应用基因芯片筛选矽肺纤维化相关差异基因表达谱的研究

Identification of differential gene expression profile related to silicosis fibrosis by cDNA

目的研究染矽尘大鼠肺组织纤维化有关基因的表达。方法将大鼠随机分为染尘组及对照组,每组8只,采用气管灌注法1次染矽尘。应用基因芯片技术,研究染尘后第28天大鼠肺组织中差异表达基因。采用实时荧光定量PCR(real-time PCR)实验进行验证。结果本次实验筛选出的有效基因22 107个,差异基因2 096个,其中fold值>5差异基因,主要有SPP1、LCN2、RETNLA、RASA1、CD68、PI4K2A。经过GO数据库分类注释的功能簇包括生物过程723个、细胞组分153个、分子功能161个。real-time PCR实验结果显示,染矽尘大鼠肺组织中GM-CSF基因相对表达量与对照组比较,4组上调,1组下调。结论染矽尘大鼠通过多种基因、多种信号共同参与生物学调控过程,最后诱导形成肺纤维化。GM-CSF是导致矽肺纤维化发生、发展的一个重要基因。

Objective To study the relevant genes of pulmonary fibrosis in silica exposed rats. Methods The rats were randomly divided into silica group and control group, with eight animals in each group. Animal models were established through intratracheal instillation under general anesthesia. Total RNA was isolated from the lung tissue at 28th days after instillation and analyzed by microarray technology. Parts of these genes were analyzed and identified by real-time PCR. Results A total of 2096 differentially expressed genes were identified in lungs of silica exposed rats, of which fold-value 〉5 included SPP1, LCN2, RETNLA, RASA1, CD68 and PI4K2A. The results of real-time PCR showed that GM-CSF was overexpressed in 4 cases and less-expressed in 1 case. Conclusions Pulmonary fibrosis in silica exposed rats is the formation of multiple signal pathways through a variety of genes involved in common biological control process. GM-CSF mRNA may participate in the development of silicosis fibrosis.