基础医学与临床 ›› 2011, Vol. 31 ›› Issue (12): 1320-1325.

• 研究论文 • 上一篇    下一篇

槐定碱抑制LPS诱导巨噬细胞株NF-κB表达

杨峰   

  1. 宁夏医科大学
  • 收稿日期:2011-01-17 修回日期:2011-05-25 出版日期:2011-12-05 发布日期:2011-12-05
  • 通讯作者: 杨峰 E-mail:zhizhuo2000-2000@163.com
  • 基金资助:
    国家重点基础研究发展计划;宁夏自然科学基金;宁夏卫生厅科研项目

Inhibiting effects of Sophoridine of NF-κB induced by lipopolysaccharide in RAW264.7 macrophages in RAW264.7 macrophages

  • Received:2011-01-17 Revised:2011-05-25 Online:2011-12-05 Published:2011-12-05

摘要: 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、槐定碱干预组。槐定碱干预组加入LPS100μg/L孵育1h,弃去LPS后加入槐定碱15.63mg/L,作用5、30、60、120min,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);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)。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。

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

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