Scielo RSS <![CDATA[Revista Brasileira de Farmacognosia]]> vol. 26 num. 5 lang. en <![CDATA[SciELO Logo]]> <![CDATA[Anatomical and histochemical analysis of <em>Dysphania ambrosioides</em> supported by light and electron microscopy]]> ABSTRACT Dysphania ambrosioides (L.) Mosyakin &amp; Clemants (syn: Chenopodium ambrosoides L.), Amaranthaceae, popularly known as “mastruz”, is an herb widely used in Brazil as anthelmintic. To contribute to the knowledge about medicinal plants, a microscopic analysis was accomplished to describe the main anatomical characters of root, stem, petiole and leaf blade of D. ambrosioides and histochemical tests were performed on the leaf blade. Cross-sections were obtained, by hand, for microscopic analysis of root, stem, petiole and leaf blade; to the leaf blade were still made paradermal sections, scanning electron microscopy analysis, maceration and histochemical tests. The main characters useful in the identification of the plant were: anomalous secondary thickening in the root and stem; presence of idioblasts containing crystal sand in the root, stem, petiole and leaf blade; in these there are also idioblasts with druses; presence of non-glandular and glandular trichomes in the stem, petiole and leaf blade; stomata on the stem, petiole and leaf blade, identified in these as anomocytic and anisocytic; dorsiventral mesophyll and collateral vascular bundles. Maceration revealed that the vessel elements are helical type. Through the histochemical tests, it was evidenced the presence of lipophilic substances, essential oils, oleoresins, phenolic compounds, starch, lignin and calcium oxalate crystals. This work provides support to the quality control of the species. <![CDATA[The structural studies on the medicinal plant <em>Haplophyllum telephioides</em>]]> ABSTRACT Haplophyllum telephioides Boiss., Rutaceae, is an endemic herb which generally grows at the steppe region in Central Turkey. Its aerial parts in flowering stage are used by local people against influenza virus due to its antimicrobial effect. The main purpose of this study was to examine the morphology, anatomy and histology of the vegetative and reproductive organs of the H. telephioides, which are used in popular medicine, thereby, contributing to the pharmacognostic evaluation of the species. The species was studied morphologically in detail. New morphological features were described for them, such as the dimension of leaf, sepal and fruit, size of filament, anther and pistil, and diameter of the ovary. In addition, some deviating features were found in relation to previous published descriptions for the species, such as plant height and petal length. The anatomy of plant parts such as stem, leaf, sepal, petal, filament and pistil, were studied using light microscopy and scanning electron microscopy. Stem has incipient secondary growth. The leaf is amphistomatic and the mesophyll is equifacial. Stomata are anomocytic and sunken. In micromorphological studies, it is determined that the leaf is coated by a thick cuticle and above epicuticular wax. Schizogenous glands were found in all vegetative and reproductive organs. The structural features herein found can assist the diagnosis of H. telephioides. <![CDATA[A validated and densitometric HPTLC method for the simultaneous quantification of reserpine and ajmalicine in <em>Rauvolfia serpentina</em> and <em>Rauvolfia tetraphylla</em>]]> ABSTRACT High performance thin layer chromatographic method (HPTLC) has been developed for the quantification of reserpine and ajmalicine in root part of two different population of Rauvolfia serpentina (L.) Benth. ex Kurz and Rauvolfia tetraphylla L., Apocynaceae, collected from Punjab and Uttarakhand. HPTLC of methanolic extract of root containing indole alkaloids, i.e., reserpine and ajmalicine, was performed on TLC Silicagel 60 F254 (10 cm × 10 cm) plates with toluene:ethyl acetate:formic acid (7:2:1), as mobile phase. Quantification of the reserpine and ajmalicine was performed in the absorption–reflection mode at 268 nm. The recovery of reserpine and ajmalicine were 99.3 and 98.7% respectively. The calibration curves were linear for both the reserpine and ajmalicine, in the range of 200–1200 ng. HPTLC densitometry has been performed for the estimation of reserpine and ajmalicine in root part of R. serpentina and R. tetraphylla for the first time. The method is simple, rapid and cost effective and can be used for routine analysis of ajmalicine and reserpine in different Rauvolfia species as well as for quality control of herbal drugs containing Rauvolfia species. <![CDATA[Validated high performance thin layer chromatography method for the quantification of bioactive marker compounds in Draksharishta, an ayurvedic polyherbal formulation]]> ABSTRACT Draksharishta is an ayurvedic polyherbal formulation with Draksha (Vitis vinifera L., Vitaceae) as chief ingredient prescribed for digestive impairment, respiratory disorders and weakness. These herbal medicines containing biologically active compounds play a significant role. Therefore it is necessary to carry out the chemical standardization of bioactive marker compounds present in the polyherbal ayurvedic formulation like Draksharishta. The aim of the present work was to develop and validate a HPTLC method for determination of gallic acid, catechin and resveratrol in commercially available marketed and in-house prepared formulations of Draksharishta. This is the first report of quantification of bioactive marker compound resveratrol using HPTLC in Draksharishta. The method employed silica gel precoated thin layer chromatography plates with F254 as the stationary phase. The respective mobile phases were used to develop the plates which separated bands according to the marker compound. Camag scanner V was used for densitometric scanning. Further, the method was validated according to International Conference of Harmonization (ICH) guidelines. The Rƒ values of the three marker compounds were measured. Correlation coefficients were calculated from the standard graph of linearity. Accuracy, precision and recovery were all within the required limits. The developed HPTLC methods for bioactive marker compounds present in in-house and marketed formulations were found to be simple, accurate, precise and robust. <![CDATA[Identification of reference genes for gene expression normalization in safflower (<em>Carthamus tinctorius</em>)]]> ABSTRACT Safflower (Carthamus tinctorius L., Asteraceae) is an important oil crop and medicinal plant. Gene expression analysis is gaining importance in the research of safflower. Quantitative PCR has become a powerful method for gene study. Reference genes are one of the major qualification requirements of qPCR because they can reduce the variability. To identify the reference genes in safflower, nine candidate genes of the housekeeping genes were selected from the EST library of safflower constructed by our lab: CtACT (actin), CtGAPDH (glyceraldehyde 3-phosphate dehydrogenase), CtE1F4A (elongation factor 1 alpha), CtTUA (alpha-tubulin), CtTUB (beta-tubulin), CtPP2A (serine/threonine-protein phosphatase), CtE1F4A (eukaryotic initiation factor 4A), CtUBI (Ubiquitin), and Ct60S (60S acidic ribosomal protein). Expression stability was examined by qPCR across 54 samples, representing tissues at different flowering stages and two chemotype of safflower lines. We assessed the expression stability of these candidate genes by employing four different algorithms (geNorm, NormFinder, ΔCt approach, and BestKeeper) and found that CtUBI and Ct60S were the highly ranked candidate genes. CtUBI and Ct60S were used as reference genes to evaluate the expression of CtFAD2-10 and CtKASII. Our data suggest CtUBI and Ct60S could be used as internal controls to normalize gene expression in safflower. <![CDATA[Potent inhibition of <em>Western Equine Encephalitis</em> virus by a fraction rich in flavonoids and phenolic acids obtained from <em>Achyrocline satureioides</em>]]> ABSTRACT Achyrocline satureioides (Lam.) DC. Asteraceae, ‘marcela del campo', possess several pharmacological properties. Previously we reported antiviral activity of an aqueous extract of A. satureioides against an alphavirus, Western Equine Encephalitis virus. Alphaviruses are highly virulent pathogens which cause encephalitis in humans and equines. There are no effective antiviral to treat its infections. The aim of this study was to evaluate in vitro cytotoxic and antiviral activities against Western Equine Encephalitis virus of five water extract chromatographic fractions from A. satureioides and identify the main compounds of the bioactive fraction. Also, it was to assess in vivo cytogenotoxic ability of the active fraction. Cytotoxicity studies revealed low toxicity of the most of fractions in Vero and in equine peripheral blood mononuclear cells. Antiviral studies showed that the water crude extract – Sephadex LH 20 – fraction 3 MeOH–H2O (Fraction 3) was active against Western Equine Encephalitis virus with Effective Concentration 50% = 5 µg/ml. Selectivity Indices were 126.0 on Vero and 133.6 on peripheral blood mononuclear cells, four times higher than aqueous extract selectivity index. Regarding the mechanism of action we demonstrated that F3 exerted its action in intracellular replication stages. Further, fraction 3 showed important virucidal action. Fraction 3 contains, in order of highest to lowest: chlorogenic acid, luteolin, 5,7,8-trimethoxyflavone, 3-O-methylquercetin and caffeic acid. Fraction 3 did not induce in vivo toxic nor mutagenic effect. Therefore, it is safe its application as antiviral potential. Further studies of antiviral activity in vivo will be developed using a murine model. <![CDATA[Anticonvulsant mechanism of saponins fraction from adventitious roots of <em>Ficus religiosa</em>: possible modulation of GABAergic, calcium and sodium channel functions]]> ABSTRACT In our previous studies, quantified saponins-rich fraction from adventitious root extract of Ficus religiosa L., Moraceae, showed anticonvulsant effect in acute, as well as chronic mice models of epilepsy. The present study was designed to reveal putative anticonvulsant mechanism of quantified saponins-rich fraction using target specific animal models. The anticonvulsant effect of quantified saponins-rich fraction was initially studied in maximal electroshock and pentylenetetrazol test at 1, 2 and 4 mg/kg; i.p. doses. Based on the results of initial anticonvulsant testing, different groups of mice were injected with vehicle or quantified saponins-rich fraction (4 mg/kg; i.p.), 30 min prior to an injection of N-methyl-D-aspartic acid (100 mg/kg; s.c.), bicuculline (5 mg/kg; i.p.), strychnine hydrochloride (2 mg/kg; i.p.), BAY k-8644 (37.5 µg; i.c.v.), veratridine (500 µg/kg; i.p.) and the convulsive episodes were studied. Treatment with the extract (1, 2 and 4 mg/kg) showed significant protection in maximal electroshock and pentylenetetrazol-induced convulsion tests, in a dose-dependent manner. Moreover, quantified saponins-rich fraction at 4 mg/kg dose showed significant increase in latency to clonic convulsions, decrease in seizure severity and increase in average wave amplitude in bicuculline, BAY k-8644 and veratridine tests, respectively, as compared to vehicle control. However, SRF treatment failed to abolish N-methyl-D-aspartic acid and strychnine-induced convulsions, indicated by insignificant change in the appearance of turning behavior and onset of tonic extension, respectively, as compared to vehicle control. From the results of present study, it is concluded that quantified saponins-rich fraction suppress maximal electroshock, pentylenetetrazol, bicuculline, BAY k-8644 and veratridine-induced convulsions, indicating its GABAergic, Na+ and Ca2+ channel modulatory effects. Further it can be correlated that quantified saponins-rich fraction causes deactivation of voltage-gated Na+ and Ca2+ channels, without effecting ligand-gated Na+ and Ca2+ channels. More studies are required at molecular levels using in vitro techniques to understand the exact molecular interactions of quantified saponins-rich fraction with these pathways. <![CDATA[Possible mechanisms of antinociception of methanol extract of <em>Melastoma malabathricum</em> leaves]]> ABSTRACT Melastoma malabathricum L., Melastomaceae, has been traditionally used to relieve diverse pain-related ailments. The objectives of the present study were to determine the antinociceptive activity of methanol extract of M. malabathricum leaves and to elucidate the possible mechanisms of antinociception involved using various rats' models. The extract (100, 250, and 500 mg/kg) was administered orally 60 min prior to subjection to the respective test. The in vivo acetic acid-induced abdominal constriction, formalin-induced paw licking, and hot plate tests were used as the models of nociception to evaluate the extract antinociceptive activity. Further studies were carried out to determine the role of opioid and vanilloid receptors, glutamate system and nitric oxide/cyclic guanosine phosphate (NO/cGMP) pathway in modulating the extract antinociceptive activity. From the results obtained, M. malabathricum exhibited significant (p &lt; 0.05) antinociceptive activity in all the chemical- and thermal-induced nociception models. Naloxone (5 mg/kg), a non-selective opioid antagonist, failed to significantly affect the antinociceptive activity of MEMM when assessed using the abdominal constriction-, hot plate- and formalin-induced paw licking-test. M. malabathricum also significantly (p &lt; 0.05) reversed the nociceptive response in capsaicin- and glutamate-induced paw licking test. Furthermore, only L-arginine (a nitric oxide precursor) alone, but not, NG-nitro-L-arginine methyl esters (L-NAME; an inhibitor of NO synthase), methylene blue (MB; an inhibitor of cGMP), or their combination thereof, significantly (p &lt; 0.05) block the antinociceptive activity of M. malabathricum. In conclusion, M. malabathricum exerted a non-opioid antinociceptive activity at the central and peripheral levels partly via the inhibition of vanilloid receptors and glutamatergic system, and activation of the NO-mediated/cGMP-independent pathway. <![CDATA[Effects of bark flour of <em>Passiflora edulis</em> on food intake, body weight and behavioral response of rats]]> ABSTRACT Effects of treatment with the bark flour of Passiflora edulis Sims, Passifloraceae, were evaluated. Adult male Wistar rats were treated for 30 days (130 mg/kg, p.o.) with the albedo flour, flavedo and full bark of P. edulis, corresponding to albedo associated with flavedo. Behavioral response observed after treatment with bark flour P. edulis showed sedative effects by the reduction of exploratory activity and increased duration of immobility in the open field test for the group of animals that received the albedo flour associated with the flavedo. Sedative effects were observed in the absence of motor incoordination or muscle relaxation. Food intake of experimental animals was not changed, but the weight gain was decreased both in animals that received only albedo flour, and in those who received the full bark flour. The full bark flour of Passiflora showed sedative effects, without anxiolytic effect detectable and muscle relaxation or motor incoordination, and reduces body weight gain. <![CDATA[Flax lignan concentrate attenuate hypertension and abnormal left ventricular contractility via modulation of endogenous biomarkers in two-kidney-one-clip (2K1C) hypertensive rats]]> ABSTRACT The present investigation was designed to study the effect of flax lignan concentrate obtained from Linum usitatissimum L., Linaceae, in two-kidney, one clip (2K1C) hypertension model in Wistar rats. 2K1C Goldblatt model rats were divided randomly into six groups: sham, 2K1C control, captopril (30 mg/kg), flax lignan concentrate (200, 400 and 800 mg/kg). Flax lignan concentrate and captopril were administered daily for eight consecutive weeks. Sham-operated, and 2K1C control rats received the vehicle. Treatment with flax lignan concentrate (400 and 800 mg/kg) significantly and dose-dependently restored the hemodynamic parameters systolic blood pressure, diastolic blood pressure, mean arterial blood pressure and left ventricular functions. The flax lignan concentrate significantly restored the elevated hepatic, renal and cardiac marker enzymes in the serum. It also restored the organs weights (kidney and heart), serum electrolyte level and histological abnormalities. Furthermore, flax lignan concentrate significantly elevated the level of biochemical markers that is enzymatic antioxidants superoxide dismutase, glutathione and decreased malondialdehyde in the heart and kidney tissues. Meanwhile, we found that plasma nitric oxide and plasma nitric oxide synthase contents were significantly increased in the flax lignan concentrate-treated group, and plasma endothelin-1 and renal angiotensin-II levels were significantly lower than 2K1C hypertensive group. In conclusion, the antihypertensive and antioxidant effect of flax lignan concentrate were dose-dependent and at the highest dose (i.e. 800 mg/kg) similar to those of captopril (30 mg/kg). It is suggested that flax lignan concentrate reduced blood pressure by reduction of renal angiotensin-II level, inhibition of plasma endothelin-1 production, induction of the nitric oxide, nitric oxide synthase and in vivo antioxidant defense system. <![CDATA[Uliginosin B, a natural phloroglucinol derivative with antidepressant-like activity, increases Na<sup>+</sup>,K<sup>+</sup>-ATPase activity in mice cerebral cortex]]> ABSTRACT Uliginosin B, a phloroglucinol isolated from Hypericum polyanthemum Klotzsch ex Reichardt, Hypericaceae, has antidepressant-like effect in the forced swimming test in rodents and inhibits monoamines neuronal reuptake without binding to their neuronal carriers. Studies showed the involvement of Na+,K+-ATPase brain activity in depressive disorders, as well as the dependence of neuronal monoamine transport from Na+ gradient generated by Na+,K+-ATPase. This study aimed at evaluating the effect of uliginosin B on Na+,K+-ATPase activity in mice cerebral cortex and hippocampus (1 and 3 h after the last administration) as well as the influence of veratrine, a Na+ channel opener, on the antidepressant-like effect of uliginosin B. Mice were treated (p.o.) with uliginosin B single (10 mg/kg) or repeated doses (10 mg/kg/day, 3 days). Acute administration reduced the immobility in the forced swimming test and tail suspension test and increased Na+,K+-ATPase activity in cerebral cortex 1 h after treating, whereas the repeated treatment induced the antidepressant-like effect and increased the Na+,K+-ATPase activity at both times evaluated. None treatment affected the hippocampus enzyme activity. Veratrine pretreatment prevented uliginosin B antidepressant-like effect in the forced swimming test, suggesting the involvement of Na+ balance regulation on this effect. Altogether, these data indicate that uliginosin B reduces the monoamine uptake by altering Na+ gradient. <![CDATA[Evaluation of mechanism for antihypertensive and vasorelaxant effects of hexanic and hydroalcoholic extracts of celery seed in normotensive and hypertensive rats]]> ABSTRACT Celery (Apium graveolens L., Apiaceae) is one of the popular aromatic vegetables and part of the daily diet around the world. In this study, aqueous-ethanolic and hexane extracts of celery seed were prepared and the amount of n-butylphthalide, as an active component, was determined in each extract. Then the effects of hexanic extract on systolic, diastolic, mean arterial blood pressure and heart rate were evaluated in an invasive rat model. The vasodilatory effect and possible mechanisms of above mentioned extracts on aorta ring were also measured. High performance liquid chromatography analysis revealed that hexanic extract contains significantly higher amounts of n-butylphthalide, compared to aqueous-ethanolic extract. The results indicated that hexanic extract significantly decreased the systolic, diastolic, mean arterial blood pressure and heart rate in normotensive and hypertensive rats. Our data revealed that celery seed extract exerts its hypotensive effects through its bradycardic and vasodilatory properties. Moreover, the active components in celery seed extracts could induce their vasodilatory properties through Ca2+ channel blocking activity in endothelial and non-endothelial pathways and particularly by interference with the extra or intracellular calcium. <![CDATA[Identification of dysregulated microRNA expression and their potential role in the antiproliferative effect of the essential oils from four different <em>Lippia</em> species against the CT26.WT colon tumor cell line]]> ABSTRACT In spite of advances in colorectal cancer treatments, approximately 1.4 million new global cases are estimated for 2015. In this sense, Brazilian plant diversity offers a multiplicity of essential oils as prospective novel anticancer compounds. This study aimed to evaluate the antiproliferative effect of the essential oils from four Lippia species in CT26.WT colon tumor cells, as a measurement of cell cycle phase distribution and microRNA expression. CT26.WT showed cell cycle arrest at G2/M phase after treatment with 100 µg/ml of Lippia alba (Mill.) N.E.Br. ex Britton &amp; P. Wilson, Lippia sidoides Cham., and Lippia lacunosa Mart. &amp; Schauer, Verbenaceae, essential oils and, at the same concentration, Lippia rotundifolia Cham. essential oil caused an augment of G0/G1 phase. The miRNA expression profiling shows change of expression in key oncogenic miRNAs genes after treatment. Our findings suggest growth inhibition mechanisms for all four essential oils on CT26.WT cells involving direct or indirect interference on cell cycle arrest and/or oncogenic miRNAs expression. <![CDATA[Hypoglycemic effect of formulation containing hydroethanolic extract of <em>Calophyllum brasiliense</em> in diabetic rats induced by streptozotocin]]> ABSTRACT Diabetes mellitus is a chronic and severe metabolic dysfunction, it's slow and progressive evolution interferes directly in the metabolism of carbohydrates, fats and proteins, causing hyperglycemia, glycosuria, polydipsia, hyperlipidaemia, among others. The aim of this study was to evaluate the antidiabetic effect of hydroethanolic extract and granulated of Calophyllum brasiliense Cambess., Clusiaceae, species in diabetic rats as well as it's biochemical parameters. The results demonstrated that both the pharmaceutical forms, hydroethanolic extract and granulated, were able to reduce significantly (p &lt; 0.001) hyperglycemia and glycosuria, in addition to improve polydipsia, polyuria, and weight loss. Treatments using hydroethanolic extract and granulated were also able to reduce significantly levels of triacylglycerides, cholesterol and low-density lipoprotein, as well as the transaminases, urea and creatinine levels. Therefore, it is concluded that these pharmaceutical forms have anti-diabetic effect and act improving the biochemical parameters, this effect is probably due to the high content of polyphenolic compounds found in the formulations. <![CDATA[Spectroscopic synthetic optimizations monitoring of silver nanoparticles formation from <em>Megaphrynium macrostachyum</em> leaf extract]]> ABSTRACT Nanobiotechnology is one of the most promising areas in modern nanoscience and technology. Metallic nanoparticles have found uses in many applications in different fields, such as catalysis, photonics, electronics, medicine and agriculture. Synthesized nanoparticles through chemical and physical methods are expensive and have low biocompatibility. In the present study, silver nanoparticles have been synthesized from Megaphrynium macrostachyum (Benth. &amp; Hook. f.) Milne-Redh., Marantaceae, leaf extract. Megaphrynium macrostachyum is a plant with large leaves found in the rainforest of West and Central Africa. Synthetic optimizations following factors such as incubation time, temperature, pH, extract and silver ion concentration during silver formation are discussed. UV–visible spectra gave surface plasmon resonance for synthesized silver nanoparticles based Megaphrynium macrostachyum peaks at 400–450 nm. X-ray diffraction revealed the average size of pure crystallites composed from Ag and AgCl. <![CDATA[Report on the Malungo expedition to the Erepecuru river, Oriximiná, Brazil. Part I: is there a difference between black and white <em>breu</em>?]]> ABSTRACT Species belonging to Burseraceae produce an oleoresin known in the north of Brazil as breu. They comprise an essential oil with a complex composition, and are used in Amazonia for smoking the environment, to caulk boats and for medicinal purposes. Depending on its organoleptic characteristics and on the breu-producing species, they are called white or black breu. In this work, we provide data about the breu-producing species occurring in the quilombola region of the Erepecuru river, the chemical composition, and whether it is possible to differentiate them based on their chemical composition and/or botanical identification. Aerial samples from breu trees and oleoresins were collected from 10 different individuals at 6 different sites on the Erepecuru river under the guidance of the quilombolas. Essential oils were extracted by hydrodistillation and characterized by GC–MS. From the analysis, 126 different substances were identified, with a large quantitative and qualitative variation. To better understand the chemical variations within the samples and to sort the variation into the categories of white or black breu as identified by the quilombola, we sorted the oil samples into five different sets according to their major compounds (A: δ-3-carene; B: p-cymene; C: γ-cadinene/p-cymene; D: limonene, β-phellandrene/α-terpineol; E: α-pinene/limonene). Essential oils from samples of white breu had the highest concentration of α-pinene, while a similarity in chemical composition could not be established for the black breu samples (sets A, B and C). Furthermore, a chemical similarity between a black breu (Protium heptaphyllum (Aubl.) Marchand) and a white breu (Protium decandrum (Aubl.) Marchand) sample was evidenced. In conclusion, it is difficult to establish definitions for white and black breu based on chemical, botanical or regional names. This designation is more cultural and regional than scientific and is based on the oleoresin production volume, its color aspect and scent. <![CDATA[Biological and therapeutic effects of honey produced by honey bees and stingless bees: a comparative review]]> ABSTRACT Honey is a natural product produced by both honey bees and stingless bees. Both types of honey contain unique and distinct types of phenolic and flavonoid compounds of variable biological and clinical importance. Honey is one of the most effective natural products used for wound healing. In this review, the traditional uses and clinical applications of both honey bee and stingless bee honey – such as antimicrobial, antioxidant, anti-inflammatory, anticancer, antihyperlipidemic, and cardioprotective properties; the treatment of eye disorders, gastrointestinal tract diseases, neurological disorders, and fertility disorders and wound healing activity are described.