lipid concentrations in liver were also not affected by dietary ergosterol. Data from the cell culture studies showed that ergosterol did not influence the conversion of vitamin D3 to 25(OH)D3. To conclude, ergosterol appears to be a modulator of vitamin D3 concentrations in the body of mice, without modulating the hydroxylation of vitamin D3 in liver.


Assuntos
Colecalciferol/farmacologia , Ergosterol/farmacologia , Vitaminas/farmacologia , 24,25-Di-Hidroxivitamina D 3/sangue , 24,25-Di-Hidroxivitamina D 3/metabolismo , Animais , Calcifediol/sangue , Calcifediol/metabolismo , Colecalciferol/sangue , Colecalciferol/farmacocinética , Células Hep G2 , Humanos , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Vitaminas/sangue , Vitaminas/farmacocinética
8.
Mar Drugs ; 17(6)2019 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-31163615

RESUMO

Sea hares of Aplysia genus are recognized as a source of a diverse range of metabolites. 5α,8α-Endoperoxides belong to a group of oxidized sterols commonly found in marine organisms and display several bioactivities, including antimicrobial, anti-tumor, and immunomodulatory properties. Herein we report the isolation of 5α,8α-epidioxycholest-6-en-3ß-ol (EnP(5,8)) from Aplysia depilans Gmelin, based on bioguided fractionation and nuclear magnetic resonance (NMR) analysis, as well as the first disclosure of its anti-inflammatory properties. EnP(5,8) revealed capacity to decrease cellular nitric oxide (NO) levels in RAW 264.7 macrophages treated with lipopolysaccharide (LPS) by downregulation of the Nos2 (inducible nitric oxide synthase, iNOS) gene. Moreover, EnP(5,8) also inhibited the LPS-induced expression of cyclooxygenase-2 (COX-2), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) at the mRNA and protein levels. Mild selective inhibition of COX-2 enzyme activity was also evidenced. Our findings provide evidence of EnP(5,8) as a potential lead drug molecule for the development of new anti-inflammatory agents.


Assuntos
Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Aplysia/química , Ésteres do Colesterol/química , Ésteres do Colesterol/farmacologia , Ergosterol/análogos & derivados , Macrófagos/efeitos dos fármacos , Adjuvantes Imunológicos/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Fracionamento Químico , Ésteres do Colesterol/isolamento & purificação , Regulação para Baixo/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Ergosterol/química , Ergosterol/isolamento & purificação , Ergosterol/farmacologia , Lipopolissacarídeos/farmacologia , Espectroscopia de Ressonância Magnética , Camundongos , Óxido Nítrico Sintase Tipo II/genética , Células RAW 264.7
9.
Org Biomol Chem ; 17(21): 5223-5229, 2019 05 29.
Artigo em Inglês | MEDLINE | ID: mdl-31025693

RESUMO

Ergosterol peroxide selectively exhibits biological activity against a wide range of diseases; however, its mode of action remains unknown. Here, we present an efficient synthesis of ergosterol peroxide chemical probes for in vitro anticancer evaluation, live cell studies and proteomic profiling. Ergosterol peroxide analogues show promising anti-proliferation activity against triple negative breast cancer cellular models, revealing information on the structure-activity relationship of this natural product in order to develop superior analogues. The combined cellular studies demonstrate that ergosterol peroxide is distributed across the cytosol with significant accumulation in the endoplasmic reticulum (ER). These chemical probes support our efforts towards uncovering the potential target(s) of ergosterol peroxide against triple negative breast cancer cell lines.


Assuntos
Antineoplásicos/química , Ergosterol/análogos & derivados , Corantes Fluorescentes/química , Imagem Óptica , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Ergosterol/síntese química , Ergosterol/química , Ergosterol/farmacologia , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/farmacologia , Humanos , Microscopia de Fluorescência , Conformação Molecular , Neoplasias de Mama Triplo Negativas/patologia
10.
Molecules ; 24(7)2019 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-30925792

RESUMO

Antrodia camphorata (AC) is a rare and unique mushroom that is difficult to cultivate. Previous studies have demonstrated the bioactivity of the compound Ergosta-7,9(11),22-trien-3ß-ol (EK100) from AC in submerged culture. The purpose of this study is to evaluate the potential beneficial effects of EK100 on fatigue and ergogenic functions following physiological challenge. Male ICR (Institute of Cancer Research) mice were randomly divided into three groups (n = 8 per group) and orally administered EK100 for six weeks at 0 (Vehicle), 10 (EK100-1X), and 20 (EK100-2X) mg/kg/day. The six-week Ek100 supplementation significantly increased grip strength (P = 0.0051) in trend analysis. Anti-fatigue activity was evaluated using 15-min. acute exercise testing and measuring the levels of serum lactate, ammonia, glucose, blood urea nitrogen (BUN), and creatine kinase (CK) after a 15-min. swimming exercise. Our results indicate that AC supplementation leads to a dose-dependent decrease in serum lactate, ammonia, BUN, and CK activity after exercise and significantly increases serum glucose and glycogen content in liver tissues. Biochemical and histopathological data demonstrated that long term daily administration of EK100 for over six weeks (subacute toxicity) was safe. EK100's anti-fatigue properties appear to be through the preservation of energy storage, increasing blood glucose and liver glycogen content, and decreasing the serum levels of lactate, ammonia, BUN, and CK. EK100 could potentially be used to improve exercise physiological adaptation, promote health, and as a potential ergogenic aid in combination with different nutrient strategies.


Assuntos
Antrodia/química , Ergosterol/farmacologia , Condicionamento Físico Animal , Administração Oral , Amônia/sangue , Animais , Biomarcadores/metabolismo , Glicemia/metabolismo , Nitrogênio da Ureia Sanguínea , Peso Corporal/efeitos dos fármacos , Creatina Quinase/sangue , Ergosterol/química , Ergosterol/toxicidade , Membro Anterior/fisiologia , Glicogênio/metabolismo , Força da Mão/fisiologia , Ácido Láctico/sangue , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos Endogâmicos ICR , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Especificidade de Órgãos/efeitos dos fármacos , Natação , Testes de Toxicidade
11.
Mar Drugs ; 17(3)2019 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-30893778

RESUMO

Five new ergostanes, penicisteroids D-H (1-5), were isolated from the liquid culture of the deep-sea-derived fungus Penicillium granulatum MCCC 3A00475, along with 27 known compounds. The structures of the new steroids were established mainly on the basis of extensive analysis of 1D and 2D NMR as well as HRESIMS data. Moreover, the absolute configurations of 1 were confirmed unambiguously by the single-crystal X-ray crystallography. Compounds 2 and 4⁻7 showed moderate antiproliferative effects selectively against 12 different cancer cell lines with IC50 values of around 5 µM. Compounds 2 and 6, potent RXRα binders with Kd values of 13.8 and 12.9 µM, respectively, could induce apoptosis by a Retinoid X Receptor (RXR)-α-dependent mechanism by regulating RXRα transcriptional expression and promoting the poly-ADP-ribose polymerase (PARP) cleavage. Moreover, they could inhibit proliferation by cell cycle arrest at the G0/G1 phase.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Organismos Aquáticos/química , Ergosterol/farmacologia , Penicillium/química , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Ergosterol/análogos & derivados , Ergosterol/química , Ergosterol/isolamento & purificação , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Sedimentos Geológicos/microbiologia , Humanos , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Receptor X Retinoide alfa/metabolismo , Transdução de Sinais/efeitos dos fármacos
12.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1864(7): 1005-1016, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30917917

RESUMO

Cells maintain physicochemical characteristics of membranes in order to allow for proper function of membrane-associated cellular processes, such as endocytosis and exocytosis. To investigate the interplay between membrane properties and biological processes, we applied lipid engineering approaches that allowed for systematic manipulation of fatty acid unsaturation and sterol biosynthesis, the main regulators of membrane fluidity. In combination with electrophysiological membrane capacitance measurements, we were able to study the dependence of the endo- and exocytic activity of Saccharomyces cerevisiae on membrane lipid composition in vivo. We found that a strong decrease in the cell's total ergosterol content leads to a severely reduced frequency of vesicle fission (endocytosis), whereas the exocytic activity remained largely unaffected. In contrast, increased lipid saturation lowered both endocytic and the exocytic activity, with the former being more severely affected. We were able to correlate the decreased ratio of endocytic/exocytic frequencies (fendo/fexo) upon lipid perturbation with the growth of yeast protoplasts, which is based on a surface enlargement resulting from a net excess of exocytic over endocytic flux. Experiments using clathrin-deficient mutants confirm a correlation between reduced endocytic activity and increased size of intact walled cells, as well as accelerated protoplast growth. These data show that lipid composition is intimately tied to membrane trafficking in yeast cells and suggest that endocytosis is particularly dependent on the lipid-defined properties of cell membrane.


Assuntos
Endocitose , Exocitose , Fluidez de Membrana/fisiologia , Saccharomycetales/fisiologia , Crescimento Celular , Ergosterol/farmacologia , Lipídeos de Membrana/metabolismo , Protoplastos
13.
Nutrients ; 11(2)2019 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-30823598

RESUMO

(1) Background: Diabetic nephropathy, a microvascular complication of diabetes, is one of the principal causes of end-stage renal disease worldwide. The aim of this study was to explore the therapeutic effects of ergosterol on diabetic nephropathy. (2) Methods: Streptozotocin (STZ)-induced C57BL/6 diabetic mice were treated with ergosterol (10, 20, 40 mg/kg/day) for 8 weeks by oral gavage. The in vitro study employed rat mesangial cells exposed to 30 mM glucose for 48 h in the presence of 10 or 20 µM ergosterol. (3) Results: Ergosterol treatment improved body weights, ameliorated the majority of biochemical and renal functional parameters and histopathological changes, and reduced extracellular matrix (ECM) deposition in diabetic mice. In vitro, ergosterol suppressed proliferation, reduced the levels of ECM proteins, and increased the expression of matrix metalloproteinase-2 and -9 in high glucose-induced mesangial cells; Furthermore, ergosterol markedly improved transforming growth factor-ß1 (TGF-ß1) expression, enhanced phosphorylation levels of drosophila mothers against decapentaplegic 2 (Smad2), and regulated the downstream factors in vivo and in vitro. (4) Conclusions: Ergosterol alleviated mesangial cell proliferation and the subsequent ECM deposition by regulating the TGF-ß1/Smad2 signaling pathway.


Assuntos
Proliferação de Células/efeitos dos fármacos , Nefropatias Diabéticas/tratamento farmacológico , Ergosterol/farmacologia , Células Mesangiais/efeitos dos fármacos , Proteína Smad2/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Células Cultivadas , Diabetes Mellitus Experimental/complicações , Relação Dose-Resposta a Droga , Ergosterol/administração & dosagem , Ergosterol/química , Matriz Extracelular/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Células Mesangiais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Estrutura Molecular , Ratos , Proteína Smad2/genética , Fator de Crescimento Transformador beta1/genética
14.
Int J Mol Sci ; 20(6)2019 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-30875866

RESUMO

The impact of fungal diseases on crop production negatively reflects on sustainable food production and overall economic health. Ergosterol is the major sterol component in fungal membranes and regarded as a general elicitor or microbe-associated molecular pattern (MAMP) molecule. Although plant responses to ergosterol have been reported, the perception mechanism is still unknown. Here, Arabidopsis thaliana protein fractions were used to identify those differentially regulated following ergosterol treatment; additionally, they were subjected to affinity-based chromatography enrichment strategies to capture and categorize ergosterol-interacting candidate proteins using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Mature plants were treated with 250 nM ergosterol over a 24 h period, and plasma membrane-associated fractions were isolated. In addition, ergosterol was immobilized on two different affinity-based systems to capture interacting proteins/complexes. This resulted in the identification of defense-related proteins such as chitin elicitor receptor kinase (CERK), non-race specific disease resistance/harpin-induced (NDR1/HIN1)-like protein, Ras-related proteins, aquaporins, remorin protein, leucine-rich repeat (LRR)- receptor like kinases (RLKs), G-type lectin S-receptor-like serine/threonine-protein kinase (GsSRK), and glycosylphosphatidylinositol (GPI)-anchored protein. Furthermore, the results elucidated unknown signaling responses to this MAMP, including endocytosis, and other similarities to those previously reported for bacterial flagellin, lipopolysaccharides, and fungal chitin.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Membrana Celular/metabolismo , Ergosterol/farmacologia , Arabidopsis/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Cromatografia de Afinidade , Cromatografia Líquida , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas Quinases/metabolismo , Transdução de Sinais/efeitos dos fármacos , Espectrometria de Massas em Tandem
15.
Phytomedicine ; 54: 291-301, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30668380

RESUMO

BACKGROUND: In our previous study the synergism of four combinations of Zuccagnia punctata (ZpE) and Larrea nitida (LnE) exudates with the reliable statistical-based MixLow method was assessed, and the markers of the most anti-C. albicans synergistic ZpE-LnE bi-herbal combination were quantified according to European Medicines Agency (EMA). PURPOSE: To study the mechanisms of action as well as the cytotoxic properties of the ZpE-LnE most synergistic combination found in the previous work. MATERIALS AND METHODS: Minimum Fungicidal Concentration (MFC) and rate of killing of ZpE-LnE were assessed with the microbroth dilution and the time-kill assays respectively. Morphological alterations were observed with both confocal and fluorescence microscopy on the yeast Schizosaccharomyces pombe. The ergosterol exogenous assay, the quantification of ergosterol, the sorbitol as well as glucan synthase (GS) and chitin synthase (ChS) assays were used to detect the effects on the fungal membrane and cell wall respectively. The capacity of ZpE-LnE of inhibiting Candida virulence factors was assessed with previously reported methods. The effect of ZpE-LnE and of ZpE or LnE alone on cell viability was determined on human hepatoma cells line Huh7. RESULTS: ZpE-Ln E was fungicidal killing C. albicans in a shorter time than amphotericin B and produced malformations in S. pombe cells. ZpE-LnE showed to bind to ergosterol but not to inhibit any step of the ergosterol biosynthesis. ZpE-LnE showed a low or moderate capacity of inhibiting GS and ChS. Regarding inhibition of virulence factors, ZpE-LnE significantly decreased the capacity of adhesion to eukaryotic buccal epithelial cells (BECs), did not inhibit the germ tube formation and inhibited the secretion of phospholipases and proteinases but not of haemolysins. ZpE-LnE demonstrated very low toxicity on Huh7 cells, much lower than that each extract alone. CONCLUSION: The fungicidal properties of ZpE-LnE against C. albicans, its dual mechanism of action targeting the fungal membrane's ergosterol as well as the cell wall, its capacity of inhibiting several important virulence factors added to its low toxicity, make ZpE-LnE a good candidate for the development of a new antifungal bi-Herbal Medicinal Product.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Fabaceae/química , Larrea/química , Extratos Vegetais/farmacologia , Anfotericina B/farmacologia , Ergosterol/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Plantas Medicinais
16.
Org Biomol Chem ; 17(7): 1624-1633, 2019 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-30357222

RESUMO

Classical steroids are long-known privileged leads in drug discovery. Their rearranged counterparts, though, have so far received less attention, although recent isolation and biological testing programmes have revealed a plethora of molecular entities that are both structurally intriguing, as well as biologically relevant. This review will highlight those natural products, and focus on ergostane-derived seco- and abeo-steroids. Their isolation, structure elucidation, and biological properties are reported. A special emphasis of this review lies in their respective (and typically proposed) biosyntheses, to help guide future bio-inspired synthetic attempts.


Assuntos
Produtos Biológicos/química , Produtos Biológicos/farmacologia , Descoberta de Drogas/métodos , Ergosterol/análogos & derivados , Química Farmacêutica , Ergosterol/química , Ergosterol/farmacologia
17.
Sci Rep ; 8(1): 17956, 2018 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-30560887

RESUMO

As part of our ongoing search for novel therapeutic structures from microorganism, the chemical examination of marine fungus Phoma sp. resulted in the isolation of ergosterol, ergosterol peroxide (EP), and 9,11-dehydroergosterol peroxide (DEP). The bioassay results demonstrated that the three isolates reduced the viability of various cancer cells, with EP being highest in human lung cancer cell line A549 cells. EP induced caspase-dependent apoptosis through mitochondrial damage in A549 cells. Additionally, EP-induced ROS generation and apoptosis were attenuated by ROS-generating enzymes inhibitors and antioxidant N-acetylcysteine, indicated that ROS played an important role in EP-mediated apoptosis in A549 cells. Furthermore, it was observed that EP induced ROS-dependent autophagy, which attenuated apoptosis in A549 cells. On the other hand, EP reduced the LPS/ATP-induced proliferation and migration of A549 cells through attenuated NLRP3 inflammasome activity. Additionally, EP showed synergistic cytotoxic effect with antitumor drug Sorafenib in A549 cell viability inhibition. Furthermore, Micro-Western Array and Western blot analyses demonstrated that the protein levels of EGFR, HSP27, MEK5, AKT1, mTOR, Smad2, Smad3, TAB1, NF-κB, and HIF1-α decreased, while the levels of p-p38α, p-ERK1/2, p-JNK, fibronectin and p27 increased. Collectively, the results of this study demonstrated that EP might be useful to develop a therapeutic candidate for lung cancer complications.


Assuntos
Apoptose/efeitos dos fármacos , Organismos Aquáticos/química , Autofagia/efeitos dos fármacos , Ergosterol/análogos & derivados , Fungos/química , Células A549 , Adenocarcinoma de Pulmão/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sinergismo Farmacológico , Ergosterol/química , Ergosterol/farmacologia , Ergosterol/toxicidade , Fungos/isolamento & purificação , Humanos , Inflamassomos , Estrutura Molecular , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sorafenibe/farmacologia , Ensaio Tumoral de Célula-Tronco
18.
Int J Mol Sci ; 19(12)2018 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-30544579

RESUMO

Isaria cicadae, a medicinal food fungus, is a fruit from Paecilomyces cicadae. In this study, we purified ergosterol peroxide (EP) from the fermentation broth of P. cicadae and investigated its effects on renal cell carcinoma (RCC) cells, in vitro. EP was purified from P. cicadae fermentation broth. The human RCC cell line 786-0 was used to analyze the anticancer mechanism of EP and inhibit its effect on cancer cell proliferation, in vitro. EP with a validated structure showed a yield rate of 20.1 mg/L and a purity of 96%. EP significantly inhibited RCC cell growth and clone formation in vitro. In addition, EP suppressed the migration and invasion, triggered the apoptosis, and modulated the cell cycle of RCC cells, in a dose-dependent manner. It also downregulated ß-catenin expression. EP could be routinely produced through P. cicadae. It fights RCC cells in vitro through multiple mechanisms, including suppressing cell growth, colonization, migration, and invasion, arresting the cell cycle, attenuating ß-catenin pathways, and triggering apoptosis.


Assuntos
Ergosterol/análogos & derivados , Paecilomyces/metabolismo , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ergosterol/metabolismo , Ergosterol/farmacologia , Fermentação/fisiologia , Humanos
19.
Artigo em Inglês | MEDLINE | ID: mdl-30126959

RESUMO

Targeting lanosterol 14α-demethylase (LDM) with azole drugs provides prophylaxis and treatments for superficial and disseminated fungal infections, but cure rates are modest for immunocompromised patients and individuals with comorbidities. The efficacy of azole drugs has also been reduced due to the emergence of drug-resistant fungal pathogens. We have addressed these problems by expressing in Saccharomyces cerevisiae functional, hexahistidine-tagged, full-length Candida albicans LDM (CaLDM6×His) and Candida glabrata LDM (CgLDM6×His) for drug discovery purposes and determining their X-ray crystal structures. Compared with S. cerevisiae overexpressing LDM6×His (ScLDM6×His), the reduced susceptibility of CgLDM6×His to all azole drugs tested correlated with its level of overexpression. In contrast, the reduced susceptibility to short-tailed (fluconazole and voriconazole) but not medium-tailed (VT-1161) or long-tailed azoles (itraconazole and posaconazole) indicates CaLDM6×His works best when coexpressed with its cognate NADPH-cytochrome P450 reductase (CaNcp1A) rather than the host reductase (ScNcp1). Overexpression of LDM or Ncp1 modified the ergosterol content of yeast and affected growth inhibition by the polyene antibiotic amphotericin B. Affinity-purified recombinant Candida LDMs bind carbon monoxide and show tight type II binding of a range of azole drugs, including itraconazole, posaconazole, fluconazole, and voriconazole. This study provides a practical basis for the phenotype-, biochemistry-, and structure-directed discovery of novel antifungals that target LDMs of fungal pathogens.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida glabrata/efeitos dos fármacos , Lanosterol/metabolismo , Esterol 14-Desmetilase/metabolismo , Anfotericina B/farmacologia , Azóis/farmacologia , Farmacorresistência Fúngica/efeitos dos fármacos , Ergosterol/farmacologia , Fluconazol/farmacologia , Proteínas Fúngicas/metabolismo , Humanos , Itraconazol/farmacologia , Testes de Sensibilidade Microbiana/métodos , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Triazóis/farmacologia , Voriconazol/farmacologia
20.
Biosci Biotechnol Biochem ; 82(10): 1803-1811, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29968517

RESUMO

Grifola frondosa is an edible mushroom consumed as a health food and/or traditional medicine in Asia. However, the anti-allergic effects of G. frondosa are not yet understood. In this study, we demonstrated the effects of G. frondosa extract (GFE) on IgE-mediated allergic responses, using antigen-stimulated RBL-2H3 cells. Three active compounds: ergosterol, 6ß-methoxyergosta-7,22-dien-3ß,5α-diol (MEDD), and 6-oxoergosta-7,22-dien-3ß-ol (6-OXO) were isolated from GFE and shown to inhibit the antigen-induced release of ß-hexosaminidase and histamine. Among the three active components, we focused on ergosterol because of its high content in GFE. Ergosterol inhibited the aggregation of high-affinity IgE receptor (FcεRI), which is the first step in the activation of mast cells and antigen-induced tyrosine phosphorylation. Furthermore, ergosterol suppressed antigen-increased IL-4 and TNF-α mRNA. Taken together, our findings suggest that G. frondosa, including ergosterol and its derivatives as active components, has the potential to be a novel functional food that prevents type I allergies.


Assuntos
Antígenos/imunologia , Degranulação Celular/efeitos dos fármacos , Degranulação Celular/imunologia , Ergosterol/farmacologia , Grifola/química , Mastócitos/efeitos dos fármacos , Receptores de IgE/efeitos dos fármacos , Animais , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Linhagem Celular , Ergosterol/química , Alimento Funcional , Liberação de Histamina/efeitos dos fármacos , Mastócitos/imunologia , Espectroscopia de Prótons por Ressonância Magnética , Ratos , Reação em Cadeia da Polimerase em Tempo Real , Receptores de IgE/imunologia , Espectrometria de Massas por Ionização por Electrospray
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