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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 investigate the effect of NAC on the expression of nuclear factor-Kβ in normal airway epithelial cell line (16HBE) and tumor airway epithelial cell line (H292) stimulated by pro-inflammatory cytokines TNF-α (NF-Кβ) expression and its mechanism of action.Methods: Western blotting was used to detect the expression of NF-КB, and the nuclear protein was isolated by SDS-PAGE gel electrophoresis and anti-NF-КB antibody P50 and P65 The presence of NF-КB protein was determined to observe the expression of NF-КB in 16HBE cells and H292 cells under the stimulation of TNF-αin different doses (100,1000u / ml) and different time (1, 2, 4, 6hrs) (4,8,16,32mM) with different doses of NAC (1000u / ml, 2hrs) stimulated NF-КB activation.Results: TNF-α can stimulate the activation of human airway epithelium NAC could inhibit NF-КB activated by TNF-αin a dose-dependent manner.Conclusion: NAC can inhibit the activation of NF-КB by epithelial cells in the asthma Inflammatory control plays a role.