目的探讨香烟提取物(cigarette smoke extract,CSE)对气道平滑肌细胞(airway smooth muscle cells,ASMCs)增殖中细胞外信号调节蛋白激酶(ERKs)及核因子-κB(NF-κB)表达的影响。方法体外培养Wistar大鼠的气道平滑肌细胞,给予不同浓度的CSE刺激24 h后,MTT法检测细胞增殖情况,采用Western blot方法检测ERKs、磷酸化细胞外信号调节蛋白激酶(p-ERK)和NF-κB p65的表达。结果1/32 CSE组、1/16 CSE组及1/8 CSE组的细胞增殖和ERKs、p-ERK、NF-κB p65的表达均逐渐增高,且与对照组相比差异有显著性意义(P〈0.05,n=4)。NF-κB p65的表达水平与p-ERK的表达水平呈明显正相关(r=0.858,P〈0.05,n=4)。结论一定浓度的CSE作用于平滑肌细胞后引起ERKs及ERKs磷酸化的增加,磷酸化激活后的ERKs可能与NF-κB的活化和ASMCs的增殖有关。
Objective To study the effect of cigarette smoke extract (CSE) on the extracellular signal-regulated kinases (ERKs) and nuclear factor-kappaB (NF-κB) in the proliferation of airway smooth muscle ceils (ASMCs). Methods The ASMCs in Wistar rats were cultured in vitro and stimulated with CSE for 24 h. Cell proliferation was determined with mitochondrial dehydrogenase activity (MTT assay). Western blotting was used to detect the activity of ERKs, pERK and NF- κBp65. Results Cell proliferation was gradually increased in 1/32, 1/16 and 1/8 CSE groups. With the increase of CSE concentration, the levels of ERKs, pERK and NF-κB p65 were gradually increased. There was significant differentce between CSE groups and control group (P〈0.05, n=4). The levels of NF-κB p65 expression had strongly positive correlation with the levels of FERK expression (r=0. 858, P〈0.05, n=4). Conclusion CSE may activate ERKs in ASMCs, and activated ERKs after phosphorylation may be associated with the activation of NF-κB and the proliferation of ASMCs.