RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Monteverdia ilicifolia (Maytenus ilicifolia, Celastraceae), known as "espinheira-santa", has been widely used in Brazil to manage mainly gastrointestinal diseases. This species has been listed in the Brazilian Pharmacopeia and in the National List of Essential Medicines (RENAME). Considering that clinical studies about M. ilicifolia are rare, our group has been performing a broader project designed to evaluate the efficacy of M. ilicifolia capsules in a clinical trial, for this reason, approaches to provide safety to those patients are relevant. AIM OF THE STUDY: We aimed to investigate the potential pharmacokinetic interaction and hepatotoxicity and intestinal toxicity of an aqueous extract of M. ilicifolia and its main phytocompounds, catequin, epicatequin, and quercetin. METHODS AND MATERIALS: Slices of liver and intestine of Wistar rats were incubated with different concentrations of M. ilicifolia extract or isolated compounds (catechin, epicatechin and quercetin). Commercial kits were used to evaluate enzyme activities of CYP2D6 and CYP3A4 isoforms, as well as cell viability (MTT) assay and intracellular enzymes leakage, specifically lactate dehydrogenase (LDH), alkaline phosphatase (AP), aspartate aminotransferase (AST), alanine aminotransferase (ALT) were studied. RESULTS: Incubation with M. ilicifolia extract, catechin, epicatechin and quercetin did not affect significantly any evaluated parameter in intestines. The intracellular enzymes leakages, CYP2D6, LDH and AST, were increased with M. ilicifolia extract and quercetin in liver slices. CONCLUSIONS: Our in vitro findings highlighted, for the first time, the potential hepatotoxicity induced by an aqueous extract of M. ilicifolia, consequently this species and its products should be avoided in liver diseases, supporting that studies of safety must be performed including in the context of traditional medicinal plants.
Assuntos
Catequina , Celastraceae , Doença Hepática Induzida por Substâncias e Drogas , Maytenus , Plantas Medicinais , Humanos , Ratos , Animais , Brasil , Extratos Vegetais/toxicidade , Quercetina , Citocromo P-450 CYP2D6 , Ratos WistarRESUMO
Due to the widespread use of Maytenus ilicifolia leaves in the treatment of gastric ulcers, herbal medicines derived from such species are distributed by the national health system in Brazil. A related species, Maytenus aquifolium, is also used for the same disorders, and both are popularly known as Espinheira-santa. Due to their popular use, the quality and efficiency of the herbal medicines derived from these species is an important public health issue. The purpose of this study was to develop and test an analytical method that could quantify the content of catechin and epicatechin in dry Maytenus spp. leaves and simultaneously obtain their chemical profile to determine authenticity of the leaf samples. Ultra-high performance liquid chromatography coupled to mass spectrometry (UHPLC-MS) was used to quantify these isomers, in the selected ion monitoring (SIM) mode, while simultaneously analyzing the extract in full-scan mode. This approach was successfully applied to the analysis of commercial and authentic samples of Maytenus spp. Fewer than half the samples presented the minimum epicatechin content of 2.8 mg per g of dry leaf mass, as specified in the 6th Brazilian Pharmacopoeia (2019) for M. ilicifolia. Furthermore, by using untargeted metabolomics, it was observed that the chemical profile of most the samples was not compatible with M. ilicifolia leaves, indicating the need for stricter quality control of this material. The method described herein could be used for this control; moreover, its concept could be adapted and used for an ample variety of medicinal plant products.
Assuntos
Catequina , Maytenus , Plantas Medicinais , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas , Maytenus/química , Extratos Vegetais/químicaRESUMO
Zika virus (ZIKV) infection is a global threat associated to neurological disorders in adults and microcephaly in children born to infected mothers. No vaccine or drug is available against ZIKV. We herein report the anti-ZIKV activity of 36 plant extracts containing polyphenols and/or triterpenes. ZIKV-infected Vero CCL-81 cells were treated with samples at non-cytotoxic concentrations, determined by MTT and LDH assays. One third of the extracts elicited concentration-dependent anti-ZIKV effect, with viral loads reduction from 0.4 to 3.8â log units. The 12 active extracts were tested on ZIKV-infected SH-SY5Y cells and significant reductions of viral loads (in log units) were induced by Maytenus ilicifolia (4.5â log), Terminalia phaeocarpa (3.7â log), Maytenus rigida (1.7â log) and Echinodorus grandiflorus (1.7â log) extracts. Median cytotoxic concentration (CC50 ) of these extracts in Vero cells were higher than in SH-SY5Y lineage. M. ilicifolia (IC50 =16.8±10.3â µg/mL, SI=3.4) and T. phaeocarpa (IC50 =22.0±6.8â µg/mL, SI=4.8) were the most active extracts. UPLC-ESI-MS/MS analysis of M. ilicifolia extract led to the identification of 7 triterpenes, of which lupeol and a mixture of friedelin/friedelinol showed no activity against ZIKV. The composition of T. phaeocarpa extract comprises phenolic acids, ellagitannins and flavonoids, as recently reported by us. In conclusion, the anti-ZIKV activity of 12 plant extracts is here described for the first time and polyphenols and triterpenes were identified as the probable bioactive constituents of T. phaeocarpa and M. ilicifolia, respectively.
Assuntos
Neuroblastoma , Triterpenos , Infecção por Zika virus , Zika virus , Animais , Criança , Chlorocebus aethiops , Humanos , Neuroblastoma/tratamento farmacológico , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Polifenóis/farmacologia , Espectrometria de Massas em Tandem , Triterpenos/farmacologia , Células Vero , Infecção por Zika virus/tratamento farmacológicoRESUMO
Monteverdia ilicifolia is a Brazilian native plant, traditionally used to treat gastric diseases that are now associated with Helicobacter pylori and are commonly associated with several human diseases. We point out the M. ilicifolia extract as active against H. pylori. The crude extract produced with acetone:water presented the best H. pylori inhibitory activity of all five extracts (MIC 64 µg/mL). The ethyl-acetate fractions from crude extracts produced with ethanol and acetone showed a MIC of 64 µg/mL. Both ethyl-acetate fractions and the crude extract produced with acetone showed an antioxidant capacity of between 14.51 and 19.48 µg/mL in the DPPH assay. In the FRAP assay, two ethyl-acetate fractions (EAF2 and EAF4) presented the antioxidant capacity of 5.40 and 5.15 mM Trolox/g of extract. According to the results obtained from the antioxidant and antibacterial assays, two fractions (EAF2 and nBF5) were analyzed by mass spectrometry and confirmed the presence of monomeric, dimeric, trimeric tannins, and glycosylated flavonoids. Some compounds were tested using bioinformatics to evaluate the best enzyme inhibitors and the molecular interaction between the enzyme and the tested ligands. The presence of these polyphenol compounds could play an important role in antioxidant and inhibitory capacities against H. pylori and can be used to assist in the treatment or prevention of infection by H. pylori.
RESUMO
Free (epi) catechin, quercetin, (epi) gallocatechin, flavonol glycosides and condensed tannins were identified according to their molecular mass, characteristic product ions and retention times in extracts obtained from leaves and branches of Maytenus ilicifolia (Congorosa) by mass spectrometry. The in vitro anthelmintic activity against cattle gastrointestinal nematodes of Congorosa extract was determined using the Egg Hatch Inhibition Assay. Additionally, commercial quercetin, gallocatechin and epicatechin were evaluated. Although total phenolics, total tannins and condensed tannins contents were lower in branches extract than in leaves extract, the EC50 were 0.065 mg/mL and 0.890 mg/mL for branches and leaves extract, respectively. Moreover, the use of polyvinylpyrrolidone as a blocking agent of tannins, did not change significantly the EC50 for branches extract, but significantly changed for leaves extract. Quercetin and gallocatechin EC50 values were in the range 0.03-0.05 mg/mL and epicatechin showed 100% inhibition of the egg hatching process at 0.004 mg/mL.
Assuntos
Anti-Helmínticos , Catequina , Maytenus , Proantocianidinas , Animais , Anti-Helmínticos/farmacologia , Catequina/análise , Bovinos , Maytenus/química , Fenóis/análise , Fenóis/farmacologia , Extratos Vegetais/química , Folhas de Planta/química , Proantocianidinas/análise , Quercetina/análise , Quercetina/farmacologia , Taninos/análiseRESUMO
Friedelin, a pentacyclic triterpene found in the leaves of the Celastraceae species, demonstrates numerous biological activities and is a precursor of quinonemethide triterpenes, which are promising antitumoral agents. Friedelin is biosynthesized from the cyclization of 2,3-oxidosqualene, involving a series of rearrangements to form a ketone by deprotonation of the hydroxylated intermediate, without the aid of an oxidoreductase enzyme. Mutagenesis studies among oxidosqualene cyclases (OSCs) have demonstrated the influence of amino acid residues on rearrangements during substrate cyclization: loss of catalytic activity, stabilization, rearrangement control or specificity changing. In the present study, friedelin synthase from Maytenus ilicifolia (Celastraceae) was expressed heterologously in Saccharomyces cerevisiae. Site-directed mutagenesis studies were performed by replacing phenylalanine with tryptophan at position 473 (Phe473Trp), methionine with serine at position 549 (Met549Ser) and leucine with phenylalanine at position 552 (Leu552Phe). Mutation Phe473Trp led to a total loss of function; mutants Met549Ser and Leu552Phe interfered with the enzyme specificity leading to enhanced friedelin production, in addition to α-amyrin and ß-amyrin. Hence, these data showed that methionine 549 and leucine 552 are important residues for the function of this synthase.
Assuntos
Alquil e Aril Transferases/metabolismo , Maytenus/enzimologia , Proteínas de Plantas/metabolismo , Triterpenos/metabolismo , Alquil e Aril Transferases/química , Alquil e Aril Transferases/genética , Substituição de Aminoácidos , Vias Biossintéticas , Ciclização , Genes de Plantas , Leucina/química , Maytenus/genética , Metionina/química , Modelos Moleculares , Mutagênese Sítio-Dirigida , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/biossíntese , Triterpenos Pentacíclicos/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidade por SubstratoRESUMO
BACKGROUND AND AIM: Maytenus ilicifolia has analgesic, healing, antioxidant and anti-inflammatory properties. This study evaluated effect of the hydroalcoholic extract of M. ilicifolia leaves on skin wound repair. EXPERIMENTAL PROCEDURE: Wounds were induced on mice and treated with the extract. The treatment was performed daily, until day 7 after wound induction. Wound closure was measured and the features of the repaired tissue were investigated, including mast cell quantification, neutrophil and macrophage activities, collagen deposition, angiogenesis, and pro-metalloproteases and metalloproteases 2 and 9 activity (pro-MMPs and MMPs). RESULTS AND CONCLUSION: The M. ilicifolia extract accelerated the closure of wounds. The extract at a concentration of 4% was found to be effective, presenting anti-inflammatory effects and hemoglobin increased, along with increased soluble, total and type III collagens in the wound. In addition, there was an increase in pro-MMP9 and MMP9 activity after day 7th of treatment. The phenolic compounds and tannins present in this plant could be associated with the anti-inflammatory and healing activities observed in this study. Therefore, the ability to modulate essential parameters for accelerated and adequate healing as shown here suggests that the use of standardised extracts of M. ilicifolia and its fractions enriched in polyphenols may represent a therapeutic strategy for the treatment of wounds.
RESUMO
Midazolam (MDZ) is routinely employed as a marker compound of cytochrome P450 3A (CYP3A) activity. Despite the many HPLC-UV methods described to quantify MDZ in plasma, all of them use acetonitrile (ACN) or a mixture of methanol-isopropanol as organic solvent of the mobile phase. Since the ACN shortage in 2008, efforts have been made to replace this solvent during HPLC analysis. A simple, sensitive, accurate and repeatable HPLC-UV method (220 nm) was developed and validated to quantify MDZ in rat plasma using methanol instead. The method was applied during a herb-drug interaction study involving Maytenus ilicifolia, a Brazilian folk medicine used to treat gastric disorders. Plasma samples were alkalinized and MDZ plus alprazolam (internal standard) were extracted with diethyl ether. After solvent removal, the residue was reconstituted with methanol-water (1:1). The analyte was eluted throughout a C18 column using sodium acetate buffer (10 mm, pH 7.4)-methanol (40:60, v/v). The precision at the lower limit of quantification never exceeded 19.40%, and 13.86% at the higher levels of quality control standards, whereas the accuracy ranged from -19.81 to 14.33%. The analytical curve was linear from 50 to 2,000 ng/ml. The activity of the hepatic CYP3A enzymes was not affected by the extract.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Interações Ervas-Drogas , Maytenus/química , Midazolam/sangue , Animais , Citocromo P-450 CYP3A/metabolismo , Modelos Lineares , Masculino , Metanol , Midazolam/administração & dosagem , Midazolam/farmacocinética , Preparações de Plantas/administração & dosagem , Preparações de Plantas/sangue , Preparações de Plantas/farmacocinética , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Objective: In this study, we aimed to determine the action of the tinctures of Hamamelis virginiana, Maytenus ilicifolia, and Casearia sylvestris on tissues. For this, we investigated the histological sections of fixed skin tissue of Wistar rats for the changes in collagen and elastic fibers, epithelial cells, conjunctive cells, epidermal attachments, pigments, and granules using the optical microscopy technique. Since in the literature and published articles, the use of in vivo models, such as Wistar rats, is predominant to evaluate the healing action of herbal medicines. Methods: The tinctures of Hamamelis virginiana, Maytenus ilicifolia, and Casearia sylvestris, and ethanol 70% (blank) were used. The tinctures were obtained at 10% (w/v) through percolation using 70% ethanol (v/v) as the extraction liquid. This study was conducted in duplicate for each tincture and different incubation times (4, 24, and 48 h) at 37 °C in an oven. The slides used in this study (Wistar rat skin) were previously processed at the histology laboratory since the waste material was donated to conduct this experiment. The Research Ethics Committee approved the use of animals of the Life Sciences Center of the Pontifical Catholic University of Campinas (PUC-CAMPINAS) under the protocol approval number 367/08. Before initiating the staining process, the slides were removed from the oven, and the tincture deposited on the slides was removed. Weigert staining was performed subsequently. For semi-quantitative analysis, the histological sections were carefully observed, and the number of collagen and elastic fibers was evaluated based on the following scale: (+) normal fiber presence, (−) slight decrease, (− −) moderate decrease, and (− − −) intense decrease. The images were digitally captured to obtain the results using a photomicroscope. Results: The degradation of collagen fibers was best evidenced upon using Hamamelis virginiana tincture, which is concordant with the existing reports in the literature on its healing action via the precipitation of dermal proteins. Maytenus ilicifolia and Casearia sylvestris tinctures exhibited low proteolytic capacity as they only caused degradation of elastic fibers that are more delicate and very different from collagen in their constitution. Therefore, the application of the latter two as healing agents (which acts through precipitating proteins) is limited. Conclusions: This experimental histological model using the optical microscopy technique exhibits the advantage of agility and high efficiency as a simple and powerful approach.
RESUMO
BACKGROUND: Maytenus ilicifolia is a Brazilian popular medicine commonly used to treat ulcer and gastritis. Despite the absence of toxicity regarding its consumption, possible interactions when co-administrated with conventional drugs, are unknown. OBJECTIVE: This study aimed to evaluate the effects of M. ilicifolia extracts on Cytochrome P450 3A (CYP3A) and P-glycoprotein (P-gp) activities. METHODS: The extracts were obtained by infusion (MI) or turbo-extraction using hydro-acetonic solvent (MT70). The content of polyphenols in each extract was determined. To assess the modulation of M. ilicifolia on P-gp activity, the uptake of fexofenadine (FEX) by Caco-2 cells was investigated in the absence or presence of MI or MT70. The effect on CYP3A activity was evaluated by the co-administration of midazolam (MDZ) with each extract in male Wistar rats. The pharmacokinetic parameters of the drug were determined and compared with those from the control group. The content of total phenolic compounds, tannins, and flavonoids on MT70 extract was about double of that found in MI. RESULTS: In the presence of the extracts, the uptake of the P-gp marker (FEX) by Caco-2 cells increased from 1.7 ± 0.4 ng.mg-1 protein (control) to 3.5 ± 0.2 ng.mg-1 protein (MI) and 4.4 ± 0.5 ng.mg-1 protein (MT70), respectively. When orally co-administrated with MDZ (substrate of CYP3A), the extracts augmented the AUC(0-∞) (Control: 911.7 ± 215.7 ng.h.mL-1; MI: 1947 ± 554.3 ng.h.mL-1; MT70: 2219.0 ± 506.3 ng.h.mL-1) and the Cmax (Control: 407.7 ± 90.4 ng.mL-1; MI: 1770.5 ± 764.5 ng.mL-1; MT70: 1987.2 ± 544.9 ng.mL-1) of the drug in rats indicating a 50% reduction of the oral Cl. No effect was observed when midazolam was given intravenously. CONCLUSION: The results suggest that M. ilicifolia can inhibit the intestinal metabolism and transport of drugs mediated by CYP3A and P-gp, respectively, however, the involvement of other transporters and the clinical relevance of such interaction still need to be clarified.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Citocromo P-450 CYP3A/metabolismo , Maytenus/química , Extratos Vegetais/farmacologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/agonistas , Animais , Células CACO-2 , Linhagem Celular , Inibidores do Citocromo P-450 CYP3A/farmacologia , Interações Medicamentosas , Humanos , Cetoconazol/farmacologia , Masculino , Midazolam/farmacocinética , Quinolinas/farmacologia , Ratos , Ratos Wistar , Terfenadina/análogos & derivadosRESUMO
BACKGROUND: Triterpenes are ubiquitous secondary metabolites present in plants. They can be found in both forms, as genins or conjugated as glycosides. Although distinct analytical methods to quantify these compounds in vegetal tissues are available in the literature, limitations like high cost, complexity on sample preparation, and selectivity are often challenging issues. This study aimed to develop and to validate a simple and rapid spectrophotometric method to detect and quantify total triterpenes in plant matrices. METHODS: The assay was conducted directly into glass tubes using vanillin, acetic acid, and sulphuric acid as reagents, and ß-sitosterol as reference standard. The samples were analyzed at 548 nm assessing the quality parameters of selectivity, linearity, precision, accuracy, limit of detection (LOD), limit of quantification (LOQ), and robustness. RESULTS: The method was selective, with precision and accuracy varying from 0.56% to 4.98% and 96.63% to 113.87%, respectively. The values of the limit of detection and quantification were 0.042 µg.mL-1 and 0.14 µg.mL-1, correspondingly. The correlation coefficient (r) at the concentration range of 3.08 µg.mL-1to 24.61 µg.mL-1 was 0.9998. The total of triterpenes found in of B. holophylla and M. ilicifolia leaves were 132.36 ± 20.36 mg EßS.g-1 of dry extract and 53.91 ± 2.6 mg EßS.g-1 of dry extract, respectively. CONCLUSION: The method was reliable to quantify total triterpenes extracted from Maytenus ilicifolia and Bauhinia holophylla. Graphical abstract.
Assuntos
Bauhinia , Celastraceae , Folhas de Planta/química , Espectrofotometria/métodos , Triterpenos/análise , Ácido Acético/química , Benzaldeídos/química , Limite de Detecção , Reprodutibilidade dos Testes , Ácidos Sulfúricos/químicaRESUMO
Triterpenes are interesting compounds because they play an important role in cell homeostasis and a wide variety exhibiting defense functions is produced by plant secondary metabolism. Those same plant secondary metabolites also exhibit biological properties with promising therapeutic potential as anti-inflammatory and antitumor agents. Friedelin is a triterpene ketone with anti-inflammatory and gastroprotective activities and it is a precursor of relevant antitumor quinonemethides. Although many triterpene synthases have been described, only two friedelin synthases were characterized and there is no information about their genomic features and alleles. In the present work, we aimed to identify the gene and new isoforms of friedelin synthase in Maytenus ilicifolia leaves to be functionally characterized in Saccharomyces cerevisiae. The gene sequence analysis elucidated the exon/intron structure and confirmed the presence of single nucleotide polymorphisms with four non-synonymous mutations outside the active site of the enzyme. Therefore, two new isoforms were observed and the heterologous production of the enzymes in yeast showed similar production of friedelin. This first description of different alleles of the gene of friedelin synthase in M. ilicifolia can guide their validation as markers for friedelin-producer specimens.
Assuntos
Maytenus/enzimologia , Oxirredutases/metabolismo , Triterpenos/metabolismo , Sequência de Aminoácidos , Éxons/genética , Genes de Plantas , Íntrons/genética , Isoenzimas/metabolismo , Maytenus/genética , Fases de Leitura Aberta/genética , Oxirredutases/química , Oxirredutases/genética , Filogenia , Polimorfismo de Nucleotídeo Único/genética , Triterpenos/químicaRESUMO
Maytenus ilicifolia Mart. ex Reissek is a plant commonly used in folklore medicine in the management of gastric diseases in South America. This study explores the effects of a supratherapeutic dose of aqueous and ethanol extracts of M. ilicifolia (1360 mg/kg) on fertility and neurobehavioral status in male and pregnant rats. A battery of sensory-motor developmental endpoints was carried out to assess impairments on pups of dams orally treated with the aqueous extract of M. ilicifolia during the organogenesis period of pregnancy (GD 9 through GD 14). The neuromotor maturation reflexes and physical developments of the offspring were not significantly different between the groups (p < 0.05). Also, the hippocampal morphology revealed no indices of cell loss in the CA1, CA2, CA3 and CA4 areas. As second protocol, some fertility aspects were investigated in young post pubertal male Wistar rats treated with the ethanol extract for 30 days. The semen quality and testicular tissue morphology of male rats treated with the ethanol extract of M. ilicifolia remained unaffected upon treatment. Thus, the results indicate that the high-dose of M. ilicifolia extracts have no neurotoxic potential on offspring and seem not to affect the sperm quality of male rats.
Assuntos
Comportamento Animal/efeitos dos fármacos , Fertilidade/efeitos dos fármacos , Maytenus/química , Medicina Tradicional/efeitos adversos , Extratos Vegetais/efeitos adversos , Gastropatias/tratamento farmacológico , Animais , Etanol/química , Feminino , Masculino , Organogênese/efeitos dos fármacos , Gravidez , Ratos , Ratos Wistar , Análise do Sêmen , América do Sul , Testículo/efeitos dos fármacos , Água/químicaRESUMO
This review article focuses on pre-clinical and clinical studies with some selected Brazilian medicinal plants in different areas of interest, conducted by research groups in Brazil and abroad. It also highlights the Brazilian market of herbal products and the efforts of Brazilian scientists to develop new phytomedicines. This review is divided into three sections. The section I describes the Brazilian large biodiversity and some attempts of Brazilian scientists to assess the pharmacological profile of most plant extracts or isolated active principles. Of note, Brazilian scientists have made a great effort to study the Brazilian biodiversity, especially among the higher plants. In fact, more than 10,000 papers were published on plants in international scientific journals between 2011 and 2013. This first part also discussed the main efforts to develop new medicines from plants, highlighting the Brazilian phytomedicines market. Despite the large Brazilian biodiversity, notably with the higher plants, which comprise over 45,000 species (20-22% of the total worldwide), and the substantial number of scientific publications on medicinal plants, only one phytomedicine is found in the top 20 market products. Indeed, this market is still only worth about 261 million American dollars. This represents less than 5% of the global Brazilian medicine market. The section II of this review focus on the use of Brazilian plant extract and/or active principles for some selected diseases, namely: central nervous systems disorders, pain, immune response and inflammation, respiratory diseases, gastrointestinal tract and metabolic diseases. Finally, section III discusses in more details some selected Brazilian medicinal plants including: Cordia verbenacea, Euphorbia tirucalli, Mandevilla velutina, Phyllanthus spp., Euterpe oleracea, Vitis labrusca, Hypericum caprifoliatum and Hypericum polyanthemum, Maytenus ilicifolia, Protium kleinii and Protium heptaphylium and Trichilia catigua. Most of these publications are preliminary and only report the effects of crude extracts, both in vitro and in vivo studies. Only very few studies have been dedicated to investigate the mechanisms of action of isolated compounds. Likewise, studies on safety (toxicology), pharmacokinetic, and especially on well-conducted clinical trials are rare. In conclusion, in spite of the abundant Brazilian biodiversity and the thousands of academic publications on plants in international peer-reviewed scientific journals, few patents and medicines have been derived from such studies. Undoubtedly, great efforts must be made to improve the development of plant-derived medicine market in Brazil, especially by involving the partnership between academia and pharmaceutical companies.
Assuntos
Descoberta de Drogas , Fitoterapia , Extratos Vegetais/uso terapêutico , Plantas Medicinais , Animais , Biodiversidade , Brasil , Doenças do Sistema Nervoso Central/tratamento farmacológico , Gastroenteropatias/tratamento farmacológico , Humanos , Inflamação/tratamento farmacológico , Doenças Metabólicas/tratamento farmacológico , Dor/tratamento farmacológico , Extratos Vegetais/economia , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Transtornos Respiratórios/tratamento farmacológicoRESUMO
In these work the in vitro antioxidant activity and the in vivo genotoxicity of M. dasyclada was compared to the reference species M. aquifolium and M. ilicifolia. M. dasyclada showed in vitro antioxidant activity comparable to M. aquifolium but lower than M. ilicifolia, being that a inverse Pearson correlation between DPPH IC50 values and total phenolic content was detected (–0.932). The carbonyl content of M. dasyclada and M. aquifolium extracts promoted a similar increase in protein oxidation in vivo, while M. ilicifolia no altered the carbonyl levels. The comet assay demonstrated that the three analyzed species promoted a low and similar level of genotoxicity; which is compatible with DNA damage induced by other medicinal plants and is partially recovered by a co-treatment with vitamin C. The data showed M. dasyclada as antioxidant activity in vitro, and that its genotoxic and pro-oxidant effects in vivo are comparable to the Maytenus reference species.
No presente trabalho a atividade antioxidante in vitro e a genotoxicidade in vivo de M. dasyclada foi comparada com as espécies de referência M.aquifolium e M. ilicifolia. M. dasyclada mostrou atividade antioxidante in vitro comparável a de M. aquifolium mas inferior a M. ilicifolia, sendo que foi detectada uma correlação de Pearson inversa entre os valores de IC50 por DPPH e o conteúdo fenólico total (–0,932). Em relação ao teor de carbonila, os extratos de M. dasyclada e M. aquifolium promoveram um aumento semelhante na oxidação das proteínas in vivo, ao passo que Maytenus ilicifolia não alterou os níveis de carbonila. O ensaio do cometa demonstrou que as três espécies analisadas promoveram um nível baixo e semelhante de genotoxicidade, o que é compatível com os danos no DNA induzidos por outras plantas medicinais e é parcialmente recuperada por um co-tratamento com a vitamina C. Os dados mostraram M. dasyclada com atividade antioxidante in vitro, e que os seus efeitos genotóxicos e pró-oxidantes in vivo são comparáveis às espécies de referência de Maytenus.
Assuntos
Animais , Masculino , Antioxidantes/farmacologia , Maytenus/química , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Antioxidantes/toxicidade , Ensaio Cometa , Maytenus/classificação , Maytenus/toxicidade , Extratos Vegetais/toxicidade , Plantas Medicinais/classificação , Plantas Medicinais/toxicidade , Ratos WistarRESUMO
In these work the in vitro antioxidant activity and the in vivo genotoxicity of M. dasyclada was compared to the reference species M. aquifolium and M. ilicifolia. M. dasyclada showed in vitro antioxidant activity comparable to M. aquifolium but lower than M. ilicifolia, being that a inverse Pearson correlation between DPPH IC50 values and total phenolic content was detected (0.932). The carbonyl content of M. dasyclada and M. aquifolium extracts promoted a similar increase in protein oxidation in vivo, while M. ilicifolia no altered the carbonyl levels. The comet assay demonstrated that the three analyzed species promoted a low and similar level of genotoxicity; which is compatible with DNA damage induced by other medicinal plants and is partially recovered by a co-treatment with vitamin C. The data showed M. dasyclada as antioxidant activity in vitro, and that its genotoxic and pro-oxidant effects in vivo are comparable to the Maytenus reference species.(AU)
No presente trabalho a atividade antioxidante in vitro e a genotoxicidade in vivo de M. dasyclada foi comparada com as espécies de referência M.aquifolium e M. ilicifolia. M. dasyclada mostrou atividade antioxidante in vitro comparável a de M. aquifolium mas inferior a M. ilicifolia, sendo que foi detectada uma correlação de Pearson inversa entre os valores de IC50 por DPPH e o conteúdo fenólico total (0,932). Em relação ao teor de carbonila, os extratos de M. dasyclada e M. aquifolium promoveram um aumento semelhante na oxidação das proteínas in vivo, ao passo que Maytenus ilicifolia não alterou os níveis de carbonila. O ensaio do cometa demonstrou que as três espécies analisadas promoveram um nível baixo e semelhante de genotoxicidade, o que é compatível com os danos no DNA induzidos por outras plantas medicinais e é parcialmente recuperada por um co-tratamento com a vitamina C. Os dados mostraram M. dasyclada com atividade antioxidante in vitro, e que os seus efeitos genotóxicos e pró-oxidantes in vivo são comparáveis às espécies de referência de Maytenus.(AU)
Assuntos
Animais , Masculino , Antioxidantes/farmacologia , Maytenus/química , Plantas Medicinais/química , Antioxidantes/toxicidade , Ensaio Cometa , Maytenus/classificação , Maytenus/toxicidade , Extratos Vegetais/toxicidade , Plantas Medicinais/classificação , Plantas Medicinais/toxicidade , Ratos WistarRESUMO
Apart from constituting the raw material used to manufacture phytomedicines, plant drugs are commonly used by people as a therapeutic resource. Thus, the market in plant natural products has become an attractive target for investments of pharmaceutical companies. The aim of this study was to test the quality of commercial plant drugs in Brazil, employing simple and low-cost methods. Anatomical and microchemical tests were performed on commercial samples of "centela" (Asian pennywort or centella), "chá verde" (green tea) and "espinheira santa", to assess their quality and check their identity. The anatomical study revealed that all 3 samples of centella consisted of Centella asiatica leaves, but some were poorly conserved. The majority of contaminants consisted of other parts of C. asiatica, leaves of Poaceae and other species, and unidentified stalks. Two samples of green tea revealed leaves of the correct species (Camellia sinensis), with twigs of the same as contaminants, while the third consisted mainly of Ilex paraguariensis (mate tea) with some Bambusoideae (Poaceae) leaves. One of the 3 samples of "espinheira santa" contained Sorocea bonplandii leaves (cincho), and the others revealed leaves and stem fragments of Maytenus ilicifolia. The 3 samples of centella showed triterpene saponins. All samples of "green tea" revealed methylxanthines, but only those with C. sinensishad flavonoids. The samples of "espinheira santa" showed condensed tannins. Thus, the proposed analytical methods provided complementary results, which may be applied to quality control of plant drugs...
Drogas vegetais constituem uma das matérias-primas utilizadas na fabricação de fitoterápicos, além de serem largamente utilizadas pela população como recurso terapêutico. O mercado de produtos derivados de matéria-prima vegetal, com isso, se tornou alvo de investimentos de empresas do setor farmacêutico. O presente trabalho objetivou verificar a qualidade de drogas vegetais comercializadas no mercado brasileiro, utilizando-se conjuntamente métodos de análise simples e de baixo custo.Para tanto , foram usadas amostras de "centela", "chá verde" e "espinheira santa", obtidas em estabelecimentos comerciais, sendo sua identidade e qualidade avaliadas por meio de análises anatômicas e microquímicas. O estudo anatômico demonstrou que as três amostras de centela continham Centella asiatica, estando algumas em mau estado de conservação. Os contaminantes desta amostra eram principalmente outras partes do corpo vegetativo de C. asiatica, além de folhas de Poaceae e de outras espécies vegetais. Duas amostras de chá verde foram identificadas como Camellia sinensis e apresentavam caules da mesma espécie como contaminantes. A terceira amostra de chá verde era constituída por Ilex paraguariensis, sendo que folhas de Bambusoideae (Poaceae) também foram encontradas. Uma das amostras de espinheira santa era constituída de Sorocea bonplandii. As demais continham folhas e fragmentos de caule de Maytenus ilicifolia. As três amostras de centela apresentaram saponinas triterpênicas. Todas as amostras de chá verde possuíam metilxantinas. Dessas, apenas aquelas constituídas por C. sinensis demonstraram a presença de flavonoides. As amostras de espinheira-santa apresentaram taninos condensados. Desse modo, as metodologias propostas forneceram resultados complementares que podem ser empregados no controle de qualidade de drogas vegetais...
Assuntos
Humanos , Camellia sinensis , Extratos Vegetais/classificação , Maytenus , Plantas Medicinais/ultraestrutura , Brasil , Controle de QualidadeRESUMO
The medicinal plant Maytenus ilicifolia is a commonly used phytomedicine for the treatment of gastritis. The high dose required and low density of these extracts make necessary a daily intake of several capsules, hindering adherence to the medication. The purpose of this work was to develop a suitable dosage form for the administration of Maytenus ilicifolia using effervescent granules. A 23 factorial design was used to study the physical characteristics of the granules (particle size distribution, repose angle, Carr index, scanning electron microscopy and disintegration time). Moisture stability was also determined. According to the experimental design, granule size is the most important factor in determining the flow characteristics of effervescent granules. In turn, the disintegration time is controlled by the content of sodium bicarbonate present in the effervescent mixture as well as the granule size. The stability of formulations when exposed to moisture is strongly influenced by the percentage of effervescent mixture present in the vegetal granules. Precautions in handling and storage should be taken to ensure the stability of these preparations. The effervescent granules produced from Maytenus ilicifolia met the pharmacopoeial quality parameters, with appropriate mechanical and physical characteristics and proved to be a promising vehicle for plant extracts.
A planta medicinal Maytenus ilicifolia é comumente empregada como fitoterápico no tratamento da gastrite. As elevadas doses requeridas e a baixa densidade dos extratos dessa planta levam à ingestão diária de várias cápsulas do medicamento, dificultando a adesão ao tratamento. A proposta desse trabalho foi desenvolver uma forma farmacêutica adequada para administração de Maytenus ilicifolia usando granulados efervescentes. Um desenho fatorial 23 foi empregado para estudar as características físicas dos granulados (distribuição dos tamanhos de partícula, ângulo de repouso, índice de Carr, microscopia eletrônica de varredura e tempo de desintegração). A higroscopicidade das preparações também foi estudada. De acordo com o desenho experimental, o tamanho de partícula é o fator mais importante para a determinação das características de fluxo dos granulados efervescentes. Em contrapartida, o tempo de desintegração é controlado pelo conteúdo de bicarbonato de sódio presente na mistura efervescente, assim como pelo tamanho do granulado. A estabilidade das formulações quando expostas à umidade é fortemente influenciada pelo percentual de mistura efervescente presente nos granulados. Precauções de manipulação e armazenamento devem ser tomadas para garantir a estabilidade dessas preparações. Os granulados efervescentes produzidos com Maytenus ilicifolia cumprem os requisitos farmacopeicos de qualidade, com adequadas características físicas e mecânicas, provando ser um veículo promissor para extratos vegetais.
Assuntos
Projetos de Pesquisa/normas , Maytenus/classificação , Extratos Vegetais/farmacocinéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Although Maytenus ilicifolia is used in Brazilian folk medicine as a diuretic drug, no study has been conducted to this date in order to evaluate this ethnopharmacological statement. So, the aim of this study was to evaluate possible mechanisms involved in acute diuretic activity of the ethanolic supernatant of the infusion (SEI) obtained from Maytenus ilicifolia and to assess its relationship with a hypotensive activity by a bioassay-guided fractionation using normotensive Wistar rats. MATERIAL AND METHODS: The preparation obtained from the infusion (SEI) and their respective fractions (Fr·H2O and Fr·EtOAc) were orally administered in a single dose to rats. The urine excretion rate, pH, density, conductivity and content of Na(+), K(+), Cl(-) and HCO3(-) were measured in the urine of saline-loaded animals. Samples of the concentration of electrolytes, urea, creatinine, aldosterone, vasopressin and angiotensin converting enzyme (ACE) activity were evaluated in collected serum. The hypotensive activity and the involvement of nitric oxide, bradykinin and prostaglandin/cAMP pathway in the hypotensive and diuretic effects were also determined. RESULTS: Water and Na(+) excretion rate were significantly increased by Fr·EtOAc and the arterial pressure was significantly reduced, while the urinary excretion of potassium and chloride were reduced. Pre-treatment with indomethacin or DDA (2',5'-dideoxyadenosine) significantly reduced the hypotensive and diuretic activity observed. All other parameters evaluated were not affected by any treatment. CONCLUSION: The present study reveals that Fr·EtOAc obtained from Maytenus ilicifolia may present compounds responsible for diuretic and hypotensive activities, and this effect, could involve the prostaglandin/cAMP pathway.
Assuntos
Anti-Hipertensivos/farmacologia , AMP Cíclico/metabolismo , Diuréticos/farmacologia , Maytenus , Extratos Vegetais/farmacologia , Prostaglandinas/metabolismo , Animais , Masculino , Ratos , Ratos Wistar , Transdução de SinaisRESUMO
Maytenus aquifolia and Maytenus ilicifolia are native plants from South America and popularly known as Espinheira-santa. Both are used as tea due to their efficiency in the treatment of ulcer, gastritis and indigestion. However, adulteration of processed Maytenus genus tea with Sorocea genus may happen due to their botanical similarity, compromising the quality of the products and opening a derogatory business opportunity that may lead to the discrediting of medicinal plant products. This study aimed to distinguish Maytenus sp and Sorocea bonplandii by PCR-RFLP of a chloroplast DNA (cpDNA) intergenic region. Three commercial products of processed tea leaves of Maytenus sp, and in natura leaves of Maytenus sp and S. bonplandii were analyzed. PCR detected unique fragments for all samples in natura. The trnH-psbA region amplicon of both M. ilicifolia and M. aquifolia was 660 bp, and for S. bonplandii was 565 bp. These PCR products can be used as markers to distinguish the two genera. Forty-five percent of the processed samples presented only Maytenus genus, without adulterations. However, the amplification of 38% of the samples suggests adulteration with S. bonplandii while 17% seem to be adulterated with another plant (fragment of 649 bp in brand A and 690 bp in brand B). Three out of the fifteen restriction enzymes were able to detect M. ilicifolia and M. aquifolia in natura and in processed leaf samples. It was concluded that PCR technique is efficient to distinguish Maytenus sp from S. bonplandii, and other adulterating plants in processed commercial products of Espinheira-santa tea. The trnH-psbA spacer of cpDNA is easily amplified and has satisfactory discriminating capacity to help in authentication processes of samples of the genera in natura and in processed plants.(AU)
Maytenus aquifolia e Maytenus ilicifolia são plantas nativas da América do Sul e conhecidas popularmente como Espinheira-santa. Ambas são usadas como chá devido à sua eficiência no tratamento de gastrite, úlcera e indigestão. No entanto adulteração de chá do gênero Maytenus com o gênero Sorocea pode acontecer devido a sua semelhança botânica e isto compromete a qualidade dos produtos, abrindo uma oportunidade para comércio depreciativo que leva ao descrédito das plantas medicinais. Este estudo teve como objetivo distinguir Maytenus sp e Sorocea bonplandii por PCR-RFLP de DNA da região intergênica do cloroplasto (cpDNA). Três produtos comerciais de chá de Maytenus sp foram analisados, e a análise foi feita também em folhas in natura de Maytenus sp e de S. bonplandii. PCR detectou fragmentos únicos para todas as amostras in natura. O amplicon da região trnH-psbA de M. ilicifolia e M. aquifolia foi de 660 pb e para S. bonplandii foi de 565 pb. Os produtos de PCR podem ser usados como marcadores para distinguir os dois gêneros. Quarenta e cinco por cento das amostras processadas apresentaram apenas o gênero Maytenus, sem adulterações. No entanto, a amplificação de 38% das amostras sugere adulteração com S. bonplandii, enquanto 17% parecem terem sido adulteradas com uma outra planta (fragmento de 649 pb na marca A e 690 pb na marca B). Três das quinze enzimas de restrição foram capazes de detectar M. ilicifolia e M. aquifolia in natura e em amostras de folhas processadas. Concluiu-se que a técnica de PCR é eficiente para distinguir Maytenus sp de S. bonplandii e outras plantas adulterantes nos produtos comerciais de chá de Espinheira-santa. O espaçador trnHpsbA de cpDNA é facilmente amplificado e tem capacidade discriminante satisfatória para ajudar em processos de autenticação dos gêneros em amostras in natura e processadas de plantas.(AU)