Brazilian Folk Medicine Research Articles

New Polyhydroxylated Steroidal Saponins from Solanum paniculatum L. Leaf Alcohol Tincture with Antibacterial Activity against Oral Pathogens.

Solanum paniculatum L. is widely used in Brazilian folk medicine for the treatment of liver and gastrointestinal disorders as well as for culinary purposes and beverage production. Fractionation of hydroalcoholic [ethanol (EtOH) 70%] tincture from S. paniculatum leaves led to the isolation of six new spirostanic saponins which included 6- O-α-l-rhamnopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 R,25 S)-3β,6α,23-trihydroxy-5α-spirostane (1), 6- O-β-d-xylopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 R,25 R)-3β,6α,23-trihydroxy-5α-spirostane (4), 3- O-α-l-rhamnopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 S,25 R)-3β,6α,23-trihydroxy-5α-spirostane (5), 3- O-β-d-xylopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 S,25 R)-3β,6α,23-trihydroxy-5α-spirostane (6), 6- O-α-l-rhamnopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,25 S)-1β,3β,6α-trihydroxy-5α-spirostane (7), and 6- O-β-d-xylopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,25 S)-3β,4β,6α-trihydroxy-5α-spirostane (8) together with two known spirostanic saponins (2, 3). The structures of these compounds were determined by one-dimensional (1D) and two-dimensional (2D) NMR experiments in addition to high-resolution electrospray ionization mass spectrometry (HRESIMS) analyses. The 70% alcohol tincture, used as phytomedicine, exhibited promising activities against oral pathogens, including, Steptococcus sanguinis, St. oralis, St. mutans, St. mitis, and Lactobacillus casei with minimal inhibitory concentration (MIC) values ranging from 6.25 to 50 μg/mL. The saponin fraction, nonetheless, showed lower activity against all the strains tested (from 100 to >400 μg/mL).

DNA Barcoding for the Identification of Phyllanthus Taxa Used Medicinally in Brazil.

Plants of the genus Phyllanthus, principally Phyllanthus amarus, Phyllanthus urinaria, Phyllanthus niruri, and Phyllanthus tenellus, are used in Brazilian folk medicine to treat kidney stones as well as other ailments, where the latter two species are listed in the Brazilian Pharmacopeia as quebra-pedra (stone-breaker). However, only P. niruri has been shown to be effective in a clinical setting. Nuclear ribosomal internal transcribed spacer (ITS1 - 5.8S rRNA-ITS2), internal transcribed spacer 2, and chloroplasts rbcL, matK, psbA-trnH, trnL, and trnL-trnF were screened for their potential as DNA barcodes for the identification of 48 Phyllanthus taxa in Brazilian medicinal plant germplasm banks and in "living pharmacies". The markers were also tested for their ability to validate four commercial herbal teas labelled as quebra-pedra. Using the criterion of high clade posterior probability in Bayesian phylogenetic analysis, the internal transcribed spacer, internal transcribed spacer 2, and chloroplast matK, psbA-trnH, trnL, and trnL-trnF markers all reliably differentiated the four Phyllanthus species, with the internal transcribed spacer and matK possessing the additional advantage that the genus is well represented for these markers in the Genbank database. However, in the case of rbcL, posterior probability for some clades was low and while P. amarus and P. tenellus formed monophyletic groups, P. niruri and P. urinaria accessions could not be reliably distinguished with this marker. Packaged dried quebra-pedra herb from three Brazilian commercial suppliers comprised P. tenellus, but one sample was also found to be mixed with alfalfa (Medicago sativa). An herb marketed as quebra-pedra from a fourth supplier was found to be composed of a mixture of Desmodium barbatum and P. niruri.

In vitro screening of cytotoxic activity of euphol from Euphorbia tirucalli on a large panel of human cancer-derived cell lines.

A large number of classic antineoplastic agents are derived from plants. Euphorbia tirucalli L. (Euphorbiaceae) is a subtropical and tropical plant, used in Brazilian folk medicine against many diseases, including cancer, yet little is known about its true anticancer properties. The present study evaluated the antitumor effect of the tetracyclic triterpene alcohol, euphol, the main constituent of E. tirucalli in a panel of 73 human cancer lines from 15 tumor types. The biological effect of euphol in pancreatic cells was also assessed. The combination index was further used to explore euphol interactions with standard drugs. Euphol showed a cytotoxicity effect against several cancer cell lines (IC50 range, 1.41–38.89 µM), particularly in esophageal squamous cell (11.08 µM) and pancreatic carcinoma cells (6.84 µM), followed by prostate, melanoma, and colon cancer. Cytotoxicity effects were seen in all cancer cell lines, with more than half deemed highly sensitive. Euphol inhibited proliferation, motility and colony formation in pancreatic cancer cells. Importantly, euphol exhibited synergistic interactions with gemcitabine and paclitaxel in pancreatic and esophageal cell lines, respectively. To the best of our knowledge, this study constitutes the largest in vitro screening of euphol efficacy on cancer cell lines and revealed its in vitro anti-cancer properties, particularly in pancreatic and esophageal cell lines, suggesting that euphol, either as a single agent or in combination with conventional chemotherapy, is a potential anti-cancer drug.

English

Croton lechleri is a native species from the Amazon and used with relative frequency in folk medicine in Brazil and other countries. Diversity and antibacterial activity of endophytic fungi associated with this plant were studied here. Samples of leaves and stems were used and 575 endophytic fungi were isolated (307 from leaves and 268 from stems), comprising 284 morphotypes distributed in 13 genera and unknown. The most frequently isolated genera were Phomopsis (30.78%), Penicillium (21.57%) and Pestalotiopsis (16.70%). Diversity and richness of species were higher in leaf tissues. Fifty-five fungi presented antibacterial activity. The fungi with the highest activity were Phomopsis (6.34%), Penicillium (3.17%), and those unknown (5.28%). Penicillium sp. 9 showed the highest antibacterial activity against Enterococcus faecalis and Phomopsis sp. 8 and Phomopsis sp. 9 against Streptococcus pneumoniae and Staphylococcus aureus. Curvularia sp. 1 and a fungus that could not be identified (Unknown sp. 9), showed the highest antibacterial activity against Klebsiella pneumoniae and Escherichia coli, respectively. Only two fungi (Penicillium sp. 9 and Curvularia sp. 1) inhibited the five tested bacteria. Endophytic fungi of C. lechleri harbor a great diversity of endophytic fungi, which have the potential for producing antibacterial compounds. Key words: Dragon’s blood, antibacterial agent, endophytic fungi, microbial interaction.

Spasmolytic and antidiarrheal activities of Lippia thymoides (Verbenaceae) essential oil

Lippia thymoides (‘alecrim-do-mato’ or ‘alecrim-do-campo’) is used in Brazilian folk medicine to treat various illnesses, including diarrhea. This work aimed to evaluate in vitro spasmolytic and in vivo antidiarrheal activities of the L. thymoides essential oil (OOS) and to correlate with the traditional use of this plant. In isolated guinea-pig ileum, OOS presented a concentration-dependent spasmolytic activity in preparations pre-contracted with KCl 40 mM [EC50 = 16.89 (11.56–24.66) μg/mL], and antagonized phasic contractions induced by 1 μM carbachol [IC50 = 42.71 (37.35–48.83) μg/mL] or histamine [IC50 = 32.38 (27.44–38.20) μg/mL]. In mice, OOS at 400 mg/kg reduced intestinal transit, at 200 and 400 mg/kg reduced total stool mass and at 400 mg/kg reduced intestinal fluid accumulation. It was shown that the antidiarrheal effect of OOS is related to the inhibition of smooth muscle contraction and may be due to the presence of major compound β-caryophyllene in this essential oil.

Incorporation of Baccharis dracunculifolia DC (Asteraceae) leaf extract into phosphatidylcholine-cholesterol liposomes improves its anti-inflammatory effect in vivo

The aerial parts of Baccharis dracunculifolia (BdE) is used in the Brazilian folk medicine to treat inflammatory conditions. Here we examined the ability of free and liposomal BdE to modulate reactive oxygen species generation in human neutrophils in vitro and zymosan-induced acute joint inflammation in Wistar rats. We prepared biocompatible liposomes of soya phosphatidylcholine and cholesterol with low diameter, homogeneous size distribution, and neutral surface charge. Free BdE decreased joint swelling, total leucocyte and neutrophil infiltration, and the synovial levels of tumour necrosis factor-α and interleukins 6 and 1β. Incorporation of BdE into liposomes preserved its capacity to inhibit the neutrophil superoxide anion and total reactive oxygen species generation, and improved its anti-inflammatory effect in vivo by decreasing the effective BdE dose by nearly sixfold. The same liposome type lowered the effective dose of caffeic acid by nearly sixteenfold. Therefore, incorporation of BdE into phosphatidylcholine-cholesterol liposomes improves its anti-inflammatory effect.

Lafoensia pacari A. St.-Hil.: Wound healing activity and mechanism of action of standardized hydroethanolic leaves extract

Ethnopharmacological importanceLafoensia pacari A. St.-Hil., (Lythraceae) is a native tree of Brazilian Cerrado and commonly known in Brazil as "mangava-brava". Its leaves are used in Brazilian folk medicine in wound healing, cutaneous mycoses, and in the treatment of gastritis and ulcers. Aim of the studyThe present study was designed to evaluate the wound healing activity and mechanism of action of the hydroethanolic extract of Lafoensia pacari A. St.-Hil. leaves (HELp), and to advance in its chemical profiling. Materials and methodsHELp was prepared by maceration in 70% hydroethanolic solution (1:10, w/v). The phytochemical analyses were investigated using colorimetry and electrospray ionization/mass spectrometric detection (ESI-MSn). Its in vitro cytotoxicity was evaluated in CHO-K1 and L929 cells, while the in vivo acute toxicity was performed in mice. The potential in vivo wound healing activity was assessed using excision and incision rat models and histopathology of the wounded skin (excision model) was carried out. The in vitro wound healing activity of HELp was demonstrated by scratch assay in L-929 cells, by measuring proliferation/migration rate and p-ERK 1/2 protein expression using western blot analysis. HELp's in vivo anti-inflammatory activity was evaluated by lipopolysaccharide (LPS) induced peritonitis in mice, along with the determination of nitric oxide (NO) and cytokines (TNF-α and IL-10) in the peritoneal lavages. Its potential in vitro antibacterial activity was performed using microbroth dilution assay, while in vitro antioxidant activities was by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and ferric reducing antioxidant power (FRAP) assays. ResultsThe phytochemical analysis of HELp revealed the presence of polyphenols with ellagic acid, punicalagin, punicalin, kaempferol, quercetin-3-O-xylopyranoside and quercetin-3-O-rhamnopyranoside being the most prominent. HELp showed no toxicity on CHO-k1 and L929 cell lines. Topical treatment with HELp (10 and 30 mg/g of gel) presented increased rates of wound contraction at all the days evaluated with complete wound re-epithelialization at 22.0 ± 1.5 (p < 0.05) and 21.7 ± 1.6 (p < 0.01) days, respectively. Topical application of HELp (10, 30 or 100 mg/g of gel) in incised wounds caused an increase in tensile break strength at all concentrations resulting in moderate re-epithelialization and neovascularization, increased cell proliferation an accelerated remodeling phase of the wound, in a manner comparable to standard drug (Madecassol®, 10 mg/g). In the scratch assay with L929 cells, HELp (0.1 and 0.03 mg/mL) and PDGF (5 ng/mL) resulted in the increased proliferation/migration rate of fibroblasts and higher expression of p-ERK 1/2 protein. In LPS-induced peritonitis, HELp (100 and 200 mg/kg p.o.) decreased total leukocyte migration, comparable to the dexamethasone (0.5 mg/kg p.o.). In RAW 264.7 macrophages activated by LPS, HELp produced anti-inflammatory activity dependent on increased concentrations of IL-10, reduction in NO production, without altering the TNF-α levels. HELp also presented potent antioxidant activity in the DPPH and FRAP, but lacks in vitro antibacterial activity. ConclusionThe present study results support the popular use of the leaves of L. pacari in the treatment of wounds. Its wound healing activity is multi-targeted and involves inhibition of the proliferative and anti-inflammatory phases, antioxidant and positive modulation of the remodeling phase that might be involved different secondary metabolites, with emphasis on the ellagic acid, punicalagin, punicalin, kaempferol, quercetin-3-O-xylopyranoside and quercetin-3-O-rhamnopyranoside.

Hepatoprotective Effect of Essential Oils fromHyptis crenatain Sepsis-Induced Liver Dysfunction

No specific therapeutics are available for the treatment of sepsis-induced liver dysfunction, a clinical complication strongly associated with the high mortality rate of septic patients. This study investigated the effect of the essential oil of Hyptis crenata (EOHc), a lamiaceae plant used to treat liver disturbances in Brazilian folk medicine, on liver function during early sepsis. Sepsis was induced by the cecal ligation and puncture (CLP) model. Rats were divided into four groups: Sham, Sham+EOHc, CLP, and CLP+EOHc. EOHc (300 mg/kg) was orally administered 12 and 24 h after surgery. The animals were sacrificed for blood collection and liver tissue samples 48 h after surgery. Hepatic function was evaluated by measuring serum bilirubin, alkaline phosphatase (ALP), aspartate aminotransferase, and alanine aminotransferase (ALT) levels. The levels of malondialdehyde and the activity of superoxide dismutase, catalase, and GSH peroxidase (GSH-Px) were measured for assessment of oxidative stress. Liver morphology was analyzed by hematoxylin and eosin staining. EOHc normalized serum ALP, ALT, and bilirubin levels and inhibited morphological changes. In addition, we observed that EOHc inhibited elevation in hepatic lipid peroxidation and reduction of the glutathione peroxidase activity induced by sepsis. Our data show that EOHc plays a protective effect against liver injury induced by sepsis.

Effectiveness of aqueous and hydroalcoholic extracts of Acanthospermum australe (Loefl.) Kuntze against diarrhea-inducing bacteria.

Leaves and roots of Acanthospermum australe (Asteraceae) have been used in Brazilian folk medicine for the treatment of various ailments including diarrhea, skin diseases, blennorrhagia, dyspepsia, parasitic worms and malaria. The aim of study was to characterize the chemical profiles of the aqueous and hydroalcoholic extracts of leaves and roots of A. australe, and to evaluate their antimicrobial activities against diarrhea-inducing bacteria (Enterococcus faecalis, Shigella dysenteriae and Yersinia enterocolitica), as well as their cytotoxic properties. Aqueous leaf extracts were obtained by infusion, while aqueous root extracts were obtained by decoction. The hydroalcoholic leaf and root extracts were prepared by maceration in 90% ethanol for 3 days. Antimicrobial activity was assessed using standard techniques and cytotoxicity was evaluated using Chinese hamster ovary cells CHO-K1. Chemical analysis revealed the presence of tannins, flavonoids, saponins and phenolic compounds in the extracts. Although root extracts were not effective against E. faecalis, leaf extracts at concentrations of 20 mg/mL exhibited bactericidal activities against this microorganism. The hydroalcoholic root extract was unique in presenting a bactericidal effect against S. dysenteriae. None of the extracts showed bacteriostatic or bactericidal activities against Y. enterocolitica. The results presented herein demonstrate that the Gram-positive E. faecalis and the Gram-negative S. dysenteriae were susceptible to A. australe extracts, although bacteriostatic/bactericidal activities were only observed at concentrations considered too high for clinical application. Our results support the ethnopharmacological use of A. australe in the treatment of gastrointestinal disorders, particularly diarrhea caused by infectious bacteria, although further studies are required to determine the anti-diarrhea effects and the toxicities of the extracts in vivo.

Hypotensive, vasorelaxant and antihypertensive activities of the hexane extract of Anacardium occidentale Linn

Anacardium occidentale is widely used in folk medicine in Brazil, India, and Africa to treat inflammation, diabetes and hypertension. The aim of the study was to assess the hypotensive, vasorelaxant and antihypertensive effects produced by the hexane extract from the leaves of A. occidentale (HEAO) in rats. The animals were anesthetized, and polyethylene catheters were embedded into the lower abdominal aorta and the inferior vena cava for blood pressure measurements and drug administration. The HEAO was injected and the mean arterial pressure and heart rate were measured. The effects of the HEAO on isolated rat mesenteric rings with and without endothelium were investigated. HEAO induced hypotension and bradycardia in normotensive rats. After pretreatment with L-NAME, indomethacin, atropine or hexamethonium, these effects were attenuated. In mesenteric rings, HEAO antagonized the contractile effects induced by phenylephrine or KCl. This effect was inhibited after removal of the vascular endothelium. Oral administration of the HEAO decreased significant mean arterial pressure in spontaneously hypertensive rats. Mass spectrometric analysis of HEAO identified malic acid and quercetin. Our findings demonstrate that HEAO produces hypotension probably due to a reduction in peripheral vascular resistance, mediated by the endothelium, in part, by a decrease in cardiac output. Bradycardia appears to be due to the indirect activation of cardiac muscarinic receptors via activation of the vagus nerve. The HEAO also induces an antihypertensive effect.

Evaluation of biological activity, toxicity, and phytochemical content of Bowdichia virgilioides (Fabaceae) aqueous extract

Background: Antibiotic resistance is a worldwide problem that poses a serious threat to human health, limiting the therapeutic options for bacterial infections. The spread of falciparum-resistant malaria is also concerning, making the patient treatment an extremely difficult task. Those facts have heightened the interest to find alternate options to treat infections caused by drug-resistant microorganisms. Objective: Considering the importance of the development of new substances with antibacterial and antimalarial properties, the present study aimed to investigate the activity of the aqueous extract of stem bark of Bowdichia virgilioides (AEBv). This plant is commonly used in Brazilian folk medicine to treat a wide range of illnesses, including signs and symptoms associated with malaria. Materials and Methods: The AEBv was assayed for toxicity against two cell lines and Artemia salina larvae. In vitro activity of the extract was screened against a panel of Gram-positive and Gram-negative bacteria, a chloroquine-resistant (W2) and a chloroquine-sensitive (3D7) Plasmodium falciparum strains. The extract was also tested as antimalarial in vivo against Plasmodium berghei. Results: The AEBv presented no significant toxicity and was found to exert in vitro growth inhibitory effect against the tested bacterial species. The lowest minimal inhibitory concentration was reported for Staphylococcus aureus (0.125 mg/ml) followed by Staphylococcus epidermidis and Staphylococcus saprophyticus (0.50 mg/ml). B. virgilioides extract showed weak in vitro antimalarial activity against P. falciparum. A preliminary phytochemical analysis revealed the presence of flavonoids, phenolic groups, terpenoids, saponins, and tannins and the absence of alkaloids. Conclusion: The AEBv showed promising activity against Gram-positive microorganisms. Abbreviation used: AEBv: Aqueous extract of stem bark of Bowdichia virgilioides; FBS: fetal bovine serum; CC50: 50% cytotoxic concentration; LC50: Median lethal concentration; ATCC: American Type Culture Collection; MIC: Minimum inhibitory concentration; MBC: Minimum bactericidal concentrations; CQR: Chloroquine resistant; CQS: Chloroquine-sensitive; HRP2 Histidine rich protein 2; ELISA: Enzyme linked immunosorbent assay; PBS T: Phosphate buffer saline with 0.05% Tween 20; ANOVA: Analysis of variance; TLC: Thin layer chromatography; Rf: Retention factor; SI: Selectivity index; MRSA: Methicillin resistant Staphylococcus aureus.

Genotoxicity assessment of Schinopsis brasiliensis Engl. (Sapindales: Anacardiaceae) in somatic cells of Drosophila melanogaster (Meigen, 1830) (Diptera: Drosophilidae)

Schinopsis brasiliensis Engl. (Sapindales: Anacardiaceae) has been used in the Brazilian folk medicine to treat several illnesses. However, the phytochemical profile of S. brasiliensis as well as its genotoxic potential are poorly understood, which compromises population safety regarding the medicinal use of this plant species. In this study, we analyzed the genotoxic effects of S. brasiliensis using the Somatic Mutation and Recombination Test (SMART) of Drosophila melanogaster. Larvae from both standard cross (ST) and high bioactivation capacity cross (HB) were exposed to different concentrations of the hydroetanolic extract and ethyl acetate fraction of S. brasiliensis. We analyzed wings from D. melanogaster according to the type and number of mutant hair. Ours results suggested no genotoxic activity of S. brasiliensis in D. melanogaster somatic cells.

Antitrypanosomal activity and evaluation of the mechanism of action of diterpenes from aerial parts of Baccharis retusa (Asteraceae)

Baccharis retusa, a medicinal Brazilian plant from Asteraceae, has been used in Brazilian folk medicine to treatment of several illnesses, including parasitic diseases. Bioactivity-guided fractionation of the n-hexane extract from the aerial parts of B. retusa resulted in the isolation and characterization of three active related diterpenes: ent-15β-senecioyl-oxy-kaur-16-en-19-oic acid (1), ent-kaur-16-en-19-oic (2) and ent-16-oxo-17-nor-kauran-19-oic (3) acids. The structures of isolated compounds were defined by spectroscopic analysis, including NMR and HRESIMS. Antitrypanosomal activity of 1–3 was performed against cell-derived trypomastigotes using the colorimetric resazurin assay. The obtained results demonstrated that isolated compounds displayed a reduced toxicity against NCTC cells and were effective against the trypomastigote forms of T. cruzi with IC50 values of 3.8μM (1), 75.3μM (2) and 44.2μM (3). Additionally, compound 3 displayed activity against amastigote forms of T. cruzi with IC50 of 83.2μM. Compound 1 displayed the highest selectivity index (SI) when considered the trypomastigote forms, and its effect in the plasma membrane of parasite was evaluated using the fluorescent probe SYTOX Green. A considerable permeabilization (57%) in the membrane of the parasite was observed when compared to the untreated trypomastigotes. These data demonstrate, for the first time, the antitrypanosomal activity and mechanism of action of 1 and related compounds 2 and 3, obtained from aerial parts of B. retusa.

Phytochemical study and evaluation of cytotoxicity, mutagenicity, cell cycle kinetics and gene expression of Bauhinia holophylla (Bong.) Steud. in HepG2 cells in vitro.

Bauhinia holophylla (Bong.) Steud. (Fabaceae) is a plant used in Brazilian folk medicine to treat diabetes and inflammation. This study evaluated the phytochemical properties, cytotoxic, apoptotic, mutagenic/antimutagenic effects and alterations in gene expression (RNAm) in HepG2 cells treated with the B. holophylla extract. The phytochemical profile highlight the presence of flavonoids isorhamentin and quercetin derivates. The MTT assay was used to evaluate the cytotoxicity of different concentrations for different treatment times. Three concentrations (7.5, 15, 30µg/mL) were chosen for assessment of apoptosis (AO/EB), mutagenicity (micronucleus), and cell cycle kinetics (flow cytometry). Thereafter, the concentration of 7.5µg/mL was chosen to evaluate the protective effects against DNA damage induced by benzo[a]pyrene (B[a]P).At concentrations higher than 7.5µg/mL (between 10 and 50µg/mL), the extract was cytotoxic, induced apoptosis, and caused antiproliferative effects. However, it did not induce micronucleus and a reduction of apoptotic and micronucleated cells was observed in treatments that included the extract and B[a]P. The protective effect is attributable to the presence of flavonoids, described as antioxidants, inhibitors of DNA adduct and activators of detoxifying enzymes. The results of the present study such as absence of cytotoxic and mutagenic effects and protective effects against known carcinogens suggest that B. holophylla has potential for use soon as herbal medicine.

Gastroprotective activity of the methanol extract from peels of Plinia edulis (Vell.) Sobral fruits and its isolated triterpenes: maslinic and ursolic acids.

According to the Brazilian folk medicine, the leaves of Plinia edulis (Vell.) Sobral (Myrtaceae), known as cambuca, are indicated in the treatment of gastric disorders. Infusions of P. edulis leaves were previously demonstrated to contain both maslinic (MA) and ursolic acids (UA). Both triterpenes have also been identified in the methanolic extract of peels from P. edulis fruit (MEPPE); however, the antiulcer effects of MEPPE have not yet been studied. This study therefore evaluates the gastroprotective potential of MEPPE, MA, and UA using ethanol/HCl- and indomethacin-induced gastric ulcers in mice. In addition, the in vitro effects of these compounds on the H+, K+-ATPase activity and on the free radical DPPH were measured. When used at concentration of 100μg/mL, both MEPPE and UA were found to reduce the DPPH radical levels by 78.66 and 60.14%, respectively. However, MA did not reduce DPPH radical levels. Our results illustrated the antiulcer effects of MEPPE, MA, and UA against experimental ulcer models when administered by either the oral or the intraperitoneal routes. In addition, MEPPE reduced the size of ethanol/HCl-induced ulcers in a dose-dependent manner (log half-maximal effective oral dose, LogED50 = 1.09). Interestingly, UA promoted gastroprotection at lower doses than MA by increasing the production of mucin levels at 692%; however, it does not alter the activity of H+, K+-ATPase. In contrast, both MEPPE and MA, when incubated at concentrations of 10 and 100μg/mL, inhibited H+, K+-ATPase activity in 61.81, 68.37, 54.04, and 70.45%, respectively. These results confirm that MEPPE, MA, and UA display gastroprotective activity through different modes of action; MA inhibits H+, K+-ATPase activity, whereas UA favour the mucus barrier.

GC×GC/qMS analyses of Campomanesia guazumifolia (Cambess.) O. Berg essential oils and their antioxidant and antimicrobial activity

The present study investigated the essential oil obtained from Campomanesia guazumifolia (Cambess.) O. Berg, an aromatic plant used in Brazilian folk medicine. The chemical composition was performed by GC×GC/qMS. The antioxidant and antimicrobial activities were evaluated by DPPH and BCB and, MIC assays, respectively. Sixty-eight compounds were identified in the oil, where the major compounds were bicyclogermacrene (15%), globulol (5%) and spathulenol (5%). Sesquiterpene hydrocarbons (29 compounds) and oxygenated sesquiterpenes (20 compounds) were the most representative classes of terpenes. DPPH (IC50 value 26.1 ± 0.5 μg/mL) and BCB (68.3 ± 1.5%) values indicated a significant antioxidant activity. The essential oil strongly inhibited Staphylococcus aureus (MIC 15 ± 0.1 μg/mL), Escherichia coli (MIC 25 ± 0.2 μg/mL) and Candida albicans (MIC 5 ± 0.1 μg/mL). The results give a deeper understanding of the chemical composition and report for the first time the antioxidant and antimicrobial potential of the C. guazumifolia essential oil.

Comparison of two Jatropha species (Euphorbiaceae) used popularly to treat snakebites in Northeastern Brazil: Chemical profile, inhibitory activity against Bothrops erythromelas venom and antibacterial activity

Ethnopharmacological relevanceJatropha species (Euphorbiaceae) are largely used in traditional medicine to treat different pathologies in Africa, Asia and Latin America. In Northeastern Brazilian folk medicine, several Jatropha species, such as Jatropha gossypiifolia L. and Jatropha mollissima (Pohl) Baill., are indistinctly used to treat snakebites. Aim of the studyTo compare two of the Brazilian most used Jatropha species for snakebites (J. gossypiifolia and J. mollissima), in relation to their ability to inhibit local edematogenic activity of Bothrops erythromelas snake venom in mice, their in vitro antibacterial activity and phytochemical profile. Material and methodsAqueous leaf extracts of J. gossypiifolia (AEJg) and J. mollissima (AEJm) were prepared by decoction. AEJg and AEJm were compared chemically, by thin layer chromatography (TLC) and high-performance liquid chromatography with diode array detection (HPLC-DAD) analysis. They were also pharmacologically compared, using the mouse model of paw edema induced by Bothrops erythromelas snake venom (BeV), and in vitro by broth microdilution and agar dilution antimicrobial tests. ResultsFlavonoids were detected as the major compounds in both extracts. However, AEJg and AEJm showed quantitatively different chemical profiles by HPLC-DAD. AEJg presented fewer peaks of flavonoids than AEJm, however, when the intensity of peaks were analyzed, these compounds were at high concentration in AEJg, even using the same concentration of both extracts. Differences were also observed in the biological activity of the two extracts. While no difference was observed when the extracts were administered by oral route (P > 0.05), by the intraperitoneal route AEJg presented anti-edematogenic activity significantly (P < 0.001) higher than AEJm. In antimicrobial assays, only AEJg presented antibacterial activity against Staphylococcus epidermidis, Staphylococcus aureus and Bacillus cereus. ConclusionsAlthough used indistinctly by folk medicine, our results suggested that AEJg is more active than AEJm in relation to its antiedematogenic and antibacterial activities. Significant differences were observed in their phytochemical profiles, especially a higher content of C-glycosylated flavonoids in the most active species, which could justify the different biological effects observed. These findings strengthen the potentiality of J. gossypiifolia species for use as complementary treatment for local effects induced by Bothrops venoms and could be helpful for distinction of the species and control quality assessment of future herbal medicines based on Jatropha plants.

Microemulsions containing Copaifera multijuga Hayne oil-resin: Challenges to achieve an efficient system for β-caryophyllene delivery

β-Caryophyllene (β-CP) is a bioactive sesquiterpene abundant in copaiba oil-resin (COR), which is widely used in Brazilian folk medicine; however, its instability due to the presence of unsaturations is an obstacle to the use of COR in its natural form. Therefore, the development of a system for β-CP delivery, like the one proposed herein for the first time, may protect it and preserve its therapeutic action. Pseudo-ternary phase diagrams were constructed in this study to obtain COR-containing nanocarriers. Two microemulsions (MEs) were obtained, namely a) ME-I containing (% w/w) 47.1 water, 8.5 Plurol Oleique, 33.8 Labrasol and 10.6 COR and b) ME-II containing (% w/w) 39.0 water, 18.3 Plurol Oleique, 36.6 Labrasol and 6.1 COR. Both MEs showed physicochemical properties suitable for a delivery system such as pH 5.25-5.74, conductivity of 79.5–120.9μScm−1 and slightly negative Zeta potential (-3.57/-2.63mV). Transmission electron microscopy revealed the nanometric size of MEs droplets, differential scanning calorimetry their oil/water nature, while refractive indices clarity and isotropicity. Both systems behaved as Newtonian fluids and exhibited low viscosity (64.42±0.667 mPa.s for ME-I and 118.78±0.503 mPa.s for ME-II). After COR incorporation in MEs, steam distillation allowed obtaining β-CP contents depending on their initial COR amount. The pharmacological effectiveness of these innovating systems for β-CP delivery was confirmed by antimicrobial and anti-inflammatory activity tests. ME-II afforded a stronger antimicrobial effect against all target microorganisms than ME-I and a much stronger one than COR. Carrageenan-induced inflammation in rat paw was visibly inhibited by MEs.

Chemical composition and toxicological evaluation of Hyptis suaveolens (L.) Poiteau (LAMIACEAE) in Drosophila melanogaster and Artemia salina

Hyptis suaveolens (L.) Poiteau, known as “Bamburral” is a plant widely used in Brazilian folk medicine, however, there are no toxicological studies to ascertain its safety. Thus, we aimed to investigate the phytochemical and toxic effects of essential oil and infusion of the dry leaves of H. suaveolens in Drosophila melanogaster and Artermia salina. The chemical composition of the essential oil was performed by CG-MS, while that of the infused extract was analyzed by HPLC-DAD. The toxicity of the essential oil-OE (3.03–30.3μg/mL air) to D. melanogaster was performed by means of volatilization; and for infusion (2.5–20mg/mL), they were submitted to ingestion and then submitted to locomotive activity by means of negative geotaxis. However, for acute toxicity, A. salina cysts were exposed for 24h to the tested samples (10–1000μg/mL). Our results demonstrated that OE had three major components: β-Caryophyllene (18.57%), sabinene (15.99%) and spathulenol (11.09%) while the leaf infusion showed caffeic acid as the major component (12.76mg/g). H. suaveolens leaves infusion was not toxic to the organisms at all the concentrations tested (p>0.05). However, OE behaved as quite toxic with LC50 of 15.5 and 49.72μg/mL in D. melanogaster and A. salina, respectively. In addition, OE caused impairment of the locomotor behavior of flies. Therefore, our study indicates that more caution should be taken regarding the dosage and frequent use of H. suaveolens leaf essential oil.

In vitro multimodal-effect of Trichilia catigua A. Juss. (Meliaceae) bark aqueous extract in CNS targets

Ethnopharmacological relevanceThe bark of Trichilia catigua A. Juss. (Meliaceae), popularly known as “big catuaba”, is traditionally used in Brazilian folk medicine for its neuroactive potential as memory stimulant, and antinociceptive and antidepressant effects. Aim of the studyTo study the aqueous extract of T. catigua bark as dual inhibitor of monoamine oxidase A (MAO-A) and acetylcholinesterase (AChE). To explore its antioxidant potential through interaction with xanthine/xanthine oxidase (X/XO) pathway, and to attempt a relationship between its phenolic profile and effects displayed. Materials and methodsPhenolic profiling was achieved by HPLC-DAD-ESI/MSn and UPLC-ESI-QTOF-MS analyses. The capacity to inhibit hMAO-A was assessed in vitro, as was that for AChE, evaluated in rat brain homogenates. The direct inhibition of the X/XO pathway and the scavenging of superoxide anion radical were the selected in vitro models to explore the antioxidant potential. The cytotoxic effects were assayed in the human neuronal SH-SY5Y cells by MTT reduction, after direct exposure (24h). ResultsTwenty-six compounds were identified and quantified (551.02 ± 37.61mg/g of lyophilized extract). The phenylpropanoid substituted flavan-3-ols were the most representative compounds (~81% of quantified mass). The extract inhibited hMAO activity in a concentration-dependent manner (IC50 = 121.06 ± 2.13μg/mL). A mixed model of inhibition of AChE activity was observed, reflected by the pronounced increase of Km values and a more discreet effect over the Vmax parameters, calculated from Michaelis-Menten fitted equations. In addition, it was demonstrated that the extract directly inhibits the X/XO pathway (IC50 = 121.06 ± 2.13μg/mL) and also imbalances the oxidative stress acting as superoxide anion radical scavenger (EC50 = 104.42 ± 10.67μg/mL), an oxidative by-product of this reaction. All these neuroprotective and neurotrophic effects were displayed within the non-toxic range of concentrations (0.063–0.500μg/mL) in SH-SY5Y cells. ConclusionsOur results validate the traditional use of T. catigua bark for its neuroactive and neuroprotective potential. A novel approach upon its application towards the management of neurodegenerative and related symptomatology was likewise demonstrated.