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目的 探讨早期生长反应因子(Egr 1)及其信号转导在矽肺发生发展中的作用。方法用细胞免疫荧光、原位杂交方法检测二氧化硅(SiO2 )刺激后Egr 1的表达定位,用报道质粒及EMSA检测其活性改变;用激酶活性分析法检测SiO2 刺激巨噬细胞后ERK1 /2活性改变,进一步用激酶抑制剂初步探讨SiO2 活化Egr 1的信号转导通路。结果 SiO2 刺激RAW264. 7细胞短时间Egr 1核蛋白表达及转录因子明显增加;且在处理后30~60min,Egr 1核蛋白结合活性明显升高(为未处理组的20倍);在刺激后15minERK1 /2活性开始升高, 30min达高峰(活性为对照组的29倍)而后渐降至基础水平;进一步用激酶阻断发现,Egr 1mRNA及蛋白表达均减少。结论 SiO2 能激活巨噬细胞中Egr 1,且此过程可能由ERK1 /2、p38介导,提示SiO2 ERK1 /2、p38 Egr 1通路可能在矽肺发生发展过程中起重要作用。
Objective To investigate the role of early growth response factor (Egr 1) and its signal transduction in the development of silicosis. Methods The expression of Egr 1 after silica (SiO2) stimulation was detected by immunofluorescence and in situ hybridization. The activity of Egr 1 was detected by the reporter plasmid and EMSA. The activity of ERK1 / 2 Activity changes, and further to explore with kinase inhibitor Egr1 activation of SiO2 signal transduction pathway. Results The expression of Egr 1 protein and the transcription factor of RAW264.7 cells in a short time after exposure to SiO2 were significantly increased, and the binding activity of Egr1 to nucleoprotein was significantly increased after 30-60 min (20-fold higher than that of the untreated group) 15minERK1 / 2 activity began to rise, 30min reached the peak (29 times the activity of the control group) and then dropped to the basal level; further blocked with kinase was found, Egr1 mRNA and protein expression were reduced. Conclusion SiO2 can activate Egr 1 in macrophages and this process may be mediated by ERK1 / 2 and p38, suggesting that SiO2 ERK1 / 2 and p38 Egr 1 pathways may play an important role in the development of silicosis.