Anti-Inflammation Assay of Black Soybean Extract and Its Compounds on Lipopolysaccharide-Induced RAW 264.7 Cell

Abstract

Abstract Inflammation response is related with various diseases. One of the useful therapeutic method to suppress inflammatory mediator synthesis is by application of compounds isolated from herbal medicine as treatment for inflammatory diseases. The aim of this study was to analyse the anti-inflammatory activity of black soybean extract (BSE), daidzein, and genistein trough in vitro analysis of inflammatory mediators such as prostaglandin 2 (PGE2) and cytokines interleukin 1β (IL-1β), and tumor necrosis factor α (TNF-α). Safety of samples was determined by viability test using MTS (3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium). Concentration tested for viability assay were 40, 200, 1000 μg/mL for BSE, daidzein, and genistein. Anti-inflammation activity of samples was determined by ELISA quantification of PGE-2, TNF-α, and IL-1β in conditioned medium (CM) of supplemented pro-inflammatory activated RAW 264.7 cell. Inflammation on cells were induced by Lipopolysaccharide (LPS). BSE 1000 ug/ml, daidzein 1000 ug/ml, and genistein 1000 ug/ml treatments shows <80% cell viability average compared to control cell, indicating the treatments have cytotoxicity effect on RAW 264.7 cells. Hence, concentration used for treatments are 40 and 200 μg/mL for each sample. Genistein with concentration of 40 μg/ml treatment result shows highest anti-inflammatory activity which indicated from PGE-2, TNF-α, and IL-1β concentration. This study suggests that BSE, daidzein, and genistein with concentration of 40 and 200 μg/ml were safe to use for RAW 264.7 cell and genistein with concentration of 40 μg/ml have the best anti-inflammatory activity compared to daidzein and BSE.