Department of Orthopaedics, Beijing Ditan Hospital, Capital Medical University, 100015, Beijing, China.
Sci Rep. 2020 Feb 28;10(1):3708. doi: 10.1038/s41598-020-60714-y.
The incidence of intervertebral disc (IVD) degeneration disease, caused by changes in the osmotic pressure of nucleus pulposus (NP) cells, increases with age. In general, low back pain is associated with IVD degeneration. However, the mechanism and molecular target of low back pain have not been elucidated, and there are no data suggesting specific biomarkers of low back pain. Therefore, the research aims to identify and verify the significant gene biomarkers of low back pain. The differentially expressed genes (DEGs) were screened in the Gene Expression Omnibus (GEO) database, and the identification and analysis of significant gene biomarkers were also performed with various bioinformatics programs. A total of 120 patients with low back pain were recruited. Before surgery, the degree of pain was measured by the numeric rating scale (NRS), which enables comparison of the pain scores from individuals. After surgery, IVD tissues were obtained, and NP cells were isolated. The NP cells were cultured in two various osmotic media, including iso-osmotic media (293 mOsm/kg HO) to account for the morbid environment of NP cells in IVD degeneration disease and hyper-osmotic media (450 mOsm/kg HO) to account for the normal condition of NP cells in healthy individuals. The relative mRNA expression levels of CCL5, OPRL1, CXCL13, and SST were measured by quantitative real-time PCR in the in vitro analysis of the osmotic pressure experiments. Finally, correlation analysis and a neural network module were employed to explore the linkage between significant gene biomarkers and pain. A total of 371 DEGs were identified, including 128 downregulated genes and 243 upregulated genes. Furthermore, the four genes (CCL5, OPRL1, SST, and CXCL13) were identified as significant gene biomarkers of low back pain (P < 0.001) based on univariate linear regression, and CCL5 (odds ratio, 34.667; P = 0.003) and OPRL1 (odds ratio, 19.875; P < 0.001) were significantly related to low back pain through multivariate logistic regression. The expression of CCL5 and OPRL1 might be correlated with low back pain in patients with IVD degeneration disease caused by changes in the osmotic pressure of NP cells.
椎间盘（IVD）退变疾病的发病率随核髓核（NP）细胞渗透压的变化而增加，与年龄有关。一般来说，腰痛与 IVD 退变有关。然而，腰痛的机制和分子靶点尚未阐明，也没有数据表明腰痛的特定生物标志物。因此，本研究旨在鉴定和验证腰痛的显著基因生物标志物。在基因表达综合数据库（GEO）中筛选差异表达基因（DEGs），并使用各种生物信息学程序进行显著基因生物标志物的鉴定和分析。共招募了 120 名腰痛患者。手术前，采用数字评分量表（NRS）测量疼痛程度，以便比较个体的疼痛评分。手术后，获取 IVD 组织，并分离 NP 细胞。NP 细胞在两种不同的渗透介质中培养，包括等渗介质（293 mOsm/kg HO）以模拟 IVD 退变疾病中 NP 细胞的病态环境，以及高渗介质（450 mOsm/kg HO）以模拟健康个体中 NP 细胞的正常状态。在体外渗透压实验中，通过定量实时 PCR 测量 CCL5、OPRL1、CXCL13 和 SST 的相对 mRNA 表达水平。最后，采用相关分析和神经网络模块探讨显著基因生物标志物与疼痛之间的关联。在渗透压实验的体外分析中，通过定量实时 PCR 测量 CCL5、OPRL1、CXCL13 和 SST 的相对 mRNA 表达水平。最后，采用相关分析和神经网络模块探讨显著基因生物标志物与疼痛之间的关联。
