Zhao Chenhao, Li Qiuwei, Shen Cailiang
Department of Orthopedics and Spine Surgery, the First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China.
Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, the First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China.
J Orthop Surg Res. 2025 Jun 13;20(1):586. doi: 10.1186/s13018-025-05999-3.
Intervertebral disc degeneration (IVDD) is a leading cause of lower back pain, imposing significant healthcare burdens globally. However, the molecular mechanisms underlying IVDD remain elusive, limiting effective therapeutic approaches. This study investigated the protective role of miR-204-5p in IVDD by targeting the Structure-specific recognition protein-1(SSRP1)/nuclear factor-kappa B (NF-κB) signaling pathway. Mendelian randomization (MR), experimental validation, and bioinformatics analysis were used.
We began with bidirectional MR analysis to explore the causal relationships between 40 microRNAs and IVDD, identifying miR-204-5p as negatively correlated with IVDD. Following this, in vitro experiments were conducted to examine the effects of miR-204-5p on lipopolysaccharide (LPS)-induced apoptosis in nucleus pulposus (NP) cells, with additional in vivo studies performed using rat models of disc degeneration. Finally, bioinformatics analysis was conducted using RNA-seq data from the GSE165722 dataset. Differential expression analysis was performed to compare SSRP1 expression between the control and degenerated tissue groups. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein-protein interaction (PPI) network analyses were used to explore the biological pathways and regulatory networks.
MR analysis identified miR-204-5p as being significantly protective against IVDD. The experimental results demonstrated that miR-204-5p reduced LPS-induced apoptosis in NP cells by downregulating SSRP1 expression and modulating the NF-κB pathway. Bioinformatics analysis further confirmed the differential expression of apoptosis-related genes, including SSRP1, BAX, and BCL2, between the control and degenerated tissues. Cluster analysis revealed distinct expression patterns of these genes, while correlation analysis showed strong interactions between SSRP1 and key extracellular matrix genes (COL2A1, ACAN) in degenerated tissues. Heatmaps and correlation matrices visualized these interactions, further supporting miR-204-5p's protective role.
This study is the first to utilize a comprehensive approach, combining MR, experimental validation, and bioinformatics analysis to uncover the protective effects of miR-204-5p in IVDD by regulating the SSRP1/NF-κB pathway. These findings provide novel insights into IVDD pathogenesis and highlight miR-204-5p as a promising therapeutic target for future interventions.
椎间盘退变（IVDD）是下腰痛的主要原因，在全球范围内造成了巨大的医疗负担。然而，IVDD潜在的分子机制仍不清楚，限制了有效的治疗方法。本研究通过靶向结构特异性识别蛋白1（SSRP1）/核因子-κB（NF-κB）信号通路，研究miR-204-5p在IVDD中的保护作用。采用了孟德尔随机化（MR）、实验验证和生物信息学分析。
我们首先进行双向MR分析，以探索40种 microRNA与IVDD之间的因果关系，确定miR-204-5p与IVDD呈负相关。在此之后，进行体外实验，以研究miR-204-5p对脂多糖（LPS）诱导的髓核（NP）细胞凋亡的影响，并使用椎间盘退变大鼠模型进行额外的体内研究。最后，使用来自GSE165722数据集的RNA-seq数据进行生物信息学分析。进行差异表达分析，以比较对照组和退变组织组之间的SSRP1表达。使用基因本体（GO）、京都基因与基因组百科全书（KEGG）富集分析和蛋白质-蛋白质相互作用（PPI）网络分析来探索生物学途径和调控网络。
MR分析确定miR-204-5p对IVDD具有显著的保护作用。实验结果表明，miR-204-5p通过下调SSRP1表达和调节NF-κB途径，减少了LPS诱导的NP细胞凋亡。生物信息学分析进一步证实了对照组和退变组织之间凋亡相关基因（包括SSRP1、BAX和BCL2）的差异表达。聚类分析揭示了这些基因的不同表达模式，而相关性分析表明退变组织中SSRP1与关键细胞外基质基因（COL2A1、ACAN）之间存在强烈的相互作用。热图和相关矩阵直观地显示了这些相互作用，进一步支持了miR-204-5p的保护作用。
本研究首次采用综合方法，结合MR、实验验证和生物信息学分析，揭示了miR-204-5p通过调节SSRP1/NF-κB途径在IVDD中的保护作用。这些发现为IVDD的发病机制提供了新的见解，并突出了miR-204-5p作为未来干预的有希望的治疗靶点。
