RESUMO
BACKGROUND: This study showed phytochemical composition and evaluates the anti-inflammatory, and analgesic activities of crude extract (CE) and fractions from E. uniflora Linn leaves. METHODS: Polyphenols present in crude extract (CE), in aqueous fraction (AqF), and ethyl acetate (EAF) treated fractions from E. uniflora Linn leaves were shown by chromatographic analysis in order to conduct a phytochemical characterization. Antibacterial activity was evaluated based on minimum inhibitory concentrations (MICs) determined using the agar dilution method. Doses of 50, 100, and 200 mg/kg of the CE and fractions were applied for conducting in vivo models (male Swiss mice, 8-10 weeks old). The peritonitis experimental model was induced by carrageenan following of Myeloperoxidase activity (MPO), Total glutathione and malondialdehyde (MDA), IL-1ß and TNF-α levels by spectroscopic UV/VIS analysis. Antinociceptive activity was evaluated based on an abdominal writhing model and hot plate test. The results were statistically evaluated using one-way analysis of variance (ANOVA), followed by Bonferroni's post-hoc test. The level of statistical significance was p < 0.05. RESULTS: High-performance liquid chromatography with photodiode array detection (HPLC-DAD) detected varying concentrations of gallic acid, ellagic acid, and myricitrin in the CE and fractions obtained from E. uniflora Linn leaves (0.05-0.87%w/w, 0.20-0.32%w/w, and 1.71-6.56%w/w, respectively). In general, the CE had lower MIC values than the fractions, including the lowest MIC against the MRSA strain. The CE and AqF also significantly reduced leukocyte migration and MPO activity (p < 0.05). In addition, AqF significantly reduced IL-1ß and TNF-α levels (p < 0.05). Furthermore, the CE and fractions exhibited an antioxidant effect (p < 0.05) and peripheral analgesic activity (p < 0.05). CONCLUSIONS: The CE and fractions from the studied E. uniflora Linn leaves exhibited antibacterial, anti-inflammatory, antioxidant, and analgesic activity in the performed assays.
Assuntos
Antibacterianos/administração & dosagem , Anti-Inflamatórios/administração & dosagem , Antioxidantes/administração & dosagem , Eugenia/química , Peritonite/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Analgésicos/administração & dosagem , Analgésicos/química , Analgésicos/isolamento & purificação , Animais , Antibacterianos/química , Antibacterianos/isolamento & purificação , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Glutationa/imunologia , Humanos , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Masculino , Malondialdeído/imunologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/fisiologia , Camundongos , Peritonite/genética , Peritonite/imunologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Folhas de Planta/química , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Vast numbers of plant species from northeastern Brazil have not yet been phytochemically or biologically evaluated. AIM OF THE STUDY: The goal of this work was to obtain, characterize and show the antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone-water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava. MATERIALS AND METHODS: The plant material (100g) was dried, and the crude extracts were obtained by using turbo-extraction (10%; w/v) with water or acetone:water (7:3, v/v) as the extraction solvent. High-performance liquid chromatography (HPLC) methods were used to screen the crude extracts for hydrolysable tannins (gallic acid) and condensed tannins (catechins). The antibacterial activity was evaluated by agar-diffusion and microdilution methods against Gram-positive strains (Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis INCQS 00016, Enterococcus faecalis ATCC 29212 and a clinical isolate of methicillin-resistant Staphylococcus aureus) as well as Gram-negative strains (Escherichia coli ATCC 25922, Salmonella enteritidis INCQS 00258, Shigella flexneri and Klebsiella pneumoniae). To evaluate the anti-inflammatory activity, a leukocyte migration model was used. Analgesic activity was determined by the hot plate test and the acetic acid-induced abdominal writhing test. Data were analyzed by analysis of variance (ANOVA) at a significance level of 5%. RESULTS: Parapiptadenia rigida presented the highest amount of total polyphenols (35.82 ± 0.20%), while the greatest catechin content was found in the acetone-water extract of Psidium guajava (EAWPg; 1.04 µg/g). The largest amounts of catechins were found in the aqueous extract of Libidibia ferrea (EALf; 1.07 µg/g) and the acetone-water extract of Parapiptadenia rigida (EAWPr; 1.0 µg/g). All extracts showed activity against Gram-positive bacteria. The aqueous and acetone-water extracts of Psidium guajava showed the greatest inhibition zones in the agar diffusion tests. In the evaluation of the minimum inhibitory concentration (MIC), the most susceptible Gram-positive bacterium was Staphylococcus epidermidis and the most susceptible Gram-negative bacterium was Shigella flexneri. EAPg and EAWPg showed the greatest MIC values. All extracts were significant inhibitors of leukocyte migration (p<0.05). Using the writhing test, significant analgesic activity was found for EAPr (50 mg/kg), EAWPr (100 mg/kg and 200 mg/kg) and EAWPg (50 mg/kg) (p<0.05). CONCLUSIONS: Thus, the appropriate extraction procedure preserves the chemical components such as gallic acid and catechin, and showed antimicrobial, anti-inflammatory and analgesic properties.
Assuntos
Caesalpinia , Mimosa , Extratos Vegetais/farmacologia , Polifenóis/química , Polifenóis/farmacologia , Psidium , Analgésicos/química , Analgésicos/farmacologia , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Brasil , Relação Dose-Resposta a Droga , Feminino , Taninos Hidrolisáveis/química , Masculino , Camundongos , Testes de Sensibilidade Microbiana , Folhas de Planta , Proantocianidinas/química , Ratos , Ratos WistarRESUMO
Objetivo: Verificar, ?in vitro?, o potencial antimicrobiano do Anapyon®, da Água Rabelo® e do Malvatricin® sobre microrganismos presentes na cavidade oral.Método: Para realização do experimento, utilizou-se o protocolo sequenciado durante quatro dias que avaliou, através da medida da densidade óptica, o potencial antimicrobiano dos fármacos nos microrganismos (Staphylococcus aureus, Candida tropicalis, Candida parapsilosis e Candida albicans), em suas formas planctônicas, apenas o Malvatricin® foi avaliado sobre as formas de biofilme por ser o único fármaco que apresentou resultados satisfatórios sobre as formas planctônicas. O estudo adotou como controle negativo a água destilada e controle positivo a Clorexidina®. Os resultados obtidos foram submetidos a uma análise estatística com os testes de Kruskal-Wallis e Mann-Whitney. Resultados: Observou-se, através da medida da densidade óptica, que apenas o Malvatricin® apresentou bons resultados na forma planctônica, sendo estes semelhantes ao controle positivo (clorexidina), considerado padrão nos ensaios antimicrobianos em Odontologia. Os resultados do Malvatricin® foram estatisticamente melhores quando comparados aos demais fármacos (Anapyon®, Água Rabelo®) e ao controle negativo. Este resultado foi semelhante para todos os microrganismos estudados: Staphylococcus aureus (p=0,002), Candida tropicalis (p=0,002), Candida parapsilosis (p=0,001) e Candida albicans (p<0,001). Desta forma, apenas o Mavatricin® foi testado para o microrganismo arranjado em biofilme. Observou-se, então, que para a C.albicans e para o S. aureus, houve diferença significativa entre clorexidina e Malvatricin® (p<0,05), com melhores resultados para o clorexidina. Para C. tropicalis, o Malvatricin® diferiu significativamente (p<0,05) da água destilada. Em relação a C.parapsilosis, nenhuma diferença foi observada em relação ao controle negativo (p=0,468). Conclusão: Apesar de alguns fármacos alternativos serem tidos como antimicrobianos, tais propriedades sobre células planctônicas e, principalmente, sobre biofilme foram observadas apenas para o Malvatricin®.
Objective: To evaluate in vitro the antimicrobial potential of Anapyon®, Água Rabelo and Malvatricin® against oral microorganisms.Method: The experiment used a four-day sequential protocol that evaluated by optical density measurements the antimicrobial potential of these products against Staphylococcus aureus, Candida tropicalis, Candida parapsilosis and Candida albicans in their planktonic forms. Only Malvatricin® was also evaluated against microbial biofilms because it was the only one to produce satisfactory results against the planktonic forms. Distilled water was used as negative control and Chlorhexidine® as positive control. The data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests. Results: According to the optical density readings, only Malvatricin® was effective against the planktonic microorganisms and its results were similar to those of the positive control (Chlorhexidine), which is the gold standard antimicrobial agent in dental research. Malvatricin® presented significantly higher efficacy than the other antiseptics (Anapyon® and Água Rabelo®) and the negative control, and this result was similar for all tested microorganisms: S. aureus (p=0.002), C tropicalis (p=0.002), C. parapsilosis (p=0.001) and C. albicans (p<0.001). For this reason, only Malvatricin® was evaluated against the microorganisms arranged as biofilms. Chlorhexidine presented significantly better results (p<0.05) than Malvatricin® against C. albicans and S. aureus. When tested against C. tropicalis, Malvatricin® differed significantly (p<0.05) from distilled water, while against C. parapsilosis no significant difference (p=0.468) was observed in comparison with the negative control. Conclusion: Although some pharmaceutical products being considered anti-microbial, such properties against planktonic cells and especially the biofilms were observed only for Malvatricin®.