RESUMO
Ethanol extract from the aerial parts of Euphorbia tirucalli L. as well as the latex of the plant suspended in water are used by the Brazilian population for the treatment of various diseases, including cancer. The purposes of this study were to determine if the ethanol extract is effective as cytotoxic agent against gastric adenocarcinoma cells (AGS) and its chemical composition by GC-MS, ESI-(-)-FT-ICR MS and (-)-ESI-LTQ-MS/MS. The results were compared with that of latex previously described by us. Hexane and aqueous fractions showed higher cytotoxicity on AGS cells. Nine triterpene compounds were detected by GC-MS in hexane fraction, including euphol and friedelin, while ellagic acid was identified as main phenolic compound in aqueous extract. Therefore, the greater cytotoxic activity of the ethanol extract of the aerial parts of Euphorbia tirucalli for gastric cancer, when compared to latex, seems to originate from the antiproliferative effects of ellagic acid and triterpenes.
Assuntos
Adenocarcinoma , Antineoplásicos , Euphorbia , Neoplasias Gástricas , Triterpenos , Humanos , Euphorbia/química , Látex/química , Hexanos , Espectrometria de Massas em Tandem , Neoplasias Gástricas/tratamento farmacológico , Ácido Elágico , Extratos Vegetais/farmacologia , Triterpenos/farmacologia , Adenocarcinoma/tratamento farmacológico , Componentes Aéreos da Planta , EtanolRESUMO
Euphorbia umbellata is used for its anti-inflammatory properties; however, there are limited data available regarding its effects on vascular function. Its bark is rich in polyphenolic compounds, which potentially improve endothelial dysfunction (ED). This study proposes to investigate the effects of E. umbellata bark extracts and its polyphenolic compounds on arginase (ARG) activity and nitric oxide (NO)-related targets. Chromatographic procedures were used for the chemical characterisation of the extracts. Furthermore, in silico (molecular docking), in vitro (ARG inhibition), in vivo (streptozotocin-induced hyperglycemia model), and ex vivo (l-arginine metabolism, vascular reactivity, western blot, and biochemical) techniques were carried out. Quercetin, gallic acid, and ellagic acid were identified in the extracts. In silico screening predicted that gallic acid and quercetin would have the most promising interactions with ARG -identified cavities. This was confirmed in vitro as both compounds had a direct inhibitory effect on ARG, as was the case regarding the extracts. Oral treatment preserved endothelium-dependent vasodilation through ARG inhibition together with an increase in l-arginine bioavailability and endothelial NO synthase expression. Biochemical parameters determined the lack of toxicity for sub-chronic treatment. E. umbellata bark extracts and its compounds can contribute to ED treatment, at least partly, through the inhibition of vascular ARG.
RESUMO
Paullinia cupana Kunth, commonly known as guarana, is a native Brazilian plant species from the Amazon area that presents various biological effects, including antimicrobial action. The aim of this study was to chemically analyse the semipurified aqueous extract (AqF) of the plant and to evaluate the activity of crude (CE), ethyl-acetate (EAF), and AqF extracts against Helicobacter pylori. The chemical profile of AqF was determined based on solid analysis 13C-NMR, direct infusion mass spectrometry (ESI-MS), and MALDI-TOF. The 13C-NMR spectrum showed characteristics of flavan-3-ol and oligomeric proanthocyanidins. ESI-MS revealed the presence of procyanidin, caffeic acid and its derivatives. MALDI-TOF analysis detected procyanidins of up to 6 units and profisetinidins of up to 5 units. Whereas CE and EAF showed inhibitory activity against H. pylori, CE, EAF, and AqF presented not high inhibitory activity against urease. The results demonstrate the potential of P. cupana to control and prevent H. pylori infection.
Assuntos
Helicobacter pylori/efeitos dos fármacos , Paullinia/química , Extratos Vegetais/química , Antioxidantes/farmacologia , Brasil , Cromatografia Gasosa-Espectrometria de Massas , Infecções por Helicobacter/prevenção & controle , Extratos Vegetais/farmacologia , Proantocianidinas/farmacologia , Urease/antagonistas & inibidoresRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia tirucalli L. is widely used by Brazilian folk medicine, mainly for its anticancer activity. However, its commercialization was banned by The Brazilian National Sanitary Surveillance Agency (ANVISA) due to the presence of some compounds considered toxic, such as the diterpene esters. AIM OF THE STUDY: Chemical and biological analyses were performed with the Brazilian Euphorbia tirucalli latex to support its wide traditional use in Brazil. MATERIAL AND METHODS: Latex was collected by using two procedures, in a solution of dichloromethane: methanol (3:1, 100 mL) and in 100 mL of distilled water. The first procedure was concentrated as a crude extract and the second one was partitioned with hexane and dichloromethane. The partitions and crude extract were subjected to phytochemical analyses using three different methods: Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS) with electrospray ionization sources in negative mode (ESI(-)) as well as in tandem mass spectrometry ESI(-) MS/MS and Atmospheric Pressure Chemical Ionization in positive mode (APCI(+)), Gas Chromatography coupled Mass Spectrometry (GC-MS) and Nuclear Magnetic Resonance (NMR) (1H-NMR and 13C-NMR). The cytotoxic potential was evaluated using the crude extract in macrophages RAW 264.7 and Gastric Adenocarcinoma (AGS) cancer cells. The evaluation of immunomodulatory activity was made through the detection of Nitric Oxide (NO) and cytokines as Tumor necrosis factor α (TNF-α) and Interleukin-6 (IL-6). RESULTS: GC-MS showed the presence of some esters of fatty acids, for instance myristic, palmitic, stearic, oleic and linoleic acid and, mainly, triterpenes such as euphol and tirucallol. With NMR, most of the signals were related to triterpenoids euphol and tirucallol. However, when the latex was analyzed with ESI(-) FT-ICR MS, a wide variety of molecules from different classes of natural products (fatty acids, diterpenes, triterpenes, steroids) were found. On the other hand, when APCI(+)FT-ICR MS was used, the ion M+. At ratio mass-charge (m/z) 426.38567, related to triterpenes euphol and tirucallol masses, presented the most intense peak, with a mass error of -0.11, indicating high accuracy. Diterpene esters from 4-deoxyphorbol and ingenol were identified only by ESI(-)FT-ICR MS and ESI(-)FT-ICR MS/MS. When evaluated biologically, the crude latex showed immunomodulatory activity, as it reduced the production of the pro-inflammatory cytokines TNF-α, IL-6 and NO, and the effect on NO reduction was more significant, obtaining in a similar result to the N(ω)-Nitro-L-Arginine Methyl Ester (L-NAME) standards, as well as significant cytotoxic activity with half inhibitory concentration (IC50) values of 69.43⯱â¯1.29⯵g/mL against AGS without damaging healthy ones. CONCLUSION: It was verified that the Brazilian Euphorbia tirucalli latex consists mainly of the triterpenes euphol and tirucallol, which may be the main cause of the anticancer activity attributed to the plant, but many other minor compounds could have been determined by the FT-ICR MS method, such as the diterpene esters. It has antitumor potential because it acts selectively against cancer cells and it also prevents the progression of tumors, because it carries an important immunomodulatory effect.
Assuntos
Antineoplásicos/análise , Antineoplásicos/farmacologia , Euphorbia , Látex/química , Látex/farmacologia , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/farmacologia , Animais , Brasil , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Medicina Tradicional , Camundongos , Células RAW 264.7 , Espectrometria de Massas em Tandem , Triterpenos/análise , Triterpenos/farmacologiaRESUMO
The long-term use of anti-inflammatory drugs is the most common cause of gastric ulcer disease, one of the major gastrointestinal disorders affecting people worldwide. Persea americana Mill. (avocado) seed is a by-product generally discarded as waste, but can be used to treat gastric disorder due to its anti-inflammatory, antioxidant and antimicrobial activities. The aim of the present study was to evaluate the potential protective effects of the ethyl acetate fraction of avocado seeds (SEAP) extracts against indomethacin-induced gastric ulcer in mice. It was found that SEAP were effective in mitigating oxidative stress through a decrease on the oxidized products levels (reduction of 90% in lipid peroxidation in plasma) and increasing superoxide dismutase enzyme (SOD) activity (4.25-fold increase compared to the indomethacin group), also preventing the rise in ulcer and lesions areas (92% of protection) and histological changes induced by indomethacin. Chemical analysis using mass spectrometry by (-)-ESI-FT-ICR MS revealed the presence of (-)-epicatechin and (+)-catechin, confirmed by HPLC-DAD, and other important phenolic compounds in avocado seeds, such as caffeoylquinic acid, flavonoids, phenylpropanoids and tannins, substances that promote inhibition of pathways involved in gastric ulcer formation. Thus, avocado seeds extract may be a suitable natural source for the prevention and treatment of gastric ulcer.
Assuntos
Indometacina/efeitos adversos , Persea/química , Extratos Vegetais/uso terapêutico , Sementes/química , Úlcera Gástrica/prevenção & controle , Animais , Antioxidantes/análise , Brasil , Catequina/análise , Cromatografia Líquida de Alta Pressão , Modelos Animais de Doenças , Flavonoides/análise , Peroxidação de Lipídeos , Masculino , Camundongos , Oxirredução , Fenóis/análise , Extratos Vegetais/química , Úlcera Gástrica/patologia , Superóxido Dismutase/sangue , Superóxido Dismutase/metabolismoRESUMO
Spondias mombin L. is used in folk medicine for the treatment of inflammation and gastrointestinal diseases. Our study investigated the antiulcer activity of S. mombin ethanolic extract (SmEE) and its majority compounds gallic acid (GA) and ellagic acid (EA). Phytochemical characterization was performed by HPLC. The SmEE was screened for in vitro antioxidant activities using phosphomolybdenum, ABTS, DPPH, and FRAP assays. The antiulcer activity of SmEE, GA, EA, or GA + EA was evaluated by gastric lesion models induced by absolute ethanol and indomethacin. Following this, it is capable of stimulating mucus production, antisecretory capacity, and the influence of -SH groups and NO in the effect of SmEE. Its healing activity was demonstrated by acetic acid-induced chronic ulcer model. Anti-Helicobacter pylori activity was assessed by determining the MIC of the SmEE (64-1024 µg/mL). The HPLC results identified the presence of gallic acid and ellagic acid in SmEE. The extract showed antioxidant activity in vitro. SmEE (50, 100, and 200 mg/kg) reduced the area of ulcerative lesions induced by ethanol in 23.8, 90.3, and 90.2%, respectively. In NSAID model, the SmEE induced protection of 36.8, 49.4, and 49.9%, respectively. GA (10 mg/kg) or EA (7 mg/kg) or the association of GA + EA (10 + 7 mg/kg) inhibited the ethanol-induced lesions in 71.8, 70.9, and 94.9%, respectively, indicating synergistic action. SmEE (100 mg/kg) decreased acid secretion and H+ concentration in the gastric contents, increased levels of mucus, and showed to be dependent of -SH groups and NO on the protection of the gastric mucosa. In chronic ulcer model, SmEE reduced the gastric area lesion. SmEE showed anti-H. pylori activity. In conclusion, our study showed that SmEE has antiulcerogenic activity. GA and EA are isolated gastric protectors and, when associated, acted synergistically to protect the gastric mucosa.
Assuntos
Anacardiaceae/química , Medicina Herbária/métodos , Fitoterapia/métodos , Folhas de Planta/química , Úlcera Gástrica/tratamento farmacológico , Animais , Camundongos , Ratos , Ratos WistarRESUMO
Na região amazônica, ainda que não conclusivo, estima-se a existência de aproximadamente 60.000 espécies de plantas contribuindo para uma vasta riqueza natural. Considerando essa estimativa, são necessários investimentos em ciência e tecnologia, proteção e valorização dos recursos naturais e, principalmente, a importância dos conhecimentos tradicionais associados à biodiversidade (Albagli, 2010). O conhecimento tradicional tem fornecido informações valiosas para a descoberta de novos agentes antimicrobianos a partir das plantas (Duraipandiyan, Ayyanar e Ignacimuthu, 2006), destacando a importância da medicina popular na busca por moléculas eficazes contra micro-organismos cada vez mais resistentes (Lopez, Hudson e Towers, 2001), como por exemplo Helicobacter pylori.(AU)
The Amazon region has an enormous diversity of plant species, which contributes to its known vast natural wealth. In this context, the aim of this work was to evaluate the antioxidant, anti-Helicobacter pylori and antiurease potentials of medicinal plant extracts popularly used in the western Pará, northern Brazil. Antimicrobial assays were carried out using spectrophotometric broth microdilution technique, the antioxidant potential by the DPPH and ABTSâ¢+ assays and anti-urease activity by inhibition of the enzyme urease. The ethanolic extract of leaves of Schnella sp. showed the best antioxidant potential in both methods, with IC50= 6.35 µg/mL (DPPH) and 1.81 µg/mL (ABTSâ¢+) and the highest percentage of inhibition of urease (45%) at 1024 µg/ml. The ethanolic extract of the resin of "breu-branco" inhibited 82.8% (512 µg/mL) of the bacterial growth. These results together show the potential of the plant species used in the western Pará, in the search for molecules with antioxidant and antimicrobial action.(AU)
Assuntos
Humanos , Anti-Infecciosos/química , Antioxidantes/química , Fabaceae/química , Fitoterapia , Plantas Medicinais/química , Ecossistema Amazônico , Brasil , Helicobacter pylori , Urease/análiseRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia umbellata (leitosinha) is used in southern Brazilian folk medicine to treat gastric problems, as well as for its analgesic and anti-inflammatory properties. AIM OF STUDY: To evaluate the anti-ulcer effects of methanolic bark fraction (MF) against in vivo and in vitro assays, as well as an antioxidant, antibacterial and chromatographic study of this fraction. MATERIALS AND METHODS: In vivo anti-ulcer activity was performed using ethanol and indomethacin models with different MF concentrations (50, 100 or 200mg/Kg). The stomachs of the animals were applied to histological evaluation, and the serum to evaluate the ABTS(â¢+) radical capture. The 200mg/Kg dose was used to analyze the mechanisms involved in antiulcerogenic properties of methanolic fraction. The in vitro activity was performed using several different antioxidant assays, in addition to anti-Helicobacter pylori and anti-urease experiments. The chromatographic study was carried out by LC-MS analysis. RESULTS: Pharmacological investigation of the MF showed an anti-ulcer potential in ethanol and indomethacin in vivo assays. The material presented a high antioxidant activity for several oxidant in vitro systems (DPPH(â¢), ABTS(â¢+), O2(â¢-), HOCl, TauCl and HRP), as well as an ABTS(â¢+) capture increasing (7.5%) by the treated animals serum (when compared to the negative control). Prostaglandins, nitric oxide/ cyclic guanosine monophosphate pathway and involvement of the protein components of the glutathione complex are some of the mechanisms related with this potential anti-ulcer action. The histological examination of the stomachs of the animals showed that the MF also prevents local action of offensive agents. Chemical analysis using LC-QTOF-MS revealed the presence of ellagic and gallic acid derivatives and flavonols. CONCLUSION: The findings provide scientific basis to the ethnopharmacological purpose of the studied plant and the biological activities of MF of E. umbellata stem bark may be due to the presence of phenolic compounds.
Assuntos
Antiulcerosos/farmacologia , Euphorbia/química , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Úlcera Gástrica/prevenção & controle , Estômago/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Antiulcerosos/química , Antiulcerosos/isolamento & purificação , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Benzotiazóis/química , Cromatografia Líquida de Alta Pressão , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Ácido Elágico/isolamento & purificação , Ácido Elágico/farmacologia , Etanol , Etnofarmacologia , Feminino , Flavonóis/isolamento & purificação , Flavonóis/farmacologia , Ácido Gálico/isolamento & purificação , Ácido Gálico/farmacologia , Mucosa Gástrica/metabolismo , Helicobacter pylori/efeitos dos fármacos , Indometacina , Metanol/química , Fitoterapia , Casca de Planta/química , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Plantas Medicinais , Polifenóis/química , Polifenóis/isolamento & purificação , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Solventes/química , Espectrometria de Massas por Ionização por Electrospray , Estômago/patologia , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/metabolismo , Úlcera Gástrica/patologia , Ácidos Sulfônicos/químicaRESUMO
Ten Brazilian medicinal plants used to treat gastritis and ulcers were carefully selected on the basis of ethnopharmacological importance and antiulcerogenic activity previously described. The antioxidant activity of the methanolic extracts was determined in analysis conditions that simulate a real biological activity on inhibition of the oxidative burst induced in neutrophils using Helicobacter pylori as activator, by a luminol-amplified chemiluminescence assay. The extracts, at low concentration (5 µ g/mL), exhibited a large variation in inhibitory effects of H. pylori-induced oxidative burst ranging from 48% inhibition to inactive, but all extracts, excluding Byrsonima intermedia, had inhibitory activity over 80% at the concentration of 100 µ g/mL. The total suppressive antioxidant capacity measured as the effective concentration, which represents the extract concentration producing 50% inhibition of the chemiluminescence induced by H. pylori, varies from 27.2 to 56.8 µ g/mL and was in the following order: Qualea parviflora > Qualea multiflora > Alchornea triplinervia > Qualea grandiflora > Anacardium humile > Davilla elliptica > Mouriri pusa > Byrsonima basiloba > Alchornea glandulosa > Byrsonima intermedia. The main groups of compounds in tested extracts are presented. Differences in the phytochemical profile, quantitatively and qualitatively, of these plants can explain and justify their protective effect on the gastric mucosa caused by the neutrophil-generated ROS that occurs when H. pylori displays its evasion mechanisms.