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目的:探讨细菌脂多糖(LPS)诱导离体鼻黏膜上皮细胞致炎后p38MAPK的活性变化及意义。方法:采用无血清原代培养法行正常鼻黏膜上皮细胞分离和培养,采用不同时间、不同浓度的LPS诱导培养的鼻黏膜上皮细胞致炎,应用WesternImmunoblotting法检测p38MAPK的活性变化及p38MAPK特异性抑制剂SB203580对其活性的抑制作用。结果:LPS(1mg/L)刺激后5～30min内和LPS浓度为0.01～1.00mg/L时,p38MAPK的活性增强;SB203580浓度为1.0～5.0μmol/L时,对p38MAPK活性的抑制作用最强。在LPS刺激之前20minSB203580可抑制p38MAPK活性;在LPS刺激后SB203580对p38MAPK活性的抑制作用甚微或无。结论:LPS刺激鼻黏膜上皮细胞致炎在30min内和浓度在1mg/L内是以时间和浓度“正依赖”方式导致p38MAPK的活化增强;特异性抑制剂SB203580对p38MAPK活性的抑制作用则是浓度在5μmol/L内以“正依赖”方式增强,且这种抑制作用仅在LPS刺激之前才发挥效应。
Objective: To investigate the changes of p38 MAPK activity induced by bacterial lipopolysaccharide (LPS) induced nasal mucosal epithelial cells in vitro and its significance. Methods: The normal nasal mucosa epithelial cells were isolated and cultured by serum-free primary culture. The nasal mucosal epithelial cells were induced by LPS at different times and concentrations. Western blotting was used to detect the changes of p38MAPK and p38MAPK Agent SB203580 inhibits its activity. RESULTS: The p38MAPK activity was enhanced within 5-30 min after stimulation with LPS (1 mg / L) and at 0.01-1.00 mg / L LPS. The inhibitory effect of p38MAPK was highest at SB203580 concentrations of 1.0-5.0 μmol / L . SB203580 inhibited p38MAPK activity 20 min before LPS stimulation; SB203580 had little or no inhibitory effect on p38 MAPK activity after LPS stimulation. CONCLUSIONS: LPS-stimulated nasal mucosal epithelial proinflammatory cytokines induced a more positive activation of p38 MAPK in a time-and concentration-dependent manner within 30 min and at a concentration of 1 mg / L. Inhibition of p38 MAPK by SB203580, a specific inhibitor, Increased in a “positive dependent” manner at 5 μmol / L, and this inhibitory effect only exerted an effect before LPS stimulation.