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利用全转录组测序鉴定和验证过敏性哮喘发病机制中差异表达的基因

Identification and validation of differentially expressed genes in allergic asthma pathogenesis using whole-transcriptome sequencing.

作者信息

Cui Fang, Liu Rui, Wang Li, He Jing, Xu Yanxia

机构信息

Department of Laboratory Medicine, Yanan University Affiliated Hospital, Yan'an, Shaanxi, China.

Department of Respiratory and Critical Care Medicine, Yanan University Affiliated Hospital, Yan'an, Shaanxi, China.

出版信息

Front Med (Lausanne). 2025 Apr 16;12:1545095. doi: 10.3389/fmed.2025.1545095. eCollection 2025.

Abstract

OBJECTIVE

This study aims to systematically identify differential gene expression profiles in patients with allergic asthma through whole-transcriptome sequencing and validate the role of these genes in asthma pathogenesis, thereby uncovering potential molecular mechanisms.

METHODS

This study recruited a cohort of 80 individuals diagnosed with allergic asthma and 40 healthy controls. RNA was extracted from both peripheral blood and airway samples, and sequencing was performed using the Illumina NovaSeq 6000 platform. Potential differential genes were confirmed through three independent techniques to validate the findings: quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunohistochemistry, and Western blot analysis. Differential gene expression was analyzed using DESeq2 software, providing a rigorous statistical framework for RNA-Seq data interpretation. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were employed to elucidate the biological significance of the differentially expressed genes, offering insights into the molecular mechanisms underlying allergic asthma.

RESULTS

Differential expression analysis identified multiple genes with significant differences between the patient and control groups. Inflammatory-related genes such as , , and were significantly upregulated in the patient group ( < 0.05), while regulatory immune factors such as and showed significantly higher expression in the control group ( < 0.05). Functional enrichment analysis indicated that the differential genes were mainly enriched in immune response, T-cell activation, and signaling pathways. Experimental validation demonstrated consistency between transcriptomic data and RNA and protein expression levels, further supporting the involvement of these genes in asthma.

CONCLUSION

Differential gene expression profiles play a crucial role in the pathogenesis of asthma. This study provides important evidence for understanding the molecular mechanisms of asthma and developing novel targeted therapeutic strategies.

摘要

目的

本研究旨在通过全转录组测序系统鉴定过敏性哮喘患者的差异基因表达谱,并验证这些基因在哮喘发病机制中的作用,从而揭示潜在的分子机制。

方法

本研究招募了80名被诊断为过敏性哮喘的个体和40名健康对照。从外周血和气道样本中提取RNA,并使用Illumina NovaSeq 6000平台进行测序。通过三种独立技术确认潜在的差异基因以验证研究结果:定量逆转录聚合酶链反应(qRT-PCR)、免疫组织化学和蛋白质印迹分析。使用DESeq2软件分析差异基因表达,为RNA测序数据解释提供严格的统计框架。采用基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路分析来阐明差异表达基因的生物学意义,深入了解过敏性哮喘的分子机制。

结果

差异表达分析确定了患者组和对照组之间存在显著差异的多个基因。炎症相关基因如 、 和 在患者组中显著上调( <0.05),而调节性免疫因子如 和 在对照组中表达显著更高( <0.05)。功能富集分析表明,差异基因主要富集在免疫反应、T细胞活化和 信号通路中。实验验证表明转录组数据与RNA和蛋白质表达水平一致,进一步支持了这些基因参与哮喘发病过程。

结论

差异基因表达谱在哮喘发病机制中起关键作用。本研究为理解哮喘的分子机制和开发新的靶向治疗策略提供了重要证据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8c1e/12041010/01437cd14312/fmed-12-1545095-g001.jpg

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