多黏菌素B对内毒素诱导大鼠肺泡巨噬细胞I-κB激酶mRNA表达的影响  

Effects of polymixin B on I-κB kinase mRNA expression in lipopolysaccharide-induced pulmonary alveolar macrophages

作　　者：尹文[1] 扬剑虹[1] 虎晓岷[1] 袁静[2] 罗雁军[1] 李月彩[1] 

机构地区：[1]第四军医大学西京医院急诊科,陕西西安710032 [2]解放军第五医院,宁夏银川750004

出　　处：《中国危重病急救医学》2003年第7期429-431,共3页Chinese Critical Care Medicine

基　　金：国家自然科学基金资助项目 ( 3 0 0 0 0 165 )

摘　　要：目的 :观察多黏菌素 B(PMB)对内毒素 (L PS)刺激大鼠的肺泡巨噬细胞 (PAM)中 IκB激酶(IKKβ)、抑制蛋白 (IκBα)和核因子 κB(NFκB)的影响 ,探讨 PMB可能的抗炎机制。方法 :分离、培养大鼠PAM,分为正常对照组、L PS刺激组、PMB+L PS干预组。在 L PS刺激后 0 .5 h采用原位杂交 (ISN)技术、酶联免疫吸附试验 (EL ISA )法及凝胶电泳迁移率改变分析法 (EMSA ) ,分别检测各组 PAM中 IKKβ m RNA、IκBα水平、NFκB活性变化。结果 :与正常对照组比较 ,L PS刺激组 PAM中 IKKβ m RNA的表达 (0 .14 7±0 .0 15 )、NFκB的活性 (0 .82 8± 0 .0 19)均明显升高 (P均 <0 .0 1) ,IκBα水平 (0 .2 2 8± 0 .0 2 1)显著降低 (P<0 .0 1) ;PMB干预后能明显下调 L PS刺激组 IKKβ m RNA表达 (0 .112± 0 .0 2 2 )和 NFκB的活性 (0 .35 8±0 .0 11) ,上调 IκBα水平 (0 .4 77± 0 .0 16 ) ,P均 <0 .0 1。结论 :L PS能激活 PAM中 IKKβ m RNA表达 ,介导IκBα降解和 NFκB活化 ;PMB通过干预 IKKβ/IκBα/NFκB传导通路发挥抗炎作用。Objective: To investigate effects of polymixin B (PMB) on IκB kinase(IKKβ), inhibitor protein(IκBα) and nuclear factorkappa B(NFκB) in lipopolysaccharide(LPS) induced pulmonary alveolar macrophages (PAM), and explore the antiinflammatory mechanism of PMB. Methods: PAM from rats collected by bronchoalveolar lavage was cultured and divided into three groups. In the control group, PAM was not stimulated with LPS and not treated with PMB. In the LPS stimulated group, PAM was stimulated with LPS. In the PMB treated group, PAM was pretreated with PMB half an hour prior to LPS stimulation. The expression of IKKβ mRNA, level of IκBα and the activity of NFκB in PAM were measured by in situ hybridization(ISH), enzyme linked immunoadsorbent assay (ELISA) and electrophoretic mobility shift assay(EMSA), respectively. Results: In the LPS stimulated group, the expression of IKKβ mRNA (0 147±0 015) and activity of NFκB (0 828±0 019) in PAM significantly increased, whereas levels of IκBα(0 228±0 021) decreased (all P <0 01) in PMB treated groups. The expression of IKKβ mRNA (0 112±0 022) and activity of NFκB (0 358±0 011) were downregulated while level of IκBα (0 477±0 016) was upregulated( P <0 01). Conclusion: LPS might induce expression of IKKβ mRNA, degradation of IκBα and activation of NFκB PMB could inhibit the expression of IKKβ, degradation of IκBα and activation of NFκB, showing marked antiinflammatory property.

关 键 词：多黏菌素B 肺泡巨噬细胞 I-κB激酶 抑制蛋白 核因子-ΚB 

分 类 号：R965[医药卫生—药理学]