RESUMO
INTRODUCTION: Species of Connaraceae are globally used in traditional medicines. However, several of these have not been studied regarding their chemical composition, and some are even at risk of extinction without proper studies. Therefore, the chemical composition and pharmacological potential of Connarus blanchetii Planch., Connarus nodosus Baker, Connarus regnellii G. Schellenb., and Connarus suberosus Planch., which were previously unknown, were analyzed. OBJECTIVE: This work aims to investigate the pharmacological potential of these four Connarus species. The chemical composition of different extracts was determined by high-resolution mass spectrometry (HRMS), with subsequent analysis by the GNPS platform and competitive fragmentation modeling (CFM). MATERIALS AND METHODS: Leaf extracts (C. blanchetii, C. nodosus, C. regnellii, and C. suberosus) and bark extracts (C. regnellii and C. suberosus) were obtained by decoction, infusion, and maceration. LC/HRMS data were submitted to the GNPS platform and evaluated using CFM in order to confirm the structures. RESULTS: The HRMS-GNPS/CFM analysis indicated the presence of 23 compounds that were mainly identified as phenolic derivatives from quercetin and myricetin, of which 21 are unedited in the Connarus genus. Thus, from the analyses performed, we can identify different compounds with pharmacological potential, as well as the most suitable forms of extraction. CONCLUSION: Using HRMS-GNPS/CFM, 21 unpublished compounds were identified in the studied species. Therefore, our combination of data analysis techniques can be used to determine their chemical composition.
Assuntos
Connaraceae , Cromatografia Líquida de Alta Pressão/métodos , Connaraceae/química , Medicina Tradicional , Fenóis/análise , Extratos Vegetais/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em TandemRESUMO
Leishmaniasis is a disease impacting public health worldwide due to its high incidence, morbidity and mortality. Available treatments are costly, lengthy and toxic, not to mention the problem of parasite resistance. The development of alternative treatments is warranted and natural products demonstrate promising activity. This study investigated the activity of Connarus suberosus extracts and compounds against Leishmania species. Several C. suberosus extracts were tested against L. amazonensis promastigotes. Active and inactive extracts were analyzed by UHPLC-MS and data evaluated using a metabolomics platform, revealing an unknown neoflavonoid (connarin, 3), isolated together with the pterocarpans: hemileiocarpin (1) and leiocarpin (2). The aforementioned compounds (1-3), together with the benzoquinones: rapanone (4), embelin (5) and suberonone (6) previously isolated by our group from the same species, were tested against: (i) L. amazonensis and L. infantum promastigotes, and (ii) L. amazonensis intracellular amastigotes, with the most active compound (3) also tested against L. infantum amastigotes. Cytotoxicity against murine peritoneal macrophages was also investigated. Compounds 2 and 3 presented an IC50 33.8 µM and 11.4 µM for L. amazonensis promastigotes; and 44.3 µM and 13.3 µM for L. infantum promastigotes, respectively. For L. amazonensis amastigotes, the IC50 of 2 was 20.4 µM with a selectivity index (SI) of 5.7, while the IC50 of 3 was 2.9 µM with an SI of 6.3. For L. infantum amastigotes, the IC50 of 3 was 7.7 µM. Compounds 2 and 3 presented activity comparable with the miltefosine positive control, with compound 3 found to be 2-4 times more active than the positive control, depending on the Leishmania species and form. The extracts and isolated compounds showed moderate toxicity against macrophages. Compounds 2 and 3 altered the mitochondrial membrane potential (ΔΨm) and neutral lipid body accumulation, while 2 also impacted plasma membrane permeabilization, culminating in cellular disorder and parasite death. Transmission electron microscopy of L. amazonensis promastigotes treated with compound 3 confirmed the presence of lipid bodies. Leiocarpin (2) and connarin (3) demonstrated antileishmanial activity. This study provides knowledge of natural products with antileishmanial activity, paving the way for prototype development to fight this neglected tropical disease.
Assuntos
Connaraceae/química , Flavonoides/farmacologia , Metabolômica/métodos , Extratos Vegetais/farmacologia , Animais , Antiprotozoários/química , Antiprotozoários/isolamento & purificação , Antiprotozoários/farmacologia , Sobrevivência Celular , Cromatografia Líquida de Alta Pressão , Flavonoides/química , Flavonoides/isolamento & purificação , Leishmania mexicana/efeitos dos fármacos , Leishmania mexicana/crescimento & desenvolvimento , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/parasitologia , Espectrometria de Massas , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificaçãoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: An interdisciplinary scientific investigation of biologically active agents is fundamental to search for natural substances with therapeutic action. This review collected the most relevant information on traditional knowledge related to the use of plants of the Connaraceae family. This work is the first to compile all the published ethnobotanical, chemical, pharmacological, and toxicological information about this important plant family. AIM OF THE STUDY: Our objective was to provide the scientific community with an up-to-date overview of the pharmacological potential of Connaraceae species. MATERIAL AND METHODS: We searched NCBI Pubmed Central, Google Scholar, Scientific Electronic Library Online (SciELO), ScienceDirect, SciFinder, and Scopus databases to review the research on ethnobotanical, chemical, pharmacognostical, pharmacological, and toxicological studies with Connaraceaes. Books that address the theme were also included. DISCUSSION AND CONCLUSION: The literature review indicated that 39 species of Connaraceaes have pharmacological potentiality. Ethnobotany reports listed 36 of the 39 species discussed. Pharmacognostical studies have been conducted with 23 species and isolates, and chemical compounds have been identified for only 15 species. At least one study has been published concerning the pharmacological activities for 20 of the 39 species analyzed. For Agelaea pentagyna, Cnestis ferruginea, Connars suberosus, and Rourea minor, pharmacological activity experiments were performed using isolated compounds, which have the highest current pharmacological potential. Studies employing a toxicological approach cover only 10 of the 39 Connaraceaes species. Thus, scientific community should conduct much more research for a broader understanding of this plant family.
Assuntos
Connaraceae , Etnobotânica , Etnofarmacologia , Fitoterapia , Extratos Vegetais/farmacologia , Animais , Connaraceae/química , Connaraceae/classificação , Connaraceae/toxicidade , Humanos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Especificidade da EspécieRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites. AIM OF THE STUDY: To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract. MATERIALS AND METHODS: AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography. RESULTS: The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50)µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked. CONCLUSION: AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.
Assuntos
Antibacterianos/farmacologia , Antivenenos/farmacologia , Connaraceae/química , Venenos de Crotalídeos/administração & dosagem , Hemorragia/tratamento farmacológico , Extratos Vegetais/farmacologia , Animais , Antibacterianos/química , Antioxidantes/farmacologia , Antivenenos/química , Bothrops , Brasil , Feminino , Hemorragia/metabolismo , Masculino , Medicina Tradicional/métodos , Camundongos , Fosfolipases A2/metabolismo , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/química , Mordeduras de Serpentes/tratamento farmacológico , Taninos/metabolismoRESUMO
Connarus suberosus is a typical species of the Brazilian Cerrado biome, and its inflorescences and young vegetative branches are densely covered by dendritic trichomes. The objective of this study was to report the occurrence of a previously undescribed glandular trichome of this species. The localization, origin and structure of these trichomes were investigated under light, transmission and scanning electron microscopy. Collections were made throughout the year, from five adult specimens of Connarus suberosus near Botucatu, São Paulo, Brazil, including vegetative and reproductive apices, leaves and fruits in different developmental stages, as well as floral buds and flowers at anthesis. Glandular trichomes (GTs) occurred on vegetative and reproductive organs during their juvenile stages. The GTs consisted of a uniseriate, multicellular peduncle, whose cells contain phenolic compounds, as well as a multicellular glandular portion that accumulates lipids. The glandular cell has thin wall, dense cytoplasm (with many mitochondria, plastids and dictyosomes), and a large nucleus with a visible nucleolus. The starch present in the plastids was hydrolyzed during the synthesis phase, reducing the density of the plastid stroma. Some plastids were fused to vacuoles, and some evidence suggested the conversion of plastids into vacuoles. During the final activity stages of the GTs, a darkening of the protoplasm was observed in some of the glandular cells, as a programmed cell death; afterwards, became caducous. The GTs in C. suberosus had a temporal restriction, being limited to the juvenile phase of the organs. Their presence on the exposed surfaces of developing organs and the chemical nature of the reserve products, suggest that these structures are food bodies. Field observations and detailed studies of plant-environment interactions, as well as chemical analysis of the reserve compounds, are still necessary to confirm the role of these GTs as feeding rewards.
Assuntos
Connaraceae/ultraestrutura , Brasil , Connaraceae/química , Connaraceae/classificação , Microscopia EletrônicaRESUMO
Connarus suberosus is a typical species of the Brazilian Cerrado biome, and its inflorescences and young vegetative branches are densely covered by dendritic trichomes. The objective of this study was to report the occurrence of a previously undescribed glandular trichome of this species. The localization, origin and structure of these trichomes were investigated under light, transmission and scanning electron microscopy. Collections were made throughout the year, from five adult specimens of Connarus suberosus near Botucatu, São Paulo, Brazil, including vegetative and reproductive apices, leaves and fruits in different developmental stages, as well as floral buds and flowers at anthesis. Glandular trichomes (GTs) occurred on vegetative and reproductive organs during their juvenile stages. The GTs consisted of a uniseriate, multicellular peduncle, whose cells contain phenolic compounds, as well as a multicellular glandular portion that accumulates lipids. The glandular cell has thin wall, dense cytoplasm (with many mitochondria, plastids and dictyosomes), and a large nucleus with a visible nucleolus. The starch present in the plastids was hydrolyzed during the synthesis phase, reducing the density of the plastid stroma. Some plastids were fused to vacuoles, and some evidence suggested the conversion of plastids into vacuoles. During the final activity stages of the GTs, a darkening of the protoplasm was observed in some of the glandular cells, as a programmed cell death; afterwards, became caducous. The GTs in C. suberosus had a temporal restriction, being limited to the juvenile phase of the organs. Their presence on the exposed surfaces of developing organs and the chemical nature of the reserve products, suggest that these structures are food bodies. Field observations and detailed studies of plant-environment interactions, as well as chemical analysis of the reserve compounds, are still necessary to confirm the role of these GTs as feeding rewards.
Connarus suberosus es una especie típica del bioma Cerrado brasileño, y sus inflorescencias junto con los ramos vegetativos jóvenes están densamente cubiertos por tricomas dendríticas. El objetivo de este estudio es presentar la ocurrencia de un tricoma glandular sin descripción previa para esta especie. La localización, origen y estructura de estos tricomas se observó mediante microscopia óptica, así como electrónica de transmisión y de barrido. Los tricomas glandulares (TGs) se producen en los órganos vegetativos y reproductivos, en sus fases jóvenes. Los TGs consisten en un pedúnculo uniseriado y multicelular, cuyas células contienen compuestos fenólicos, así como un sitio en el cual las células glandulares acumulan compuestos pécticos, almidón y lípidos. La acumulación de aceite en el citosol es impresionante, pues ocupa una gran parte del volumen celular. La porción glandular presenta células con paredes delgadas, un denso citoplasma y un núcleo grande con nucleolo visible. El citoplasma contiene gran cantidad de mitocondrias, plástidos y dictiosomas. El almidón presente en los plástidos se hidroliza durante la fase de síntesis, de tal modo que reduce la densidad del estroma plastidial. Algunos plástidos se fusionan con las vacuolas, además hay pruebas de la conversión de los plástidos en vacuolas. Durante las etapas finales de la actividad de los TGs, se observó un oscurecimiento del protoplasma en algunas de las células glandulares lo que sugiere la existencia de la muerte celular programada; después de lo anterior, los TGs se convierten en caducas. Por lo tanto la aparición de TGs en las superficies expuestas de los órganos en desarrollo en C. suberosus y la naturaleza química de los productos de reserva sugieren que estas estructuras son cuerpos alimenticios. Hay evidencias sobre el papel de estos tricomas en las interacciones planta-hormiga, especialmente en el bioma Cerrado.