Table5_A microarray data analysis investigating the pathogenesis and potential biomarkers of autophagy and ferroptosis in intervertebral disc degeneration.XLS

Background: Intervertebral disc degeneration (IDD) entails complex pathological changes and causes lower back pain (LBP). However, there is still a lack of understanding of the mechanisms involved in IDD, particularly regarding the roles of autophagy and ferroptosis. The current study used microarray data to investigate the pathogenesis of IDD and potential biomarkers related to autophagy and ferroptosis in IDD.Methods: Differentially expressed genes (DEGs) were identified by analyzing the mRNA and miRNA expression profiles of IDD patients from the Gene Expression Omnibus (GEO). The protein-protein interaction network, Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and gene set enrichment analysis (GSEA) were utilized. The Human Autophagy Database (HADb) and Ferroptosis Database were used in conjunction with hub genes to identify autophagy- and ferroptosis-related genes. The Transcription Factor -hub gene-miRNA network was constructed. Lastly, the expression of DEGs in normal and degenerated nucleus pulposus cells (NPCs) was investigated via the quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results: A total of 362 DEGs associated with IDD were identified. GO and KEGG analyses indicated that oxidative stress, extracellular matrix, PI3K-AKT signaling pathway, and ferroptosis were key factors in IDD occurrence. GSEA indicated that IDD was associated with changes in autophagy, iron ion homeostasis, extracellular matrix, and oxidative stress. Eighty-nine hub genes were obtained, including five that were autophagy-related and three that were ferroptosis-related. Of these, TP53 and SESN2 were the intersections of autophagy- and ferroptosis-related genes. In qRT-PCR analysis, CANX, SLC38A1, and TP53 were downregulated in degenerative NPCs, whereas GNAI3, SESN2, and VAMP3 were upregulated.Conclusion: The current study revealed aspects of autophagy- and ferroptosis-related genes involved in IDD pathogenesis, warranting further investigation.

背景：椎间盘退行性变（IDD）涉及复杂的病理改变，并导致下背痛（LBP）。然而，对于IDD中涉及的机制，尤其是自噬和铁死亡的作用，目前仍缺乏理解。本研究利用微阵列数据调查IDD的发病机制，以及与自噬和铁死亡相关的潜在生物标志物。方法：通过分析基因表达综合数据库（GEO）中IDD患者的mRNA和miRNA表达谱，确定了差异表达基因（DEGs）。运用蛋白质-蛋白质相互作用网络、基因本体（GO）富集、京都基因与基因组百科全书（KEGG）分析和基因集富集分析（GSEA）等方法。结合人类自噬数据库（HADb）和铁死亡数据库，利用枢纽基因识别自噬和铁死亡相关基因。构建转录因子-枢纽基因-miRNA网络。最后，通过定量逆转录聚合酶链反应（qRT-PCR）研究正常和退变的纤维环细胞（NPCs）中DEGs的表达。结果：共鉴定出与IDD相关的362个DEGs。GO和KEGG分析表明，氧化应激、细胞外基质、PI3K-AKT信号通路和铁死亡是IDD发生的关键因素。GSEA表明，IDD与自噬、铁离子稳态、细胞外基质和氧化应激的变化相关。获得89个枢纽基因，包括5个与自噬相关和3个与铁死亡相关。其中，TP53和SESN2是自噬和铁死亡相关基因的交集。在qRT-PCR分析中，CANX、SLC38A1和TP53在退变的NPCs中下调，而GNAI3、SESN2和VAMP3上调。结论：本研究揭示了自噬和铁死亡相关基因在IDD发病机制中的作用，值得进一步研究。