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In this study, a novel compound was isolated, identified, and its chemical structure was determined from the extract of the roots of Senna velutina. In addition, we sought to evaluate the anticancer potential of this molecule against melanoma and leukemic cell lines and identify the pathways of cell death involved. To this end, a novel anthraquinone was isolated from the barks of the roots of S. velutina, analyzed by HPLC-DAD, and its molecular structure was determined by nuclear magnetic resonance (NMR). Subsequently, their cytotoxic activity was evaluated by the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) method against non-cancerous, melanoma, and leukemic cells. The migration of melanoma cells was evaluated by the scratch assay. The apoptosis process, caspase-3 activation, analysis of mitochondrial membrane potential, and measurement of ROS were evaluated by flow cytometry technique. In addition, the pharmacological cell death inhibitors NEC-1, RIP-1, BAPTA, Z-VAD, and Z-DEVD were used to confirm the related cell death mechanisms. With the results, it was possible to elucidate the novel compound characterized as 2′-OH-Torosaol I. In normal cells, the compound showed no cytotoxicity in PBMC but reduced the cell viability of all melanoma and leukemic cell lines evaluated. 2′-OH-Torosaol I inhibited chemotaxis of B16F10-Nex2, SK-Mel-19, SK-Mel-28 and SK-Mel-103. The cytotoxicity of the compound was induced by apoptosis via the intrinsic pathway with reduced mitochondrial membrane potential, increased levels of reactive oxygen species, and activation of caspase-3. In addition, the inhibitors demonstrated the involvement of necroptosis and Ca2+ in the death process and confirmed caspase-dependent apoptosis death as one of the main programmed cell death pathways induced by 2′-OH-Torosaol I. Taken together, the data characterize the novel anthraquinone 2′-OH-Torosaol I, demonstrating its anticancer activity and potential application in cancer therapy.

Cancer is a complex disease, considered a major public health problem worldwide. Among the types of cancer, melanoma, and leukemias present high mortality rates in Brazil and worldwide. Currently, conventional cytotoxic treatments cause severe side effects by the non-selectivity between normal cells and cancer cells. Therefore, molecules of natural origin with more efficient anticancer properties and that present fewer adverse effects are of extreme importance for cancer therapy and improvement in patients’ quality of life. This study isolated, identified, and characterized the chemical structure of a new anthraquinone present in the extract of the roots of Senna velutina. In addition, we sought to evaluate the anticancer potential of this molecule against melanoma and leukemic cell lines and identify the pathways of cell death involved. To this end, a novel anthraquinone was isolated from the barks of the roots of S. velutina, analyzed by HPLC-DAD, and its molecular structure was determined by NMR. Subsequently, their cytotoxic activity was evaluated by the MTT method against non-cancerous, melanoma, and leukemic cells. The migration of melanoma cells was evaluated by the scratch assay. By flow cytometry technique, the apoptosis process, caspase-3 activation, analysis of mitochondrial membrane potential, and measurement of ROS were evaluated. In addition, the pharmacological cell death inhibitors NEC-1, RIP-1, BAPTA, Z-VAD, and Z-DEVD were used to confirm the related cell death mechanisms. With the results, it was possible to elucidate the novel compound characterized as 2'-OH-Torosaol I. In normal cells, the compound showed no cytotoxicity in PBMC but reduced the cell viability of all melanoma and leukemic cell lines evaluated. 2'-OH-Torosaol I inhibited chemotaxis of B16F10-Nex2, SK-Mel-19, SK-Mel28 and SK-Mel-103. The cytotoxicity of the compound was induced by apoptosis via the intrinsic pathway with reduced mitochondrial membrane potential, increased levels of reactive oxygen species, and activation of caspase-3. In addition, the inhibitors demonstrated the involvement of necroptosis and CA+ in the death process and confirmed caspase-dependent apoptosis death as one of the main programmed-cell death pathways induced by 2'-OH-Torosaol I. Taken together, the data characterize the novel anthraquinone 2'-OH-Torosaol I, demonstrating its anticancer activity and potential application in cancer therapy.

Ocimum gratissimum L. has attracted substantial consideration from researchers because of its anti-inflammatory uses in ethnomedicine in Sub-Saharan Africa and Asia. This study investigated the effect of flavonoid-rich ethylacetate fraction of O. gratissimum (EAFOg) in apoptosis induction of activated macrophages and inflammatory response in LPS-induced air pouch in rats. Apoptotic effect of EAFOg in LPS-stimulated RAW 264.7 cells was evaluated using flow cytometry after staining with annexin-V and 7-aminoactinomycin D. Its effects on inflammatory cells and mediators were investigated utilizing 6 day old subcutaneous air pouch-rats. Sterile saline (0.9%) or LPS (100 ng/mL) was injected into the air pouch on 6th day after EAFOg (25, 50 and 100 mg/kg) pretreatment. Rectal body temperature was recorded hourly for 5 h after LPS injection. Thereafter, the neutrophil count, nitrite, TNF-α, PGE2, nitrite, malondialdehyde and reduced glutathione (GSH) levels were determined in the pouch lavage. The activities of myeloperoxidase and superoxide dismutase (SOD) as well as immunohistochemical staining for cyclooxygenase-2 were also performed. EAFOg (10, 30 and 100 µg/mL) induced apoptosis in LPS-stimulated RAW 264.7 macrophages. The EAFOg reduced hyperthermia and decreased neutrophil counts, TNF-α, PGE2, nitrite, myeloperoxidase as well as cyclooxygenase-2 expression evoked by LPS in rats. It also reduced malondialdehyde, and increased GSH and SOD levels in LPS-induced air pouch in vivo. The results of this study suggest that the EAFOg induces apoptosis in activated macrophages and reduces inflammatory responses provoked by LPS via down regulation of cyclooxygenase-2 expression in rats.

Preparations of Helicteres sacarolha (Malvaceae) leaves and roots are used in the form of decoction, infusion or maceration, to treat gastrointestinal disturbances, among others. Studies supporting some of its ethnomedicinal uses are still incipient. The present study aimed to investigate it potential effect on chronic ulcer, ulcerative colitis and possible prokinetic activities as part of its mechanism of action. The powdered leaves of Helicteres sacarolha (HEHs) was prepared by maceration in 70 % hydroethanolic solution. Its qualitative phytochemical constituents were investigated by direct flow injection analysis coupled to atmospheric pressure chemical ionization ion trap tandem mass spectrometry (FIA-APCI-IT-MS

Objetivo: Verificar a intensidade do basquete em cadeira de rodas (BCR) em pessoas com deficiência (PCD) durante 8 semanas de intervenção. Métodos: Estudo experimental longitudinal com 13 PCD foi avaliado o monitoramento da FC, freqüência cardíaca média (FCMéd) para cálculo da frequência cardíaca mínima (FCMin) e % frequência cardíaca de treinamento (% FCT) foram baseados na FCpico. Estes foram então analisados para possível correlação com o tipo, nível e tempo da lesão, bem como os valores antropométricos obtidos por bioimpedância. Resultados: Após análise estatística, nas variáveis independentes verificou-se que o percentual de gordura corporal (p 0,05) quando correlacionadas com a FCpico. Conclusões: A maioria dos participantes obteve a % FCT acima de 60% da FCpico e indicando que a intervenção auxiliou na promoção da saúde cardiovascular, além dos benefícios mencionados dos níveis de hidratação e % gordura corporal de cada indivíduo. O presente estudo pode servir de base científica para o educador físico e afins, principalmente no que diz respeito ao uso do basquete em cadeira de rodas na reabilitação ou treino de aptidão física para PCD.

Cochlospermum regium (Bixaceae) is a native shrub of Brazil and its xylopodium (infusion/decoction) is being used for the treatment of gastritis, ulcers, arthritis, intestinal infections, gynaecological infections, skin diseases, among others. The aim of the present study was to evaluate the gastroprotective/antiulcer activity and the mechanism of action of hydroethanolic extract of C. regium xylopodium (HECr), using in vitro and in vivo models. Additionally, phytochemical constituents were identified by high-performance liquid chromatography (HPLC).C. regium xylopodium was macerated with ethanol/water to obtain the HECr. The phytochemical characterisation was carried out by HPLC. The antiulcer efficacy of HECr (25, 100 and 400 mg/kg, p.o.) was evaluated using acute acidified ethanol (HCl/EtOH), piroxicam and water immersion-induced experimental ulcer models. Chronic gastric ulcer healing activity of HECr was evaluated through acetic acid (99.8%) - induced model. Histological analysis and myeloperoxidase (MPO), glutathione (GSH), catalase (CAT) activities were also evaluated in chronic ulcer induced gastric tissues. The plausible mode of action of the HECr was assessed by estimation of gastric wall mucus production and the role of gastric secretion in pylorus ligature. The animals were also pre-treated with various inhibitors which includes indomethacin (10 mg/kg, p.o.) a selective inhibitor of cyclooxygenase, L-NAME (10 mg/kg, i.p.), an inhibitor of nitric oxide synthase, glibenclamide, a ATP-sensitive potassium channels (KThe HPLC analysis data revealed the presence of gallic acid, rutin, myricetin, morin and kaempferol. HECr promoted protective effect against acute ulcers induced by HCl/EtOH with inhibitions of 47.52% (p 0.01) and 62.69% (p 0.001) at 100 and 400 mg/kg, and in piroxicam by 34.11% (p 0.05), 49.14% (p 0.01) and 61.34% (p 0.001), at 25, 100 or 400 mg/kg, respectively, and in water restraint stress by 78.26% inhibition, p 0.001, at the dose of 400 mg/kg when compared to the vehicle control group respectively. In the chronic gastric ulcer model, HECr (25, 100 and 400 mg/kg p.o.) significantly (p 0.001) decreased the injured area by 58.80%, 77.87% and 71.10% respectively. Histological examination indicated that oral treatment of HECr promoted healing of gastric lesions by regenerating gastric mucosa layer with less inflammatory cells. HECr augmented the GSH, CAT activities and reduced MPO level. The pre-treatment with HECr increased the gastric wall mucus production. It also significantly altered the gastric secretion parameters by causing the reduction in the gastric juice volume, elevated the pH level and reduced the total acidity at all doses tested when compared with the vehicle group. HECr at the most active dose (100 mg/kg) reversed completely the reduction of PGs, NO production, closure of KThe HECr presented preventive and curative effects in the experimental gastric ulcer models, besides good anti-Helicobacter pylori activity, which supports the traditional medicinal use of the xylopodium of this plant for gastrointestinal diseases. The underlying mechanisms of this antiulcerogenic/antiulcer action involve, at least, augmentation of mucus production, inhibition of gastric secretion, stimulation of PGs and NO synthesis. And that it involves activation of K

Made available in DSpace on 2019-10-06T16:06:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-03-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Ethnopharmacological relevance: Virola elongata is a tree species belonging to the Myristicaceae family, distributed in the North and Midwest regions of Brazil, in the phytogeographic domain of the Amazon. The aqueous infusion or the hydroethanolic macerate of the stem bark of V. elongata are used in Brazilian and Ecuadorian indigenous folk medicine for several ethnopharmacological purposes, principally, in the treatment of stomach pain, indigestions, and gastric ulcers. This study was aimed to investigate the gastroprotective activity of this plant in order to support its popular use with scientific evidence. Materials and methods: The stem bark hydroethanolic extract of the plant (HEVe) was prepared by maceration. Its qualitative and quantitative phytochemical constituents were investigated by classical colorimetric techniques, HPLC, and electrospray ionization-multiple stage fragmentation (ESI-MSn). The gastroprotective and antiulcer activity of HEVe at doses of 100, 300 and 900 mg/kg p.o. were tested using three acute (acidified ethanol, piroxicam, and in-water-restrain stress), and one chronic (acetic acid) animal ulcer models. The probable mode of action of the HEVe was evaluated by analyzing gastric acid secretion, mucus content, nitric oxide effect, and its antioxidant properties (on catalase, myeloperoxidase, and GSH content) in experimental rodents. The direct extract's activity on the growth of Helicobacter pylori was also investigated. Results: Total phenolic content in the HEVe was of 146.20 ± 1.07 mg, being flavonoids about 50% (71.79 ± 0.70 mg) of it. Comparative HPLC fingerprint analysis revealed the presence of known phenolic antiulcer compounds, such as gallic acid, catechin, and rutin. Also, methanol/water fractionation and ESI-MSn analysis of the HEVe reveals the presence of quinic acid, 3,3′,4-trihydroxystilbene, juruenolid D, one catechin dimer, one C-glycosyl flavonoid, one polyketide and two neolignans as the major components of the extract. The HEVe attenuated gastric ulceration in all the different models of acute gastric ulcer, by enhancing gastroprotection through its antioxidant properties in vivo, and reducing also considerably the gastric secretion and total acidity. The HEVe also presented healing properties against the induced chronic ulceration process. On the other hand, the HEVe did not exhibit direct activity against H. pylori. Conclusion: The HEVe exhibited significant gastroprotective/antiulcer effects and contain a relative high proportion of phenolic compounds, especially flavonoids, that could likely account, at least in part, for its pharmacological properties. The results justify its traditional usage and provided scientific evidence for its potential as a new herbal medicine to treat gastric ulcers. Área de Farmacologia Departamento de Ciências Básicas em Saúde Universidade Federal de Mato Grosso (UFMT), Av. Fernando Correa da Costa, no. 2367 Curso de Farmácia Faculdade Noroeste do Mato Grosso Associação Juinense de Ensino Superior (AJES) Laboratório de Pesquisa de Produtos Naturais Faculdade de Medicina Universidade Federal de Tocantins (UFT) Laboratório de Bioprospecção de Produtos Naturais. Instituto de Biociências Universidade Estadual Paulista (UNESP) - Campus do Litoral Paulista Departamento de Química Orgânica Instituto de Química Universidade Estadual Paulista (UNESP) - Campus Araraquara Laboratório de Bioprospecção de Produtos Naturais. Instituto de Biociências Universidade Estadual Paulista (UNESP) - Campus do Litoral Paulista Departamento de Química Orgânica Instituto de Química Universidade Estadual Paulista (UNESP) - Campus Araraquara CNPq: 132286/015-7 CAPES: 23038.000731/2013-56

Ethnopharmacological relevance Sphenodesme involucrata var. paniculata (C. B. Clarke) Munir is native as well as endemic to South India. Its leaves are used in folklore medicine to treat pain and rheumatism. Objective This study was aimed to investigate the chemical characterization, anti-nociceptive and mode of action underlying the anti-inflammatory effects of methanol extract of S. involucrata leaves (MESi). Methods Phytoconstituents of MESi was analyzed using colorimetric and liquid chromatography–mass spectrometry (LC-MS) methods, and the oral acute toxicity was evaluated in mice up to 2000 mg/kg. The anti-nociceptive effect was evaluated in hot plate and writhing tests; whereas the anti-inflammatory effect was investigated using carrageenan, cotton pellet and lipopolysaccharide (LPS)-induced peritonitis models at doses of 100, 200 and 400 mg/kg. Additionally nitric oxide (NO) and inflammatory cytokines levels were also evaluated. Results MESi exhibited the high content of phenolics and flavonoids as well as compounds like austricine, benzylglucosinolate, gossypin, justicidin B and cirsimarin were detected in LC-MS. In the acute toxicity study, oral administration of MESi did not cause any toxic effect and mortality up to 2000 mg/kg body weight in mice. In the anti-nociceptive tests, MESi augmented the latency period at higher dose (400 mg/kg), on the other hand attenuated writhings at the dose of 400 mg/kg by 87.87% (p Conclusion The results conclude that MESi has no acute toxic effect and it demonstrated potent anti-nociceptive and anti-inflammatory activities. Its anti-nociceptive activities are probably mediated through peripheral and central mechanisms. The anti-inflammatory effect of MESi involved the inhibition of neutrophils migration and the modulation of Th1 and Th2 cytokines, besides the attenuation of production of PGE2 and NO. LC-MS analysis revealed the predominant presence of the austricine, benzylglucosinolate, gossypin, justicidin B and cirsimarin compounds, which are possibly involved in the anti-nociceptive and anti-inflammatory effects of MESi. The current study provided supportive evidence for the folklore use of S. involucrata in the treatment of pain and inflammatory conditions.

Ethnopharmacological relevance Copaifera malmei Harms (Fabaceae), known mainly as oleo-mirim, is a native and endemic plant found in the states of Mato Grosso and Goias of Brazil. The plant's leaves infusion is popularly used by riverine communities of the northern Araguaia microregion, Mato Grosso, Brazil, for the treatment of gastric ulcers and inflammatory diseases of the respiratory tract. The gastric antiulcer activity of the standardized leaves infusion extract of Copaifera malmei (SIECm) in rodents has been reported. The objective of this study was to advance the investigation of the safety profile of SIECm by evaluating the genotoxicity and subchronic toxicity using in vitro and in vivo experimental models. Materials and methods SIECm was prepared by infusion, by incubating the powdered dried leaves material in boiled water for 15 min. In vitro genotoxicity of SIECm (10, 30 or 100 μg/mL) was assessed by micronucleus and comet tests using Chinese hamster ovary (CHO-k1) epithelial cells. The evaluation of subchronic toxicity profile was performed by daily oral administration of SIECm (100, 400 or 1000 mg/kg) to Wistar rats for 30 days. Clinical observations of toxicological related parameters were done every 6 days. After the treatment period, blood was collected for hematological and biochemical analysis, and some organs were removed for macroscopic and histopathological analysis. Results In the micronucleus assay, SIECm demonstrated anti-mutagenic activity. In the comet assay, SIECm presented anti-genotoxic effect preventing DNA damage at all the three concentrations tested with pre-treatment, while the same effect was only observed in the co-treatment at the lowest concentration. Post-treatment with SIECm increased the genetic damage induced by hydrogen peroxide (H 2 O 2 ) at the highest concentration. In the subchronic toxicity test, few changes were observed, such as increase in feed consumption in the group of animals treated with 100 mg/kg of the SIECm, which reversed after 6 days. There were no macroscopic, histological and relative weights changes in the organs of animals treated with SIECm. No toxicologically relevant changes were observed in the hematological analysis. Subchronic administration of SIECm reduced levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in animals treated with 100 mg/kg and serum triglyceride levels at 400 and 1000 mg/kg. However, the hematological and biochemical changes observed are within the physiological ranges for this animal species. Conclusion The results demonstrate that SIECm is not genotoxic, and does not present toxicity when used orally for up to 30 days. In addition, it showed protection to the genetic damage induced by H 2 O 2. The SIECm therefore has a high safety margin for therapeutic use.

Ethnopharmacological relevance Ocimum gratissimum L. is a herbaceous plant that has been reported in several ethnopharmacological surveys as a plant readily accessible to the communities and widely used for the treatment of inflammatory diseases. The main goal of this study was to investigate the in vitro and in vivo anti-inflammatory activity and mechanism of action of the ethylacetate fraction of O. gratissimum leaf (EAFO g ) and to chemically characterize this fraction. Materials and methods EAFO g was obtained from a sequential methanol extract. The safety profile was evaluated on RAW 264.7 cells, using the alamarBlue® assay. Phenolic contents were determined by spectrophotometry, and metabolites quantified by high performance liquid chromatography. The anti-inflammatory activity of EAFO g and its ability to acts on leucocytes infiltration, inflammatory mediators as NO, IL-1β, TNF-α, and IL-10 in lipopolysaccharide-induced peritonitis in mice and LPS-stimulated RAW 264.7 macrophage were evaluated. In addition, the anti-inflammatory activity of EAFOg was also investigated in arachidonic acid-related enzymes. Results Total phenolic and flavonoid contents of EAFO g were 139.76±1.07 mg GAE/g and 109.95±0.05 mg RE/g respectively. HPLC analysis revealed the presence of rutin, ellagic acid, myricetin and morin. The fraction exhibited no cytotoxic effects on the RAW 264.7 cells. The EAFO g (10, 50 and 200 mg/kg) significantly reduced ( p g (5, 20 and 100 µg/mL) produced significant reduction in NO, IL-1β, and TNF-α in RAW 264.7 cells. However, IL-10 level was not affected by the EAFOg, and it preferentially inhibits COX-2 (IC 50 =48.86±0.02 µg/mL) than COX-1 and 15-LO (IC 50 >100 µg/mL). Conclusion The flavonoid-rich fraction of O. gratissimum leaves demonstrated anti-inflammatory activity via mechanisms that involves inhibition of leucocytes influx, NO, IL-1β, and TNF-α in vivo and in vitro, thus supporting its therapeutic potential in slowing down inflammatory processes in chronic diseases.

Objectives: To evaluate the effect of a probiotic supplement containing two genera and five species of bacteria versus placebo on the quality of life (QoL) in female university students with intest...

Ethnopharmacological relevance Leonotis nepetifolia (L) R. Br., Lamiaceae, a pantropical shrub, popularly known in Brazil as “cordao-de-frade”, “rubim”, is reportedly used in Brazilian ethnomedicine as well as in different countries in the treatments of ailments such as infections, inflammations, wounds, stomach disorders, among others. Aim of the study To evaluate its potential cytotoxicity and antibacterial mode of action of the hydroethanolic extract of L. nepetifolia (HELn) leaves, including phytochemical analysis. Materials and methods The cytotoxicity of HELn was investigated by Alamar blue assay, using CHO–K1 cells. Antibacterial activity of HELn was tested by broth microdilution methods against a panel of bacteria of clinical interest. The mode of action of L. nepetifolia was studied by targeting bacterial membranes. Phytochemical analysis was performed by determining total secondary metabolites with spectrophotometric assays and HPLC. Results HELn is not cytotoxic in the in vitro evaluation (IC50>200 μg/mL). It demonstrated a good spectrum of antibacterial activity with major activity against Shigella flexneri, Enterococcus faecalis, Staphylococcus aureus and Bacillus subtilis with MIC=6.25 µg/mL, Helicobacter pylori with MIC of 25 µg/mL and Streptococcus pyogenes with MIC of 50 µg/mL. Its mode of action is associated, at least partly, with changes in the permeability of bacterial membranes, as evidenced by the increased entry of hydrophobic antibiotics in Shigella flexneri and intense efflux of K+ and nucleotide leakage in E. faecalis and Shigella flexneri. In addition, the presence of phenols, flavonoids and carotenoids, described in the literature to possess antibacterial effects, were detected in the composition of HELn, with high phenol content (11.55%), especially the flavonoids (6.47%). Conclusion The results indicate that HELn has low cytotoxicity and potent antibacterial activity. It is bacteriostatic in nature, possibly acting at the level of bacterial membranes, especially on the cytoplasmic membrane and outer membrane, thus supporting its popular use in infectious processes. In addition, the presence of phenols, flavonoids, carotenoids, fatty acids and steroids, described in the literature as possessing antimicrobial activity, were detected in the composition of HELn.

Background: The constant pursuit of improved athletic performance characterizes high-performance sport and the use of medicinal plants as dietary supplements is becoming widespread among athletes to enhance long-term endurance performance. Aim: The present study evaluated the toxicity of Heteropterys tomentosa (HEHt) and its acute adaptogenic effects. Methods: The in vitro safety profile was evaluated on CHO-k1 cells using the alamar Blue assay, at concentrations ranging from 3.125 to 200 µg/mL. In vivo acute oral toxicity was conducted in male and female mice with oral administration of graded doses of HEHt from 400 to 2000 mg/kg. A subchronic oral toxicity study was completed by oral administration of HEHt (50, 200 or 1000 mg/kg) and vehicle for 30 days in male Wistar rats. Clinical observations and toxicological related parameters were determined. Blood was collected for biochemical and hematological analyses, while histological examinations were performed on selected organs. Thereafter, an adaptogenic test consisting of progressive loads until exhaustion was conducted in rats ( n = 5/group) orally pre-treated with the vehicle and HEHt (25, 100 or 400 mg/kg). Results: HEHt exhibited no cytotoxic effects on the CHO-k1 cells and, apparently, no acute toxicity in mice and no subchronic toxicity in rats. An ergogenic effect was observed only at the dose of 25 mg/kg compared with the vehicle in relation to time to exhaustion and exercise load ( p = .011 and .019, respectively). HEHt is safe at up to 400 mg/kg, contains astilbin and taxifolin as the major phytochemical compounds, and exhibited a potential adaptogenic effect. Conclusions: These results justify its anecdotal usage as a tonic, show that the hydroethanolic maceration of the root does not cause toxicity, and provide scientific evidence of its potential as a source of new adaptogenic substance(s).

Ethnopharmacological relevance Simaba ferruginea A. St.-Hil., Simaroubaceae, popularly known as “calunga” is a typical subtropical shrub used in Central Brazil mainly for infection, anti-inflammatory, analgesic and gastric duodenal-ulcers. It presents in its composition the alkaloid canthin-6-one, an alkaloid indole β-carboxylic. Aim This study aims to investigate the toxicity, antimicrobial activities of methanol extract of Simaba ferruginea (MESf) and canthin-6-one by using different experimental models. Methods The present study evaluated the phytochemical analysis by high performance liquid chromatography (HPLC), toxicological potential of MESf and canthin-6-one, using the cytotoxicity, genotoxicity assays with CHO-K1 cells and in vivo acute test in mice. Antimicrobial activity was evaluated by the broth microdilution assays, while the antimicrobial mechanism of action was also assessed using different in vitro bacterial and fungal models. Results The HPLC analysis of MESf revealed the presence of canthin-6-one, kaempferol and morin. Differential in vitro toxicities were observed between MESf and canthin-6-one. In the cytotoxicity assay, MESf presented toxicity against CHO-K1, while canthin-6-one did not. In the case of in vitro genotoxicity, both showed to be potentially genotoxic. In the in vivo toxicity study, both MESf (up to 1000 mg/kg) and cantin-6-one (up to 100 mg/kg) caused no toxicologically relevant alterations and are thus considered not to be toxic. MESf was shown to be relatively safe with NOAEL (100 mg/kg) when administrate in mice. Both MESf and canthin-6-one also showed differential antimicrobial activities. On one hand, MESf demonstrated good spectrum of antibacterial action against Staphylococcus aureus (MIC 12.5 μg/mL) and Escherichia coli (MIC 25 μg/mL) and moderate activity against Enterococcus faecalis and Shigella flexneri (MIC 200 μg/mL) but no antifungal effect. On the hand, canthin-6-one showed no antibacterial activity, except against Staphylococcus aureus (100 μg/mL), but potent in vitro fungicidal activity against clinically important Aspergillus niger and Candida species at MFC intervals ranging from 3.12 to 25 μg/mL. Both MESf and canthin-6-one were bacteriostatic in action. MESf antimicrobial mechanism of actions are associated with changes in the permeability of bacterial membranes, evidenced by the increased entry of hydrophobic antibiotic in Shigella flexneri, intense K+ efflux (Shigella flexneri, Staphylococcus aureus) and nucleotides leakage (Staphylococcus aureus). In the antifungal mode of action, canthin-6-one inhibited Saccharomyces cerevisiae growth and including alteration in the cell membrane of Neurospora crassa. Conclusion The results of this work demonstrated the differential antimicrobial activities of MESf and its alkaloid isolate, canthin-6-one with antibacterial and antifungal activities, respectively. The present study support the popular use of Simaba ferruginea in combatting afflictions related to bacterial infections, and demonstrate that canthin-6-one as a promising antifungal agent. Both MESf and canthin-6-one are considered non-toxic based on the in vitro toxicological study.

Vitexin is an important component of various medicinal plants frequently used to treat asthma, such as Crataegus spp., Vitex spp., Passiflora spp., and Echinodorus spp. However, there is no information about the vitexin potential as anti-asthmatic. The aim of the present work was to evaluate the anti-hypersensitive activity of vitexin in a murine ovalbumin (OVA)-induced allergic asthma model. Mice were sensitized to OVA by i.p. injection on days 1st and 10th, followed by a daily challenge with OVA using a nebulizer, from days 19th to 24th. Vitexin or dexamethasone were orally administered 1 h before each OVA challenge. Vitexin attenuates migration induced by OVA-hypersensitivity of eosinophil, neutrophil, and mononuclear cells in bronchoalveolar lavage fluid (BALF). Histological analysis of the lungs shows that vitexin suppressed leukocyte infiltration, mucus production and pulmonary edema. Increases in Th2 cytokines in BALF in OVA-induced asthma is also attenuated by vitexin, as well as plasma levels of IgE. Overall, these results suggest that vitexin can suppress OVA-induced allergic inflammation in mice and provide a strong rationale for further developing vitexin as a candidate treatment for allergic hypersensitivity. These data corroborate the popular use of vitexin-rich plants for asthma treatment.

Gallesia integrifolia is a Brazilian Amazon tree whose bark decoction is popularly used to treat peptic ulcer. The essential oil from the inner stem bark of G. integrifolia (EOGi) was chemically characterized by GC/MS. The in vitro cytotoxicity and genotoxicity were evaluated in CHO-K1 cells, while the in vivo oral acute toxicity was performed in mice. The gastroprotective effect of EOGi was assessed in acidified ethanol and piroxicam and ulcer healing on acetic acid -induced ulcer models in rodents. Anti-secretory, mucus, K+-ATP channels, prostaglandins (PGs), nitric oxide (NO), TNF-α, IL-1β, IL-10, catalase (CAT) and myeloperoxidase (MPO) activities and in vitro Helicobacter pylori action by EOGi were evaluated. EOGi exhibited cytotoxic effects only at 72h and no acute toxicity. EOGi showed gastroprotective and ulcer healing effects. EOGi gastroprotection was attenuated by indomethacin pre-treatment. Gastric volume and total acidity were reduced, while gastric pH was elevated. EOGi increased mucus and NO productions and CAT activity, and inhibited MPO activity, TNF-α and IL-1β concentrations and augmented IL-10. EOGi was not active against H. pylori. These results indicated that EOGi is safe and exerts preventive and curative gastric ulcer effects by multitarget actions. Twenty compounds were identified and (-)-alpha-santalene was the main compound.

Ethnopharmacological importance Currently, in many traditional communities, such as the riverine community in the North Araguaia microregion (Mato Grosso, Brazil), plant knowledge and use represent the main, if not the only, therapeutic resource for the maintenance of health and/or treatment of diseases. This study aimed to identify and document species of medicinal plants used by local experts from riverine communities in the North Araguaia microregion in Mato Grosso State, and to further chemical and pharmacological studies on species selected based on searches in the relevant literature. Materials and methods This is a cross-sectional ethnobotanical study, with non-probabilistic sampling (n =60), that applied the snowball method to select local riverine experts who understand medicinal plant use. Socio-demographic, ethnobotanical and ethnopharmacological data (vernacular name, uses, geographical origin, habit, method of preparation and part used) on medicinal plants were collected during semi-structured interviews. The results were analyzed by descriptive and quantitative means: indices of use-report (UR) were used to select plant species with therapeutic potential. Results In total, 309 plant species belonging to 86 botanical families were cited; 73% were native to Brazil, and Fabaceae was the most representative family (11.3%). Arboreal was the predominant life form (37.2%). The leaf was the most used part (28.9%). Infusion was the most commonly reported method of preparation (31.3%). The plants reported in the survey were indicated for 18 of the 22 ICD-10 disease categories. The disease categories most commonly cited were the infectious and parasitic diseases (IPD, 718 UR), digestive system diseases (DSD, 565 UR) and respiratory system diseases (RSD, 504 UR), representing 16.6%, 13.1% and 11.7%, respectively of the total UR. Dysphania ambrosioides L. was the most sighted in the IPD category 50 UR. Copaifera langsdorffii Desf. (133), Lafoensia pacari A. St.-Hil. (131), and Cecropia pachystachya Trecul (126) were the species with the highest UR. Bidens pilosa L., Vernonia ferruginea Less, and L. pacari , respectively, were the most cited native plants used to treat such diseases. Of the 8 investigated native plants, C. langsdorffii, and Brosimum gaudichaudii are the most prominent: in addition to having been widely studied, in terms of phytochemical and pharmacological, these species have been marketed as pharmaceutical products, with associated patent deposits. Conclusions Local riverine experts from the North Araguaia microregion use a wide variety of medicinal plants in self-care health, especially those species used to treat IPD. The therapeutic potential of some of these plants has been scientifically validated; however, there are other species whose pharmacological effects and safety remain to be properly investigated. Thus, the present study, aside from being a basis for future chemical, pharmacological and agronomic bioprospecting studies, may contribute to the development of the management, conservation and sustainable use of medicinal flora in the microregion studied.

Ethnopharmacological relevance Piper umbellatum L. (Piperaceae) is a shrub found in the Amazon, Savannah and Atlantic Forest region of Brazil. It is widely used in folk medicine in many countries primarily for the treatment of gastric disorders. The aim of this study was to evaluate the gastroprotective and anti-ulcer effects of hydroethanolic extract of P. umbellatum (HEPu) leaves in experimental rodents. In addition, the anti- Helicobacter pylori activity of the extract was assessed. Materials and methods The leaves of P. umbellatum were macerated in 75% (1:3 w/v) hydroethanolic solution to obtain HEPu. The gastroprotective and ulcer healing activities of HEPu were evaluated using acidified ethanol (acute) and acetic acid (chronic) gastric ulcer models in rodents. The anti- H. pylori activity was evaluated by in vitro broth microdilution assay using H. pylori cagA + and vacA + strain. The probable mechanism of action of HEPu was evaluated by determining gastric secretory parameters, antioxidant enzyme (catalase), non-protein sulfhydryl (glutathione) and malondialdehyde levels in gastric tissue, including pro-inflammatory (IL-1β, TNF-a, IL -17, RANTES, IFN-γ and MIP-2) and anti-inflammatory (IL-10) cytokines. Results HEPu demonstrated potent gastroprotection against acute ulcer induced by acidified ethanol and excellent healing effect of the chronic ulcer induced by acetic acid. The gastroprotective activity in acidified ethanol is partly attributed to the antioxidant mechanisms, while anti-secretory, anti-inflammatory and regeneration of the gastric mucosa are evoked as part of its antiulcer mechanism of action. The gastric ulcer healing of HEPu also involves restoration of the altered cytokines levels to near normal. However, it has no in vitro anti- H. pylori activity. Conclusion The results of this study showed that HEPu possesses preventive and curative effects in experimental models of gastric ulcers in animals. These effects are partially dependent on antioxidant, antisecretory, anti-inflammatory and mucosa regeneration. It is independent of anti- H. pylori activity, with substances probably responsible for the pharmacological activity being flavonoids, quercetin and rutin. These results support the popular use of P. umbellatum leaves in the treatment of peptic ulcers.

A B S T R A C T Introduction: Ficus trichopoda is a ficus species growing in wet places - swamp forest, river banks and swamp grassland. It is well used in the management of inflammation - related conditions locally. This present work was undertaken to investigate anti-inflammatory and analgesic properties of aqueous ethanolic extract of Ficus trichopoda bark in rats. Methods: The anti-inflammatory effect was investigated by using the acute inflammatory model of carrageenan - induced paw oedema and its analgesic activity using formalin test and tail flick test in rats. Results: The study for preliminary phytochemical secondary metabolites revealed the presence of saponins, tannins, alkaloids and free amines/amino acids. Oral administration of the aqueous ethanolic extract of F. trichopoda in doses up to 5000 mg/kg body weight did not produce any mortality and any visible signs of toxicity. Ficus trichopoda extract at the doses (125 - 500 mg/kg) showed significant (p < 0.05) dose dependent inhibition of oedema formation in the carregeenan induced model. The extract showed significant dose dependent inhibition of the inflammatory (late) phase but not the neurogenic (early) phase of the formalin test in rats. The analgesic activity in tail flick method showed significant (p < 0.05) elevation in pain latency threshold from 30, 60, 90 and 120 minutes after pretreatment. Conclusion: The results obtained suggest marked analgesic and anti-inflammatory activity of ethanolic extract (125- 500 mg/kg). This finding supports that the stem bark is useful in inflammatory and painful conditions.

Ethnopharmacological relevance Gallesia integrifolia (Phytolaccaceae) is commonly known as “pau-d’alho” in Brazil or “garlic plant” due to the strong scent of garlic peculiar to all parts of the plant. The bark decoction is used for the treatment of microbial infections among other diseases by different ethnic groups in Brazil, Peruvian Amazonians, Bolivia and Mosetene Indians. This study aimed to advance in the antibacterial activity and characterize the mode of action of the hydroethanolic extract of the inner stem bark of G. integrifolia (HEGi) using in vivo and in vitro experimental models. Materials and methods The qualitative and quantitative phytochemical analyzes of HEGi were carried out using colorimetric and HPLC technique. The cytotoxic potential of HEGi was evaluated against CHO-K1 cells by Alamar blue assay and its acute toxicity was assessed by the Hippocratic screening test using Swiss-Webster mice. The antibacterial activity was evaluated by micro- dilution method against ten strains of Gram-positive and Gram-negative bacteria. The mode of action of HEGi was investigated by outer membrane permeability, nucleotide leakage and potassium efflux assays. In vivo infection model was established by using Staphylococcus aureus infection model Wistar rats. Results Qualitative phytochemical analysis of HEGi revealed the presence of saponins, alkaloids, phenolic compounds and flavonoids. Phytochemical quantification of HEGi showed that higher total phenolic (80.10±0.62 mg GAE/g) and flavonoid (16.10±0.03 mg RE/g) contents. HPLC fingerprint analysis revealed the presence of gallic acid, rutin, and morin. In the Alamar blue assay no cytotoxic effect of HEGi in CHO-K1 cells was observed up to 200 µg/mL, and no signs or symptoms of acute toxicity were observed in mice of both sexes at higher doses of up to 2000 mg/kg, p.o. HEGi demonstrated bacteriostatic effect against selected Gram positive and Gram negative bacterial pathogens. Its mode of action is associated, at least partly, with changes in the permeability of bacterial membranes, evidenced by the increased entry of hydrophobic antibiotic in Pseudomonas aeruginosa , intense K + efflux and nucleotides leakage in Shigella flexneri , Streptococcus pyogenes and S. aureus . HEGi attenuated the experimental blood borne S. aureus infection in rats at all the tested doses levels (10, 50 and 250 mg/kg). Conclusion HEGi is safe at the dose tested when used acutely, and it presented broad antibacterial effect, which support its traditional use in the treatment of bacterial infections. It contains well known important phytochemicals, recognized to be active against bacterial pathogens in vitro and might be collectively responsible for the antibacterial activity of HEGi. It is bacteriostatic in nature, with membrane perturbation being one of it mode of action. HEGi represent a potential phytotherapic antibacterial agent.

Background Vitexin is a flavonoid found in plants of different genus such as Vitex spp. and Crataegus spp. Despite being an important molecule present in phytomedicines and nutraceuticals, the mechanisms supporting its use as anti-inflammatory remains unclear. Purpose To investigate the cellular and molecular mechanisms involved in acute anti-inflammatory effect of vitexin with regard to neutrophil recruitment and macrophages activation. Methods Anti-inflammatory properties of vitexin were evaluated in four models of neutrophil recruitment. The regulation of inflammatory mediators release was assessed in vivo and in vitro. Vitexin (5, 15 and 30 mg/kg p.o) effects on leukocytes migration to peritoneal cavity induced by zymosan (ZY), carrageenan (CG), n-formyl-methionyl-leucyl-phenylalanine (fMLP) and lipopolysaccharide (LPS) were evaluated in Swiss-Webster mice and the effects on the levels of TNF-α, IL-1β and IL-10 cytokines, and NO concentration were in the LPS-peritonitis. RAW 264.7 macrophages viability were determined by Alamar Blue assay as well as the capacity of vitexin in directly reducing the concentrations of TNF-α, IL-1β, IL-10, NO and PGE2. Additionally, vitexin effects upon the transcriptional factors p-p38, p-ERK1/2 and p-JNK were evaluated by western blotting in cells activated with LPS. Results Vitexin was not cytotoxic (IC50 > 200 µg/ml) in RAW 264.7 and at all doses tested it effectively reduced leukocyte migration in vivo, particularly neutrophils in the peritoneal lavage, independently of the inflammatory stimulus used. It also reduced TNF-α, IL-1β and NO releases in the peritoneal cavity of LPS-challenged mice. Vitexin had low cytotoxicity and was able to reduce the releases of TNF-α, IL-1β, NO, PGE2 and increase in IL-10 release by LPS activated RAW 264.7 cells. Vitexin was also able to regulate transcriptional factors for pro-inflammatory mediators, reducing the expression of p-p38, p-ERK1/2 and p-JNK in LPS-elicited cells. Conclusions Vitexin presented no in vitro cytotoxicity. Inhibition of neutrophil migration and pro-inflammatory mediators release contributes to the anti-inflammatory activity of vitexin. These effects are associated with the inactivation of important signaling pathways such as p38, ERK1/2 and JNK, which act on transcription factors for eliciting induction of inflammatory response.

Ethnopharmacological relevance Copaifera malmei Harms (Fabaceae) is a plant that occurs in the central region of Brazil, where the plant's leaves infusion is popularly used to treat gastric ulcer and inflammatory diseases. This study was aimed to investigate the gastroprotective activity and mode of action of the plants' leaves infusion in order to establish the scientific basis for such usage, and to assess its potential as a source of an anti-ulcer agent. Materials and methods Leaves infusion extract of the plant (SIECm) was prepared, freeze dried and lyophilised. Its qualitative and quantitative phytochemical constituents were investigated using TLC and HPLC techniques. The safety profile was evaluated on CHO-k1 epithelial cells viability using the Alamar blue assay, and by acute toxicity test in mice. The gastroprotection and anti-ulcer efficacy of the SIECm (25, 100 and 400 mg/kg, p.o.) were tested using acute (acidified ethanol, piroxicam and water restrain stress), and chronic (acetic acid) experimental ulcer models. The plausible mode of action of the SIECm was assessed using gastric secretion, gastric barrier mucus, nitric oxide, and its antioxidant (myeloperoxidase and catalase) effects in mice and rats. The histopathological analyses of the ulcerated tissues as well as the extract’s activity on Helicobacter pylori were also investigated. Results Phytochemical tests indicated the presence of mainly phytosterols, phenolics and flavonoids. The SIECm exhibited no cytotoxic effects on the CHO-k1 cells, and no oral acute toxicity in mice. It prevented against the acute induced ulcerations by enhancing gastroprotection through gastric mucus production, NO modulation, antioxidant, reduced gastric secretion and enhanced chronic ulcers healing process, as shown by reduction/prevention of epithelial and vascular damage, in addition to reduction in leucocyte infiltration. The SIECm however did not exhibit activity against H. pylori. Conclusion The SIECm is safe, contain useful phytochemicals and exhibited significant gastroprotective/anti-ulcer effects. The results justify its folkloric usage, and provided scientific evidence of its potential as a source of new phytodrug to treat gastric ulcers.

Ethnopharmacological importance The use of medicinal plants for treatment, cure and prevention of diseases has been described by many people since time immemorial. Because of this use, commercial and scientific interests have emerged, making it necessary to realize ethnobotanical surveys of medicinal plants species, which is important for subsequent chemical and pharmacological bioprospections. Aim of the study This study aimed at surveying, identifying, cataloging and documenting the medicinal plants species used in the Valley of Juruena, Northwestern Mato Grosso, Legal Amazon Brazil for the treatment of various human diseases, as well as assessed the species of interest for bioprospecting potential. Materials and methods Informants were interviewed using semi-structured form to capture information on socio-demographic and ethnopharmacological data of medicinal plants such as vernacular name, uses, geographic origin, habit, form of preparation and part used. Results were analyzed using descriptive and quantitative means: indices of use-report (Ur) and informant consensus factor (ICF), for the selection of plant species with therapeutic potential. Results Three hundred and thirty two (332) plants species belonging to 90 families were reported for medicinal purposes and totaling 3973 use-reports were reported by 365 (92.9%) of the people interviewed. Asteraceae (32.2%), Fabaceae (26.7%) and Lamiaceae (24.4%) families were the most represented, with majority being species native (64.45%) to Brazil. Leaves (64.5%) were the part of the plant most used and infusion (45.7%) was the most utilized form. Gastrointestinal disorders followed by respiratory complaints topped the list of use-reports. The native or naturalized plants with the highest use reports in the order of decreasing absolute frequency per each emic-category are Cymbopogon citratus (DC.) Stapfc (104), Mentha pulegium L. (94), Arrabidaea chica (Humb. & Bonpl.) B. Verl. (97), Alternanthera brasiliana (L.) Kuntze (71), Baccharis crispa Spreng (57), Phyllanthus niruri L. (48), Gossypium barbadense L. (44), Solidago microglossa DC. (40) and Bauhinia forficata L. (20). And the most cited exotics are: Chenopodium ambrosioides L. (151), Aloe vera (L.) Burm. f., (89) and Rosmarinus officinalis L. (72). In some cases, high ICF values were found, which reflects high degree of homogeneity of consensus among informants in this region on medicinal plants. Conclusion The population of Valle of Juruena makes use of a wide array of medicinal plants distributed in all use categories with predominance of those use in the treatments of gastrointestinal and respiratory ailments. The therapeutic potential of some of the species of medicinal importance extensively utilized by the population of the region have been scientifically validated, and are therefore promising prototype of new drugs. However, there are some of these species whose ethnomedicinal uses are yet to be scientifically verified and thus constitute an unexplored terrain for future biological/pharmacological studies.

Objective: The purpose of this study was to evaluate selected Brazilian plants from Juruena valley region of Mato Grosso, for their in vitro antimicrobial and antioxidant activities. Methods: The powder obtained from different parts of the twenty-six (26) plants were macerated in hydroethanolic solution to obtain the extracts. The hydroethanolic extracts were tested for their in vitro antimicrobial activity by determining the MIC using broth microdilution. The 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical, ferric reducing antioxidant power (FRAP) and nitric oxide (NO) methods were used for the determination of antioxidant activities. Correlation between classes of secondary metabolites and antioxidant activity was assessed. Results: Phanera glabra extract (HEPg) showed broad antibacterial spectrum, presenting the best activity against Klebsiella pneumoniae . Hevea microphylla extract (HEHm) presented a narrow spectrum of antibacterial activity with strong effect against Shigella flexneri . The only plant with broad spectrum antifungal activity was Bertholletia excelsa (HEBe), with moderate activity against strains of Aspergillus and Candida . The following extracts were prominent regarding their activities in the DPPH and FRAP assays-HEBe, Cariniana rubra (HECr) and in the FRAP assay alone, Cedrela odorata (HECo) and HEPg. None of the extracts was active in the NO assay. A significant association was observed between DPPH activity and the total phenolic contents. Conclusion: Our results justified the use of some of the investigated plants in the Brazilian ethnomedicine. The antibacterial activities of these plants are bacteriostatic in nature. These findings support that a number of investigated plants could be a valuable source of new antioxidant and antimicrobial compounds that can potentially deliver novel mechanisms of actions.However, further studies are required.

Ethnopharmacological relevance Helicteres sacarolha A. St.- Hil. et al. popularly known in Brazil as ‘semente-de-macaco’, is widely employed in the popular medicine in many of parts of Brazil in the alleviation of symptoms of ailments such as peptic ulcer and inflammation. Up to the present, there is no study addressing the gastroprotective activity of the hydroethanolic extract of H. sacarolha and its possible mechanism of actions. Materials and methods The hydroethanolic (70%) extract of H. sacarolha (HEHs) was obtained by maceration. The gastroprotective activity was assessed using gastric ulcer models induced by acidified ethanol, piroxicam, and water restraint stress in mice and rats at doses of 20, 50 and 250 mg/kg p.o. Mechanistic studies involved the antisecretory assay evaluated with pylorus ligation in rats and pre-treatments with appropriate antagonists/inhibitors such as yohimbine, glibenclamide, indomethacin and l -NAME, effect on catalase and myeloperoxidase activities and gastric mucus determination using acidified ethanol- induced ulcer in mice. Results HEHs at all doses tested demonstrated potent gastroprotective activities in the acute ulcer models. The gastroprotective activity of HEHs was attenuated by pre-treatments with yohimbine, glibenclamide, indomethacin and l -NAME. HEHs effectively reduced basal gastric juice production without any effect on the free and total acidity. The gastroprotective action of HEHs involved increasing the antioxidant enzyme catalase and mucus secretion and inhibition of neutrophyl infiltration as reflected by the reduction in the myeloperoxidase activity. Conclusion The results of this study gave a scientific support for the popular use of the leaves of H. sacarolha in the treatment of gastric ulcers and that it has a multi-targeted action.

Ethnopharmacological relevance Cedrela odorata L. (Meliaceae) is a native plant of the Amazon region and its inner stem bark is used in the treatment of diabetes in the form of maceration in Brazilian popular medicine. Until now, there is no scientific study on this activity. The present study was aimed at evaluating the anti-hyperglycemic activity, anti-diabetic, toxicity, antioxidant and potential mechanism of action of hydroethanolic extract of the inner stem bark of Cedrela odorata . Material and methods The inner stem bark extract of Cedrela odorata was prepared by maceration in 70% ethanol for 7 days to obtain hydroethanolic extract of Cedrela odorata (HeECo). The preliminary phytochemical analysis was performed according to procedures described in the literature. Selected secondary metabolites detected were quantified by high performance liquid chromatography (HPLC). Acute toxicity of HeECo was investigated in male and female mice with oral administration of graded doses of HeECo from 10 to 5000 mg/kg. Subchronic oral toxicity study was done by oral administration of HeECo (500 mg/kg) and vehicle for 30 days to both sexes of Wistar rats. Clinical observations and toxicological related parameters were determined. Blood was collected for biochemical and hematological analyses, while histological examinations were performed on selected organs. Anti-hiperglycemic and antidiabetic effects were evaluated in streptozotocin-induced diabetic rats. In acute evaluation, the animals received pretreatment with 250 and 500 mg/kg of HeECo, before carbohydrate overload. For subchronic effect, the antidiabetic activity of HeECo was evaluated using the same doses for 21 days. At the end of the treatments, the levels of triacylglycerols, malondialdehyde, total antioxidant status, superoxide dismutase and glutathione peroxidase activities were evaluated in the plasma. Results The extract showed low acute toxicity. HeECo exhibited inhibitory activity against α-glucosidase and caused a lowering in the peak levels of blood glucose in animals that received glucose overload by 36.7% and 24.1% in the area under the glucose curve (AUC). When the overload was sucrose, HeECo reduced the blood glucose level by 44.4% without affecting AUC. Treatment with HeECo of the blood glucose of the diabetic animals for 21 days did not lead to improvement in weight gain and regularization of the blood glucose level, but reduced the triacylglycerol and malondialdehyde levels by 36.6% and 48.1%, respectively. The activity of the antioxidant enzymes, superoxide dismutase and glutathione peroxidase were significantly increased when compared to diabetic control rats. HPLC analysis showed the presence of polyphenols, such as gallic acid, (−)- gallocatechin and (+)- catechin, the latter is present in higher quantity. Conclusions Collectively, these data showed that HeECo could blunt the postprandial glycemic surge in rats; possibly through inhibition of alpha-glucosidase and positive modulation of antioxidant enzymes. Our findings confirmed the anti-hiperglycemic activity of HeECo in STZ- diabetic rats. Cedrela odorata is effective in diminishing glucose levels in vitro and in vivo and in ameliorating oxidative damage that occurs in diabetes and/or due to hyperglycemia in rats. According to our results, the efficacy of Cedrela odorata preparation could be due to the presence of active principles with different mode of actions at the molecular level, including α-glycosidases and glucose transporter inhibitors and antioxidant property.

Ethnopharmacological relevance Helicteres sacarolha , popularly known in Brazil as ‘rosquinha’, ‘sacarolhas’, ‘semente-de-macaco’, is widely distributed in different phytogeographic zones in Brazil. Preparations from its roots and leaves are employed in popular Brazilian medicine in the treatments of ailments such as peptic ulcer, hypertension among others. Cytotoxicity, acute oral and subchronic toxicity of the hydroethanolic extract of Helicteres sacrolha was investigated as well as the classes of phytochemical present in the extract. Materials and methods Hydroethanolic (70%) extract of Helicteres sacarolha (HEHs) was prepared by maceration. Potential cytotoxicity was evaluated in CHO-k1 cells. Acute administration of HEHs was done in mice as a single dose up to 5000 mg/kg and subchronic oral toxicity study for 30 days in Wistar rats at daily oral doses of 0, 250 and 750 mg/kg b.w. Clinical observations and toxicological related parameters were determined every 6 days. Blood was collected for biochemical and hematological analyses, while histological examinations were performed on selected organs. Selected secondary metabolites detected were quantified by UV-spectrophotometry and high performance liquid chromatography (HPLC). Results The extract was non-cytotoxic to CHO-k1 cells. In acute oral toxicity, there was no mortality or clinical alterations in the female mice, at all doses, except for the transient diarrhea observed at 5000 mg/kg acute. Doses up to 2000 mg/kg caused no mortality or treatment-related clinical manifestations in the male mice, but treatment-related alterations were however observed at 4000 mg/kg, with mortalities recorded at 5000 mg/kg. During the subchronic oral toxicity study, no mortality or treatment-related clinical signs were observed. Differences in relative organ weight, hematological parameters and histopathology observations between the treated and the control groups were considered not to be treatment-related. Spectrophotometric analysis revealed the presence of relatively high content of phenolics and flavonoids in HEHs. HPLC analysis confirmed the presence of the quantified compounds and demonstrated the presence of ellagic acid, morin and naringin. Conclusion Our results confirmed that HEHs have a broad safety margin for therapeutic use.

Ethnopharmacological relevance Dilodendron bipinnatum Radlk., Sapindaceae, is popularly known as ‘mulher-pobre’. Its stem bark macerate or decoction is popularly used mainly to treat uterine inflammation and bone fractures. Acute oral and subchronic toxicities of the hydroethanolic extract of Dilodendron bipinnatum (HEDb) were investigated as well as the classes of phytochemical present in the extract. Materials and methods Acute toxicity of HEDb was investigated by hippocratic screening in Swiss mice, cytotoxicity and potential genotoxic effect were evaluated with micronucleus test in the CHO-k1cells. Subchronic oral toxicity of HEDb was assessed in Wistar rats with graded doses of HEDb repeatedly administered for 30 days, in order to evaluate the behavioral changes, weight gain, water and feed consumption, urine and feces excretion, hematological and biochemical parameters and histopathological examinations of vital organs. Results In hippocratic screening doses up to 5000 mg/kg p.o. did not cause any changes in female mice, while signs of reduction in motility, increased respiratory rate and tail erection were observed in male mice but were all reversed within an hour. In cytotoxicity assay, IC50 was 118±1.55 µg/mL in CHO-k1 cells. Micronucleus test demonstrated that the plant extract is non-genotoxic. In subchronic toxicity studies, minor change was observed as an increase in the urine output in the last 18 days of treatment with 500 mg/kg of HEDb, while other parameters remained unchanged. All alterations observed were discrete and minor in nature and were not accompanied by any relevant clinical signs or any toxicologically significant biochemical, hematological and histopathological changes. Preliminary phytochemical analysis revealed the presence of aurones, chalcones, coumarins, flavonones, flavononoids, phenols, and saponins. Conclusion The results demonstrate high safety profile of HEDb in both experimental animals and cell models assays. However, there may be risk of hepatotoxicity in the use of the extract at high doses for a prolonged period.

Macrosiphonia longiflora (Desf.) Müll. Arg (Apocynaceae), popularly known as 'velame' and 'velame branco', is a native subshrub that grows in the Brazilian Cerrado. This plant is widely used in traditional medicine in the form of decoction and infusion, particularly as anti-inflammatory, depurative, anti-rheumatic, antisyphilitic and antiulcer remedy. There is no available information in the literature that has addressed its pharmacological activity and phytochemical analysis.This study aimed to evaluate the anti-inflammatory pharmacological profile of the hydroethanolic extract of Macrosiphonia longiflora, using in vivo and in vitro acute inflammation experimental models, as well as investigate the roles of cytokines and nitric oxide in its mechanism of action, and including phytochemical analysis constitution of its hydroethanolic extract.Hydroethanolic (70%) extract of Macrosiphonia longiflora (HEMl) was prepared by maceration. The preliminary phytochemical analysis was performed according to procedures described in the literature. Selected secondary metabolites detected were quantified by spectrophotometry and high performance liquid chromatography (HPLC). Its cytotoxic potential in Chinese hamster ovary (CHO-k1) epithelial cell lines was evaluated using Alamar Blue. in vivo anti-inflammatory activity was evaluated with carrageenan- and dextran-induced paw edemas, carrageenan-induced pleurisy in rats and lipopolysaccharide (LPS)-induced peritonitis in mice. The in vitro anti-inflammatory activity was evaluated using RAW 264.7 cells stimulated with LPS and interferon (INF)-γ. Effects of HEMl on the inflammatory cytokines (IL-1β, IL-10, IL-17, INF-γ and TNF-α) concentrations in the peritoneal lavage were evaluated using commercial ELISA kits, while the Griess method was employed to determine nitric oxide (NO) concentrations in the peritoneal lavage, as well as in the supernatants of RAW 264.7 cells.Preliminary phytochemical analysis, revealed the presence of phenolics compounds, terpenoids, alkaloids and flavonoids. Spectrophotometric analysis revealed the presence of relatively high content of phenolics and flavonoids in HEMl. HPLC analysis confirmed the presence of the quantified compounds and demonstrated the presence of ellagic acid in the detected matrix of compounds. HEMl appeared to be non-cytotoxic. It effectively inhibited (p0.05) paw edema induced by carrageenan and dextran. Furthermore, HEMl also significantly reduced exudates volume and leukocyte migration in the carrageenan-induced pleurisy and LPS-induced peritonitis, neutrophils counts in LPS-induced peritonitis. HEMl also acts by effectively inhibiting the following inflammatory cytokines: IL-1β and IL-10 levels in the peritoneal lavage, but had no effect on IL-17 level in the peritonitis model. In addition, HEMl had no effect on the levels of tumor necrosis factor alpha (TNF-α) present in the peritoneal lavage and cells supernatants. The concentration of NO, as assessed by measurement of nitrite (NO2(-)), showed that pretreatment with HEMl reduced NO significantly in the peritoneal lavage and in RAW 264.7 cells co-stimulated with LPS and INF-γ.The results obtained in this study indicate that HEMl possesses very low cytotoxic potential. In addition, it demonstrated a potent anti-inflammatory activity in both the in vivo and in vitro models of acute inflammation. The anti-inflammatory effect is partly related to the inhibition of IL-1β, IL-10, and nitric oxide releases, but independent of TNF-α and IL-17 modulation. Phytochemical analysis revealed the predominant presence of the flavonoids (naringin, rutin, myricetin, morin, quercetin, (±)-naringenin, and luteolin) and phenols (ellagic acid), which are possibly involved in the anti-inflammatory effect of HEMl. The current study provided supportive evidence for the popular use of HEMl in the treatment of inflammatory conditions, and shed more light on the possible roles of the inflammatory cytokines in its mechanisms of action as anti-inflammatory agent.

Ellagic acid (EA), a plant-derived polyphenol, exhibits antioxidant, anti-inflammatory, and gastroprotective effects. Its gastroprotective mechanisms have not been fully elucidated nor have its effects on chronic ulcer previously been described. Toward these ends, the antiulcer activities of EA were evaluated in acute (ethanol and indomethacin) and chronic (acetic acid) ulcer models in Wistar rats. In this study, oral administration of EA significantly prevented the gastric ulceration caused by ethanol, indomethacin, and acetic acid treatments. Its gastroprotective mechanism in ethanol-induced ulcer were partly due to intensification in the endogenous production of nitric oxide, an antioxidant effect by replenishing depletion of endogenous nonprotein sulfhydryls and attenuation of tumor necrosis factor-α increase, whereas in indomethacin ulcer, it is partly due to a reduction in the plasma level of leukotriene B(4). In acetic acid ulcer, promotion of ulcer-healing effects was partly due to attenuation of the elevated levels of the inflammatory cytokines TNF-α, interferon-γ, and interleukins-4 and -6. These findings suggest that ellagic acid exerts its antiulcer activity by strengthening the defensive factors and attenuating the offensive factors.

Relevance Simaba ferruginea A. St-Hil. (Simaroubaceae) is a subshrub typical of the Brazilian Cerrado, whose rhizomes are popularly used as infusion or decoction for the treatment of gastric ulcers, diarrhea and fever. Aim of the study To evaluate the pharmacological mechanism(s) of action of the antiulcer effects of the methanol extract of Simaba ferruginea and its alkaloid canthin-6-one. Materials and methods Rhizome of Simaba ferruginea was macerated with methanol to obtain the methanol extract (MESf) from which was obtained, the chloroform fraction. Canthin-6-one alkaloid (Cant) was purified and then isolated from the chloroform fraction (CFSf). The isolated Cant was identified by HPLC. Anti-ulcer assays were determined using ethanol and indomethacin-induced ulcer models in mice and rats respectively. In order to determine the probable mechanisms of actions of MESf and Cant animals were pretreated with l -NAME prior to anti-ulcer agent treatments and ulcer induction and nitric oxide (NO) level determined in order to assess NO involvement in the gastroprotective effects. Assays of malondialdehyde (MDA), myeloperoxidase (MPO), pro-inflammatory cytokines: interleukin 8 (IL-8) and tumor necrosis factor-alpha (TNF-α) and prostaglandin E2 (PGE2) were also carried out according to previously described methods. Results The results indicate that the antiulcerogenic effects of MESf and Cant in ethanol-induced ulcer is mediated in part through increase in the production of protective endogenous NO as the antiulcerogenic activity of MESf and Cant was reduced in animals pre-treated with l -NAME. In indomethacin-induced ulcer pre-treatment with MESf and Cant showed reduction in the levels of MPO and MDA in the gastric tissue, thus indicating the participation of the antioxidant mechanisms on the gastroprotective effects. The plasma levels of IL-8 in ulcerated rats with indomethacin were also reduced by Cant, but not by MESf, indicating that inhibition of this cytokine contributes to the gastroprotective effect of Cant. However MESf and Cant had no effect on the mucosal membrane levels of PGE2, indicating that the gastroprotective effects of these agents is independent of PGE2 modulation. Conclusion The results obtained in this study with MESf and Cant added insights into the pharmacological mechanisms involved in their mode of antiulcer action. The results indicate that Cant is one of the compounds responsible for these effects. Such findings are of extreme importance in the strive for future development of potent, safer and effective antiulcer agent. The efficacy of MESf and Cant in gastroprotection shows that Simaba ferruginea might be a promising antiulcer herbal medicine, in addition to confirming the popular use of this plant against gastric ulcer models utilised in this study.

Ethnopharmacological relevanceDilodendron bipinnatum Radlk. (Sapindaceae), popularly known as “mulher-pobre”, is a native tree of the Pantanal of Mato Grosso, Brazil. The stem bark of Dilodendron bipinnatum is used by the population, in the forms of decoction and maceration in the treatment of inflammatory conditions. There is no information in the literature demonstrating the anti-inflammatory activity of Dilodendron bipinnatum and its respective mechanism of action. This study aimed to evaluate the anti-inflammatory activity and mechanism of action of the hydroethanolic extract of the stem bark of Dilodendron bipinnatum (HEDb) using in vivo and in vitro experimental models.Materials and methodsThe stem bark of Dilodendron bipinnatum was macerated in 70% hydroethanolic solution (1:3, w/v) for 7 days, filtered, concentrated on a rotary evaporator and the residual solvent removed in oven at 40°C, thus obtaining HEDb. Cytotoxicity of HEDb in RAW 264.7 was assessed by the Alamar blue assay. in vivo anti-inflammatory activity of HEDb was evaluated with carrageenan and dextran-induced paw edemas and lipopolysaccharide (LPS)-induced peritonitis in mice. Effects of HEDb on the inflammatory cytokines (TNF-α, IL-1β and IL-10) concentrations in the peritoneal fluid were evaluated using commercial ELISA kits. The in vitro anti-inflammatory activity was evaluated using RAW 264.7 cells stimulated with LPS and/or INF-γ, while a Griess method was employed to determine nitric oxide (NO) concentrations in the peritoneal lavage and in the supernatants of RAW 264.7 cells. Preliminary phytochemical analysis was carried out using classical methods and secondary metabolites detected on HEDb were analyzed and confirmed by high performance liquid chromatography (HPLC).ResultsHEDb showed very low cytotoxicity with IC50>200±0.38μg/mL. HEDb effectively inhibited paw edema by carrageenan in the 2nd hour at 20mg/kg (36%, p