关键词: 关键词：槐定碱, LPS, RAW264.7细胞, NF-κB

Abstract: Abstract:Objective This paper aimed at observing the effect of Sophoridine on the expression of IKKβ, IκBα, NF-κB P65 and TNF-α induced by LPS in RAW264.7 macrophages and explore its anti-endotoxin mechanism. Method: The macrophage line RAW264.7 was cultivated and divided into following four groups as control group, Sophoridine control group, LPS group and Sophoridine intervention group. We first put LPS 100μg/L into Sophoridine intervention group for a hour , then remove LPS and add Sophoridine 15.63 mg/L to incubate for 5, 30, 60 and 120minutes respectively to obtain the cells and culture solution. Use RT-PCR technology and Western Blot to detect the mRNA and protein expression . Measure of TNF –α content of supernatant by using Radioimmunoassay (RIA). Result: IKKβ , pIκBα , NF-κB P65 mRNA and /or protein and TNF–α expression in LPS group were higher than control group (P < 0.01) . Sophoridine intervene group above factor mRNA and /or protein and TNF–α expression are dropped significantly compared with LPS group (P < 0.01). Conclusion: It is one of the Sophoridine anti-endotoxin mechanisms that Sophoridine regulate the expression of IKKβ、IκBα、NF-κB P65 of NF-κB pathway , activated by LPS in macrophages，, then inhibit the secretion of downstream TNF – α..

Key words: Key words: Sophoridine, Lipopolysaccharide, RAW264.7 macrophage, NF-κB