ESTUDO DO EFEITO DA Malva sylvestris EM FIBROBLASTOS ESTIMULADOS POR LPS

• Autor(a) principal: Benso, Bruna
• Data de Publicação: 2012
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Biblioteca Digital de Teses e Dissertações da UEPG
• Texto Completo: http://tede2.uepg.br/jspui/handle/prefix/1780
• topic: Malva sylvestris; doença periodontal; inflamação; Malva sylvestris; periodontal disease; inflammation; CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA

The objective of this study was to develop biomonitoring of the hydroalcoholic extract of Malva sylvestris to evaluate the phytochemical profile beyond the effect of the hydroalcoholic extract of the plant on the release of IL-1β, TNF-α, LPS-induced nitric oxide, and finally, to assess possible toxicological effects of the extract. The activity of the various forms of extraction (soxhlet, maceration, turbolise and infusion) were tested using the ear edema model in mice, induced by 2-Otetradecanoylphorbol 13-acetate (TPA) and subsequently evaluated to determine which ethanol concentration presented the most anti-edematous activity. ESI-MS and LC-ESI-MS analyses were performed to exploit the ions that were representative of patterns of malvidin 3-glucoside, quercetin and scopoletin. Using in vitro tests, fibroblasts from the stock cell line 3T3 were grown in Dulbecco’s modified eagle medium (DMEM). The cells were analyzed by experimental tests after four passages. Stimulation with LPS in 3T3 fibroblasts resulted in an increased production of nitric oxide released in the cell culture supernatant. A method based on the use of fluorogenic probe (DAF-FM DA) was used to quantify the amount of nitric oxide. After sensitization with LPS, the release of IL-1 and TNF-α were measured, and subsequently separately evaluated by ELISA. To identify substances with genotoxic activity the Comet test was employed, using the technique of electrophoresis on microgel. The results demonstrated that at the highest dose tested (3.0 g/ml), the three forms of extraction were able to reduce ear edema by 44-46% and that the extract of Malva sylvestris in 100% ethanol concentration reached a higher inhibition (50.69%). In the in vitro tests, the treatment with hydroalcoholic extract was able to prevent the formation of nitric oxide by 80 ± 14% at the highest concentration used, in comparison to the control group, and the concentration of 1.0 g/ml was able to inhibit 86 ± 2% levels of IL-1. The levels of TNF-α were reduced by 84 ± 4%. The hydroalcoholic extract obtained by maceration at 100% concentration suggests an anti-inflammatory potential when used topically in a skin inflammation model. In assessing cell culture it suggests the interference of the extract on the IL-1 and TNF-α cytokines. This creates the need for further methods to investigate the cellular and molecular mechanisms on which the plant has an anti-inflammatory effect.