1. Citotoxicidade e a????o anti-inflamat??ria in vitro do extrato hidroalco??lico de Libidibia ferrea e do ??cido g??lico
- Author
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Lins, M??rcia Arruda, Toda, Carina, Bandeira, Maria Fulg??ncia Costa Lima, Conde, Nikeila Chacon de Oliveira, and Vasconcellos, Marne Carvalho de
- Subjects
Mat??ria m??dica vegetal ,Agentes anti-infecciosos ,Citotoxicidade ,Inflama????o ,Plantas medicinais ,Juc?? ,??xido n??trico ,ODONTOLOGIA [CI??NCIAS DA SA??DE] ,??cido g??lico - Abstract
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No. of bitstreams: 4 Carta de encaminhamento_autodeposito.pdf: 448021 bytes, checksum: e58b6f5ec34f4ffe1b4d5f16daf4a459 (MD5) Ata Ma??rcia-Assinada (1).pdf: 243431 bytes, checksum: bce5f3a3b5a3d94d31291395015a5c98 (MD5) Disserta????o_M??rciaArrudaLins.pdf: 1351776 bytes, checksum: 680cb6c618e06a6fda3179cfd0276ac2 (MD5) Ficha cartologr??fica_Disserta????oM??rcia.pdf: 1889 bytes, checksum: cf3ac91769290171f14d76a30e277634 (MD5) Previous issue date: 2020-08-28 FAPEAM - Funda????o de Amparo ?? Pesquisa do Estado do Amazonas Popularly known as '' juc?? '' or '' pau-ferro '', a Libidibia ferrea L. is a plant species widely used in traditional medicine for the treatment of various diseases, therefore, demonstrating several therapeutic properties such as antimicrobial, antioxidant activity and anti-inflammatory. Phytochemical studies highlight the presence of one of the possible responses for some of these biological activities, the gallic acid, polyphenol found in several plants, which is used as one of the main markers for control studies of Libidibia ferrea L. objective to evaluate in vitro the cytotoxicity and anti-inflammatory activity of a 7.5% (w / v) extract of Libidibia ferrea L. on RAW 264.7 macrophages and human peripheral blood monocytes. The evaluation was carried out through the MTT colorimetric assay (3- [4,5-dimethylthiazolyl-2] -2,5-diphenyltetrazolium bromide) and indirect quantification of nitric oxide (NO) by the Griess method. The culture of RAW 264.7 macrophages and the primary cells of human peripheral blood monocytes were handled in culture media, which were subsequently challenged with Escherichia coli LPS (Lipopolysaccharide), and treated with the hydroalcoholic extract of Libidibia ferrea L. and gallic acid for 24h. The assays were first performed on RAW 264.7 macrophages, to obtain the minimum and non-cytotoxic practices possible to inhibit NO production. They were divided into six groups: extract of Libidibia ferrea L. should from 1.56 ??g / mL to 100 ??g / mL, gallic acid 3.125 ??M to 100 ??M, positive control (LPS), negative control (culture medium), and the standard drug (20 ??g / mL dexamethasone). Once the minimal non-cytotoxic and inhibitory options were obtained in the macrophages, the same assays were performed in human peripheral blood monocytes with the same six groups, but with L.ferrea extract in the 50 and 100 ??g / mL procedures, and the 6.25 ??M gallic acid. An analysis of variance of the data was performed using the ANOVA tests, followed by the Tukey and Dunnett test, with a statistically significant difference when p??0.05. In macrophages, the tested concentrations of 1.56 ??g / mL to 100 ??g / mL of L.ferrea extract, none compromised cell viability, whereas in the tested concentrations of gallic acid from 3.25 ??M to 100 ??M, only concentrations of 3.125 ??M and 6.25 ??M kept the cells viable. In the quantification of NO, the extract of L.ferrea at 50 and 100 ??g / mL and the gallic acid at 6.25 ??M, obtained the most satisfactory results in the inhibition of NO production, considering minimum non-cytotoxic and anti-inflammatory concentrations . In the monocytes of primary culture, the results obtained in the macrophages RAW 264.7 were maintained, highlighting these samples on an anti-inflammatory potential. It can be concluded that the extract of Libidibia ferrea L. and gallic acid presented non-cytotoxic concentrations and capable of inhibiting the production of nitric oxide when tested in different strains, in which it is noteworthy that in both cell lines, tested samples obtained more satisfactory results when compared to the standard anti-inflammatory drug, dexamethasone. Popularmente denominada como ??????juc???????? ou ??????pau-ferro??????, a Libidibia ferrea L. ?? uma esp??cie vegetal extensamente utilizada na medicina tradicional para o tratamento de diversas enfermidades, portanto, demonstrando diversas propriedades terap??uticas como: atividade antimicrobiana, antioxidante e anti-inflamat??ria. Estudos fitoqu??micos destacam a presen??a de um dos poss??veis respons??veis por algumas destas atividades biol??gicas, o ??cido g??lico, polifenol encontrado em diversas plantas, que ?? utilizado como um dos principais marcadores para estudos controle da Libidibia ferrea L. O presente trabalho teve como objetivo avaliar in vitro a citotoxicidade e a atividade anti-inflamat??ria de um extrato de Libidibia ferrea L. a 7,5% (m/v) sobre macr??fagos RAW 264.7 e mon??citos de sangue perif??rico humano. A avalia????o foi realizada atrav??s do ensaio colorim??trico de MTT (3-[4,5-dimethylthiazolyl-2]-2,5-diphenyltetrazolium bromide) e quantifica????o indireta de ??xido nitr??co (NO) pelo m??todo de Griess. A cultura de macr??fagos RAW 264.7 e a c??lulas prim??rias de mon??citos de sangue perif??rico humano foram manuseadas em meios de cultura, que posteriormente foram desafiadas com LPS (Lipopolissacarideo) de Escherichia coli, e tratadas com o extrato hidroalco??lico de Libidibia ferrea L. e ??cido g??lico por 24h. Os ensaios foram primeiramente realizados em macr??fagos RAW 264.7, para obten????o das concentra????es n??o citot??xicas e m??nimas capazes de inibir a produ????o de NO. Foram divididos em seis grupos: extrato do Libidibia ferrea L. nas concentra????es de 1,56 ??g/mL a 100 ??g/mL, ??cido g??lico 3,125 ??M a 100 ??M, controle positivo (LPS), controle negativo (meio de cultura), e a droga padr??o (dexametasona a 20??g/mL). Uma vez obtida as concentra????es n??o citot??xicas e inibit??rias m??nimas nos macr??fagos, os mesmos ensaios foram realizados em m??nocitos de sangue perif??rico humano com os mesmos seis grupos, por??m com o extrato de L.ferrea nas concentra????es de 50 e 100 ??g/mL, e o ??cido g??lico a 6,25 ??M. Foi realizada a an??lise de vari??ncia dos dados pelos testes ANOVA, seguido do Teste de Tukey e Dunnett, com diferen??a estatisticamente significativa quando p??0,05. Nos macr??fagos, as concentra????es testadas de 1,56 ??g/mL a 100 ??g/mL do extrato de L.ferrea, nenhuma comprometeu a viabilidade celular, enquanto que nas concentra????es testadas de ??cido g??lico de 3,25 ??M a 100 ??M, apenas as concentra????es 3,125 ??M e 6,25 ??M mantiveram as c??lulas vi??veis. Na quantifica????o de NO, o extrato de L.ferrea a 50 e 100 ??g/mL e o ??cido g??lico a 6,25 ??M, obtiveram os resultados mais satisfat??rios na inibi????o da produ????o do NO, sendo consideradas concentra????es n??o citot??xicas e anti-inflamat??rias m??nimas. Nos mon??citos de cultura prim??ria, houve a manunten????o dos resultados obtidos nos macr??fagos RAW 264.7, destacando estas amostras sobre um potencial anti-inflamat??rio. Pode-se concluir que, o extrato da Libidibia ferrea L. e o ??cido g??lico apresentaram concentra????es n??o citot??xicas e capazes de inibirem a produ????o de ??xido n??trico quando testadas em linhagens diferentes, no qual ressalta-se ainda que em ambas as linhagens celulares, as amostras testadas obtiveram resultados mais satisfat??rios quando comparados a droga anti-inflamat??ria padr??o, a dexametasona.
- Published
- 2020