RESUMEN
Bacillus subtilis species complex is known as lipopeptide-producer with biotechnological potential for pharmaceutical developments. This study aimed to identify lipopeptides from a bacterial isolate and evaluate their antifungal effects. Here, we isolated and identified a lipopeptide-producing bacterium as a species of Bacillus subtilis complex (strain UL-1). Twenty lipopeptides (six iturins, six fengycins, and eight surfactins) were identified in the crude extract (CE) and fractions (F1, F2, F3, and F4), and the highest content of total lipopeptides was observed in CE and F2. The chemical quantification data corroborate with the hemolytic and antifungal activities that CE and F2 were the most hemolytic and inhibited the fungal growth at lower concentrations against Fusarium spp. In addition, they caused morphological changes such as shortening and/or atypical branching of hyphae and induction of chlamydospore-like structure formation, especially in Fusarium solani. CE was the most effective in inhibiting the biofilm formation and in disrupting the mature biofilm of F. solani reducing the total biomass and the metabolic activity at concentrations ≥ 2 µg/mL. Moreover, CE significantly inhibited the adherence of F. solani conidia on contact lenses and nails as well as disrupted the pre-formed biofilms on nails. CE at 100 mg/kg was nontoxic on Galleria mellonella larvae, and it reduced the fungal burden in larvae previously infected by F. solani. Taken together, the lipopeptides obtained from strain UL-1 demonstrated a potent anti-Fusarium effect inducing morphological alterations and antibiofilm activities. Our data open further studies for the biotechnological application of these lipopeptides as potential antifungal agents. KEY POINTS: ⢠Lipopeptides inhibit Fusarium growth and induce chlamydospore-like structures. ⢠Lipopeptides hamper the adherence of conidia and biofilms of Fusarium solani. ⢠Iturins, fengycins, and surfactins were associated with antifungal effects.
Asunto(s)
Antifúngicos , Bacillus subtilis , Bacillus subtilis/metabolismo , Antifúngicos/química , Esporas Fúngicas/metabolismo , Biopelículas , Lipopéptidos/metabolismo , Péptidos Cíclicos/metabolismo , Enfermedades de las Plantas/microbiologíaRESUMEN
Machaerium hirtum (Vell.) Stellfeld (M.hirtum) is a plant known as 'jacarandá-bico-de-pato' whose bark is commonly used against diarrhea, cough and cancer. The aim of this study was to phytochemically characterise the hydroethanolic extract of this plant, investigate its antimutagenic activities using the Ames test and evaluate its effects on cell viability, genomic instability, gene expression and cell protection in human hepatocellular carcinoma cells (HepG2). Antimutagenic activity was assessed by simultaneous pre- and post-treatment with direct and indirect mutagens, such as 4-nitro-o-phenylenediamine (NPD), mitomycin C (MMC), benzo[a]pyrene (B[a]P) and aflatoxin B1 (AFB1), using the Ames test, cytokinesis blocking micronucleus and apoptosis assays. Only 3 of the 10 concentrations evaluated in the MTT assay were cytotoxic in HepG2 cells. Micronucleated or apoptotic cells were not observed with any of the tested concentrations, and there were no mutagenic effects in the bacterial system. However, the Nuclear Division Index and flow cytometry data showed a decrease in cell proliferation. The extract showed an inhibitory effect against direct (NPD) and indirect mutagens (B[a]P and AFB1). Furthermore, pre- and post-treated cells showed significant reduction in the number of apoptotic and micronucleated cells. This effect is not likely to be associated with the modulation of antioxidant genes, as shown by the RT-qPCR results. Six known flavonoids were identified in the hydroethanolic extract of Machaerium hirtum leaves, and their structures were elucidated by spectroscopic and spectrophotometric methods. The presence of the antioxidants apigenin and luteolin may explain these protective effects, because these components can inhibit the formation of reactive species and prevent apoptosis and DNA damage. In conclusion, the M.hirtum extract showed chemopreventive potential and was not hazardous at the tested concentrations in the experiments presented here. Moreover, this extract should be investigated further as a chemopreventive agent.
Asunto(s)
Antimutagênicos/farmacología , Fabaceae/química , Extractos Vegetales/farmacología , Antimutagênicos/química , Antimutagênicos/toxicidad , Apoptosis/efectos de los fármacos , Apoptosis/genética , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Línea Celular Tumoral , Daño del ADN/efectos de los fármacos , Flavonoides/química , Flavonoides/farmacología , Flavonoides/toxicidad , Citometría de Flujo , Expresión Génica , Humanos , Micronúcleos con Defecto Cromosómico , Pruebas de Micronúcleos , Pruebas de Mutagenicidad , Mutación/efectos de los fármacos , Extractos Vegetales/química , Extractos Vegetales/toxicidad , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genéticaRESUMEN
Harvestmen have a pair of scent glands that open through ozopores. The literature suggests a link between the morphology of the ozopore area and the emission of a defensive secretion. A previous study on a species that aggregates in open areas, where individuals are probably more easily spotted by predators, showed that this defensive secretion causes conspecifics to flee. However, it is unknown whether this behavior occurs in species that aggregate in sheltered areas, where prey are harder to find. Herein, we describe the morphology of the ozopore area, the mode of emission of the defensive secretion, and its chemical composition in the harvestman Discocyrtus pectinifemur. We also tested if the defensive secretion is used as an alarm pheromone. We found that D. pectinifemur releases the defensive secretion in different ways, one of them being as a jet. Emission as a jet contrasts with that known for all congeners previously studied, and is in accord with the expected morphology of the ozopore. We found that the defensive secretion of D. pectinifemur does not function as an alarm pheromone. The composition of the defensive secretion, a mixture of quinones, is congruent with those already described for the clade that includes Discocyrtus. Our results support the link between the morphology of the scent glands area and the emission behavior of the defensive secretion, and they suggest that the alarm pheromone function in harvestmen may be dependent on ecological factors.
Asunto(s)
Arácnidos/fisiología , Animales , Arácnidos/anatomía & histología , Arácnidos/química , Femenino , Masculino , Feromonas/análisis , Glándulas Odoríferas/anatomía & histología , Glándulas Odoríferas/química , Glándulas Odoríferas/fisiologíaRESUMEN
In this study, we specifically focused on the crude methanolic leaf extract of Byrsonima coccolobifolia, investigating its antifungal potential against human pathogenic fungi and its antiviral activity against COVID-19. Through the use of high-performance liquid chromatography coupled with electrospray ionization ion trap tandem mass spectrometry, direct infusion electrospray ionization ion trap tandem mass spectrometry, and chromatographic dereplication procedures, we identified galloyl quinic acid derivatives, catechin derivatives, proanthocyanidins, and flavonoid glycosides. The broth dilution assay revealed that the methanolic leaf extract of B. coccolobifolia exhibits antifungal activity against Cryptococcus neoformans (IC50 = 4 µg/mL). Additionally, docking studies were conducted to elucidate the interactions between the identified compounds and the central residues at the binding site of biological targets associated with COVID-19. Furthermore, the extract demonstrated an in vitro half-maximum effective concentration (EC50 = 7 µg/mL) and exhibited significant selectivity (>90%) toward SARS-CoV-2.
Asunto(s)
COVID-19 , Extractos Vegetales , Humanos , Extractos Vegetales/farmacología , Extractos Vegetales/química , Antifúngicos , Estructura Molecular , SARS-CoV-2 , Espectrometría de Masa por Ionización de Electrospray/métodos , Metanol , Antivirales/farmacología , Cromatografía Líquida de Alta Presión/métodosRESUMEN
The identification of new drugs with few or no adverse effects is of great interest worldwide. In cancer therapy, natural products have been used as chemopreventive and chemotherapeutic agents. Plants from the Brazilian savannah belonging to the Byrsonima genus are popularly known as muricis and have attracted much attention due to their various pharmacological activities. However, there are currently no data on these plants concerning their use as chemopreventive or chemotherapeutic agents in human cell lines. The present study assessed the potential of B. correifolia, B. verbascifolia, B. crassifolia, and B. intermedia extracts as natural alternatives in the prevention and/or treatment of cancer. The chemical constituents present in each extract were analyzed by electrospray ionization-mass spectrometry (ESI-MSN). The mutagenic/antimutagenic (micronucleus assay), genotoxic/antigenotoxic (comet assay), apoptotic/necrotic (acridine orange/ethidium bromide uptake), and oxidative/antioxidative (CM-H2DCFDA) effects of the extracts and their influence on gene expression (RTqPCR) were investigated in nonmetabolizing gastric (MNP01) and metabolizing hepatocarcinoma (HepG2) epithelial cells to evaluate the effects of metabolism on the biological activities of the extracts. The genotoxicity, mutagenicity, and apoptotic effects observed in HepG2 cells with B. correifolia and B. verbascifolia extracts are probably associated with the presence of proanthocyanidins and amentoflavone. In MNP01 cells, none of the four extracts showed mutagenic effects. B. crassifolia and B. intermedia extracts exhibited strong antimutagenicity and enhanced detoxification in HepG2 cells and antioxidant capacities in both types of cells, possibly due to the presence of gallic and quinic acids, which possess chemopreventive properties. This study identifies for the first time B. correifolia and B. verbascifolia extracts as potential agents against hepatocarcinoma and B. crassifolia and B. intermedia extracts as putative chemopreventive agents.
Asunto(s)
Anticarcinógenos , Antimutagênicos , Humanos , Extractos Vegetales/farmacología , Extractos Vegetales/química , Brasil , Plantas , Antioxidantes/farmacología , Mutágenos/toxicidad , Inestabilidad Genómica , Antimutagênicos/farmacologíaRESUMEN
Byrsonima crassa Niedenzu (Malpighiaceae) is used in Brazilian folk medicine for the treatment of diseases related mainly to gastric ulcers. In a previous study, our group described the gastric protective effect of the methanolic extract from the leaves of B. crassa. The present study was carried out to investigate the effects of methanolic extract and its phenolic compounds on the respiratory burst of neutrophils stimulated by H. pylori using a luminol-based chemiluminescence assay as well as their anti-H. pylori activity. The suppressive activity on oxidative burst of H. pylori-stimulated neutrophils was in the order of methyl gallate > (+)-catechin > methanol extract > quercetin 3-O-α-l-arabinopyranoside > quercetin 3-O-ß-d-galactopyranoside > amentoflavone. Methyl gallate, compound that induced the highest suppressive activity with IC(50) value of 3.4 µg/mL, did not show anti-H. pylori activity. B. crassa could be considered as a potential source of natural antioxidant in gastric ulcers by attenuating the effects on the damage to gastric mucosa caused by neutrophil generated reactive oxygen species, even when H. pylori displays its evasion mechanisms.
Asunto(s)
Antioxidantes/farmacología , Helicobacter pylori/patogenicidad , Malpighiaceae/química , Fenoles/farmacología , Estallido Respiratorio/efectos de los fármacos , Antioxidantes/química , Ácido Gálico/análogos & derivados , Ácido Gálico/química , Ácido Gálico/aislamiento & purificación , Ácido Gálico/farmacología , Mucosa Gástrica/citología , Mucosa Gástrica/microbiología , Mucosa Gástrica/patología , Helicobacter pylori/efectos de los fármacos , Malpighiaceae/metabolismo , Metanol/química , Neutrófilos/inmunología , Neutrófilos/microbiología , Fenoles/química , Fenoles/aislamiento & purificación , Extractos Vegetales/química , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Byrsonima fagifolia Niedenzu (Malpighiaceae) and other Byrsonima species are popularly employed in Brazilian traditional medicine in the form of preparations as cicatrizing, anti-inflammatory, and antimicrobial. AIM OF THE STUDY: To characterize the phytochemical profile of the hydromethanolic extract obtained from B. fagifolia leaves (BF extract) and to evaluate the toxicity and the antifungal activity. MATERIALS AND METHODS: The compounds from BF extract were isolated by HPLC and the structures were elucidated based on extensive analyses of 1D and 2D NMR spectra (HMQC, HMBC and COSY) data. The antifungal effect was determined by the broth microdilution method and the toxicity was evaluated on erythrocytes from sheep's blood and Galleria mellonella larvae. RESULTS: Phytochemical investigation of the BF extract led to the isolation and characterization of pyrogallol, n-butyl gallate, 3,4-di-O-galloylquinic acid, 3,5-di-O-galloylquinic acid, 3,4,5-tri-O-galloylquinic acid, and 1,3,4,5-tetra-O-galloylquinic acid. The BF extract showed high content of galloylquinic acid derivatives reaching more than twenty-times the quercetin derivatives content, according to the quantification by HPLC. These galloylquinic acid derivatives, obtained during this study, and quercetin derivatives, previously isolated, were submitted to the antifungal assays. The BF extract inhibited yeast growth mainly against Cryptococcus spp., at concentrations of 1-16 µg/mL, comparable to isolated compounds galloylquinic acid derivatives. However, the quercetin derivatives as well as quinic acid, gallic acid, and methyl gallate showed lower antifungal effect compared with galloylquinic derivatives. In addition, the BF extract had no hemolytic effect and no toxicity on G. mellonella. CONCLUSION: The phytochemical analysis revealed that galloylquinic acid derivatives are the major compounds in the leaves of B. fagifolia and they are associated to anti-cryptococcal activity and presented reduced toxicity.
Asunto(s)
Antifúngicos , Malpighiaceae , Animales , Antifúngicos/toxicidad , Malpighiaceae/química , Fitoquímicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/toxicidad , Hojas de la Planta , Quercetina , OvinosRESUMEN
The analysis by HPLC-PDA of the hydroalcoholic extract of the leaves of M. eriocarpum together with the injection of the fractions containing the already identified metabolites allowed the detection of at least 5 flavonoids, of which two are derived from apigenin and three from luteolin. After isolating larger amounts of isovitexin (I), assays were performed to evaluate the allelopathic activity together with the crude extract. The results show that the initial inhibition indexes were very similar to those observed in the treatments with F17 (Fraction enriched in isovitexin) and F18 (isovitexin), mainly in the concentrations of 500 and 1000 mg L-1. The index of the number of lateral roots, an increase of the inhibitory effect is observed with the increase of the concentration of M. eriocarpum extract.
Asunto(s)
Alelopatía , Fabaceae/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Apigenina/aislamiento & purificación , Apigenina/farmacología , Cromatografía Líquida de Alta Presión , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Luteolina/química , Extractos Vegetales/química , Raíces de PlantasRESUMEN
:Machaerium hirtum (Vell.) Stellfeld (Fabaceae) known in Brazil as "jacaranda de espinho" or "espinheira santa nativa" is a medicinal plant commonly used in folk medicine to treat ulcers, cough and diarrhea. This study aimed to investigate the anti-inflammatory and antinociceptive effects of hydroalcoholic extracts from M. hirtum twig (HEMh) using in vivo experimental models of nociception through the involvement of transient receptor potential channels, acid-sensing ion channel (ASIC), nitrergic, opioidergic, glutamatergic, and supraspinal pathways. Our results revealed an antinociceptive effect of HEMh mediated by the opioidergic, L-arginine-nitric oxide and glutamate systems, as well as by interactions with TRPA1/ASIC channels. The anti-inflammatory effect of HEMh evaluated with a xylene-induced ear edema and by the involvement of arachidonic acid and prostaglandin E2 (PGE2) showed involvement of the COX pathway, based on observed decreases in PGE2 levels. A phytochemical investigation of the HEMh led to the isolation of α-amyrin, ß-amyrin, allantoin, apigenin-7-methoxy-6-C-ß-D-glucopyranoside, and apigenin-6-C-ß-D-glucopyranosyl-8-C-ß-D-xylopyranoside. In conclusion, the acute oral administration of HEMh inhibits the nociceptive behavioral response in animals through the nitrergic, opioid, glutamatergic pathways, and by inhibition of the TRPA1 and ASIC channels, without causing locomotor dysfunction. In addition, its anti-inflammatory effect is associated with the COX pathway and decreased PGE2 levels.
Asunto(s)
Dolor Agudo/tratamiento farmacológico , Fabaceae/química , Inflamación/tratamiento farmacológico , Dolor Agudo/complicaciones , Analgésicos Opioides/metabolismo , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Ácido Araquidónico , Arginina/metabolismo , Peso Corporal/efectos de los fármacos , Dinoprostona/metabolismo , Edema/tratamiento farmacológico , Etanol , Femenino , Formaldehído , Glutamatos/metabolismo , Indometacina/efectos adversos , Inflamación/complicaciones , Canales Iónicos/metabolismo , Masculino , Ratones , Actividad Motora/efectos de los fármacos , Óxido Nítrico/metabolismo , Nocicepción/efectos de los fármacos , Fitoquímicos/química , Fitoquímicos/farmacología , Fitoquímicos/uso terapéutico , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Extractos Vegetales/toxicidad , Pruebas de Toxicidad Aguda , Úlcera/inducido químicamente , Úlcera/complicaciones , Úlcera/tratamiento farmacológico , XilenosRESUMEN
BACKGROUND: Several in vitro studies have looked at the effect of medicinal plant extracts against Helicobacter pylori (H. pylori). Regardless of the popular use of Byrsonima crassa (B. crassa) as antiemetic, diuretic, febrifuge, to treat diarrhea, gastritis and ulcers, there is no data on its effects against H. pylori. In this study, we evaluated the anti-H. pylori of B. crassa leaves extracts and its effects on reactive oxygen/nitrogen intermediates induction by murine peritoneal macrophages. METHODS: The minimal inhibitory concentration (MIC) was determined by broth microdilution method and the production of hydrogen peroxide (H2O2) and nitric oxide (NO) by the horseradish peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. RESULTS: The methanolic (MeOH) and chloroformic (CHCl3) extracts inhibit, in vitro, the growth of H. pylori with MIC value of 1024 microg/ml. The MeOH extract induced the production H2O2 and NO, but CHCl3 extract only NO. CONCLUSION: Based in our results, B. crassa can be considered a source of compounds with anti-H. pylori activity, but its use should be done with caution in treatment of the gastritis and peptic ulcers, since the reactive oxygen/nitrogen intermediates are involved in the pathogenesis of gastric mucosal injury induced by ulcerogenic agents and H. pylori infections.
Asunto(s)
Antibacterianos/farmacología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/efectos de los fármacos , Malpighiaceae , Extractos Vegetales/farmacología , Animales , Antibacterianos/uso terapéutico , Infecciones por Helicobacter/tratamiento farmacológico , Peróxido de Hidrógeno/metabolismo , Macrófagos/metabolismo , Malpighiaceae/química , Ratones , Óxido Nítrico/metabolismo , Extractos Vegetales/uso terapéutico , Hojas de la PlantaRESUMEN
The purpose of the present study was to investigate antitumour and anti-inflammatory activities of flavonoids isolated from Byrsonima crassa, Davilla elliptica and Mouriri pusa. The antitumour activity was measured by the MTT assay in murine mammary tumour cells (LM2) and the IC50 values of the flavonoids tested ranged from (31.5 +/- 2.97) to (203.1 +/- 5.9) microg/ml. The flavonoids 1 (myricetin-3-O-alpha-L-rhamnopyranoside) and 3 (quercetin-3-O-galactopyranoside) from D. elliptica were the most active ones against the tumour cells. The same samples were tested to determine the inhibition of the release of nitric oxide (NO) and of the tumour necrosis factor-alpha (TNF-alpha) in murine macrophages by the Griess and ELISA sandwich assay, respectively. Almost all the samples showed inhibitory activity to the release of NO but not of TNF-alpha. Of all substances tested, flavonoids 2 (quercetin) and 6 (myricetin) may show promising activity in the treatment of murine breast cancer by immunomodulatory and antiproliferative activities.
Asunto(s)
Antineoplásicos/farmacología , Flavonoides/farmacología , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Animales , Antineoplásicos/aislamiento & purificación , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cisplatino/farmacología , Femenino , Flavonoides/aislamiento & purificación , Malpighiaceae , Neoplasias Mamarias Experimentales/patología , Ratones , Ratones Endogámicos BALB C , MyrtaceaeRESUMEN
Byrsonima intermedia is a species of bush popularly used to treat gastrointestinal disorders, such as gastric ulcers, gastritis, and diarrhea. Previous studies have revealed that the methanolic crude extract of B. intermedia leaves has gastroprotective and healing properties. In this new study, we specifically investigated two purified partitions, ethyl acetate (EtOAc) and water (AcoAq), obtained from the crude extract to characterize the antiulcer effects of these two partitions and the mechanisms of action of this medicinal plant. The healing effects of these partitions on the gastric and duodenal mucosa were assessed after ischemia-reperfusion (I/R) or acetic acid-induced injury. The involvement of tumor necrosis factor-alpha (TNF-alpha), interleukin 1ß (IL-1ß), interleukin 10 (IL-10), and myeloperoxidase (MPO) activity and glutathione (GSH) levels were determined. The antibacterial activity against Helicobacter pylori was evaluated using microdilution methods. The phytochemical analysis of AcoAq revealed a predominance of oligomeric proanthocyanidins and galloyl quinic esters, whereas EtOAc was found to contain concentrated flavonoids. Both partitions led to a significant reduction in gastric lesions, but AcoAq was more effective than EtOAc with regard to anti-Helicobacter pylori activity in addition to protecting the gastric mucosa against ethanol, non-steroidal anti-inflammatory drugs (NSAIDs) and duodenal mucosal damage induced by cysteamine. Additionally, both partitions were associated with a significant increase in gastric and duodenal healing and increased gastric mucosal GSH content after damage induced by acetic acid. On the other hand, after 6 days of treatment, EtOAc was more effective than AcoAq in ameliorating gastric damage upon initiation of the gastric I/R, which was accompanied by a significant reduction in the activity of gastric mucosal MPO, IL 1-ß and TNF-alpha, as well as an elevation in IL-10 and GSH content. These results demonstrate that the oligomeric proanthocyanidins and galloyl quinic esters present in AcoAq were more effective in the prevention of gastric and duodenal ulcers due to the antioxidant effects of these compounds, whereas the flavonoids present in EtOAc were more effective due to their anti-inflammatory activity on the gastric and duodenal tissue. All these results confirm that the rich phytochemical diversity of B. intermedia contributes to the pharmacological actions of this medicinal plant on the gastrointestinal tract in addition to its activity against H. pylori.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Malpighiaceae/química , Úlcera Péptica/tratamiento farmacológico , Animales , Antiulcerosos/farmacología , Flavonoides/farmacología , Mucosa Gástrica/efectos de los fármacos , Mucosa Gástrica/metabolismo , Gastritis/tratamiento farmacológico , Gastritis/metabolismo , Glutatión/metabolismo , Masculino , Medicina Tradicional/métodos , Úlcera Péptica/metabolismo , Fitoquímicos/farmacología , Fitoterapia/métodos , Extractos Vegetales/farmacología , Hojas de la Planta/química , Plantas Medicinales/química , Ratas , Ratas Wistar , Estómago/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacosRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ethnopharmacological survey indicated leaves of Byrsonima fagifolia Nied. (Malpighiaceae) against gastrointestinal disorders. AIM OF THE STUDY: The methanolic extract from the leaves of Byrsonima fagifolia (denominated BF) was evaluated for toxic, mutagenic, gastroprotective, antidiarrheal, antibacterial and immunomodulatory activities. MATERIALS AND METHODS: The preventive and healing action of BF against gastric ulcer was evaluated in experimental models in rodents. We evaluated immunomodulatory (by murine peritoneal macrophages), antidiarrheal (by induced diarrhea with castor oil and intestinal motility) and antibacterial action of BF against standard strain of Escherichia coli, Staphylococcus aureus and Helicobacter pylori. The safety of use of BF was also evaluated by mutagenic (Ames assay) and by analyses of toxicity parameters. RESULTS: Phytochemical BF profile indicated the presence of phenolic compounds with antioxidant and radical-scavenging properties. BF significantly inhibited gastric lesions induced by ethanol and HCl/ethanol and endogenous mucosal sulphydryl groups (SHs) participated efficaciously in BF gastroprotection. BF blocked development of inflammation process and also has antidiarrheal actions. This extract accelerated the healing of the gastric ulcerated mucosa by stimulating proliferative factors and by increasing production of gastric mucus with no toxic action. The substances responsible for the protective action are concentrated in the ethyl acetate fraction that demonstrated no mutagenic action in vitro. CONCLUSIONS: Byrsonima fagifolia presents gastroprotective, healing and antidiarrheal activities supporting previous claims that its traditional use by Brazilians can treat these gastrointestinal ailments.
Asunto(s)
Antioxidantes/farmacología , Malpighiaceae/química , Extractos Vegetales/farmacología , Animales , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Antibacterianos/toxicidad , Antiulcerosos/aislamiento & purificación , Antiulcerosos/farmacología , Antiulcerosos/toxicidad , Antidiarreicos/aislamiento & purificación , Antidiarreicos/farmacología , Antidiarreicos/toxicidad , Antioxidantes/aislamiento & purificación , Antioxidantes/toxicidad , Brasil , Modelos Animales de Enfermedad , Depuradores de Radicales Libres/aislamiento & purificación , Depuradores de Radicales Libres/farmacología , Depuradores de Radicales Libres/toxicidad , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Factores Inmunológicos/toxicidad , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Masculino , Medicina Tradicional , Ratones , Pruebas de Mutagenicidad , Extractos Vegetales/toxicidad , Hojas de la Planta , Ratas , Ratas Wistar , Úlcera Gástrica/tratamiento farmacológico , Pruebas de ToxicidadRESUMEN
Byrsonima basiloba A. Juss. species is a native arboreal type from the Brazilian "cerrado" (tropical American savanna), and the local population uses it to treat diseases, such as diarrhea and gastric ulcer. It belongs to the Malpighiaceae family, and it is commonly known as "murici." Considering the popular use of B. basiloba derivatives and the lack of pharmacological potential studies regarding this vegetal species, the mutagenic and antimutagenic effect of methanol (MeOH) and chloroform extracts were evaluated by the Ames test, using strains TA97a, TA98, TA100, and TA102 of Salmonella typhimurium. No mutagenic activity was observed in any of the extracts. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine, sodium azide, mitomycin C, aflatoxin B(1), benzo[a]pyrene, and hydrogen peroxide. Both the extracts evaluated showed antimutagenic activity, but the highest value of inhibition level (89%) was obtained with the MeOH extract and strain TA100 in the presence of aflatoxin B(1). Phytochemical analysis of the extracts revealed the presence of n-alkanes, lupeol, ursolic and oleanolic acid, (+)-catechin, quercetin-3-O-alpha-L-arabinopyranoside, gallic acid, methyl gallate, amentoflavone, quercetin, quercetin-3-O-(2"-O-galloyl)-beta-D-galactopyranoside, and quercetin-3-O-(2"-O-galloyl)-alpha-L-arabinopyranoside.
Asunto(s)
Antimutagênicos/farmacología , Malpighiaceae/química , Mutágenos/farmacología , Extractos Vegetales/farmacología , Hojas de la Planta/química , Cloroformo , Flavonoides/análisis , Metanol , Pruebas de Mutagenicidad , Salmonella typhimuriumRESUMEN
Byrsonima intermedia is a native species of the cerrado formation (tropical American savannah). In Brazil, this plant has been used for the treatment of fever, in ulcers, as a diuretic, as antiasthmatics and in skin infections. Members of the genus Byrsonima (Malpighiaceae) are employed not only in the folk medicine but also as food to make juice, jellies and liquor. The aim of this work was to evaluate the mutagenic effects of Byrsonima intermedia, common name 'murici'. Phytochemical analysis of methanol extract furnished (+)-catechin, (-)-epicatechin, quercetin-3-O-beta-d-galactopyranoside, methyl gallate, gallic acid, quercetin-3-O-alpha-l-arabinopyranoside, amentoflavone, quercetin, quercetin-3-O-(2''-O-galloyl)-beta-galactopyranoside and quercetin-3-O-(2''-O-galloyl)-alpha-arabinopyranoside. Methanol, hydromethanol and chloroform extracts were evaluated in mutagenic assay with Salmonella typhimurium (Ames test) and mice (Micronucleus test). The methanolic extract presented signs of mutagenic activity for the strains TA98 and TA100 in the Ames assay. Mutagenicity was not observed in vivo.
Asunto(s)
Mutágenos/toxicidad , Plantas/química , Plantas/toxicidad , Animales , Cloroformo , Ciclofosfamida/toxicidad , Metanol , Ratones , Pruebas de Micronúcleos , Extractos Vegetales/toxicidad , Hojas de la Planta/química , Ratas , Reticulocitos/efectos de los fármacos , Reticulocitos/ultraestructura , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , SolventesRESUMEN
Byrsonima crassa is a plant pertaining to the Brazilian central savannah-like belt of vegetation and popularly used for the treatment of gastric dysfunctions and diarrhoea. The methanol extract contains catechin, tannins, terpenes and flavonoids; both mutagenic potential and antioxidant properties have been ascribed to flavonoids. The mutagenicity of some flavonoids is believed to be associated with the formation of reactive oxygen species and seems to depend on the number and position of hydroxyl groups. In the present study the mutagenic activity of the methanol, chloroform and 80% aqueous methanol extracts, as well as acetate and aqueous sub-fractions, of this medicinal plant were evaluated by Salmonella typhimurium assay, using strains TA100, TA98, TA102 and TA97a, and in mouse reticulocytes. The results showed mutagenic activity of the methanolic extract in the TA98 strain without S9, but no mutagenicity to mouse cells in any of the extracts. The acetate fraction showed strong signs of mutagenicity without S9, suggesting that in this enriched fraction were concentrated the compounds that induced mutagenic activity. The aqueous fraction showed no mutagenic activity. The TLC and HSCCC analyses of the acetate fraction with some standard compounds permitted the isolation of the quercetin-3-O-beta-D-galactopyranoside, quercetin-3-O-alpha-L-arabinopyranoside, amentoflavone, methyl gallate and (+)-catechin, of which only the amentoflavone exhibited positive mutagenicity to TA98 (+S9, -S9).
Asunto(s)
Biflavonoides/toxicidad , Malpighiaceae/química , Salmonella typhimurium/efectos de los fármacos , Animales , Catequina/toxicidad , Femenino , Galactósidos/toxicidad , Ácido Gálico/análogos & derivados , Ácido Gálico/toxicidad , Glicósidos/toxicidad , Masculino , Metanol/química , Ratones , Pruebas de Micronúcleos , Mutación , Extractos Vegetales/toxicidad , Plantas Medicinales/química , Quercetina/análogos & derivados , Quercetina/toxicidad , Salmonella typhimurium/genéticaRESUMEN
Brazilian flora biodiversity has been widely investigated to identify effective and safe phytotherapeutic compounds. Among the investigated plant species, the Byrsonima genus exhibits promising biological activities. This study aimed at evaluating the cytotoxicity of B. correifolia, B. verbascifolia, B. fagifolia and B. intermedia extracts using different assays in two cell lines (primary gastric and HepG2 cells). The different extract concentrations effects on cell viability were assayed using the MTT, aquabluer, neutral red and LDH assays. Non-cytotoxic concentrations were selected to generate cell proliferation curves and to assess cell cycle kinetics by flow cytometry. Byrsonima extracts differentially affected cell viability depending on the metabolic cellular state and the biological parameter evaluated. B. fagifolia and B. intermedia extracts exhibited lower cytotoxic effects than B. correifolia and B. verbascifolia in all assays. The results obtained with LDH and flow cytometry assays were more reliable, suggesting that they can be useful in the screening for herbal medicine and to further characterize these extracts as phytotherapeutic compounds.
RESUMEN
The methanolic extract of the leaves of the medicinal plant Byrsonima crassa (Malpighiaceae) contain flavonoids with antioxidant activity. They were separated in a preparative scale using high-speed counter-current chromatography. The optimum solvent system used was composed of a mixture of ethyl acetate-n-propanol-water (140:8:80 (v/v/v)) and led to a successful separation between monoglucosilated flavonoids (quercetin-3-O-alpha-L-arabinoside, quercetin-3-O-beta-D-galactoside) and the biflavonoid amentoflavone in only 3.5 h. The purities of quercetin-3-O-alpha-L-arabinoside (95 mg), quercetin-3-O-beta-D-galactoside (16 mg) and the biflavonoid amentoflavone (114 mg) were all isolated at purity over 95%. Identification was performed by 1H NMR, 13C NMR and UV analyses.
Asunto(s)
Distribución en Contracorriente/métodos , Flavonoides/aislamiento & purificación , Malpighiaceae/química , Hojas de la Planta/química , Espectrofotometría UltravioletaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: An ethnopharmacological survey indicated that the leaves of Byrsonima intermedia A. Juss. (Malpighiaceae), a medicinal species commonly found in the Brazilian Cerrado, can be used against gastroduodenal disorders, such as gastric ulcers and diarrhea. AIM OF THE STUDY: The objective of this study was to evaluate the effects of a methanolic extract of Byrsonima intermedia (MBI) leaves on gastric and duodenal ulcers and to assess the antimicrobial and antidiarrheal effects of this extract. MATERIAL AND METHODS: The anti-ulcerogenic effect of MBI was investigated with different ulcerogenic agents in rodents (mice and rats), including non-steroidal anti-inflammatory drug (NSAID), HCl/ethanol, pyloric ligature, absolute ethanol, cysteamine and ischemia-reperfusion. The gastroprotective effect of MBI was assessed by analysing the volume of gastric juice, pH, total acidity, mucus, NO, sulfhydryl compound, vanilloid receptor, glutathione (GSH) levels, and myeloperoxidase (MPO) activity in the gastric and duodenal mucosa. The gastric and duodenal healing effects of MBI were also evaluated during 7 or 14 days of treatment. The antidiarrheal action (measured by intestinal motility and diarrhea induced by castor oil) and anti-bacterial action of MBI against Staphylococcus aureus, Escherichia coli and Helicobacter pylori were also evaluated by microdilution methods. RESULTS: The phytochemical profile from MBI indicated the presence of phenolic acids, flavan-3-ols, oligomeric proanthocyanidins, and flavonoids. MBI (500mg/kg, p.o.) significantly inhibited totally gastric and duodenal lesions (69%) and healed gastric (49% on 14 days) and duodenal lesions (45% on 7 and 14 days). The MBI exert gastroprotective action by participation of endogenous sulfhydryl compounds, vanilloid receptors and increase in GSH level to effective gastric and duodenal protection. MBI also displayed curative (42%) and preventive (49%) antidiarrheal effects by involvement of opiate receptors and also antimicrobial effects in vitro. CONCLUSIONS: Byrsonima intermedia leaves present gastroprotective, healing and antidiarrheal activities, supporting previous claims that its traditional use can treat gastrointestinal disorders.
Asunto(s)
Antibacterianos/uso terapéutico , Antiulcerosos/uso terapéutico , Antidiarreicos/uso terapéutico , Malpighiaceae/química , Úlcera Péptica/prevención & control , Fitoterapia , Polifenoles/uso terapéutico , Animales , Antibacterianos/análisis , Antibacterianos/farmacología , Antiulcerosos/análisis , Antiulcerosos/farmacología , Antidiarreicos/análisis , Antidiarreicos/farmacología , Bacterias/efectos de los fármacos , Brasil , Diarrea/inducido químicamente , Diarrea/tratamiento farmacológico , Diarrea/prevención & control , Modelos Animales de Enfermedad , Motilidad Gastrointestinal/efectos de los fármacos , Glutatión/metabolismo , Masculino , Ratones , Ratones Endogámicos , Úlcera Péptica/tratamiento farmacológico , Úlcera Péptica/etiología , Extractos Vegetales/análisis , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Hojas de la Planta , Polifenoles/análisis , Polifenoles/farmacología , Ratas , Ratas Endogámicas , Receptores Opioides/metabolismo , Compuestos de Sulfhidrilo/metabolismo , Canales Catiónicos TRPV/metabolismoRESUMEN
ABSTRACT Operculina macrocarpa (L.) Urb., Convolvulaceae, is used by the population as a laxative. In this work we described the isolation of the three phenolic acids present in the hydroethanolic extract of the O. macrocarpa roots. The quantification of the caffeic, chlorogenic acids and of the new caffeic dimer in the hydroethanolic and infusion extracts was performed by high-performance liquid chromatography coupled photodiode array detector. These analyses showed the higher content of the chlorogenic, caffeic and the new 3,4'-dehydrodicaffeic acid in hydroethanolic and hydroethanolic extracts without resin in which infusion. The acid found in greater quantity is caffeic acid followed by the 3,4'-dehydrodicaffeic acid. The laxative activity was evaluated by different experimental models of intestinal transit with the hydroethanolic and infusion extracts, and the resin fraction, caffeic, chlorogenic and ferulic acids. The results showed all extracts and compounds tested had significant activity in the experimental model tested. These results obtained are essential for the future development of a pharmaceutical product with safety and efficacy.