槐定碱抑制脂多糖诱导巨噬细胞系NF-κB表达  

Sophoridine inhibits NF-κB expression induced by lipopolysaccharide in RAW264.7 macrophage line

作　　者：杨峰[1] 周娅[1] 曹相原[1] 王艳荣[1] 赵建宁[1] 王宁萍[1] 

机构地区：[1]宁夏医科大学基础医学院病原生物学与免疫学系,宁夏银川750004

出　　处：《基础医学与临床》2011年第12期1320-1325,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30660227);宁夏自然科学基金(NZ0783);银川市科技计划项目基金(银财发[2009]308)

摘　　要：目的观察槐定碱对LPS诱导的RAW264.7巨噬细胞IKKβ,IκBα,NF-κB P65及TNF-α表达的影响,探讨其抗内毒素机制。方法培养RAW264.7巨噬细胞,分为对照组、槐定碱对照组、LPS组及槐定碱干预组。槐定碱干预组加入LPS 100μg/L孵育1 h,弃去LPS后加入槐定碱15.63 mg/L,作用5、30、60和120 min,分别获取细胞与培养液。RT-PCR与Western blot分别检测NF-κB等的mRNA与蛋白表达,放免法检测培养液中TNF-α含量。结果LPS组各时间点IKKβ、pIκBα、NF-κB P65 mRNA和/或蛋白表达及TNF-α含量均显著高于同时间点对照组(P<0.01),IκBαmRNA低于同时间点对照组(P<0.01或P<0.05);槐定碱干预组以上因子mRNA和/或蛋白表达及TNF-α含量均较同时间点LPS组显著回落(P<0.01或P<0.05),而IκBαmRNA则显著升高(P<0.01或P<0.05)。结论槐定碱调控LPS激活的巨噬细胞NF-κB通路IKKβ、IκBα、NF-κB P65等分子的表达,进而抑制下游TNF-α分泌是其抗内毒素作用机制之一。Objective To observe the effect of Sophoridine on the expression of IKKβ, IκBα, NF-KB P65 and TNF-α induced by LPS in RAW264. 7 macrophages and to explore its anti-endotoxin mechanism. Methods The macrophage line RAW264.7 was cultivated and divided into four groups as control group, Sophoridine control group, LPS group and Sophoridine intervention group. We first put LPS 100 μg/L into Sophoridine intervention group for a hour, then removed LPS and add Sophoridine 15.63 mg/L to incubate for 5, 30, 60 and 120 minutes respectively to obtain the cells and culture solution. RT-PCR technology and Western blot were used to detect the mRNA and protein expression. Measure of TNF-α content of supernatant by using Radioimmunoassay （RIA）. Resuits IKK/3, plκBα, NF-KB P65 mRNA and/or protein and TNF-αexpression in LPS group were all higher than that in control group （ P 〈 0. 01 ） at each time-control-point, but IKBa mRNA was lower than that of control group （P 〈0. 01 or P 〈 0. 05 ）. In sophoridine intervention group mRNA and/or protein and TNF-α expression were dropped significantly compared with LPS group （P 〈 0. 01 or P 〈 0. 05） at each time-control-point, and IκBa mRNAwas higher than that of LPS group （P 〈 0. 01 or P 〈 0. 05 ）. Conclusions It might be the one of the Sophoridine anti-endotoxin mechanisms that Sophoridine regulates the expression of IKKβ ,IκBot,NF-αB P65 of NF-κB pathway activated by LPS in macrophages, then inhibited the secretion of downstream TNF-α.

关 键 词：槐定碱 LPS RAW264.7细胞 NF-ΚB 

分 类 号：R965[医药卫生—药理学]