与退行性腰椎间盘疾病中铁死亡相关的PDK4的验证及其作用

Validation and the role of PDK4 relevant to ferroptosis in degenerative lumbar disc disease.

作者信息

Chen Wenhao, Yang Wanliang, Meng Bin, Wang Xingkun, Duan Heng, Xu Qian, Li Hao

机构信息

Department of Orthopaedics, Qilu Hospital of Shandong University, No.107, Wenhuaxi Road, Lixia District, Jinan, Shandong Province, 250012, China.

Cheeloo College of Medicine, Shandong University, No.107, Wenhuaxi Road, Lixia District, Jinan, Shandong Province, 250012, China.

出版信息

J Orthop Surg Res. 2025 Jan 10;20(1):30. doi: 10.1186/s13018-024-05293-8.

DOI:10.1186/s13018-024-05293-8

PMID:39794775

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11724506/

Abstract

BACKGROUND

Ferroptosis was involved in the pathogenesis of intervertebral disc degeneration (IVDD). However, the exact mechanism of IVDD associated with ferroptosis still required deeper studies.

METHOD

The differentially expressed genes (DEGs) in rat lumbar disc tissue between the control and IVDD group treated with IL-1β were detected by RNA sequencing (RNA-seq). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on DEGs. We further screened the differential expressed ferroptosis-related genes (DEFRGs). Besides, a protein-protein interaction (PPI) network of DEFRGs was constructed by STRING database. The Cytoscape database identified significant modules and the hub genes. The loss function of PDK4 by siRNA inference was investigated in NPCs by CCK8 assay, ELISA assay, and the analysis of ferroptosis indicators.

RESULT

DEGs were identified using RNA-seq. KEGG pathway analysis showed that these genes were mainly involved in Parkinson's disease, oxytocin signaling pathway, calcium ion signaling pathway, AMPK signaling pathway, and glucagon signaling pathway. Eight hub genes (including LDHA, PKM, EP300, EGFR, EGLN1, SCD, PDK4, and FABP4) were found by the PPI network and Cytoscape on a total of 25 ferroptosis-related genes that were identified in rat lumbar disc tissue after IVDD treatment. PDK4 silencing promoted NPCS proliferation, decreased the levels of the proinflammatory factors, and suppressed ferroptosis.

CONCLUSION

The study suggested the potential roles of ferroptosis-related genes in IVDD and further revealed the role of PDK4 in the progression of IVDD.

摘要

背景

铁死亡参与了椎间盘退变（IVDD）的发病机制。然而，与铁死亡相关的IVDD的确切机制仍需要更深入的研究。

方法

通过RNA测序（RNA-seq）检测对照组和经白细胞介素-1β处理的IVDD组大鼠腰椎间盘组织中的差异表达基因（DEGs）。对DEGs进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。我们进一步筛选了差异表达的铁死亡相关基因（DEFRGs）。此外，利用STRING数据库构建了DEFRGs的蛋白质-蛋白质相互作用（PPI）网络。通过Cytoscape数据库确定了显著模块和枢纽基因。通过CCK8检测、ELISA检测和铁死亡指标分析，研究了siRNA干扰对神经前体细胞中PDK4的功能缺失作用。

结果

通过RNA-seq鉴定出DEGs。KEGG通路分析表明，这些基因主要参与帕金森病、催产素信号通路、钙离子信号通路、AMPK信号通路和胰高血糖素信号通路。通过PPI网络和Cytoscape在IVDD处理后的大鼠腰椎间盘组织中鉴定出的总共25个铁死亡相关基因中发现了8个枢纽基因（包括LDHA、PKM、EP300、EGFR、EGLN1、SCD、PDK4和FABP4）。PDK4沉默促进神经前体细胞增殖，降低促炎因子水平，并抑制铁死亡。