RESUMO
Considering the drawbacks elicited by the conventional antidepressants, the interest in natural products for the management of major depressive disorder has increased in the last years. Therefore, this study investigated the phenolic profile of Maclura tinctoria leaf aqueous extract (MtAE) and its possible antidepressant-like effect in mice. The LC-MS/MS analysis demonstrated MtAE has epicatechin as the major phenolic, followed by catechin, gallic acid, quercetin, syringaldehyde, ferulic acid, and syringic acid. Moreover, the acute treatment of MtAE elicited an antidepressant-like response in mice. Importantly, this antidepressant-like effect produced by MtAE was reinforced in the chronic corticosterone (20 mg/kg p.o.) administration model. MtAE treatment was also effective to protect hippocampal and cerebrocortical slices against glutamatergic excitotoxicity. Our results indicated that MtAE displayed antidepressant-like and neuroprotective effects and these responses could be associated with the presence of the phenolic compounds identified.
Assuntos
Catequina , Transtorno Depressivo Maior , Maclura , Fármacos Neuroprotetores , Animais , Antidepressivos/farmacologia , Antidepressivos/uso terapêutico , Catequina/análise , Cromatografia Líquida , Transtorno Depressivo Maior/tratamento farmacológico , Camundongos , Fármacos Neuroprotetores/farmacologia , Fenóis/análise , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Folhas de Planta/química , Espectrometria de Massas em TandemRESUMO
Our objective was to isolate peptidases from the latex of Maclura pomifera fruits and use them to hydrolyze food proteins, as well as to purify and characterize the main peptidase. Two partially purified proteolytic extracts were prepared by ethanol (EE) and acetone (AE) precipitation from an aqueous suspension of exuded fruit latex. EE was used to hydrolyze food proteins with a ratio of 0.19 caseinolytic units (Ucas) per mg of substrate. Different values of hydrolysis degree were observed for hydrolysates of egg white, soy protein isolate, and casein at 180 min (9.3%, 31.1%, and 29.1%, respectively). AE was employed to purify a peptidase which exhibited an isoelectric point (pI) of 8.70 and whose abundance in AE was 28.3%. This enzyme was purified to homogeneity using a single-step procedure by cation-exchange chromatography, achieving an 8.1-fold purification and a yield of 16.7%. The peptidase was named pomiferin I and showed a molecular mass of 63,177.77 Da. Kinetic constants (KM 0.84 mM, Vmax 27.50 uM s-1, kcat 72.37 s-1, and kcat/KM 86.15 mM-1 s-1) were determined employing N-α-carbobenzoxy-L-alanyl-p-nitrophenyl ester as substrate. Analysis by PMF showed only partial homology of pomiferin I with a serine peptidase from a species of the same family.
Assuntos
Proteínas Alimentares/química , Frutas/enzimologia , Maclura/enzimologia , Peptídeo Hidrolases , Proteínas de Plantas , Hidrolisados de Proteína/química , Cromatografia por Troca Iônica , Peptídeo Hidrolases/química , Peptídeo Hidrolases/isolamento & purificação , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificaçãoRESUMO
BACKGROUND: The Atlantic Forest biome extends along the entire Brazilian coast and is home to approximately 20,000 plant species, many of which are endemic; it is considered one of the hotspot regions of the planet. Several of these species are sources of natural products with biological activities that are still unknown. In this study, we evaluated the antimicrobial activity of 90 extracts derived from native Atlantic Forest tree species against Staphylococcus aureus, an important human and veterinary pathogen. METHODS: Extracts from native Atlantic Forest tree species were evaluated for their antimicrobial activity against S. aureus by in vitro standard methods. Phytochemical fractionation of the extract from Maclura tinctoria was performed by liquid-liquid partitioning. LC-DAD-ESI-MS was used for identification of constituents in the most active fraction. Damage of cells and alterations in the permeability of cell membrane were determined by atomic force microscopy (AFM) and crystal violet uptake assay, respectively. In vivo antimicrobial activity was evaluated using Galleria mellonella larvae infected with S. aureus with survival data collected using the Kaplan-Meier method. RESULTS: Among the organic or aqueous extracts tested here, 26 showed biological activity. Eight species showed relevant results, with a minimum inhibitory concentration (MIC) below 1 mg/mL. Antibacterial activity was registered for three species for the first time. An organic extract from Maclura tinctoria leaves showed the lowest MIC (0.08 mg/mL). Fractionation of this extract by liquid-liquid partitioning led to obtaining fraction 11FO d with a MIC of 0.04 mg/mL. This fraction showed strong activity against veterinary S. aureus isolates and contributed to the increased survival of Galleria mellonella larvae infected with S. aureus ATCC 29213. The bacterial surface was not altered by the presence of 11FO d, and no cell membrane damage was detected. The LC-DAD-ESI/MS analyses identified prenylated flavonoids as the major constituents responsible for the antibacterial activity of this active extract. CONCLUSION: A fraction enriched in prenylated isoflavones and flavanones from M. tinctoria showed in vitro and in vivo efficacy as antistaphylococcal agents. These findings justify the need for further research to elucidate the mechanisms of action of these compounds.
Assuntos
Antibacterianos/administração & dosagem , Flavonoides/administração & dosagem , Maclura/química , Mariposas/microbiologia , Extratos Vegetais/administração & dosagem , Substâncias Protetoras/administração & dosagem , Infecções Estafilocócicas/prevenção & controle , Staphylococcus aureus/efeitos dos fármacos , Animais , Antibacterianos/química , Brasil , Modelos Animais de Doenças , Flavonoides/química , Humanos , Larva/crescimento & desenvolvimento , Larva/microbiologia , Testes de Sensibilidade Microbiana , Mariposas/crescimento & desenvolvimento , Extratos Vegetais/química , Substâncias Protetoras/química , Infecções Estafilocócicas/microbiologiaRESUMO
The presence of articulated laticifers in the Moraceae family was recently discovered, which means that the location of pectinase and cellulase activities must be of great importance for their growth. Thus, the present study aimed to determine the role of these enzymes in the laticifer growth in Ficus montana and Maclura tinctoria. Reproductive meristems were collected and fixed in Karnovsky. Pectinase and cellulase labeling was performed in part of the samples, while another part was processed for usual TEM analyses. Pectinase and cellulase activities were detected in the vacuole and close to the middle lamella in both species. The presence of cellulases in the laticifers supports their articulated origin. Therefore, the occurrence of pectinase and cellulase activity in the laticifers points out that these enzymes could act in the dissolution of the transverse walls and in the processes of intrusive growth (through the dissolution of the middle lamella) and cell elongation (through the partial disassembly of components of the wall making it more plastic). Both enzymes are synthesized in the endoplasmic reticulum and transported to the cell wall by exocytosis or stored in the vacuole. The species studied showed a diverse subcellular composition, which is probably related to the species and not to the laticifer type (they present the same type) and to the composition of the latex (they show similar latex composition). We conclude that the presence of pectinases and cellulases can be used as a diagnostic condition for the laticifer types (articulated vs. non-articulated).
Assuntos
Celulases/metabolismo , Ficus/metabolismo , Maclura/metabolismo , Poligalacturonase/metabolismo , Parede Celular/metabolismo , Parede Celular/ultraestrutura , Ficus/citologia , Látex/metabolismo , Maclura/citologia , Meristema/metabolismo , Microscopia Eletrônica de Transmissão , Células Vegetais/metabolismo , Células Vegetais/ultraestrutura , Vacúolos/metabolismoRESUMO
BACKGROUND AND AIMS: Maclura (ca. 12spp., Moraceae) is a widespread genus of trees and woody climbers found on five continents. Maclura pomifera, the Osage orange, is considered a classic example of an anachronistic fruit. Native to the central USA, the grapefruit-sized Osage oranges are unpalatable and have no known extant native dispersers, leading to speculation that the fruits were adapted to extinct megafauna. Our aim was to reconstruct the phylogeny, estimate divergence dates, and infer ancestral ranges of Maclura in order to test the monophyly of subgeneric classifications and to understand evolution and dispersal patterns in this globally distributed group. METHODS: Employing Bayesian and maximum-likelihood methods, we reconstructed the Maclura phylogeny using two nuclear and five chloroplast loci from all Maclura species and outgroups representing all Moraceae tribes. We reconstructed ancestral ranges and syncarp sizes using a family level dated tree, and used Ornstein-Uhlenbeck models to test for significant changes in syncarp size in the Osage orange lineage. KEY RESULTS: Our analyses support a monophyletic Maclura with a Paleocene crown. Subgeneric sections were monophyletic except for the geographically-disjunct Cardiogyne. There was strong support for current species delineations except in the widespread M. cochinchinensis. South America was reconstructed as the ancestral range for Maclura with subsequent colonization of Africa and the northern hemisphere. The clade containing M. pomifera likely diverged in the Oligocene, closely coinciding with crown divergence dates of the mammoth/mastodon and sloth clades that contain possible extinct dispersers. The best fitting model for syncarp size evolution indicated an increase in both syncarp size and the rate of syncarp size evolution in the Osage orange lineage. CONCLUSIONS: We conclude that our findings are consistent with the hypothesis that M. pomifera was adapted to dispersal by extinct megafauna. In addition, we consider dispersal rather than vicariance to be most likely responsible for the present distribution of Maclura, as crown divergence post-dated the separation of Africa and South America. We propose revised sectional delimitations based on the phylogeny. This study represents a complete phylogenetic and biogeographic analysis of this globally distributed genus and provides a basis for future work, including a taxonomic revision.
Assuntos
Frutas/genética , Maclura/classificação , Maclura/genética , Filogenia , África , Teorema de Bayes , Núcleo Celular/genética , Cloroplastos/genética , Frutas/anatomia & histologia , Frutas/classificação , Genes de Plantas/genética , Funções Verossimilhança , Maclura/anatomia & histologia , Filogeografia , América do SulRESUMO
Four chalcone glycosides (1-4), including three new natural products, and three flavanones (5-7) were isolated from the methanol extract of stem bark of Maclura tinctoria. The new compounds have been characterized as 4'-O-beta-D-(2' '-p-coumaroyl)glucopyranosyl-4,2',3'-trihydroxychalcone (1), 4'-O-beta-D-(2' '-p-coumaroyl-6' '-acetyl)glucopyranosyl-4,2',3'-trihydroxychalcone (2), and 3'-(3-methyl-2-butenyl)-4'-O-beta-D-glucopyranosyl-4,2'-dihydroxychalcone (3); the known derivatives were elucidated as 4'-O-beta-D-(2' '-acetyl-6' '-cinnamoyl)glucopyranosyl-4,2',3'-trihydroxychalcone (4), eriodictyol 7-O-beta-D-glucopyranoside (5), naringenin (6), and naringenin 4'-O-beta-D-glucopyranoside (7). Their structures were determined by 1D and 2D NMR and ESIMS. The antioxidant activity of all the isolated compounds was determined by measuring free-radical-scavenging effects using two different assays, namely, the Trolox Equivalent Antioxidant Capacity (TEAC) assay and the coupled oxidation of beta-carotene and linoleic acid (autoxidation assay). The results showed that compound 3 was the most active in both antioxidant assays.
Assuntos
Antioxidantes/isolamento & purificação , Chalcona/isolamento & purificação , Flavanonas/isolamento & purificação , Glicosídeos/isolamento & purificação , Maclura/química , Plantas Medicinais/química , Antioxidantes/química , Antioxidantes/farmacologia , Bolívia , Chalcona/química , Chalcona/farmacologia , Espectroscopia de Ressonância de Spin Eletrônica , Flavanonas/química , Flavanonas/farmacologia , Glicosídeos/química , Glicosídeos/farmacologia , Estrutura Molecular , Oxirredução , Casca de Planta/química , Caules de Planta/química , Estereoisomerismo , beta Caroteno/metabolismoRESUMO
We report on the yeast community associated with sap fluxes of Maclura tinctoria, family Moraceae, in the dry forest of the Area de Conservación Guanacaste, Costa Rica. Eleven samples yielded seven hitherto undescribed ascomycetous yeasts in the genera Candida and Myxozyma. We describe the two most abundant as new species. Candida galis utilizes very few carbon compounds limited to some alcohols and acids. Analysis of rDNA sequences suggests that it occupies a basal position with respect to the Pichia anomala clade, with no obvious sister species. Candida ortonii is also restricted in nutritional breadth, and growth is generally very slow. It is a sister species to Candida nemodendra. The type cultures are: C. galis, strain UWO(PS)00-159.2=CBS 8842; and C. ortonii, strain UWO(PS)00-159.3=CBS 8843.