Abstract in English:ABSTRACT The Lauraceae family is widely distributed in tropical and subtropical climates, has more than 2500 species and in the taxonomic point of view, it is one of the most difficult families to identify species. The aim of this study was to analyze the leaf anatomy of three species of Lauraceae (Ocotea indecora (Schott) Mez, Nectandra barbellata Coe-Teix. and Endlicheria paniculata (Spreng.) J.F.Macbr.) and identify the chemical profile of essential oil extracted from the leaves by hydrodistillation method. The leaves of the three species were obtained in “Parque Estadual Intervales”, Atlantic Forest, São Paulo state, Brazil. Samples of leaves were fixed, dehydrated, embedded in synthetic resin and sectioned for mounting histological slides for anatomical description of leaf tissues. The essential oil extracted by hydrodistillation method from dried leaves was analyzed by gas chromatography to establish its chemical profile. The leaves are hypostomatic, the epidermis in E. paniculata and N. barbellata present regular cells walls and irregular cells walls in O. indecora in both sides of epidermis. The three species present a dorsiventral mesophyll. Histochemical analyses presented lipid substances in secretory cavity and cuticle; starch, phenolic compounds and mucilage were observed in parenchyma cells of midrib and mesophyll. Ultra structural analyses demonstrated that trichomes in the species E. paniculata and O. indecora are shown only on the abaxial leaf face and species N. barbellata presented trichomes on both sides of the epidermis (abaxial and adaxial). Ocotea indecora essential oil revealed as main compound the bicyclogermacrene and N. barbellata the δ-cadinene. The species showed different morphological characters and different compounds of the essential oil, being these data useful for the differentiation of the species.
Abstract in English:ABSTRACT Piper caldense C. DC., Piperaceae, commonly known as “pimenta-d’água”, “pimenta-darda” or “paguarandy” in Brazil, is a shrub that grows mainly in humid and shaded habitats. The present study investigates the anatomy of the leaves and stems of P. caldense by light and scanning electron microscopy in order to provide supporting data for correct identification of the species. The leaves are hypostomatic, have a 2-layered hypodermis, and posses pearl glands. The midrib shows a ‘U’-shaped stele comprised of about ten collateral vascular bundles. The main anatomical marker of the stem is the presence of a continuous sclerenchymatous sheath in the pith. Two forms of calcium oxalate crystals, namely crystal sand and raphides, are observed in this species.
Abstract in English:ABSTRACT Spiranthera odoratissima A. St.-Hil., Rutaceae, known as “manacá” is a shrub native of the Brazilian Cerrado. Their leaves and roots are popularly used to treat rheumatism, infection and abdominal pain. This study analyzed the chemical composition of volatile oils from leaves of S. odoratissima and verified the seasonal variability of its chemical composition. The volatile oils were obtained by hydrodistillation using a Clevenger type apparatus and analyzed by gas chromatography coupled to mass spectrometry. The main chemical components found in samples of volatile oils were β-caryophyllene, bicyclogermacrene, δ-cadinene, amorphous-4,7(11)-diene, α-epi-muurolol, α-cadinol, α-muurolol and γ-cadinene. The hierarchical clustering identified three groups: the first was characterized by α-epi-muurolol, the second by amorphous-4,7(11)-diene and the third group was characterized by α-muurolol. The discriminant canonical analysis was used to differentiate between clusters on the basis of oil composition. The results suggest that the rainfall presented a relationship with the chemical composition of the volatile oil. This is the first study conducted on the seasonal behavior of the chemical constituents in volatile oil from leaves of S. odoratissima.
Abstract in English:ABSTRACT The aim of this paper is to provide an overview on the chemical composition of triterpenes in widespread used folk medicine species, through the development and validation of eleven compounds using HPLC-UV detection. The compounds were separated using isocratic elution, on a reverse phase column (Kinetex C18, 250 mm × 4.6 mm, 5 µm) with mobile phase consisted of acetonitrile:tetrahydrofuran (90:10, v/v), flow-rate of 0.5 ml/min and detection in 210 nm. Diverse validation parameters were successfully evaluated. The samples of Bauhinia variegata L., B. variegata var. candida Voigt, Fabaceae, Cecropia palmata Willd. and C. obtusa Trécul, Urticaceae, collected in 2012, 2013 and 2014 from Amazon were treated with two different solvents (ethyl acetate and chloroform) and analyzed by the proposed method. Stigmasterol, lupeol, β-sitosterol, β-amirin and α-amirin were found in all the studied plants. Highlighting the presence of oleanolic acid, maslinic acid in C. obtusa and C. palmata extracts, erythrodiol only in C. palmata, stigmasteol in B. variegata and α-amirin in B. variegata var. candida. Overall, ethyl acetate showed better performance as the extractor solvent than chloroform. Moreover, it could be used for the quality control of medicinal plants and to assess potential marker compounds.
Abstract in English:ABSTRACT Screening of medicinal plants from Iranian flora against human cancer cell-lines have shown that an hexane extract from roots of Salvia sahendica Boiss. & Buhse, Lamiaceae, is active against human cervical cancer (HeLa) and colorectal adenocarcinoma (Caco-2) cell-lines at the test concentration of 100 µg/ml (100% inhibition). Cytotoxicity of the extract was localized with the aid of HPLC-time-based activity profiling adapted to the tetrazolium colorimetric bioassay. Four abietane-type diterpenoids in active time-windows were identified as cytotoxic compounds namely: sahandone (1), sahandol (2), 12-deoxy-salvipisone (3) and sahandinone (4). Compound 1 showed the highest toxicity against HeLa cells (IC50 = 5.6 ± 0.1 µg/ml), which was comparable with betulinic acid (IC50 = 4.3 ± 1.2 µg/ml), the positive control. Compound 2 was active against the HeLa cells (IC50 = 8.9 ± 0.7 µg/ml) but not the Caco-2 cell-line. Compounds 1, 3 and 4 exhibited moderate activity (IC50 = 22.9–41.4 µg/ml) against the Caco-2 cells. This study reveals that the HeLa cells are more sensitive to all tested compounds than the Caco-2 cells. In silico molecular docking study showed a rigid binding of the compounds to tyrosine kinase pp60src, and proved their cytotoxic activity.
Abstract in English:ABSTRACT Acetylcholinesterase is an important target for control of neurodegenerative diseases causing cholinergic signaling deficit. Traditionally, galanthamine has been used as an Amaryllidaceae-derived acetylcholinesterase inhibitor, although new Amaryllidaceae plants could serve as source for better acetylcholinesterase inhibitors. Therefore, the objective of this study was to characterize the alkaloid composition from bulbs of Rhodolirium andicola (Poepp.) Traub, a native Chilean Amaryllidaceae specie, and assess their inhibitory activity on acetylcholinesterase by in vitro and in silico methodologies. Alkaloidal extracts from R. andicola exhibited an inhibitory activity with IC50 values between 11.25 ± 0.04 and 57.78 ± 1.92 µg/ml that included isolated alkaloid, galanthamine (2.3 ± 0.18 µg/ml), Additionally, 12 alkaloids were detected using gas chromatography–mass spectrometry and identified by comparing their mass fragmentation patterns with literature and database NIST vs.2.0. To better understand the bioactivity of isolated compounds and alkaloidal extracts against acetylcholinesterase, a molecular docking approach was performed. Results suggested that alkaloids such as lycoramine, norpluvine diacetate and 6α-deoxy-tazettine expand the list of potential acetylcholinesterase inhibitors to not only galanthamine. The role of R. andicola as a source for acetylcholinesterase inhibitors is further discussed in this study.
Abstract in English:ABSTRACT Curcuma zanthorrhiza Roxb., Zingiberaceae, a species from Indonesia with xanthorrhizol as the major metabolite, has been used as a folk medicine in several of pharmacological activities. This work aimed to evaluate the xanthorrhizol contents, α-glucosidase inhibition, and cytotoxic activities in ethyl acetate fraction from accessions of C. zanthorrhiza. High-performance liquid chromatography investigated xanthorrhizol content with the standard. Pharmacological activities were evaluated by inhibition of α-glucosidase, the brine shrimp lethality test, and anticancer activity. The ethyl acetate fraction yield varied from 8.24% (Karanganyar) to 13.13% (Sukabumi). The xanthorrhizol contents were found to be in the range 43.55% to 47.99% with Ngawi and Wonogiri promising accessions having the lowest and highest value, respectively. IC50 value for α-glucosidase inhibition ranged from 339.05 µg/ml (Karanganyar) to 455.01 µg/ml (Ngawi). LC50 value for cytotoxic activities ranged from 33.25 µg/ml (Ngawi) to 42.28 µg/ml (Karanganyar) in brine shrimp lethality test, 3.10 µg/ml (Karanganyar) to 9.85 µg/ml (cursina-III) in Vero cell, and 1.17 µg/ml (Ngawi) to 6.83 µg/ml (Sukabumi) in MCF-7 cell. In this study, C. zanthorrhiza accessions have a high in xanthorrhizol contents and cytotoxic activities that showed a high potential of studied accessions for breeding programs on a commercial scale.
Abstract in English:ABSTRACT Cissampelos sympodialis Eichler, Menispermaceae, a Brazilian medicinal plant and its alkaloid warifteine present immunomodulatory activity on asthma experimental model by reducing antigen-specific IgE levels, eosinophil infiltration and lung hyperactivity. Allergic rhinitis is a chronic inflammatory disorder of the nasal tissue that affect the quality of life and it is a risk factor for asthma exacerbation. This study evaluated the effect of inhaled warifteine in an allergic ovalbumin rhinitis model. Inhaled warifteine (2 mg/ml) treatment of ovalbumin-sensitized BALB/c mice significant decreased total and differential number of cells on the nasal cavity and decreased ovalbumin-specific IgE serum levels. Hematoxylin & eosin staining of histological preparations of ovalbumin nasal tissues showed changes such as congestion and a massive cell infiltration in the perivascular and subepithelial regions characterizing the nasal inflammatory process. However, inhaled warifteine or dexamethasone treatment decreased cell infiltration into the perivascular regions and it was observed an intact nasal tissue. Periodic acidic staining of nasal epithelium of ovalbumin animals demonstrated high amount of mucus production by goblet cells and inhaled warifteine or dexamethasone treatment modulated the mucus production. In addition, toluidine blue staining of the nasal epithelium of ovalbumin animals demonstrated an increase of mast cells on the tissue and inhaled warifteine or dexamethasone treatment decreased in average of 1.4 times the number of these cells on the nasal epithelium. Taken these data together we postulate that warifteine, an immunomodulatory alkaloid, can be a medicinal molecule prototype to ameliorate the allergic rhinitis conditions.
Abstract in English:ABSTRACT This study evaluated the anti-inflammatory and antioxidant activities of Impatiens noli-tangere L., Balsaminaceae, and of Stachys officinalis L., Lamiaceae, polyphenolic-rich extracts obtained by nanofiltration process. Results showed the great potential and efficiency of the nanofiltration process to concentrate the herbal extract's main polyphenolic compounds (over 91% phenolic acids and flavonoids retention). S. officinalis polyphenolic-rich extracts had high antioxidant activities (IC50 2.5 µg/ml) compared to I. noli-tangere polyphenolic-rich extracts (IC50 19.3 µg/ml) and similar with that of ascorbic acid. Polyphenolic-rich extracts were investigated to determine the pro-inflammatory enzymes lipoxygenase, cyclooxygenase-1 and cyclooxygenase-2 and their inhibitory activity. Furthermore, high inhibitory activity of the examined extracts was reported for the first time, for both lipoxygenase (IC50 2.46 and 1.22 µg/ml for I. noli-tangere and S. officinalis polyphenolic-rich extracts, respectively), cyclooxygenase-1 (IC50 18.4 and 10.1 µg/ml for I. noli-tangere and S. officinalis polyphenolic-rich extracts, respectively) and cyclooxygenase-2 (IC50 = 1.9 and 1.2 mg/ml for I. noli-tangere and S. officinalis polyphenolic-rich extracts, respectively). Additionally, the in vivo studies showed that S. officinalis polyphenolic-rich extract has a higher anti-inflammatory effect, the hind-paw volume employed for both models determined that I. noli-tangere polyphenolic-rich extract and is also higher than that of diclofenac. It was noticed that their anti-inflammatory effect persists for more than 24 h. The I. noli-tangere and S. officinalis polyphenolic-rich extracts exert anti-inflammatory and antioxidant activities and these properties can be at least partly assigned to the presence of ursolic acid, caffeic acid, rosmarinic acid, quercetin and also anthocyanidins (genistin). The obtained results indicate the anti-inflammatory potential of the studied herbal extracts.
Abstract in English:ABSTRACT The current study explored hepatoprotective effect of Aegle marmelos (L.) Corrêa, Rutaceae, leaves extract. Potentiation of A. marmelos hepatoprotective effect with piperine co-administration was also explored. Wistar rats were randomly divided into seven groups: (i) normal control, (ii) paracetamol group, (iii) silymarin group, (iv) extract-25 group (25 mg/kg body), (v) extract-50 group: (50 mg/kg), (vi) extract-100 group (100 mg/kg) and (vii) extract-25 + piperine group. Hepatotoxicity was induced by administering paracetamol orally in a dose of 400 mg/kg for seven days. The drugs were administered 30 min prior to paracetamol administration and continued for seven days. Animals were ‘sacrificed’ at the end of treatment and serum was collected for evaluating alkaline phosphatase, bilirubin, lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase IL-10 and TNF-α levels. Liver homogenates were used for determination of oxidative stress (malondialdehyde, reduced glutathione, superoxide dismutase, catalase, glutathione reductase, GSH-S-transferase, glutathione peroxidase and glucose-6-phosphate dehydrogenase). Serum biochemical markers were significantly higher in paracetamol group as compared to normal control group. Significant increase in oxidative stress parameters and inflammatory mediators was also observed. Treatment with A. marmelos curtailed the toxic effects of paracetamol in a dose dependent fashion. 100 mg/kg dose of A. marmelos was found to be most hepatoprotective. The results of extract-100 group were comparable to silymarin group. Low dose of A. marmelos i.e., 25 mg/kg was combined with piperine to evaluate potentiation of hepatoprotective effects of A. marmelos. Piperine co-administration potentiated the hepatoprotective effects, because the combination group results were comparable to high dose A. marmelos group. A. marmelos exerts hepatoprotective activity through its antioxidant and anti-inflammatory properties which was enhanced by piperine.
Abstract in English:ABSTRACT Moringa oleifera Lam., Moringaceae, and Telfairia occidentalis Hook. f., Curcubitaceae, leaves are two tropical vegetables of medicinal properties. In this study, the inhibitory activities and the radical scavenging potentials of these vegetables on relevant enzymes of type 2-diabetes (α-amylase and α-glucosidase) were evaluated in vitro. HPLC-DAD was used to characterize the phenolic constituents and Fe2+-induced lipid peroxidation in rat's pancreas was investigated. Various radical scavenging properties coupled with metal chelating abilities were also determined. However, phenolic extracts from the vegetables inhibited α-amylase, α-glucosidase and chelated the tested metals (Cu2+ and Fe2+) in a concentration-dependent manner. More so, the inhibitory properties of phenolic rich extracts from these vegetables could be linked to their radical scavenging abilities. Therefore, this study may offer a promising prospect for M. oleifera and T. occidentalis leaves as a potential functional food sources in the management of type 2-diabetes mellitus.
Abstract in English:ABSTRACT Celtis iguanaea (Jacq.) Sarg., Cannabaceae, is popularly used in the treatment of diabetes mellitus. However, chemical and pharmacological investigations are lacking. In this study, we investigated the effects of the hydroalcoholic extract from C. iguanaea on markers of cardiovascular diseases and the glucose metabolism in cholesterol-fed rats. Therefore, hypercholesterolemic rats (1% cholesterol) were orally treated with C. iguanaea extract (C-150, CI-300, or CI-600 mg/kg) or simvastatin (4 mg/kg) (n = 6) once a day (30 days) with a hypercholesterolemic diet. A control group (C) was given saline. C. iguanaea extract showed significant decreases in serum levels of total cholesterol, LDL-cholesterol, HMG-CoA-reductase, interleukin-1 and 6, TNF-α and IFN-γ when compared to group C (p < 0.001). Hypoglycemic effects were observed along with a decrease of the activity of sucrase (CI-600), maltase (CI-150, CI-300), and an increase in muscle glycogen levels (CI-300). Antioxidant effects were observed in plasma by the decrease of TBARS and increase of nonprotein thiols levels (CI-600). The histopathological analysis showed a significant decrease in the liver fat area for C. iguanaea extract compared to group C (p < 0.001). Our results suggest that the biological effects of C. iguanaea extract could be related to the flavonoids that possibly exert antioxidant, enzymatic inhibitory, and insulin-mimetic effects.
Abstract in English:ABSTRACT Eugenia uniflora L., Myrtaceae, popularly known as “pitanga”, is used in traditional medicine due the properties attributed to its chemical content, these being mainly hydrolysable tannins and flavonoids. This study provides a qualitative and quantitative evaluation of chemical profile from leaves of E. uniflora. The HPLC analysis was carried out on a C18 column (4.6 mm × 250 mm, 5 µm) by gradient elution with methanol and water (acidified with trifluoracetic acid); and silica gel Plates 60-F 254 with 10–12 µm and 5–6 µm particles, respectively for TLC and High HPTLC analysis. The chromatographic data obtained from HPLC, TLC and HPTLC presented bands and peaks related to flavonoids (myricitrin and derivatives) and tannins (gallic and ellagic acids), which were observed from different samples. The chromatographic similarities enabled the building of a typical fingerprint for the herbal material. The similarity analysis of the sample data by Pearson correlation showed R values >0.9 among peaks (HPLC) and bands (HPTLC). In addition, the analytical methodology developed by HPLC enabled the satisfactory quantification of marker substances [ellagic acid = 0.22% and 0.20% (m/m); gallic acid = 0.20% and 0.43%; myricitrin = 0.42 and 1.74% (m/m) in herbal drug and crude extract, respectively]. The procedure was also validated in accordance with the assays required by Brazilian legislation. Thus, the HPTLC and HPLC methods developed in this study provide helpful and simple tools for the quality evaluation both qualitatively and quantitatively of raw materials and extractives from leaves of E. uniflora.
Abstract in English:ABSTRACT Natural latex from Hevea brasiliensis (Wild. ex A.Juss) Müll.Arg., Euphorbiaceae, showed angiogenic action and Casearia sylvestris Sw., Salicaceae, leaf derivatives presented anti-inflammatory and wound healing activities. Therefore, an association of these effects was interesting for wound healing applications. The aims of this study were the development of membranes of natural latex incorporated with C. sylvestris leaf derivatives (ethanolic extract, diterpene concentrated fraction and casearin J), their chemical and physical characterization, and the evaluation of in vitro skin permeation and retention of C. sylvestris bioactive secondary metabolites (diterpenes and phenolic compounds). The membranes were developed mixing hydroethanolic solutions of C. sylvestris derivatives with latex and drying them in a desiccator. They were characterized by infrared spectroscopy, scanning electron microscopy, water vapor permeability and mechanical resistance assays, demonstrating that all membranes were permeable, resistant and homogeneous in surfaces. The permeation and retention assays demonstrated dermal penetration of phenolic compounds for ethanolic extract membrane and of casearin-like clerodane diterpenes for all membranes, indicating that these membranes have great potential for therapeutical application as a topical system for C. sylvestris components releasing.
Abstract in English:ABSTRACT The aim of this paper was to study and optimize the dynamic maceration process to obtain Matricaria chamomilla L., Asteraceae, inflorescences extracts with optimum flavonoid content and antioxidant activity using a multivariate approach. Hydroalcoholic extracts were obtained by dynamic maceration in lab scale and the influence of extraction temperature, ratio of plant to solvent, ethanol strength; extraction time and stirring speed on the flavonoid content and antioxidant activity were unveiled using a fractional factorial design. The ethanol strength, ratio of plant to solvent and temperature were the three factors that influenced most the extract properties and were studied by a central composite design. Total flavonoid content and antioxidant activity were affected by the ethanol strength and ranged from 1.49 to 3.95% and 13.3 to 36.2 µg/ml, respectively. The desirability functions resulted in an optimal dynamic maceration condition using 1 h extraction at stirring speed of 900 rpm, ethanol 74.7%, temperature of 69 °C and using 36.8% of plant in solvent (w/v). Under this set of conditions, the extract had total flavonoid content of 4.11 ± 0.07%, in vitro antioxidant activity with IC50 of 18.19 µg/ml and apigenin and apigenin-7-glycoside contents of 2.0 ± 0.1 mg/g and 20.1 ± 0.9 mg/g, respectively. The results showed a low solvent consumption compared to previous works. The model was able to predict extract properties with maximum deviation of 12% and the extraction process developed herein showed to be reliable, efficient and scalable for M. chamomilla inflorescences, enriched with flavonoids, apigenin and apigenin-7-glycoside and high antioxidant activity.
Abstract in English:ABSTRACT A validation study of a reverse-phase high-performance liquid chromatographic assay for the quantification of two furanocoumarins (psoralen and bergapten) in soft extract obtained from Brosimum gaudichaudii Trécul, Moraceae, roots was conducted. The developed method was sensitive, rapid, reproducible, easy and precise, and showed linearity (r > 0.99) in the range of 10–64 µg/ml for psoralen, and 9–56 µg/ml for bergapten. It also showed a good efficiency for the photodegradation analysis of psoralen and bergapten in the soft extract. The photostability results showed that the Higuchi model presented the best fitting to the obtained data. Both chemical markers showed stability over 2.6 days, suggesting potential applications of the extract in obtaining intermediate products from this plant material. Furanocoumarins take around 30 min to be activated by UV light, reaching the maximum biological potential. Thus, the results obtained to the Higuchi model, corresponding to 2.6 days of stability, shows feasibility with future applications of these chemical markers.