Song Chao, Zhou Daqian, Cheng Kang, Liu Fei, Cai Weiye, Mei Yongliang, Chen Jingwen, Huang Chenyi, Liu Zongchao
Department of Orthopedics and Traumatology (Trauma and Bone-setting), Laboratory of Integrated Chinese and Western Medicine for Orthopedic and Traumatic Diseases Prevention and Treatment The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University Luzhou Sichuan China.
RuiKang Hospital affiliated to Guangxi University of Chinese Medicine Nanning Guangxi China.
JOR Spine. 2023 Dec 22;7(1):e1311. doi: 10.1002/jsp2.1311. eCollection 2024 Mar.
Intervertebral disc degeneration (IVDD) is a common chronic disease in orthopedics, and its molecular mechanisms are still not well explained.
This study's objective was to bioinformatics-based discovery of IVDD biomarkers and immune-inflammatory infiltrates.
The IVDD illness gene collection was gathered from GeneCards, DisGeNet, and gene expression profiles were chosen from the extensive Gene Expression Omnibus database (GSE124272, GSE150408, and GSE153761). The STRING database was used to create a network of protein-protein interactions, while the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) databases were used for functional enrichment analysis. Using hub genes, the immune cell infiltration between IVDD patient samples and control tissues was examined. Finally, quantitative polymerase chain reaction and Western blot experiments were used to verify the expression of hub genes.
A total of 27 differentially expressed hub genes were identified by bioinformatics. According to GO and KEGG analyses, hub genes were prominent in immunological responses, chemokine-mediated signaling pathways, and inflammatory responses, with the key signaling pathways engaged in cellular senescence, apoptosis, Th1 and Th2 cell differentiation, and Th17 cell differentiation. Immune cell infiltration research revealed that T cells, lymphocytes, B cells, and NK cells were decreased in IVDD patients while monocytes, neutrophils, and CD8 T cells were increased. The expression levels of the senescence hub genes SP1, VEGFA, IL-6, and the apoptosis key gene CASP3 were considerably greater in the IVDD model group than in the control group, according to in vitro validation.
In conclusion, the cellular senescence signaling pathway, the apoptosis signaling pathway, and associated hub genes play significant roles in the development and progression of IVDD, this finding may help direct future research on the senescence signaling route in IVDD.
椎间盘退变（IVDD）是骨科常见的慢性疾病，其分子机制仍未得到充分阐释。
本研究旨在基于生物信息学发现IVDD生物标志物及免疫炎症浸润情况。
从GeneCards、DisGeNet收集IVDD疾病基因集，并从庞大的基因表达综合数据库（GSE124272、GSE150408和GSE153761）中选取基因表达谱。利用STRING数据库构建蛋白质-蛋白质相互作用网络，同时使用京都基因与基因组百科全书（KEGG）和基因本体论（GO）数据库进行功能富集分析。通过枢纽基因检测IVDD患者样本与对照组织之间的免疫细胞浸润情况。最后，采用定量聚合酶链反应和蛋白质印迹实验验证枢纽基因的表达。
通过生物信息学共鉴定出27个差异表达的枢纽基因。根据GO和KEGG分析，枢纽基因在免疫反应、趋化因子介导的信号通路和炎症反应中显著富集，关键信号通路涉及细胞衰老、凋亡、Th1和Th2细胞分化以及Th17细胞分化。免疫细胞浸润研究显示，IVDD患者的T细胞、淋巴细胞、B细胞和NK细胞减少，而单核细胞、中性粒细胞和CD8 T细胞增加。体外验证结果显示，衰老枢纽基因SP1、VEGFA、IL-6以及凋亡关键基因CASP3在IVDD模型组中的表达水平显著高于对照组。
总之，细胞衰老信号通路、凋亡信号通路及相关枢纽基因在IVDD的发生发展中起重要作用，这一发现可能有助于指导未来关于IVDD衰老信号通路的研究。
