RESUMO
Species of Piperaceae are known by biological properties, including antiparasitic such as leishmanicidal, antimalarial and in the treatment of schistosomiasis. The aim of this work was to evaluate the antileishmania activity, cytotoxic effect, and macrophage activation patterns of the methanol (MeOH), hexane (HEX), dichloromethane (DCM) and ethyl acetate (EtOAc) extract fractions from the leaves of Piper cabralanum C.DC. The MeOH, HEX and DCM fractions inhibited Leishmanina amazonensis promastigote-like forms growth with a half maximal inhibitory concentration (IC50) of 144.54, 59.92, and 64.87 µg/mL, respectively. The EtOAc fraction did not show any relevant activity. The half maximal cytotoxic concentration (CC50) for macrophages were determined as 370.70, 83.99, 113.68 and 607 µg/mL for the MeOH, HEX and DCM fractions, respectively. The macrophage infectivity was concentration-dependent, especially for HEX and DCM. MeOH, HEX and DCM fractions showed activity against L. amazonensis with low cytotoxicity to murine macrophages and lowering infectivity by the parasite. Our results provide support for in vivo studies related to a potential application of P. cabralanum extract and fractions as a promising natural resource in the treatment of leishmaniasis.
Assuntos
Antiprotozoários/química , Piper/química , Extratos Vegetais/química , Animais , Antiprotozoários/isolamento & purificação , Antiprotozoários/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Hexanos/química , Leishmania/efeitos dos fármacos , Leishmania/crescimento & desenvolvimento , Estágios do Ciclo de Vida/efeitos dos fármacos , Extração Líquido-Líquido , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Cloreto de Metileno/química , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Fagocitose/efeitos dos fármacos , Piper/metabolismo , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Folhas de Planta/química , Folhas de Planta/metabolismoRESUMO
Underutilized marine food products such as cephalopods' ink could be sources of bioactive compounds providing health benefits. This study aimed to assess the anti-proliferative and anti-inflammatory effects from Octopus vulgaris ink extracts (hexane-, ethyl acetate-, dichloromethane- (DM), and water extracts) using human colorectal (HT-29/HCT116) and breast (MDA-MB-231) cancer cells, and LPS-challenged murine RAW 264.7 cells. Except by ethyl-acetate, all of the extracts exhibited anti-proliferative effects without being cytotoxic to ARPE-19 and RAW 264.7 cells. Among DM fractions (F1/F2/F3), DM-F2 showed the highest anti-proliferative effect (LC50 = 52.64 µg/mL), inducing pro-apoptotic morphological disruptions in HCT116 cells. On RAW 264.7 cells, DM-F2 displayed the lowest nitrites reduction and up-regulation of key-cytokines from the JAK-STAT, PI3K-Akt, and IL-17 pathways. Compared to control, DM-F2 increased IL-4 and decreased NF-κB fluorometric expression in peripheral blood mononuclear cells (PBMCs). Metabolomic analysis of DM-F2 highlighted hexadecanoic acid and 1-(15-methyl-1-oxohexadecyl)-pyrrolidine as the most important metabolites. These compounds also exhibited high in silico binding affinity (-4.6 to -5.8 kcal/mol) to IL-1α, IL-1ß, and IL-2. Results suggested the joint immuno-modulatory and anti-proliferative effect derived from selected compounds of underutilized marine food products such as ink. This is the first report of such biological activities in extracts from O. vulgaris ink.
Assuntos
Anti-Inflamatórios/farmacologia , Proliferação de Células/efeitos dos fármacos , Octopodiformes/química , Animais , Citocinas/metabolismo , Células HCT116 , Humanos , Mediadores da Inflamação/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Metabolômica , Cloreto de Metileno/química , Camundongos , Nitritos/metabolismo , Células RAW 264.7 , Transdução de SinaisRESUMO
Dichloromethane (DCM) fraction and sub-fractions obtained from Smilax brasiliensis leaves were examined in order to determine their phytotoxic and antioxidant effects. The dichloromethane fraction was submitted to a preparative layer chromatography leading to seven sub-fractions (DCM1-DCM7). Gas chromatography-mass spectrometry (GC-MS) was performed on the dichloromethane sub-fractions. The DCM sub-fractions presented phytotoxic potential; at a concentration of 125 µg per plate, DCM6 and DCM4 showed the strongest results on Lactuca sativa and Allium cepa, respectively. The DCM fraction and DCM4 sub-fraction were more effective than 2,6-di-tert-butyl-4-methylphenol (BHT) at scavenging the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. Analysis by GC-MS showed the presence of methyl palmitate (33.05%) in DCM4 and methyl palmitate (17.29%) and methyl oleate (50.96%) in DCM6, suggesting that the activities exhibited by the sub-fractions may be attributed, at least partially, to these major compounds. These results indicate that the DCM sub-fractions of S. brasiliensis could be used as natural herbicides and antioxidants.
Assuntos
Antioxidantes/farmacologia , Extratos Vegetais/farmacologia , Smilax/química , Antioxidantes/química , Cromatografia Gasosa-Espectrometria de Massas , Lactuca/efeitos dos fármacos , Cloreto de Metileno/química , Cebolas/efeitos dos fármacos , Palmitatos/análise , Extratos Vegetais/química , Extratos Vegetais/toxicidade , Folhas de Planta/químicaRESUMO
Caatinga flora which are found in a poor Brazilian region contain a substantial number of endemic taxa with biomedical and social importance for regional communities. This study examined the antioxidant and cytotoxic potential of 35 samples (extracts/fractions) from 12 Caatinga species and determined the antiproliferative and genotoxic action of dichloromethane fraction from Mimosa caesalpiniifolia stem bark (DC-Mca) on human and vegetal cells. Samples were assessed for chemopreventive ability, toxic effects on Artemia salina shrimp as well as cytotoxicity on tumor cell lines and erythrocytes. DC-Mca was also tested with respect to antiproliferative and genotoxic effects upon normal leukocytes and meristematic cells from A. cepa roots. Some extracts reduced free radical levels >95% and 7 samples exhibited a lethal concentration (LC) 50 < 100 µg/ml upon Artemia salina larvae. Eight samples displayed in vitro antitumor effects and three produced hemolysis. Data also demonstrated the pharmacological significance of bioactive extracts from Brazilian semi-arid region. There was no significant relationship between antioxidant, toxic, and antiproliferative activities, and that these properties were dependent upon the extractant. DC-Mca contained betulinic acid as main compound (approximately 70%), which showed higher (1) cytotoxic activity on cancer cell lines and dividing leukocytes, (2) reduced mitotic index of Allium cepa roots, and (3) induced cell cycle arrest and chromosomal bridges, thereby providing native promising sources for phytotherapy development. ABBREVIATIONS: ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); AcOH: ethyl acetate; ANOVA: analysis of variance; SUS: Brazilian Unified Health System; DC-Mca: dichloromethane fraction from Mimosa caesalpiniifolia stem bark; DMSO: dimethylsulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EC50: effective concentration 50%; EtOAc: ethyl acetate; FDA: Food and Drug Administration; GC-Qms: gas chromatograph quadrupole mass spectrometer; GI: genotoxic index; HCT-116: colon carcinoma line; HL-60: promyelocytic leukemia line; HPLC: high-performance liquid chromatography; HRAPCIMS: high resolution atmospheric pressure chemical ionization mass spectrum; IC50: inhibitory concentration 50%; LC50: lethal concentration 50%; MeOH = methyl alcohol; MI: mitotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide; MutI: mutagenic index; OVCAR-8 = ovarian carcinoma line; PBMC: peripheral blood mononuclear cells; RPMI-1640: Roswell Park Memorial Institute medium; SF-295: glioblastoma line; TEAC: trolox equivalent antioxidant capacity; TLC: thin-layer chromatography; Trolox: 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Plantas Medicinais/química , Antineoplásicos Fitogênicos/química , Antioxidantes/química , Brasil , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , Citotoxinas/química , Citotoxinas/farmacologia , Dano ao DNA , Ecossistema , Ecotoxicologia , Humanos , Cloreto de Metileno/química , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Plantas Medicinais/classificação , Plantas Medicinais/toxicidadeRESUMO
Banana inflorescences are a byproduct of banana cultivation consumed in various regions of Brazil as a non-conventional food. This byproduct represents an alternative food supply that can contribute to the resolution of nutritional problems and hunger. This product is also used in Asia as a traditional remedy for the treatment of various illnesses such as bronchitis and dysentery. However, there is a lack of chemical and pharmacological data to support its consumption as a functional food. Therefore, this work aimed to study the anti-inflammatory action of Musa acuminata blossom by quantifying the cytokine levels (NOx, IL-1ß, TNF-α, and IL-6) in peritoneal neutrophils, and to study its antiparasitic activities using the intracellular forms of T. cruzi, L. amazonensis, and L. infantum. This work also aimed to establish the chemical profile of the inflorescence using UPLC-ESI-MS analysis. Flowers and the crude bract extracts were partitioned in dichloromethane and n-butanol to afford four fractions (FDCM, FNBU, BDCM, and BNBU). FDCM showed moderate trypanocidal activity and promising anti-inflammatory properties by inhibiting IL-1ß, TNF-α, and IL-6. BDCM significantly inhibited the secretion of TNF-α, while BNBU was active against IL-6 and NOx. LCMS data of these fractions revealed an unprecedented presence of arylpropanoid sucroses alongside flavonoids, triterpenes, benzofurans, stilbenes, and iridoids. The obtained results revealed that banana inflorescences could be used as an anti-inflammatory food ingredient to control inflammatory diseases.
Assuntos
1-Butanol/farmacologia , Anti-Inflamatórios/farmacologia , Cloreto de Metileno/farmacologia , Musa/química , Tripanossomicidas/farmacologia , 1-Butanol/química , Animais , Anti-Inflamatórios/química , Sobrevivência Celular/efeitos dos fármacos , Flores/química , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Leishmania/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Cloreto de Metileno/química , Camundongos , NADPH Oxidases/metabolismo , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Células THP-1 , Tripanossomicidas/química , Fator de Necrose Tumoral alfa/metabolismoRESUMO
This study aimed to evaluate the antimicrobial activity of the dichloromethane fraction (DCMF) from the stem bark of Mimosa caesalpiniifolia and its effect on the activity of conventional antibiotics against Staphylococcus aureus strains overexpressing specific efflux pump genes. DCMF showed activity against S. aureus, Staphylococcus epidermidis and Candida albicans. Addition of DCMF at subinhibitory concentrations to the growth media enhanced the activity of norfloxacin, ciprofloxacin and ethidium bromide against S. aureus strains overexpressing norA suggesting the presence of efflux pump inhibitors in its composition. Similar results were verified for tetracycline against S. aureus overexpressing tetK, as well as, for ethidium bromide against S. aureus overexpressing qacC. These results indicate that M. caesalpiniifolia is a source of molecules able to modulate the fluoroquinolone- and tetracycline-resistance in S. aureus probably by inhibition of NorA, TetK and QacC respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: Drug resistance is a common problem in patients with infectious diseases. Dichloromethane fraction from the stem bark of Mimosa caesalpiniifolia showed antimicrobial activity against Gram-positive bacterium Staphylococcus aureus and against Candida albicans, but did not show activity against Gram-negative specie Escherichia coli. Moreover, this fraction was able to potentiate the action of norfloxacin, ciprofloxacin and tetracycline against S. aureus strains overexpressing different efflux pump genes. Thus, Mimosa caesalpiniifolia is a source of efflux pump inhibitors which could be used in combination with fluoroquinolones or tetracycline in the treatment of infectious diseases caused by S. aureus strains overexpressing efflux pump genes.
Assuntos
Antibacterianos/farmacologia , Candida albicans/efeitos dos fármacos , Mimosa/química , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Antiporters/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Ciprofloxacina/farmacologia , Resistência a Múltiplos Medicamentos/genética , Etídio/farmacologia , Fluoroquinolonas/farmacologia , Humanos , Cloreto de Metileno/química , Testes de Sensibilidade Microbiana , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Norfloxacino/farmacologia , Casca de Planta/química , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/genética , Tetraciclina/farmacologia , Resistência a Tetraciclina/genéticaRESUMO
Conventional methods for determination of polycyclic aromatic compounds (PACs) in sediments usually require large sample sizes (grams) and solvent volumes (at least 100â¯mL) through the employment of Soxhlet extraction, which is both time (hours) and energy consuming, among other disadvantages. We developed a new analytical protocol for the determination of PACs in sediments using microextraction, which requires small sample masses (25â¯mg), 500⯵L of acetonitrile-dichloromethane mix and sonication for 23â¯min, followed by GC-MS analysis. The method was validated using the certified reference material SRM 1941b - NIST organic marine sediment, as well as internal deuterated standards. Seventeen PAHs, seven nitro-PAHs and one quinone were detected and quantified. The mean concentrations were 90.4â¯ngâ¯g-1 for PAHs, 179.2â¯ngâ¯g-1 for nitro-PAHs and 822.5â¯ngâ¯g-1 for quinones. The proposed method showed good sensitivity, linearity, precision and accuracy for the determination of PAC in sediments samples.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Sedimentos Geológicos/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Químicos da Água/análise , Acetonitrilas/química , Brasil , Fracionamento Químico/métodos , Cloreto de Metileno/química , Nitratos/química , Oxigênio/química , Hidrocarbonetos Policíclicos Aromáticos/química , Quinonas/análise , Reprodutibilidade dos Testes , Rios , Poluentes Químicos da Água/químicaRESUMO
Peperomia hispidula (Sw.) A. Dietr. is used in Mexican traditional medicine for treating respiratory illnesses such as asthma. The latter disorder results from an excessive and inappropriate constriction of airway smooth muscle. The aim of the present study was to evaluate the relaxant activity of P. hispidula on isolated rat tracheal rings contracted with carbachol. The methyleugenol was identified as the main active constituent in the dichloromethane extract. To explore the possible mechanism of action, concentration-response curves were constructed in the presence and absence of propranolol (3 µM), indomethacin (10 µM), glibenclamide (1 µM), and L-NAME (300 µM), finding that neither reduced methyleugenol-induced smooth muscle relaxation. In conclusion, P. hispidula herein displayed relaxant activity on rat tracheal rings. The effect of methyleugenol, was probably not related to the activation of ß2-adrenoceptors, prostaglandins, K+ATP channels or nitric oxide.
Peperomia hispidula (Sw.) A. Dietr. es utilizada en la medicina tradicional mexicana para tratar enfermedades respiratorias como el asma. Este uÌltimo trastorno es el resultado de una contraccioÌn excesiva e inapropiada del muÌsculo liso de las viÌas respiratorias. El objetivo del presente estudio fue evaluar la actividad relajante de P. hispidula sobre anillos aislados de traÌquea de rata contraiÌdos con carbacol. El metileugenol fue identificado como el principal constituyente activo en el extracto de diclorometano. Para explorar el posible mecanismo de accioÌn, se construyeron curvas concentracioÌn-respuesta en presencia y ausencia de propranolol (3 µM), indometacina (10 µM), glibenclamida (1 µM), y L-NAME (300 µM), encontrando que ninguno redujo la relajacioÌn del muÌsculo liso inducida por metileugenol. En conclusioÌn, P. hispidula muestra actividad relajante en anillos de traÌquea de rata. El efecto de metileugenol, al parecer no estaÌ implicado con la activacioÌn de los receptores ß2-adreneÌrgicos, prostaglandinas, canales de K+ATP u oÌxido niÌtrico.
Assuntos
Animais , Masculino , Ratos , Traqueia/efeitos dos fármacos , Eugenol/análogos & derivados , Eugenol/farmacologia , Extratos Vegetais/farmacologia , Peperomia , Asma/metabolismo , Estenose Traqueal/induzido quimicamente , Eugenol/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas em Tandem , Cloreto de Metileno/química , Relaxamento Muscular/efeitos dos fármacosRESUMO
The present study aimed to characterize the chemical composition of pyroligneous acid (PA) obtained from slow pyrolysis of the clone GG100 of Eucalyptus urophylla × Eucalyptus grandis. The efficiency of extraction of organic compounds by using different solvents-dichloromethane (DCM), diethyl ether (DE) and ethyl acetate (EA)-was evaluated. Wood discs were collected and carbonized at a heating rate of 1.25 °C/min until 450 °C. Pyrolysis gases were trapped and condensed, yielding a crude liquid product (CLP), which was refined to obtain pure PA. Then liquid-liquid extraction was carried out. Each extracted fraction was analyzed by GC-MS and the chemical compounds were identified. Experimental results showed that a larger number of chemical compounds could be extracted by using DCM and EA in comparison to diethyl ether DE. A total number of 93 compounds were identified, with phenolic compounds being the major group, followed by aldehydes and ketones, furans, pyrans and esters. Higher contents of guaiacol, phenol, cresols and furfural seem to explain the antibacterial and antifungal activity shown by PA, as reported previously in the literature. Experimental data indicated that the organic phase extracted from GG100 PA consists of a mixture of compounds similar to liquid smokes regularly used in the food industry.
Assuntos
Eucalyptus/química , Fenóis/química , Solventes/química , Terpenos/química , Acetatos/química , Aldeídos/química , Ésteres/química , Éter/química , Furanos/química , Cromatografia Gasosa-Espectrometria de Massas , Cetonas/química , Cloreto de Metileno/química , Fenóis/isolamento & purificação , Piranos/química , Terpenos/isolamento & purificação , Madeira/químicaRESUMO
Phoradendron mucronatum and P. microphyllum are plants that found in tropical and subtropical areas, used in traditional medicine and popularly known as mistle-thrush. The aim of this study was to identify the chemical constituents of different leaf extracts from P. mucronatum and P. microphyllum and assess cytotoxic activity against strains from a human tumour cells. Extracts obtained with hexane, dichloromethane, chloroform and ethyl acetate from the leaves were analysed by gas chromatography coupled with mass spectrometry (GC-MS) and the cytotoxicity was assessed by the MTT method (bromide (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)). The tested human tumour cells were NCI-H292 (human pulmonar mucoepidermoid carcinoma), MCF-7 (human breast adenocarcinoma) and HEp-2 (epidermoid carcinoma of the larynx). Analysis by GC/MS of the extracts from leaves of P. microphyllum and P. mucronatum detected 51 different compounds, such as alkaloids, diterpenes, triterpenes, sterols, alcohols, aldehydes, fatty acids and hydrocarbons. In the cytotoxic evaluation, hexane and ethyl acetate extracts from the leaves P. microphyllum inhibited cell growth of NCI-H292 strains (72.97%) and HEp-2 (87.53%), respectively. The extracts of P. mucronatum species showed an inhibitory effect towards NCI-H292 (83.19%/hexane), MCF-7 (88.69%/dichloromethane) and HEp-2 (93.40%/hexane). The extracts showed cytotoxic activity against the tested strains, especially the P. mucronatum, which presented the highest percentages of inhibition of cell growth.