ABSTRACT
Objective To investigate the inhibitory effect and mechanism of soybean-derived Bowman-Birk inhibi-tor (BBI)on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells.Methods Cytotoxicity effect of LPS and BBI on intestinal epithelial cells was analyzed by MTT assay.Intestinal epithelial cells were pre-treated with BBI,followed by LPS stimulation,expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1)was detected by quantitative real-time polymerase chain reaction;the activation of NF-κB was measured by pNF-κB-luc system and Western Blot.Results The maximum concentration of LPS (10000 ng/mL)and BBI (1000 μg/mL)had no cytotoxicity effect on intestinal epithelial cells.LPS could potently up-regulate the expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1 ),the up-regulation was positively correlated to the concentration of LPS;LPS-induced expression of inflammatory cytokines in intestinal epithelial cells could achieve the highest level,then decreased over time.The up-regulation of LPS on inflammatory cytokines in intestinal epi-thelial cells had dose-time effect;when intestinal epithelial cells were pretreated by BBI for 6 hours,the inhibitory effect of BBI on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells was most obvious, and had dose-time effect.Conclusion BBI can potently inhibit LPS-induced expression of inflammatory cytokines through inhibiting NF-κB in intestinal epithelial cells.
ABSTRACT
Objective To investigate the inhibitory effect and mechanism of soybean-derived Bowman-Birk inhibi-tor (BBI)on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells.Methods Cytotoxicity effect of LPS and BBI on intestinal epithelial cells was analyzed by MTT assay.Intestinal epithelial cells were pre-treated with BBI,followed by LPS stimulation,expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1)was detected by quantitative real-time polymerase chain reaction;the activation of NF-κB was measured by pNF-κB-luc system and Western Blot.Results The maximum concentration of LPS (10000 ng/mL)and BBI (1000 μg/mL)had no cytotoxicity effect on intestinal epithelial cells.LPS could potently up-regulate the expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1 ),the up-regulation was positively correlated to the concentration of LPS;LPS-induced expression of inflammatory cytokines in intestinal epithelial cells could achieve the highest level,then decreased over time.The up-regulation of LPS on inflammatory cytokines in intestinal epi-thelial cells had dose-time effect;when intestinal epithelial cells were pretreated by BBI for 6 hours,the inhibitory effect of BBI on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells was most obvious, and had dose-time effect.Conclusion BBI can potently inhibit LPS-induced expression of inflammatory cytokines through inhibiting NF-κB in intestinal epithelial cells.