N-乙酰半胱氨酸对气道上皮细胞核因子-κВ表达的影响的实验研究

Effect of N - acetylcysteine on Nuclear Factor - кB Expression in Human Airway Epithelial Cell

目的:探讨抗氧化剂N-乙酰半胱氨酸(NAC)对炎前性细胞因子TNF-α刺激正常人气道上皮细胞株(16HBE)和肿瘤病人气道上皮细胞株(H292)的核因子-Кβ(NF-Кβ)表达的影响及作用机理.方法:采用Western-Blot免疫印迹电泳法:提取胞核蛋白,经SDS-PAGE凝胶电泳分离,以抗NF-КB抗体P50 ,P65作探针来确定NF-КB蛋白的存在.观察16HBE细胞和H292细胞的NF-КB在不同剂量(100,1000u/ml)和不同作用时间(1,2,4,6hrs)的TNF-α刺激下的表达;以及在相同TNF-α(1000u/ml,2hrs)刺激条件下,不同剂量NAC(4,8,16,32mM)对NF-КB活化抑制作用.结果:TNF-α在体外能刺激激活人气道上皮细胞16HBE和H292的NF-КB表达;同时发现:NAC能以一种剂量依赖方式抑制由TNF-α刺激激活的NF-КB.结论:NAC通过抑制上皮细胞激活NF-КB的途径,可能在哮喘的炎症控制中起着一定作用.

Objective:To assess the effect and mechanism of antioxidant (N - acetylcysteine, NAC) on nuclear factor-κB(NF-κB)expression induced by tumor necrosis factor-α(TNF-α)in airway epithelial cell strain of normal subject (16HBE) and tumor patient (H292). Methods: Western - Blot analyis was used: Nuclear protein was extracted and separated with SDS - PAGE,and the existence of NF - κB was determined with antibody (P50,P65) as probe. We observed the NF - κB activation induced by TNF - α in different doses (100, 1000u/ml)and time (l,2,4,6hrs) in 16HBE and H292,and also the NF - κB inhibition of NAC in different doses(4,8,16,32mM)under the same condition of TNF-α(1000u/ml,2hrs) . Result: It was found that the TNF - αa could stimulate the NF -κB activation, and that TNF - α induced NF -κB activation could be inhibited by NAC in a dose - dependent effect. Conclusion: NAC may play a role in the anti - inflammatory treatment of asthma through inhibiting NF - κB activation in airway epithelium cells.