基于泛素化机制的肺纤维化关键基因筛选与鉴定

Sc reening and identification of key genes of pulmonary fibrosis based on ubiquitination mechanism

目的通过基因表达综合(GEO)数据库微阵列分析,筛选出重要的肺纤维化相关泛素化基因,探讨其在肺纤维化大鼠肺组织中表达情况及作用。方法采用GEO数据库中GSE47460和GSE70866两个数据集,利用R语言limma包获取肺纤维化组和正常对照组间差异表达基因(DEGs)。从DEGs中选出泛素化相关基因,利用Cytoscape软件筛选关键(Hub)基因,并对后者进行GO和KEGG富集分析、Kaplan-Meier分析差异表达基因生存率。同时,构建博来霉素(BLM)诱导肺纤维化大鼠模型,通过蛋白免疫印迹(WB)检测肺纤维化大鼠肺组织Hub基因表达情况。结果筛选出38个泛素化相关的DEGs,5个是泛素化相关的Hub基因,其中UCHL1是肺纤维化较好的生存预测基因,其在肺纤维化组大鼠肺组织中的表达也较正常组升高(P<0.05),抑制UCHL1可明显改善肺纤维化。结论泛素化相关Hub基因可能参与肺纤维化进程,其中UCHL1在肺纤维化过程中发挥重要作用,有望成为肺纤维化治疗的分子靶点和诊断性生物标志物。

Objective To screen out important ubiquitination genes associated with pulmonary fibrosis through Gene Expression Omnibus(GEO)database microarray analysis,and to explore their expression in pulmonary fibrosis rat lung tissue.Methods Two GEO datasets GSE47460 and GSE70866 were used to obtain differentially expressed genes(DEGs)between fibrosis group and the control group used the limma package.Ubiquitination-related genes were selected from DEGs,and the Hub genes were screened by using Cytoscape software.GO and KEGG enrichment analysis and Kaplan-Meier analysis were performed on the survival rate of differentially expressed genes.At the same time,the Bleomycin-induced pulmonary fibrosis rat model was constructed,and the expression of Hub gene in the pulmonary tissue of the fibrotic rats was detected by Western blotting(WB).Results 38 ubiquitination-related DEGs were screened,and 5 were ubiquitination-related Hub genes,among which UCHL1 was a better predictor of pulmonary fibrosis,and its expression in lung tissues of pulmonary fibrosis group was higher than thr control group,with statistical significance(P<0.05).The inhibition of UCHL1 could significantly improve pulmonary fibrosis.Conclusion Ubiquitination-related Hub genes may be involved in the pathogenesis of pulmonary fibrosis,among which UCHL1 may play an important role in the process of pulmonary fibrosis,and is expected to be a molecular target and diagnostic biomarker for the treatment of pulmonary fibrosis.