Atividade citotóxica e anti-inflamatória do extrato glicólico de Camellia sinensis (L.) Kuntze em macrófagos (RAW 264.7) estimulados por LPS

Abstract

Atividade citotóxica e anti-inflamatória do extrato glicólico de Camellia sinensis (L.) Kuntze em macrófagos (RAW 264.7) estimulados por LPS / Cytotoxic and antiinflamatory activity of the glycolic extract of Camellia sinensis (L.) Kuntze in LPS-stimulated machrophages (RAW 264.7) ABSTRACT Plant extracts can be a source of diverse biologic activities, as anti-inflammatory action, which is an interesting characteristic for mouthwashes, toothpastes and intracanal medication. This study aimed to evaluate cytotoxic and anti-inflammatory activity of Camellia sinensis (L.) Kuntze (green tea) glycolic extract in mouse macrophages (RAW 264.7). Cytotoxic activity was measured by the metabolic activity of MTT test, macrophages were distributed in 96 wells and exposed to 11 serial dilutions of each extract (200 mg/mL to 0.20 mg/mL). After 5 min and 24 h of contact, cell viability was assessed. Then, the extract at concentrations of 3.13 mg / mL and 12.5 mg / mL, were chosen to verify antiinflammatory activity. For this, after exposure time of 5 min or 24 h to the extract, supernatants of LPS-stimulated RAW 264.7 cultures were collected to quantify pro-inflammatory cytokines IL-1β and TNF-α by immunoenzymatic test (ELISA). The results were evaluated with statistical analyses (ANOVA and Turkey test), with p ≤ 0.05. Results: In MTT test green tea promoted increase cell viability in all concentrations at 5 min. The cell viability was greater than 100% in concentration of 0,20mg/mL to 12,5 mg/mL, at the time of exposure of 24h. The concentration of 12.5 mg/mL was the highest concentration less cytotoxic. The extract showed anti-inflammatory potential evidenced by the production decrease of IL-1β and TNF-α with better results at a concentration of 12.5 mg/mL 1096 for both exposure times. These results indicate promising immunomodulatory features of green tea. Therefore, this plant extract showed to be an interesting alternative to be inserted in medical or oral products or even as a source of active compounds. RESUMOExtratos de plantas podem ser fonte de diversas atividades biológicas. Dentre essas, a ação antiinflamatória se apresenta como uma das características de interesse para enxaguantes bucais, pastas dentais e medicações intracanais. O objetivo deste estudo foi avaliar a atividade citotóxica e antiinflamatória do extrato glicólico de Camellia sinensis (L.) Kuntze (chá verde) em macrófagos de camundongos (RAW 264.7). A atividade citotóxica foi medida pela atividade metabólica do teste de MTT, os macrófagos foram distribuídos em 96 poços e expostos a 11 diluições seriadas de cada extrato (200 mg / mL a 0,20 mg / mL). Após 5 min e 24 h de contato, a viabilidade celular foi avaliada. Então, o extrato nas concentrações de 3,13mg/mL e 12,5 mg / mL , foram escolhidas para verificar a atividade anti-inflamatória. Para isso, após a exposição do extrato por 5 minutos ou 24 horas, os sobrenadantes das células RAW 264.7 foram coletados para quantificação das citocinas pró-inflamatórias IL-1β e TNF-α pelo teste imunoenzimático ELISA...