摘要
目的 探讨前炎因子肿瘤坏死因子 (TNF α)对气道上皮细胞核因子κB的激活情况及糖皮质激素对核因子κB的影响。方法 在人气道上皮细胞株 (16HBE)加入不同浓度的TNF α及地塞米松 ,于不同时间提取RNA和细胞核蛋白 ,利用逆转录聚合酶链反应 (RT PCR)和电泳迁移率实验检测白细胞介素 8(IL 8)mRNA和核因子κB的表达。结果 核因子κB活性表达水平在TNF α刺激后 1小时明显增加 ,2小时达到高峰 (143± 11) ,与对照组 (6 4± 10 )比较差异有显著性 (P <0 0 5 ) ,4小时时下降。随着TNF α量的增加 ,核因子κB活性水平也随之升高 ,同时IL 8mRNA表达明显升高 (0 48±0 0 3)。加入地塞米松后核因子κB活性水平和IL 8mRNA的表达均降低 (12 6 38± 2 5 5 ;0 34± 0 0 5 )。亚单位p5 0和p6 5抗体均参与核因子κB的组成。结论 TNF a激活人气道上皮细胞核因子κB ,进而通过调节IL 8的表达参与炎症进程 ,糖皮质激素通过抑制核因子κB的激活而发挥抗炎的作用。
Objective To observe the expression of NF κB activation and the effect of dexamethasone on the NF κB activity in the human airway epithelial cell line 16HBE after TNF α stimulation Methods After 16HBE was treated with different concentration of TNF α(10 U/ml,100 U/ml,1 000 U/ml)and dexamethasone(100 nmol/L), total RNA and cellular nuclear protein were extracted at 1 hour,2 hour,4 hour ,respectively RT PCR and electrophoresis mobility shift assay(EMSA) were used to detect the expression of IL 8 mRNA and NF κB activation Results The activity of NF κB activation became stronger at 1 hour in the TNF α stimulated group than the control, peaked at 2 hours and then decreased at 4 hours Supershift assay confirmed that both p50 and p65 were components of active NF κB At the same time, IL 8 mRNA expression was elevated at 4 hours After dexamethasone treatment,the expression of NF κB activation and IL 8 mRNA became lower Conclusions It is suggested that activated NF κB played a key role in the inflammatory process of respiratory diseases through regulating the expression of some important factors (cytokines). Glucocorticoid inhibited the activation of NF κB and showed antiinflammatory effect
出处
《中华结核和呼吸杂志》
CSCD
北大核心
2000年第5期296-299,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
