肺纤维化与铁死亡的相关性分析与机制探讨

Correlation analysis and mechanism study of ferroptosis with pulmonary fibrosis

目的探讨铁死亡与肺纤维化的相关性与机制。方法从基因表达综合数据库(Gene Expression Omnibus)和FerrDb数据库中获得2019年1月—2023年12月肺纤维化组织测序数据。采用生物信息学方法分析正常对照组与肺纤维化组之间的差异表达基因,提取肺嗜铁相关的差异表达基因。通过富集分析、蛋白质相互作用(protein-protein interaction,PPI)分析和随机森林算法筛选枢纽基因。制作小鼠肺纤维化模型进行运动干预,并通过实时荧光定量聚合酶链反应验证枢纽基因的表达。结果对103例特发性肺纤维化肺组织和103例正常肺组织的比较,鉴定出13个上调基因和7个下调基因为铁死亡相关差异表达基因。PPI结果显示这些铁死亡相关基因之间存在相互作用。京都基因与基因组数据库途径富集和基因本体论富集分析表明,铁死亡相关基因参与有机阴离子转运、缺氧反应、氧水平降低反应、缺氧诱导因子-1信号通路、肾细胞癌和花生四烯酸代谢信号通路。通过PPI分析和随机森林算法确定的基因包括CAV1、NOS2、GDF15、HNF4A和CDKN2A。小鼠模型纤维化肺组织的实时荧光定量聚合酶链反应结果显示,与运动组比较,博来霉素组的NOS2、PTGS2和GDF15 mRNA水平上调(P<0.05),CAV1和CDKN2A mRNA水平下调(P<0.05);与博来霉素组比较,博来霉素+运动组CAV1和CDKN2A的表达增加(P<0.05),NOS2、PTGS2和GDF15的表达减少(P<0.05)。结论生物信息学分析鉴定出与肺纤维化中铁死亡相关的20个潜在基因。CAV1、NOS2、GDF15和CDKN2A通过调节铁死亡影响肺纤维化的发生。跑步机训练可以减少肺纤维化组织中的铁死亡,从而减少肺部炎症。

Objective To explore the correlation and mechanism of ferroptosis with pulmonary fibrosis.Methods Pulmonary fibrosis tissue sequencing data were obtained from Gene Expression Omnibus and FerrDb databases from January 2019 to December 2023.Differentially expressed genes(DEGs)between the normal control group and the pulmonary fibrosis group were analyzed by bioinformatic method,and DEGs related to pulmonary iron addiction were extracted.The hub genes were screened by enrichment analysis,protein-protein interaction(PPI)analysis and random forest algorithm.The mouse model of pulmonary fibrosis was made for exercise intervention,and the expression of hub genes was verified by real-time quantitative reverse transcription polymerase chain reaction.Results A comparison of 103 patients with idiopathic pulmonary fibrosis and 103 normal lung tissues showed that 13 up-regulated genes and 7 down-regulated genes were identified as ferroptosis-related DEGs.PPI results showed that there was an interaction between these ferroptosis-related genes.The Kyoto Encyclopedia of Genes and Genomes pathway enrichment and Genome Ontology enrichment analysis showed that ferroptosis-related genes were involved in organic anion transport,hypoxia response,oxygen level reduction response,hypoxia-inducible factor-1 signaling pathway,renal cell carcinoma,and arachidonic acid metabolic signaling pathway.Genes identified by PPI analysis and random forest algorithm included CAV1,NOS2,GDF15,HNF4A,and CDKN2A.Real-time fluorescence quantitative polymerase chain reaction results of mouse fibrotic lung tissue showed that compared with the exercise group,the mRNA levels of NOS2,PTGS2 and GDF15 were up-regulated and the mRNA levels of CAV1 and CDKN2A were down-regulated in the bleomycin group(P<0.05);compared with the bleomycin group,the expression of CAV1 and CDKN2A increased and the expression of NOS2,PTGS2 and GDF15 decreased in the bleomycin+exercise group(P<0.05).Conclusions Bioinformatic analysis identifies 20 potential genes associating with ferroptosis in pulmonary fibrosis.CAV1,NOS2,GDF15,and CDKN2A influence the development of pulmonary fibrosis by modulating ferroptosis.Treadmill training can reduce ferroptosis in fibrotic tissues,thereby reducing lung inflammation.