醒脑静注射液对内毒素诱导大鼠肺泡巨噬细胞核转录因子-kB激活和细胞因子产生的影响  

Effects of Xingnaojing injection (醒脑静注射液) on lipopolysaccharide-induced nuclear factor-kB activation and cytokine production in rat alveolar macrophages

作　　者：王进[1] 杨光田[1] 乔礼芬[2] 李永胜[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院急诊科,湖北武汉430030 [2]华中科技大学同济医学院附属同济医院呼吸科,湖北武汉430030

出　　处：《中国中西医结合急救杂志》2008年第4期212-215,共4页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：湖北省科技攻关项目(2003AA301C51)

摘　　要：目的探讨内毒素诱导大鼠肺泡巨噬细胞(AMs)核转录因子-kB(NF-kB)的激活和细胞因子的释放以及醒脑静注射液(XNJ)的干预作用。方法通过支气管肺泡灌洗获取大鼠AMs。先用XNJ(10ml/L)孵育AMs 2 h后,加入内毒素(10μg/L)分别刺激2、4和6 h。用逆转录-聚合酶链反应(RT-PCR)检测AMs中肿瘤坏死因子-α(TNF-α)基因表达水平;用酶联免疫吸附法(ELISA)检测培养上清液中TNF-α和白细胞介素-8 (IL-8)含量;蛋白质免疫印迹法(Western blotting)检测AMs中NF-kB抑制蛋白-α(kB-α)、磷酸化IkB-α(pIkB-α)和NF-kB的水平变化。结果内毒素能增加AMs中TNF-α基因表达水平和培养上清液中TNF-α、IL-8的水平,同时能促进IkB-α降解和NF-kB激活。与内毒素组比较,XNJ能减少AMs中TNF-α基因表达水平和培养上清液中TNF-α和IL-8的水平;抑制内毒素诱导的IkB-α降解和NF-kB激活(P均<0.05)。结论XNJ通过抑制AMs中IkB-α的降解,减少了NF-kB的激活,减少了内毒素诱导大鼠AMs细胞因子的产生。Objective To study the effects of Xingnaojing injection （XNJ,醒脑静注射液） on lipopolysaccharide （LPS）-indueed nuclear factor kappa B （NF-κB） activation and cytokine release in rat alveolar macrophages （AMs）. Methods AMs were collected from bronchoalveolar lavage fluid in rats. The AMs were incubated for 2 hours with XNJ （10 ml/L）, and then stimulated for 2, 4 and 6 hours by LPS （10 μg/L）. Reverse transcription-polymerase chain reaction （RT-PCR） was used to detect the gene levels of tumor necrosis factor-α （TNF-α）. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein levels of TNF-α and interleukin-8 （IL-8） in the supernatant fluid of the culture. Western blotting was used to detect the levels of NF-κB inhibitory protein-α （boB-α）, phosphorylated-bcB-α （IκB-α） and NF-κB in the AMs. Results LPS increased the gene expression level of TNF-α in AMs and TNF-α and IL-8 contents in the supernatant fluid of the AMs culture; in the mean time LPS promoted the degradation of IκB-α and NF-κB activation. Compared with LPS group, XNJ decreased the gene expression level of TNF-α in the AMs and the TNF-α and IL-8 release in the supernatant fluid ; XNJ inhibited IκB-α degradation and NF-κB activation induced by LPS （all P 〈 0.05）. Conclusion XNJ can attenuate LPS -induced cytokine production in rat AMs by inhibiting IκB-α degradation and NF-κB activation in AMs.

关 键 词：核转录因子-κB 醒脑静注射液 细胞因子 肺泡巨噬细胞 

分 类 号：R285.5[医药卫生—中药学] Q786[医药卫生—中医学]