Scielo RSS <![CDATA[Revista Brasileira de Farmacognosia]]> vol. 28 num. 6 lang. en <![CDATA[SciELO Logo]]> <![CDATA[Microscopic characterization and HPTLC of the leaves, stems and roots of <em>Fadogia agrestis</em> – an African folk medicinal plant]]> ABSTRACT Fadogia agrestis Schweinf. ex Hiern (Vangueria agrestis (Schweinf. ex Hiern) Lantz), Rubiaceae, is an African traditional medicinal plant also used as a dietary supplement in the US. The present paper is the first report of the pharmacognostic study of the leaf, stem and root of F. agrestis by microscopy, HPTLC and total phenolic/flavonoid content analyses. Noteworthy microscopic features that can help in identification and quality control are septate and lignified non-glandular trichomes on leaf and stem epidermises, paracytic stomata on leaf abaxial epidermis, numerous cells containing yellow substances of presumably phenolic compounds in leaf and stem, calcium oxalate druses and prismatic crystals in leaf and styloids in stem, primary phloem fibers in stem, brachysclereids in stem and root, spherical starch grains in root, and vessels with vestured pits and simple perforated end walls. In addition to microscopy, a total phenolic/flavonoid content determination and an HPTLC method were also developed for rapid chemical fingerprint analyses of Fadogia samples and dietary supplements. <![CDATA[Anatomy and microscopic characteristics of <em>Picris japonica</em>]]> ABSTRACT Picris japonica Thunb., Asteraceae, is an herbal medicine used to dispel heat, reduce swelling and alleviate pain in traditional Mongolian medicine. Its dried whole plant is mainly used to treat flu and mammary abscesses. Given the potential applications of such an herb, detailed pharmacognostic research on P. japonica is needed. This study attempted to fill this need by producing permanent and semi-permanent slides of different organs (root, stem, leaf, pollen grain, fruit and powder of the whole plant) using safranine staining, safranine-fast green double staining and common methods. Furthermore, several featured microscopic structures of P. japonica are described herein. The results obtained provide us with valuable information for botanical quality control and species identification and enable us to detect adulterations in commercial samples of Picris or in laboratory samples. <![CDATA[Combined OPLS-DA and decision tree as a strategy to identify antimicrobial biomarkers of volatile oils analyzed by gas chromatography–mass spectrometry]]> ABSTRACT Bioguided isolation to discriminate antimicrobial compounds from volatile oils is a time- and money-consuming process. Considering the limitations of the classical methods, it would be a great improvement to use chemometric techniques to identify putative biomarkers from volatile oils. For this purpose, antimicrobial assays of volatile oils extracted from different plant species were carried out against Streptococcus mutans. Eight volatile oils that showed different antimicrobial effects (inactive, weakly active, moderately active and very active) were selected in this work. The volatile oils' composition was determined by GC–MS-based metabolomic analysis. Orthogonal projection to latent structures discriminant analysis and decision tree were carried out to access the metabolites that were highly correlated with a good antimicrobial activity. Initially, the GC–MS metabolomic data were pretreated by different methods such as centering, autoscaling, Pareto scaling, level scaling and power transformation. The level scaling was selected by orthogonal projection to latent structures discriminant analysis as the best pretreatment according to the validation results. Based on this data, decision tree was also carried out using the same pretreatment. Both techniques (orthogonal projection to latent structures discriminant analysis and decision tree) pointed palmitic acid as a discriminant biomarker for the antimicrobial activity of the volatile oils against S. mutans. Additionally, orthogonal projection to latent structures discriminant analysis and decision tree predicted as "very active" the antimicrobial activity of volatile oils, which did not belong to the training group. This predicted result is in agreement with our experimental result (MIC = 31.25 µg ml−1). The present study can contribute to the development of useful strategies to help identifying antimicrobial constituents of complex oils. <![CDATA[Isocryptolepine, an indoloquinoline alkaloid from <em>Cryptolepis sanguinolenta</em> promotes LDL uptake in HepG2 cells]]> ABSTRACT About 31 percent of deaths worldwide result from atherosclerotic cardiovascular disease. Hyperlipidemia remains the major risk factor for this disease and therefore, it is necessary to identify antihyperlipidemic compounds for drug development. The crude ethanolic extract of Cryptolepis sanguinolenta (Lindl.) Schltr., Apocynaceae, has demonstrated antihyperlipidemic properties. However, the chemical constituents responsible for this action are unknown. Hence, to identify chemical constituent(s) of C. sanguinolenta with anti-hyperlipidemic effect, five indoloquinoline alkaloids were isolated and evaluated in 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate labeled low density lipoprotein uptake assay using HepG2 cells. The minor alkaloid, isocryptolepine, showed strong activity in promoting low lipid lipoprotein uptake by 1.85-fold. Isocryptolepine may, therefore, serve as a lead compound for future studies in the development of novel antihyperlipidemic drugs. <![CDATA[Application of high performance liquid chromatography and Fourier-transform infrared spectroscopy techniques for evaluating the stability of <em>Orthosiphon aristatus</em> ethanolic extract and its nano liposomes]]> ABSTRACT Orthosiphon aristatus (Blume) Miq., Lamiaceae, is a medicinal plant from Southeast Asia. Pharmacological effects of O. aristatus are attributed to the presence of lipophilic flavones. This study aimed to carry out accelerated stability studies on O. aristatus ethanolic extract and its nano liposomes. The extracts were exposed to four different temperatures at 30, 40, 50 and 60 °C for 6 months. The samples were analyzed at 0, 1, 2, 3, 4, 5 and 6 months by high performance liquid chromatography using rosmarinic acid, 3′-hydroxy-5,6,7,4′-tetramethoxyflavone, sinensetin and eupatorin as markers. Different chemical kinetic parameters of the markers were evaluated by Arrhenius equation to predict shelf life (t90) at different storage conditions and at room temperature. Moreover, the stability of O. aristatus ethanolic extract and O. aristatus nano liposomes were analyzes by chemical fingerprinting using FTIR spectroscopy, principal component analysis and hierarchical clustering analysis. The degradation of markers in both O. aristatus ethanolic extract and O. aristatus nano liposomes followed the first order degradation reaction (dependening on their initial concentration). The loss of marker compounds in O. aristatus ethanolic extract, stored at 30, 40, 50 and 60 °C for six months were up to 25, 52, 72 and 89% for all compounds, respectively. However, in O. aristatus nano liposomes 16, 71, 85 and 100% of compounds were lost during 6 months of storage at 30, 40, 50 and 60 °C, respectively. Therefore, the markers in O. aristatus nano liposomes seems to be more stable at a temperature below 30 °C compared to O. aristatus ethanolic extract. However, markers present in O. aristatus ethanolic extract are more stable at a higher temperature (above 30 °C). principal component analysis or hierarchical clustering analysis analyses were applied to the FTIR results in order to demonstrate the discrimination between extracts based on the storage conditions. The results show that the functional group of the components in the extracts and their chemistry relationship is influenced by the temperature setup indicating the extracts are not stable during the storage conditions. <![CDATA[Development and validation of reverse phase high performance liquid chromatography method for the determination of delta-9-tetrahydrocannabinol and cannabidiol in oromucosal spray from cannabis extract]]> ABSTRACT A simple, fast, precise, accurate and responsive high performance liquid chromatography method was developed and validated carefully for determining an amount of delta-9-tetrahydrocannabinol and cannabidiol. A reverse phase Zorbax C-18 column 4.6 mm × 100 mm, 3.5 µm was eluted by using a mixture (85:15) of methanol and water as the mobile phase in an isocratic system with a flow rate of 1.0 ml/min and the injection volume was 10 µl, at a wavelength of 220 nm. The method was developed and validated through linearity, accuracy, precision and detection and quantitation limits studies. A good linear relationship (R2) of delta-9-tetrahydrocannabinol and cannabidiol were 0.9998 and 0.9995, respectively. The obtained % recoveries were found to be 98.3% and 96.5%. The relative standard deviations values of peak areas were found to range from 0.20% to 2.55% and 0.30% to 3.28% for delta-9-tetrahydrocannabinol and cannabidiol, respectively. Delta-9-tetrahydrocannabinol and cannabidiol presented limits of detection of 0.12, 0.23 µg/ml and limits of quantitation of 0.40, 0.76 µg/ml. The developed method could be employed for quantitative analysis of cannabis extract and oromucosal spray formulation. <![CDATA[In vitro activities of glycoalkaloids from the Solanum lycocarpum against Leishmania infantum]]> ABSTRACT Leishmania infantum is an etiologic agent of visceral leishmaniasis. This disease is a neglected disease that can be fatal if not treated and additionally, the few therapeutic option present several drawbacks, including difficult route of administration and toxicity, which turn the search for new therapeutic alternatives necessary. Herein, we evaluated the leishmanicidal in vitro activity of the solanum extract from Solanum lycocarpum A. St.-Hil., Solanaceae, and the isolated alkaloids solasodine, solamargine and solasonine against promastigotes and intracellular amastigotes of L. infantum. Solasodine (IC50-pro = 4.7 µg/ml; IC50-ama = 10.8 µg/ml) and solamargine (IC50-pro = 8.1 µg/ml; IC50-ama = 3.0 µg/ml) exhibited interesting leishmanicidal ativity. Solasonine was approximately four-times (Selective Index 3.7) more selective to the parasite than to the host cells. This data suggest that solasonine might be considered as a potential drug candidate for leishmaniasis treatment. <![CDATA[<em>Caryocar brasiliense</em> induces vasorelaxation through endothelial Ca<sup>2+</sup>/calmodulin and PI3K/Akt/eNOS-dependent signaling pathways in rats]]> ABSTRACT Caryocar brasiliense Cambess., Caryocaraceae (pequi) is a typical Brazilian Cerrado tree. A previous study showed that the butanolic fraction of pequi leaves promotes endothelium-dependent relaxation mediated by nitric oxide and that it causes reversible hypotension in rats. In the present study, we investigated the cell signaling pathways associated with the butanolic fraction-induced nitric oxide release, and we characterized the chemical composition of its fraction. Vascular reactivity tests, a western blotting analysis, and a chemiluminescence assay were used to investigate the signaling pathways involved in the vasorelaxant effect of the butanolic fraction. Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry was used to characterize the butanolic fraction chemical composition. Vasorelaxation was mediated through the activation of the calmodulin and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathways, leading to subsequent endothelial nitric oxide synthase phosphorylation and nitric oxide production, as evidenced by western blotting and chemiluminescence assays, respectively. The chemical characterization of the butanolic fraction revealed the presence of 72 oxygenated compounds, whose molecular formulae are compatible with phenolic compounds, suggesting a potential contribution of these compounds for the butanolic fraction vasorelaxant effect. These findings show that the calmodulin and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathways are involved in the butanolic fraction-induced endothelial nitric oxide synthase activation and are promoted by polyphenol compounds present in the C. brasiliense leaves. <![CDATA[Virtual screening of secondary metabolites of the genus <em>Solanum</em> with potential antimicrobial activity]]> ABSTRACT Infectious diseases are a health problem today and have high mortality rates with a wide diversity of potentially pathogenic microorganisms. Research that is based either on the search for new drugs from plants or on the improvement of phytotherapeutics is prominent and continues to play an important role nowadays. From this perspective, use of in silico studies to carry out investigations of new molecules potentially active for methicillin-resistant Staphylococcus aureus and Escherichia coli using an in-house database with 421 different secondary metabolites selected from the literature from Solanum genus was performed. We also realized an in vitro study with strains of S. aureus and E. coli and compared the results. Two databases from ChEMBL were selected, the first one with activity against methicillin-resistant S. aureus and another against E. coli. The compounds were classified according to the pIC50 values to generate and validate the model using a "Random Forest". The "Random Forest" prediction model for methicillin-resistant S. aureus obtained an accuracy of 81%, area under the Receiver Operating Characteristic curve of 0.885, selecting eight molecules with an active potential above 60%. The prediction model for E. coli obtained an accuracy rate of 88%, area under the Receiver Operating Characteristic curve of 0.932, selecting four molecules with potential probability above 84%. Rutin proved to be potentially active in the in silico study for S. aureus and E. coli. Microbiological tests have shown that rutin has activity only for E. coli. An interaction study with strains of S. aureus ATCC 25923, a standard strain sensitive to all antibiotics, and SAM-01, a multidrug-resistant strain, was designed. There was interaction only between rutin and oxacillin, one of the three antibiotics studied in the interaction, for the strain SAM-01, reducing the resistance of this strain. <![CDATA[Extract from byproduct <em>Psidium guajava</em> standardized in ellagic acid: additivation of the <em>in vitro</em> photoprotective efficacy of a cosmetic formulation]]> ABSTRACT The fruits of guava, Psidium guajava L., Myrtaceae, are cultivated as food and used in agroindustries, generating byproducts or waste that represent environmental problems and require adequate destination. However, these byproducts present high levels of secondary metabolites and have been awakened interest regarding to its reusing. The extract was standardized in ellagic acid concentration by high performance liquid chromatography. The additivation capacity in the in vitro photoprotective efficacy of guava byproduct extract standardized in ellagic acid was verified as a result of its incorporation in cosmetic formulations, comparing it with a standard product. The extract presented synergy with the chemical UV filter (ethylhexyl methoxycinnamate), enhancing the solar protection factor of the phytocosmetic in 17.99%. Besides that, it was possible to show its antioxidant activity and the presence of secondary metabolites such as phenols and flavonoids. According to the results, it is possible to claim that the extract from the guava's agroindustrial byproducts present potential to be studied and reused, applying on the development of innovative products intended to the photoprotection care. <![CDATA[Antifungal activity of extracts and phenolic compounds from <em>Deguelia duckeana</em>]]> ABSTRACT Candida spp. is associated with almost 80% of all nosocomial fungal infections and is considered a major cause of blood stream infections. In humans, Cryptococcosis is a disease of the lungs caused by the fungi Cryptococcus gattii and Cryptococcus neoformans. It can be potentially fatal, especially in immune-compromised patients. In a search for antifungal drugs, Deguelia duckeana extracts were assayed against these two fungi and also against Candida albicans, which causes candidiasis. Hexane branches and CH2Cl2 root extracts as well as the substances 4-hydroxylonchocarpine, 3,5,4′-trimethoxy-4-prenylstilbene and 3′,4′-methylenedioxy-7-methoxyflavone were assayed to determine the minimal inhibitory concentration. Phytochemical study of CH2Cl2 root and hexane branch extracts from D. duckeana A.M.G. Azevedo, Fabaceae, resulted in the isolation and characterization of nine phenolic compounds: 4-hydroxyderricine, 4-hydroxylonchocarpine, 3′,4′,7-trimethoxy-flavonol, 5,4′-dihydroxy-isolonchocarpine, 4-hydroxyderricidine, derricidine, 3,5,4′-trimethoxy-stilbene, 3′,4′,7-trimethoxyflavone and yangambin. The only active extract was a CH2Cl2 root showing minimal inhibitory concentration 800 µg/ml against C. gattii, and the investigation of compounds obtained from this extract showed that 4-hydroxylonchocarpine was active against all three fungi (C. neoformans, C. gattii and C. albicans). These results suggest that D. duckeana extracts have potential therapeutic value for the treatment of pathogenic fungi. <![CDATA[Antiproliferative potential of solidagenone isolated of <em>Solidago chilensis</em>]]> ABSTRACT Plants are considered among the main sources of biologically active chemicals. The species Solidago chilensis Meyen, Asteraceae, is native to the southern parts of South America, where the aerial parts of the plant are commonly used for the treatment of inflammatory conditions. However, the effects of S. chilensis on human cancer cells remain to be elucidated. In this study, we evaluated the antiproliferative effects of the hydroalcoholic and dichloromethane extracts of S. chilensis, as well as their chemical constituents quercitrin and solidagenone against the five human tumor cell lines in vitro. The dichloromethane extract showed a promisor antiproliferative effects in vitro, especially against glioma cell line. Besides, the hydroalcoholic extract and quercitrin were inactive. The diterpene solidagenone showed highly potent antiproliferative effects against breast (MCF-7), kidney (786-0), and prostate cancer (PC-3) cells (total growth inhibition: TGI &lt; 6.25 µg/ml). Solidagenone meets the theoretical physico-chemical criteria for bioavailability of drugs, according to the "Rule of Five" and, by theorical studies, the observed biological effects were probably related to the interaction of the molecule with nuclear receptors and as an enzymatic inhibitor. This study contributes to chemical study and to the identification of antiproliferative molecules in S. chilensis. <![CDATA[Nutritional and biological evaluation of <em>Phoenix canariensis</em> pollen grains]]> ABSTRACT The nutritional value of pollens of Phoenix canariensis Chabaud, Arecaceae, was evaluated. HPLC analysis of vitamins revealed the predominant presence of vitamin C (109.13 ppm), followed by vitamin A (53.71 ppm) and vitamin E (40.60 ppm) and the total carbohydrates (28.12%), proteins (17.10%). In addition, 16 amino acids of which nine are essential (75.07%), and seven are non-essential (24.93%) were determined. On the other hand, two steroidal saponins (dioscin and methyl protodioscin) were isolated from the pollens, their structures were elucidated by spectroscopic techniques, including 1H NMR and 13C NMR. Their content in the 70% ethanolic extract was quantified using reversed phase HPLC and found to be 0.013% and 19.35%, respectively. The modulatory effect of the isolated compounds on the prevention of benign prostatic hyperplasia induced in rats and their moderate curative effect on stressed testicular tissue were studied. Being a good source of carbohydrates, proteins, vitamins, and amino acids pollens of P. canariensis can be used as a promising source of dietary supplement. Meanwhile pollens can act as prophylactic agent against benign prostatic hyperplasia. <![CDATA[Lupin gamma conglutin protein: effect on <em>Slc2a2, Gck</em> and <em>Pdx-1</em> gene expression and GLUT2 levels in diabetic rats]]> ABSTRACT Recently, lupin seed (Lupinus albus L., Fabaceae) products have emerged as a functional food due to their nutritional and health benefits. Numerous reports have demonstrated the hypoglycemic effects of lupin's gamma conglutin protein; nonetheless, its mechanism of action remains elusive. To understand the role of this protein on glucose metabolism, we evaluated the effect of administering L. albus' gamma conglutin on Slc2a2, Gck, and Pdx-1 gene expression as well as GLUT2 protein tissue levels in streptozotocin-induced diabetic rats. While consuming their regular diet, animals received a daily gamma conglutin dose (120 mg/kg per body weight) for seven consecutive days. Serum glucose levels were measured at the beginning and at the end of the experimental period. At the end of the trial, we quantified gene expression in pancreatic and hepatic tissues as well as GLUT2 immunopositivity in Langerhans islets. Gamma conglutin administration lowered serum glucose concentration by 17.7%, slightly increased Slc2a2 and Pdx-1 mRNA levels in pancreas, up-regulated Slc2a2 expression in the liver, but it had no effect on hepatic Gck expression. After gamma conglutin administration, GLUT2 immunopositivity in Langerhans islets of diabetic animals resembled that of healthy rats. In conclusion, our results indicate that gamma conglutin up-regulates Slc2a2 gene expression in liver and normalizes GLUT2 protein content in pancreas of streptozotocin-induced rats. <![CDATA[Effect of the treatment of <em>Copaifera duckei</em> oleoresin (copaiba) in streptozotocin-induced diabetic rats]]> ABSTRACT Diabetes mellitus is a syndrome that reaches more than 382 million people worldwide. It interferes with the metabolism of carbohydrates, causing chronic hyperglycemia. The objective of this study was to evaluate the effect of the Copaifera duckei, Dwyer, Fabaceae, oleoresin on streptozotocin-induced (STZ) diabetic rats. This study was based on the induction of diabetes mellitus by streptozotocin (55 mg/kg, i.p.) in Wistar rats and treated with doses of C. duckei oleoresin (250 and 500 mg/kg, p.o.). Subsequently, the clinical, biochemical and histopathological of the pancreas parameters were evaluated. Gas chromatographic analysis indicated that β-bisabolene (22.29%), β-caryophyllene (21.25%) and α-farnesene (15.58%) sesquiterpenes were the major components of the C. duckei oleoresin. In streptozotocin-induced diabetes mellitus, it was possible to observe that the C. duckei oleoresin treatment had a significant effect (p &lt; 0.001) on the clinical parameters, and that there was a positive improvement. This was attenuated by the urea, creatinine, and transaminases alterations (p &lt; 0.001) observed in animals with diabetes mellitus, as well as the significantly reduced (p &lt; 0.001) values of total cholesterol, triacylglycerides, and glucose. In the histopathological analyses of the pancreas, it was observed that the C. duckei oleoresin was able to restore β-cells and to significantly increase the quantity and diameter of the Langerhans islets (p &lt; 0.05), when compared to the diabetic group. The treatment with C. duckei oleoresin, employed under the conditions of this study, presented antidiabetic activity and can improve the complications found in this syndrome. <![CDATA[Effect of FPZ, a total flavonoids ointment topical application from <em>Pouzolzia zeylanica</em> var. <em>microphylla</em>, on mice skin infections]]> ABSTRACT The present study was designed to investigate the effect of FPZ, a total flavonoids ointment topical application from Pouzolzia zeylanica var. microphylla (Wedd.) Masam, Urticaceae, on skin infections in mice. FPZ ointment anti-infective effect was investigated on Staphylococcus aureus-induced skin abscess and skin ulcers in mice by evaluating the variation in abscess volume, histopathology of skin tissue and healing rate. Secondary, it is topical anti-inflammatory activities on carrageenan-induced hind paw edema in mice was estimated. Besides, FPZ ointment fingerprint was performed by using ultra-performance liquid chromatography and FPZ ointment chemical constituents were isolated and identified by repeated column chromatograph and spectroscopic methods. The results revealed that FPZ ointment topical application at the concentration of 2.5–10% could attenuate skin abscess and ulcers and accelerate wound healing, as compared with control group treated with vehicle (p &lt; 0.05). The histological analysis indicated that FPZ ointment acted via inflammation inhibition, granulation promotion and epidermis formation. Moreover, FPZ ointment effectively inhibited carrageenan-induced paw edema in a dose-dependent manner, especially 10% FPZ which showed superior activities in comparison with dexamethasone used as reference drug. FPZ ointment topical application showed a significant anti-infective effect against pyogenic bacterial skin infection in mice. <![CDATA[Fabaceae medicinal flora with therapeutic potential in Savanna areas in the Chapada do Araripe, Northeastern Brazil]]> ABSTRACT Fabaceae is one of the largest families of ethnopharmacological importance. From this botanical group, important chemical constituents that act in the treatment and/or healing of various bodily systems arise. The objective of this study was to evaluate the most versatile Fabaceae species and the agreement of use among the informants, in the Chapada do Araripe Savanna. The research included five rural communities located in the municipalities of Nova Olinda, Crato, Barbalha, Moreilândia and Exu, covering the states of Ceará and Pernambuco. We conducted semi-structured interviews with 126 informants, adopting the snowball technique and using a standardized form. The relative importance and the Informant Consensus Factor were analyzed for the selection of species with therapeutic potential. Twenty-six medicinal species, distributed across 22 genera, were associated with seventy therapeutic purposes. As for the habitat of the species, the arboreal habitat predominated (76.92%). In relation to the plant parts, the bark (28.57%) and stem inner bark (26.53%) were the most used structures. Among the species, four showed great versatility in relation to their uses, with relative importance (RI&gt;1), these being: Copaifera langsdorffii Desf. (1.70), Stryphnodendron rotundifolium Mart. (1.52), Bowdichia virgiloides Kunth (1.10) and Amburana cearenses (Allemand) A. C. Sm. (1.02). These species had eight to thirty curative properties and ranged from five to twelve body systems. The therapeutic indications cited were grouped into sixteen categories of body systems, of which Sensory System Disorders and Mental and Behavioral Diseases presented maximum values for the Informant Consensus Factor. For the Sexual Impotence category there was no agreement, whereas for the remaining systems the Informant Consensus Factor ranged from 0.33 to 0.91. The study evidences that the information of use and/or knowledge are shared among the people within the community, proving the great importance of Fabaceae in the use of medicinal plants.