Anxiolytic-like effect of Citrus limon (L. ) Burm f. essential oil inhalation on mice / Efeito ansiolítico da inalação do óleo essencial de Citrus limon (L. ) Burm f. em camundongos.

ABSTRACT Experimental in vivo study aimed to characterize the anxiolytic-like effect of the Citrus limon fruit peel’s essential oil (CLEO) in animal models of anxiety, besides evaluating the viability J774.A1 cells in vitro through the MTT reduction method at the concentrations of 10 and 100 µg/mL. The anxiolytic behavior was evaluated in Swiss mice (n = 8) using the methodology of Elevated Plus-Maze (EPM) and Open-Field (OF). CLEO was tested by inhalation at the doses of 100, 200, and 400 µL, and as control, animals were subjected to inhalation of the vehicle (saline solution 0.9% + Tween80®) and intraperitoneal administration of diazepam (1.5 mg/kg). In the cell viability assay, it was observed that none of the concentrations showed cytotoxicity. OF test showed significant anxiolytic activity at all tested doses of OECL, compared to the control group, without changing the motor performance of the animals. Corroborating OF data, the EPM test confirmed anxiolytic activity in at least two doses of the tested oil (200 and 400 µL), justified by the number of entries and increase in the percentage of time in the open arms. The data analysis of this study evidenced that inhalation of OECL was able to induce an anxiolytic behavior in mice; however, further studies are required to ensure its safe use by the population.

RESUMO O estudo experimental in vivo objetivou caracterizar o efeito ansiolítico do óleo essencial das cascas do fruto de Citrus limon (OECL) em modelos animais de ansiedade, além de avaliar in vitro a viabilidade de células J774.A1 através do método de redução do MTT nas concentrações de 10 e 100 µg/mL. O comportamento ansiolítico foi avaliado em camundongos Swiss (n = 8), utilizando a metodologia do Labirinto em Cruz Elevado (LCE) e do Campo Aberto (CA). O OECL foi testado por inalação nas doses de 100, 200 e 400 µL, e como controle, os grupos foram submetidos à inalação do veículo (Solução fisiológica 0,9% + Tween80®) e administração intraperitoneal de diazepam (1,5 mg/kg). No ensaio de viabilidade celular, observou-se que nenhuma das concentrações apresentou citotoxicidade. O teste de CA demonstrou atividade ansiolítica significativa em todas as doses testadas do OECL, comparada ao grupo controle, sem alterar a performance motora dos animais. Corroborando os dados do CA, o teste do LCE confirmou a atividade ansiolítica em pelo menos duas das doses do óleo testadas (200 e 400 µL), justificada pelo número de entradas e aumento na percentagem de tempo nos braços abertos. A análise dos dados deste estudo evidenciou que a inalação do OECL foi capaz de induzir um comportamento ansiolítico em camundongos; entretanto, outros estudos são necessários para garantir o seu uso seguro pela população.

Avaliação da atividade antibacteriana, citotóxica e antioxidante da espécie vegetal Opuntia cochenillifera (L. ) Mill / Evaluation of antibacterial, cytotoxic, and antioxidant activity of the plant species Opuntia cochenillifera (L. ) Mill

RESUMO O presente estudo teve como objetivo avaliar a atividade antibacteriana, antioxidante e citotóxica da espécie Opuntia cochenillifera (L.) Mill. Foi realizada a prospecção fitoquímica e espectroscopia de absorção de infravermelho (IV) dos extratos etanólicos brutos e frações dos cladódios grande e pequeno. A atividade antioxidante foi avaliada pelo método da capacidade sequestradora de radicais livres utilizando o radical sintético 2,2-difenil-1-picrilhidrazila (DPPH). A atividade citotóxica foi obtida através do método colorimétrico do Metiltetrazolium (MTT). Já a atividade antibacteriana foi avaliada pelo método de microdiluição em caldo para determinar a concentração inibitória mínima (CIM) frente às estirpes bacterianas Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa e Escherichia coli. A prospecção fitoquímica revelou principalmente a presença de fenóis, esteroides livres, alcaloides, alcanos, além de outras classes químicas. O IV apresentou grupos funcionais como alcanos, carbonilas, grupos de metila, duplas ligações de carbono, grupamentos alquilamina, entre outros. Sobre a citotoxicidade na concentração de 100 μg/mL, os dois extratos brutos, todas as frações do cladódio grande e as frações de clorofórmio e metanol do cladódio pequeno não apresentaram toxicidade. Os extratos brutos e frações do cladódio grande e pequeno, não demonstraram atividade antibacteriana e nem antioxidante. Esses resultados podem fornecer suporte para pesquisas futuras, visando outras atividades biológicas da presente espécie vegetal.

ABSTRACT The purpose of this study was to evaluate the antibacterial, antioxidant, and cytotoxic activity of Opuntia cochenillifera (L.) Mill. A phytochemical screening and infrared (IR) absorption spectroscopy were performed in the crude ethanolic extracts and fractions of large and small cladodes. The antioxidant activity was evaluated through the qualitative method of free-radical scavenging capacity using the synthetic radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). The cytotoxic activity was obtained by the cell viability assay using methyl thiazolyl tetrazolium (MTT). Antibacterial activity was evaluated by broth microdilution method to determine the minimum inhibitory concentration (MIC) against the bacterial strains Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Escherichia coli. The phytochemical screening mainly revealed the presence of phenols, flavonoids, free steroids, alkaloids, alkanes, and other chemical classes. The IR spectroscopy presented functional groups such as alkanes, carbonyls, methyl groups, carbon double bonds, and alkylamino groups, among others. Regarding cytotoxicity in the concentration of 100 μg/mL, neither the crude extracts, the fractions of the large cladode, nor the chloroform and methanol fractions of small cladode presented toxicity. The crude ethanolic extracts and fractions of large and small cladode showed no antibacterial or antioxidant activity. These results may provide support for future research aimed at other biological activities of this plant species.

Modulation of B lymphocyte function by an aqueous fraction of the ethanol extract of Cissampelos sympodialis Eichl (Menispermaceae)

Cissampelos sympodialis Eichl species are used in folk medicine for the treatment of asthma, arthritis and rheumatism. In the present study, we investigated the immunomodulatory effect of an aqueous fraction of a 70 percent (v/v) ethanol extract of C. sympodialis leaves on B lymphocyte function. The hydroalcoholic extract inhibited the in vitro proliferative response of resting B cells induced by LPS (IC50 = 17.2 æg/ml), anti-delta-dextran (IC50 = 13.9 æg/ml) and anti-IgM (IC50 = 24.3 æg/ml) but did not affect the anti-MHC class II antibody-stimulated proliferative response of B cell blasts obtained by stimulation with IL-4 and anti-IgM. Incubation with the hydroalcoholic extract used at 50 æg/ml induced a 700 percent increase in intracellular cAMP levels. IgM secretion by resting B cells (obtained from normal mice) and polyclonally activated B cells (obtained from Trypanosoma cruzi-infected animals) was inhibited by the hydroalcoholic extract. The latter were more sensitive to the hydroalcoholic extract since 6.5 æg/ml induced a 20 percent inhibition in the response of cells from normal mice while it inhibited the response of B cells from infected animals by 75 percent. The present data indicate that the alcoholic extract of C. sympodialis inhibited B cell function through an increase in intracellular cAMP levels. The finding that the hydroalcoholic extract inhibited immunoglobulin secretion suggests a therapeutic use for the extract from C. sympodialis in conditions associated with unregulated B cell function and enhanced immunoglobulin secretion. Finally, the inhibitory effect of the hydroalcoholic extract on B cells may indicate an anti-inflammatory effect of this extract.

Cissampelos sympodialis Eichl (Menispermaceae) leaf extract induces interleukin-10-dependent inhibition of Trypanosoma cruzi killing by macrophages

The aqueous fraction of the ethanolic extract (AFL) of Cissampelos sympodialis Eichl (Menispermaceae), popularly known as milona, has been shown to have both immunosuppressive and anti-inflammatory effects. In the present study we investigated the modulation of macrophage antimicrobicidal activity by in vitro treatment with the extract from C. sympodialis. Normal and thioglycolate-elicited mouse peritoneal macrophages were infected in vitro with the protozoan Trypanosoma cruzi DM28c clone. We observed that the AFL (used at doses ranging from 13 to 100 æg/ml) increased T. cruzi growth and induced a 75 percent reduction in nitric oxide production. This inhibition could be mediated by the stimulation of macrophage interleukin-10 (IL-10) secretion since the in vitro treatment with the AFL stimulated IL-10 production by T. cruzi-infected macrophages. These results suggest that the anti-inflammatory effect of the AFL from C. sympodialis could be, at least in part, mediated by the inhibition of macrophage functions and that the inhibition of macrophage microbicidal activity induced by the C. sympodialis extract may be mediated by the decrease in macrophage function mediated by interleukin-10 production