Mônica Freiman de Souza Ramos

Pharmacological study of anti-allergic activity of Syzygium cumini (L.) Skeels

Myrtaceae is a plant family widely used in folk medicine and Syzygium and Eugenia are among the most important genera. We investigated the anti-allergic properties of an aqueous leaf extract of Syzygium cumini (L.) Skeels (SC). HPLC analysis revealed that hydrolyzable tannins and flavonoids are the major components of the extract. Oral administration of SC (25-100 mg/kg) in Swiss mice (20-25 g; N = 7/group) inhibited paw edema induced by compound 48/80 (50% inhibition, 100 mg/kg; P <or= 0.05) and, to a lesser extent, the allergic paw edema (23% inhibition, 100 mg/kg; P <or= 0.05). SC treatment also inhibited the edema induced by histamine (58% inhibition; P <or= 0.05) and 5-HT (52% inhibition; P <or= 0.05) but had no effect on platelet-aggregating factor-induced paw edema. SC prevented mast cell degranulation and the consequent histamine release in Wistar rat (180-200 g; N = 7/group) peritoneal mast cells (50% inhibition, 1 microg/mL; P <or= 0.05) induced by compound 48/80. Pre-treatment of BALB/c mice (18-20 g; N = 7/group) with 100 mg/kg of the extract significantly inhibited eosinophil accumulation in allergic pleurisy (from 7.662 +/- 1.524 to 1.89 +/- 0.336 x 10(6)/cavity; P <= 0.001). This effect was related to the inhibition of IL-5 (from 70.9 +/- 25.2 to 12.05 +/- 7.165 pg/mL) and CCL11/eotaxin levels (from 60.4 +/- 8.54 to 32.8 +/- 8.4 ng/mL) in pleural lavage fluid, using ELISA. These findings demonstrate an anti-allergic effect of SC, and indicate that its anti-edematogenic effect is due to the inhibition of mast cell degranulation and of histamine and serotonin effects, whereas the inhibition of eosinophil accumulation in the allergic pleurisy model is probably due to an impairment of CCL11/eotaxin and IL-5 production.

Trans-β-Caryophyllene: An Effective Antileishmanial Compound Found in Commercial Copaiba Oil (Copaifera spp.)

This study investigated the leishmanicidal activity against Leishmania amazonensis of four commercial oils from Copaifera spp. named as C1, C2, C3, and C4, the sesquiterpene and diterpene pools obtained from distilling C4, and isolated β-caryophyllene (CAR). Copaiba oils chemical compositions were analyzed by gas chromatography and correlated with biological activities. Diterpenes-rich oils C2 and C3 showed antipromastigote activity. Sesquiterpenes-rich C1 and C4, and isolated CAR presented a dose-dependent activity against intracellular amastigotes, with IC50s of 2.9 µg/mL, 2.3 µg/mL, and 1.3 µg/mL (6.4 µM), respectively. Based on the highest antiamastigote activity and the low toxicity to the host cells, C4 was steamdistillated to separate pools of sesquiterpenes and diterpenes. Both pools were less active against L. amazonensis and more toxic for the macrophages than the whole C4 oil. The leishmanicidal activity of C3 and C4 oils, as well as C4 fractions and CAR, appears to be independent of nitric oxide production by macrophages. This study pointed out β-caryophyllene as an effective antileishmanial compound and also to its role as potential chemical marker in copaiba oils or fractions derived thereof, aiming further development of this rainforest raw material for leishmaniasis therapy.

Análise química quantitativa para a padronização do óleo de copaíba por cromatografia em fase gasosa de alta resolução

Quantitative GC-FID was evaluated for analysis of methylated copaiba oils, using trans-(-)-caryophyllene or methyl copalate as external standards. Analytical curves showed good linearity and reproducibility in terms of correlation coefficients (0.9992 and 0.996, respectively) and relative standard deviation (< 3%). Quantification of sesquiterpenes and diterpenic acids were performed with each standard, separately. When compared with the integrator response normalization, the standardization was statistically similar for the case of methyl copalate, but the response of trans-(-)-caryophyllene was statistically (P < 0.05) different. This method showed to be suitable for classification and quality control of commercial samples of the oils.

Essential oils from oleoresins of Protium spp. of the Amazon region.

The essential oils of six species of Protium (Burseraceae), collected near Manaus, Amazon, were extracted by hydrodistillation and analysed by GC – MS. Resins from four species were collected yearly, and two species were represented by more than one tree. All six oils were found to be entirely monoterpenoid in constitution and were characterized by the predominance of p-cymene and α- and β-phellandrene. The composition of fresh exudates is associated with increasing content of α-pinene and unsaturated p-menthanes. Copyright

Anti-inflammatory activity of essential oils from Syzygium cumini and Psidium guajava

Abstract Context: Despite the many biological activities reported for essential oils, their anti-inflammatory ability is relatively underexplored considering the wide variation in plant sources and in their volatile composition. Oils from Syzygium cumini Skells (SC) and Psidium guajava L. (PG) (Myrtaceae) have been described as having diverse pharmacological activities. Objective: The current study seeks to evaluate the anti-inflammatory activity of the essential oils from the leaves of SC and PG, as well as some of their terpene-enriched fractions (+V = more volatile and −V = less volatile) obtained by vacuum distillation. Both the pharmacological responses and chemical compositions were correlated. Materials and methods: The relative contents of the oils and their fractions were evaluated by gas chromatography. Individual constituents in the oils were characterized by gas chromatography coupled to mass spectrometry. Anti-inflammatory activity was accessed in the lipopolysaccharide-induced pleurisy model, by measuring the inhibition of total leukocyte, neutrophil and eosinophil migration in the mice pleural lavage, after oil treatment with the oils at 100 mg/kg. Results: Eosinophil migration was inhibited by SC (67%), SC (+V) (63%), PG (76%), PG (+V) (67%) and PG (−V) (74%). This efficacy was correlated with the presence of β-pinene and β-caryophyllene in the oils, a result that was reinforced by evaluating both these pure components (38 and 50% inhibition, respectively). Synergistic effects associated with the presence of α-pinene were speculated. Discussion and conclusion: Essential oils from SC and PG may be useful to treat inflammatory diseases by mechanisms that include the inhibition of eosinophil migration.

Application of High Temperature High Resolution Gas Chromatography to Crude Extracts of Propolis

The underivatized acetone and hexane fractions from propolis samples (predominant flora Citrus spp. and Vernonia polyanthes) were analyzed by HT-HRGC (high temperature high resolution gas chromatography) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS). Several compounds, including flavonoid aglycones, phenolic acids, and high molecular weight compounds were characterized in crude extracts by HT-HRGC-MS. HT-HRGC and HT-HRGC-MS were shown to be quick and informative tools for rapid analysis of crude extracts without need for prior derivatization and purification.

Essential Oils From Myrtaceae Species of the Brazilian Southeastern Maritime Forest (Restinga)

Abstract The leaf oils from five Myrtaceae species growing in the restinga forest of Carapebus, on the southeastern Brazilian coast, were obtained by hydrodistillation and analyzed by GC/MS. Eugenia punicifolia oil contained only sesquiterpenes, with predominance of β-elemene (22.1%) followed by β-caryophyllene (8.5%) and components of the selinane (24.8%) and cadinane (14.0%) skeletal-types. Eugenia sulcata oil had the highest content of monoterpenes, among which α-pinene (12.2%), β-pinene (10.8%) and 1,8-cineole (19.6%) predominated. This profile was completed with 23.5% of caryophyllane and humulane derivatives. α-Pinene also appeared significantly (18.8%) in Eugenia rotundifolia, followed by small amounts of α-terpineol (4.4), while limonene (4.7%) and 1,8-cineole (6.8%) were the main monoterpenes in Myrciaria floribunda. The chemical profile of Neomitranthes obscura was also dominated by selinane-type sesquiterpenes: α-cadinene (23.4%), selina-3,7(11)-diene (13.9%), β-selinene (8.5%) and α-selinene (5.5%), followed by traces of pinenes, limonene and 1,8-cineole (2.4%). Total cyclic sesquiterpene alcohols, esters and epoxides decreased from 55.5% in E. rotundifolia to 8.6% in M. floribunda. However, the latter species contained 26.9% of (E)-nerolidol in the oil composition. Traces of ketones (up to 3.1%) are present in E. punicifolia, N. obscura and M. floribunda.

Genotoxicity assessment of Copaiba oil and its fractions in Swiss mice

Copaiba oil-resin, extracted from the trunk of Copaifera, and traditionally used in folk medicine in the treatment of various disorders, has been shown to be an effective antiinflamatory, antitumor, antitetanus, antiseptic and anti-blenorrhagea agent. As, there are few studies evaluating its genotoxicity, this aspect of the commercial oil-resin, and its volatile and resinous fractions, were evaluated in mice by comet assay and micronucleus (MN) test. A single dose of oil resin, volatile or resin fractions (500; 1,000 or 2,000 mg/kg b.w.) was administered by gavage. The chemical compositions of Copaiba oil resin and its fractions was analyzed by gas chromatography. According to comet assaying, treatment with either one did not increase DNA damage, and as to MN testing, there was no alteration in the incidence of micronucleated polychromatic erythrocytes. Chromatographic analysis of the oil-resin itself revealed sesquiterpenes, diterpenic carboxylic acid methyl esters and high levels of β-caryophyllene. Thus, it can be assumed that the oil resin and volatile and resinous fractions from the commercial product are not genotoxic or mutagenic.

Chemical composition of South American Burseraceae non-volatile oleoresins and preliminary solubility assessment of their commercial blend.

INTRODUCTION Non-volatile oleoresins from neotropical Burseraceae are traditionally used for craft, technological and medicinal purposes. The crude resin is usually sold in popular markets of the forest communities. Adding value to this rainforest raw material requires establishing its composition. OBJECTIVE To analyse the resin composition from different Burseraceae species and establish a minimally reproducible profile by gas chromatography, in order to parameterise its quality control. METHODOLOGY Crude oleoresin samples of 10 Protium and Trattinnickia species and a commercial blend were subjected to hydrodistillation to remove volatile compounds. The chloroform-soluble residues were methylated, analysed by GC-FID (flame ionisation detection), and individual components were identified by analysing their mass fragmentation pattern in GC-MS and comparison with data from the literature. The blend solubility was assayed in 30 non-chlorinated solvents at three different proportions. RESULTS The resins consisted exclusively of triterpenes, showing a common predominance of four major compounds in all the samples, corresponding to α-amyrin, β-amyrin, α-amyrenone and β-amyrenone. This profile was complemented with minor amounts of the tetracyclic β-elemolic and α-elemolic acids, maniladiol, brein and other oxidised trace compounds. The better solvents for the resin were those chemically bearing more than four carbon atoms, as n-butyl acetate, 2-ethoxyethanol and isopropanol. The crude resin blend sold contained around 10% of insoluble material that was constituted by up to 70% inorganic residues mixed with humic acid derivatives, as attested by ash analysis and IR spectroscopy, respectively. CONCLUSION The experimental results, complemented by a general inspection of the literature, demonstrated a systematically reproducible triterpene profile in Protium and Trattinnickia species.

Fish oil attenuates persistent inflammatory pain in rats through modulation of TNF-α and resolvins.

UNLABELLED Fish oil (FO), source of omega-3 fatty acids (FA), has been widely studied in the treatment of inflammatory diseases and inflammatory pain (IP). Omega-3 FA give rise to eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, metabolized to eicosanoids and converted to resolvins with important anti-inflammatory action. AIMS This study investigates the effects of oral doses of omega-3 FA from FO and concentrated fish oil (CFO) in a model of sub-chronic IP, induced by Complete Freunds Adjuvant (CFA). MAIN METHODS IP was induced by intraplantar injection of CFA into the right hind paw of Wistar rats. Three groups were pre-treated with omega-3 FA: two groups received CFO (460mg of EPA/360mg of DHA and 690mg of EPA/540mg of DHA) and one group received natural FO (460mg EPA/300mg DHA), for 7days before IP induction (pre-treatment) and 5days after induction (treatment). KEY FINDINGS TNF-α levels were reduced by CFO 690 (67.9%; p<0.01), CFO 460 (57.7%; p<0.01), FO 460 (26.2%), compared to the augment promoted by CFA (549.7%; p<0.001). Resolvin levels were increased in treated groups with respect to the CFA control group (CFO 690=3196.3%, p<0.01; CFO 460=3347.1%, p<0.01; FO=1653.5%). SIGNIFICANCE The results indicate that the tested doses reduced inflammatory pain effectively in a short pre-treatment period, through modulation of TNF-α and resolvins and that CFO presented better results than FO. Therefore, Ω-3 FA from FO can be proposed for use as complementary medicine in the treatment of painful and inflammatory diseases.