摘要
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30～60 min,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15 min ERK1/2活性开始升高,30 min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 discuss the role of early growth response factor (Egr)-1 and it′s upstream signaling pathway in the development of silicosis. Methods The expression and localization of Egr-1 were analyzed by immunofluorescence and in-situ hybridization. The activity of Egr-1 was observed in treated cells by using a reporter plasmid and EMSA, the activity of ERK1/2 in RAW264.7 incubated with SiO 2 by using a kinase assay, and further by using a kinase inhibitor assay to investigate the role of upstream kinase in the signal pathway of the activation of Egr-1. Results The obvious increase of expression and transcription of Egr-1 was observed shortly after being treated by silica and its activity increased abruptly. There was an increase of the activity of ERK1/2 in RAW264.7 cells treated, which reached a peak at 30 minutes. The expression and transcription of Egr-1 decreased maniferstly after using kinase inhibitors. Conclusion Egr-1 expression can be induced by silica dioxide in RAW264.7 cells, and the ERK1/2、p38 kinases may take part in this process which suggest the pathway of SiO 2, ERK1/2、p38 and Egr-1 may play an important role in the development of silicosis.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2005年第5期293-296,共4页
Chinese Journal of Pathology
基金
国家自然科学基金资助项目(30170399)
