Antinociceptive and anti-inflammatory potential, and chemical characterization of the dichloromethane fraction of Solanum lycocarpum (Solanaceae) ripe fruits by LC-DAD-MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Solanum lycocarpum A. St. Hil. (Solanaceae) is a species from the Brazilian Cerrado, exhibiting several medicinal properties, being used by the population in the treatment of ulcers, bronchitis, asthma and hepatitis, which involve inflammatory processes. AIM OF THIS STUDY: This study aimed to chemically characterize the dichloromethane fraction (DCM), as well as verify its antinociceptive, anti-inflammatory and antioxidant potential. MATERIALS AND METHODS: The DCM fraction was obtained by partitioning the ethanol extract. The chemical constituents of the DCM fraction were characterized by LC-DAD-MS. The DPPH and FRAP assays were used to evaluate the antioxidant potential. The carrageenan-induced paw edema model was used to assess the anti-inflammatory effects, and the inflammatory infiltrate was evaluated by qualitative and quantitative histological analyses. The antinociceptive action of the DCM fraction was evaluated by acetic acid-induced abdominal writhing test, formalin-induced nociception and hot-plate test. RESULTS: Steroidal alkaloids solasonine, solasodine and solamargine, as well as the alkaloid peiminine/imperialine and caffeoylquinic acids, were annotated in DCM fraction by LC-DAD-MS. The DCM fraction showed antioxidative action in the in vitro DPPH and FRAP tests, as well as an anti-inflammatory effect for the three evaluated doses of 30, 100 and 300 mg/kg in the fourth and sixth hours after the administration of carrageenan. The histological analyses evidenced considerably reduction in leukocyte migration and the number of polymorphonuclear leukocytes. The study also demonstrated antinociceptive activity for the DCM fraction, which reduced abdominal writhing at three concentrations evaluated, as well as a decrease in paw licking in the formalin-induced nociception test both in the neurogenic phase and the inflammatory phase, with greater effectiveness compared to the anti-inflammatory indomethacin. The DCM fraction also increased the latency time of the animals in the hot plate test 60 min after treatment, although it did not seem to involve the opioidergic system. CONCLUSION: This work evidenced that the dichloromethane fraction of S. lycocarpum fruit possesses antinociceptive and anti-inflammatory potential, which supports its use in folk medicine for management inflammatory conditions.

The Cardiorenal Effects of Piper amalago Are Mediated by the Nitric Oxide/Cyclic Guanosine Monophosphate Pathway and the Voltage-Dependent Potassium Channels.

Piper amalago L. is used in Brazilian traditional medicine to treat inflammation, chest pain, and anxiety. This study aimed to investigate the safety and the renal and cardiovascular effects of the volatile oil (VO) and the aqueous (AE) and hydroalcoholic (HE) extracts from P. amalago. The gas chromatography-mass spectrometry analyses identified 47 compounds in the VO, with ß-cyclogermacrene, spathulenol, ß-phellandrene, and α-pinene standing out. Among the 47 compounds also found in AE and HE by liquid chromatography-mass spectrometry, glycosylated flavones, organic acids, amino acids, and amides were highlighted. Some examples of these compounds are methoxy-methylenedioxy cis-cinnamoyl pyrrolidine, methoxy-methylenedioxy trans-cinnamoyl pyrrolidine, and cyclobutene-2,4-bis-(1,3-benzodioxol-5-methoxy-6-yl)-1,3-dicarboxapyrrolidide. The acute toxicity experiments were conducted on female rats (n = 5). The cardiorenal assays (n = 8) and evaluations of vasodilatory effects on the mesenteric vascular bed (n = 5) were conducted on male rats. In either extract or VO, there were no mortality or changes in relative weights or histopathological analysis of the organs. Urinary volume and renal electrolyte excretion were elevated significantly during repeated dose 7-day treatment with different preparations from P. amalago. None of the preparations induced hypotension or changes in cardiac electrical activity. Only HE promoted significant vasodilatory effects in rats' isolated mesenteric vascular beds. These effects were completely abolished in the presence of L-NAME plus 4-aminopyridine. Therefore, P. amalago leaves are safe and present diuretic activity after acute and repeated dose administration over 7 days. Moreover, the HE induced significant vasodilator response in rats' mesenteric vascular beds by NO/cGMP pathway and voltage-dependent K+ channels activation.

Spondias purpurea L. Bark Extract Protects against Oxidative Stress and Reduces Hypercholesterolemia in Mice Fed High-Fat Diet.

Oxidative stress plays a key role in the initiation and progression of metabolic diseases, including obesity. Preventing the accumulation of reactive oxygen species and oxidative damage to macromolecules is a beneficial strategy for reducing comorbidities associated with obesity. Fruits from the Spondias genus are known for their antioxidant activity, but they are not available year-round due to their seasonality. In this context, we investigated the antioxidant activity and identified the chemical constituents of the aqueous extract of the stem bark of Spondias purpurea L. (EBSp). Additionally, we evaluated the effect of EBSp consumption on metabolic parameters in mice with obesity induced by a high-fat diet. Chemical analyses revealed 19 annotated compounds from EBSp, including flavan-3-ols, proanthocyanidins, methoxylated coumarin, and gallic and ellagic acids, besides other phenolic compounds. In vitro, EBSp showed antioxidant activity through the scavenging of the free radicals and the protection of macromolecules against oxidative damage. Cellular assays revealed that EBSp reduced the levels of malondialdehyde produced by erythrocytes exposed to the oxidizing agent AAPH. Flow cytometry studies showed that EBSp reduced reactive oxygen species levels in human peripheral blood mononuclear cells treated with hydrogen peroxide. Obese mice treated with EBSp (400 mg.kg-1) for 60 days showed reduced levels of malondialdehyde in the heart, liver, kidneys, and nervous system. The total cholesterol levels in mice treated with EBSp reached levels similar to those after treatment with the drug simvastatin. Together, the results show that the combination of the different phenolic compounds in S. purpurea L. bark promotes antioxidant effects in vitro and in vivo, resulting in cytoprotection in the context of oxidative stress associated with obesity and a reduction in hypercholesterolemia. From a clinical perspective, the reduction in oxidative stress in obese individuals contributes to the reduction in the emergence of comorbidities associated with this metabolic syndrome.

Baccharis dracunculifolia (Asteraceae) essential oil toxicity to Culex quinquefasciatus (Culicidae).

The control of mosquitoes by means of chemical insecticides has been a problem, mainly due to the possibility of resistance developed by insects to xenobiotics. For this reason, demand for botanical insecticides has increased. In this sense, the present work aims to verify the susceptibility and morphological and biochemical alterations of Culex quinquefasciatus larvae after exposure to essential oil (EO) of leaves of Baccharis dracunculifolia. To observe the larvicidal action, larvae were exposed to EO at concentrations of 25, 50, 100, and 200 mg/L, until their emergence to adults. The control group was exposed to deionized water and dimethyl sulfoxide. Morphological analyses were also carried out using hematoxylin and eosin, mercury bromophenol blue, Nile blue, and periodic acid Schiff. Biochemical analyses of total glucose, triacylglyceride (TAG), protein, and acetylcholinesterase levels were performed. The phytochemical analysis of the EO showed (E)-nerolidol as the major compound (30.62%). Larvae susceptibility results showed a LC50 of 34.45 mg/L for EO. Morphological analysis showed that there were histological changes in midgut. For biochemical analyses, the glucose level in the larvae exposed to EO for 24 h decreased significantly, unlike the TAG levels, which increased. The total protein level of the larvae also increased after exposure for 24 h, and acetylcholinesterase levels decreased significantly. Taking all our data into account, we can conclude that EO causes destabilization in larva, leading to histological changes, metabolic deregulation and, consequently, their death.

The role of tannins as antiulcer agents: a fluorescence-imaging based study

Abstract Condensed tannins have been used for many years in folk medicine to treat gastric problems. The mechanism of action that explains why tannins improve gastritis symptoms is based on their ability to chelate metals, antioxidant activity, and their complexation power with other molecules. Even though these uses are well-known, the requirements to become an herbal medicine are much more complex. Herein, we analyzed Stryphnodendron rotundifolium Mart., Fabaceae, extract using MALDI for tannin characterization and carried out a fluorescence-imaging study to prove the gastroprotective effects of tannins as coating agents. Through these methods we show that condensed tannins form a gastroprotective layer. Moreover, we revise and discuss other possible mechanisms of action for phenolic-rich plant extracts and their potential in the development of herbal medicines to treat ulcers and gastritis.

Leaf and Root Extracts from Campomanesia adamantium (Myrtaceae) Promote Apoptotic Death of Leukemic Cells via Activation of Intracellular Calcium and Caspase-3.

Phytochemical studies are seeking new alternatives to prevent or treat cancer, including different types of leukemias. Campomanesia adamantium, commonly known as guavira or guabiroba, exhibits pharmacological properties including antioxidant, antimicrobial, and antiproliferative activities. Considering the anticancer potential of this plant species, the aim of this study was to evaluate the antileukemic activity and the chemical composition of aqueous extracts from the leaves (AECL) and roots (AECR) of C. adamantium and their possible mechanisms of action. The extracts were analyzed by LC-DAD-MS, and their constituents were identified based on the UV, MS, and MS/MS data. The AECL and AECR showed different chemical compositions, which were identified as main compounds glycosylated flavonols from AECL and ellagic acid and their derivatives from AECR. The cytotoxicity promoted by these extracts were evaluated using human peripheral blood mononuclear cells and Jurkat leukemic cell line. The cell death profile was evaluated using annexin-V-FITC and propidium iodide labeling. Changes in the mitochondrial membrane potential, the activity of caspases, and intracellular calcium levels were assessed. The cell cycle profile was evaluated using propidium iodide. Both extracts caused concentration-dependent cytotoxicity only in Jurkat cells via late apoptosis. This activity was associated with loss of the mitochondrial membrane potential, activation of caspases-9 and -3, changes in intracellular calcium levels, and cell cycle arrest in S-phase. Therefore, the antileukemic activity of the AECL and AECR is mediated by mitochondrial dysfunction and intracellular messengers, which activate the intrinsic apoptotic pathway. Hence, aqueous extracts of the leaves and roots of C. adamantium show therapeutic potential for use in the prevention and treatment of diseases associated the proliferation of tumor cell.

New cascarosides from Rhamnus purshiana and fragmentation studies of the class by ion trap mass spectrometry.

RATIONALE: Anthrone and oxanthrone are important anthraquinone derivatives present in medicinal plants which are used in therapeutics as laxatives. Some of these plants need to be stored at least one year before they can be used in order to oxidize anthrones into oxanthrones, so to avoid severe diarrhea and dehydration. Therefore, this work aimed to characterize fragmentation reactions between these anthraquinones to provide an easy way to differentiate between the two classes, since it is necessary and important to discriminate and identify these derivatives in laxative plants and phytotherapic drugs. METHODS: Anthrone (cascarosides A-D) and oxanthrone (10-hydroxycascaroside A and B) derivatives were isolated and identified by NMR (1 H, 13 C, DEPT, NOESY) and used for fragmentation study by direct infusion on an electrospray ionization (ESI) ion trap mass spectrometer (AmazonSL, Bruker) in positive and negative mode. RESULTS: The additional hydroxyl at C-10 in oxanthrones allowed McLafferty-type rearrangements to form the quinone group in positive mode, while in negative mode the second sugar loss infringed the odd-electron rule and formed a radical fragment. No differences in fragmentation reactions were found between diastereoisomeric pairs, although the additional oxygen at C-10 of oxanthrones allowed a different fragmentation pattern. CONCLUSIONS: The proposed fragmentation patterns can be used to differentiate anthrones from oxanthrones in both ion modes. In addition, they can be applied to differentiate these compounds in anthraquinone-rich plants and phytotherapic drugs. Finally, herein, the strategy applied allowed us to identify new natural products. Copyright © 2017 John Wiley & Sons, Ltd.

Apigenin-7-O-glucoside oxidation catalyzed by P450-bioinspired systems.

Apigenin-7-O-glucoside (A7G) is the main flavonoid of Bidens gardneri Bak., a Brazilian plant with wide application in folk medicine. Despite the popular use of this plant, its biological effects are not completely known. This work tested the 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin iron(III) and manganese(III) chloride (Fe(TFPP)Cl and Mn(TFPP)Cl), and Jacobsen's catalyst as P450-bioinspired catalysts for A7G oxidation by different oxidants (PhIO, H2O2, m-CPBA, and t-BuOOH). Up to nine different products were detected by HPLC analysis; Reactions with metalloporphyrin/PhIO systems afforded high catalytic conversions (58-89%). In spite of providing smaller product yields, the metalloporphyrin/H2O2 systems led to superior product distribution. Fe(TFPP)Cl yielded the highest A7G conversion rates (79-93%) with the four different oxidants tested herein. In the presence of PhIO, the oxidative profile of the manganese catalysts was very close to the oxidative profile of Fe(TFPP)Cl. However, in medium containing peroxide, the reactivity of the manganese catalysts was lower as compared to the reactivity of Fe(TFPP)Cl. Reactions with Fe(TFPP)Cl/oxidant systems were analyzed by UPLC-MS; up to thirteen compounds were detected. A7G oxidation catalyzed by Fe(TFPP)Cl yielded seven compounds. Three other compounds had m/z profile compatible with the profile of the A7G metabolites. The A7G oxidation assays performed in the presence of P450-bioinspired catalysts demonstrated their great catalytic potential toward A7G. The present results may be useful to many areas of knowledge and to the research and development of numerous chemical and phamarcological processes, especially in terms of drug design, biological assays, and applications in medicinal chemistry.

Antibacterial activity of Limonium brasiliense (Baicuru) against multidrug-resistant bacteria using a statistical mixture design.

ETHNOPHARMACOLOGICAL RELEVANCE: Limonium brasiliense (Boiss.) Kuntze (Plumbaginaceae) is commonly known as "baicuru" or "guaicuru" and preparations of its dried rhizomes have been popularly used in the treatment of premenstrual syndrome and menstrual disorder, and as an antiseptic in genito-urinary infections. This study evaluated the potential antibacterial activity of rhizome extracts against multidrug-resistant bacterial strains using statistical mixture design. MATERIALS AND METHODS: The statistical design of four components (water, methanol, acetone, and ethanol) produced 15 different extracts and also a confirmatory experiment, which was performed using water:acetone (3:7, v/v). The crude extracts and their ethyl-acetate fractions were tested against vancomycin-resistant Enterococcus faecium (VREfm), methicillin-resistant Staphylococcus aureus (MRSA) and Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae, all of which have been implicated in hospital and community-acquired infections. The dry residue, total polyphenol, gallocatechin and epigallocatechin contents of the extracts were also tested and statistical analysis was applied in order to define the fit models to predict the result of each parameter for any mixture of components. The principal component and hierarchical clustering analyses (PCA and HCA) of chromatographic data, as well as mass spectrometry (MS) analysis were performanced to determine the main compounds present in the extracts. RESULTS: The Gram-positive bacteria were susceptible to inhibition of bacterial growth, in special the ethyl-acetate fraction of ternary extracts from water:acetone:ethanol and methanol:acetone:ethanol against, respectively, VREfm (MIC=19µg/mL) and MRSA (MIC=39µg/mL). On the other hand, moderate activity of the ethyl-acetate fractions from primary (except water), secondary and ternary extracts (MIC=625µg/mL) was noted against KPC. The quadratic and special cubic models were significant for polyphenols and gallocatechin contents, respectively. Fit models to dry residue and epigallocatechin contents were not possible. PCA and HCA of the chromatographic fingerprints were disturbed by displacement retention time of some peaks, but the ultraviolet spectra indicated the homogeneous presence of flavan-3-ols characteristic of tannins. The MS confirmed the presence of gallic acid, gallocatechin, and epigallocatechin in extracts, and suggested the presence of monomers and dimers of B- and A-type prodelphinidins gallate, as well as a methyl gallate. CONCLUSION: Our results showed the antibacterial potential of L. brasiliense extracts against multidrug-resistant Gram-positive bacteria, such as VREfm and MRSA. The statistical design was a important tool to evaluate the biological activity by optimized form. The presence of some phenolic compounds was also demonstrated in extracts.

Simultaneous Characterization of Intravenous and Oral Pharmacokinetics of Lychnopholide in Rats by Transit Compartment Model.

The pharmacokinetic properties of a new molecular entity are important aspects in evaluating the viability of the compound as a pharmacological agent. The sesquiterpene lactone lychnopholide exhibits important biological activities. The objective of this study was to characterize the pharmacokinetics of lychnopholide after intravenous administration of 1.65 mg/kg (n = 5) and oral administration of 3.3 mg/kg (n = 3) lychnopholide in rats (0.2 ± 0.02 kg in weight) through nonlinear mixed effects modeling and non-compartmental pharmacokinetic analysis. A highly sensitive analytical method was used to quantify the plasma lychnopholide concentrations in rats. Plasma protein binding of this compound was over 99 % as determined by a filtration method. A two-compartment body model plus three transit compartments to characterize the absorption process best described the disposition of lychnopholide after both routes of administration. The oral bioavailability was approximately 68 %. The clearance was 0.131 l/min and intercompartmental clearance was 0.171 l/min; steady-state volume of distribution was 4.83 l. The mean transit time for the absorption process was 9.15 minutes. No flip-flop phenomenon was observed after oral administration. The pharmacokinetic properties are favorable for further development of lychnopholide as a potential oral pharmacological agent.

Natural Green coating inhibits adhesion of clinically important bacteria.

Despite many advances, biomaterial-associated infections continue to be a major clinical problem. In order to minimize bacterial adhesion, material surface modifications are currently being investigated and natural products possess large potential for the design of innovative surface coatings. We report the bioguided phytochemical investigation of Pityrocarpa moniliformis and the characterization of tannins by mass spectrometry. It was demonstrated that B-type linked proanthocyanidins-coated surfaces, here termed Green coatings, reduced Gram-positive bacterial adhesion and supported mammalian cell spreading. The proposed mechanism of bacterial attachment inhibition is based on electrostatic repulsion, high hydrophilicity and the steric hindrance provided by the coating that blocks bacterium-substratum interactions. This work shows the applicability of a prototype Green-coated surface that aims to promote necessary mammalian tissue compatibility, while reducing bacterial colonization.

Leishmanicidal evaluation of tetrahydroprotoberberine and spirocyclic erythrina-alkaloids.

Leishmaniasis is one of the World's most problematic diseases in developing countries. Traditional medicines to treat leishmaniasis have serious side effects, as well as significant parasite resistance problems. In this work, two alkaloids 1 and 2 were obtained from Corydalis govaniana Wall and seven alkaloids 3-9, were obtained from Erythrina verna. The structures of the compounds were confirmed by mass spectrometry and 1D- and 2D-NMR spectroscopy. The leishmanicidal activity of compounds 1-9 against Leishmania amazonensis was tested on promastigote forms and cytotoxicity against J774 (macrophage cell line) was assessed in vitro. Compound 1 showed potent activity (IC50 = 0.18 µg/mL), compared with the standard amphotericin B (IC50 = 0.20 µg/mL). The spirocyclic erythrina-alkaloids showed lower leishmanicidal activity than dibenzoquinolizine type alkaloids.

Variação química no óleo essencial das folhas de seis indivíduos de Duguetia furfuracea (Annonaceae) / Chemical variability of the essential oil of the leaves from six individuals of Duguetia furfuracea (Annonaceae)

Duguetia furfuracea, Annonaceae, uma invasora de pastagens, é também usada na medicina tradicional em diversos estados no Brasil. Devido à significante diferença de odor entre dois grupos de indivíduos desta planta em um remanescente de cerrado em Campo Grande, Mato Grosso do Sul, seis amostras foram selecionadas de acordo com a intensidade de odor em suas folhas: In-01, In-03 e In-05: odor pronunciado (Perfil A); In-02, In-04 e In-06: odor fraco ou ausência de odor (Perfil B). As folhas foram coletadas e submetidas a hidrodestilação para obtenção dos óleos essenciais, os quais foram analisados por CG/EM. Uma boa compatibilidade foi encontrada entre os resultados obtidos por CG/EM e a análise olfativa adotada para as amostras: espécimes odor pronunciado apresentaram uma alta porcentagem de monoterpenos e alguns sesquiterpenos: In-01 (β-felandreno, 42,2 por cento; mirceno, 6,8 por cento; α-felandreno, 4,6 por cento); In-03 (terpin-4-ol, 21,6 por cento; sabineno, 17,3 por cento; p-cimeno, 5,6 por cento); In-05 (sabineno, 25,1 por cento; terpin-4-ol, 16,2 por cento; p-cimeno, 8,3 por cento). Apenas sesquiterpenos foram encontrados nos espécimes que apresentaram odor fraco ou ausência do mesmo (com constituinte majoritário biciclogermacreno: 21,4 por cento, 24,0 por cento, and 29,1 por cento, respectivamente, para os In-02, In-04 e In-06).

Duguetia furfuracea, Annonaceae, a pasture weed, is also used in folk medicine in several Brazilian states. Because of the significant difference in odor between two groups of this plant in a remnant patch of savanna in Campo Grande county, Mato Grosso do Sul, six samples were selected according to the odor intensity in the leaves (In-01, In-03, and In-05: pronounced odor; In-02, In-04, and In-06: weak odor or its absence). The leaves were collected and subjected to steam distillation for extraction of essential oils, which were analyzed by GC-MS. A good agreement was found between CG/MS results and olfactory evaluation of the samples: specimens exhibiting leaf scent had a high percentage of monoterpenes and some sesquiterpenes, as follows: In-01 (β-phellandrene, 42.2 percent; myrcene, 6.8 percent; α-phellandrene, 4.6 percent); In-03 (terpin-4-ol, 21.6 percent; sabinene, 17.3 percent; p-cymene, 5.6 percent); In-05 (sabinene, 25.1 percent; terpin-4-ol, 16.2 percent; p-cymene, 8.3 percent). Only sesquiterpenes were found in the specimens having weakly scented or scentless leaves (main constituent bicyclogermacrene: 21.4 percent, 24.0 percent, and 29.1 percent, respectively, for In-02, In-04 and In-06).