Xu Mu, Wang Zhijian, Xu Gang, Zhu Mengye, Zhang Daying, Yan Yi
Department of Pain Medicine, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi, China.
Front Mol Neurosci. 2024 Feb 6;16:1261217. doi: 10.3389/fnmol.2023.1261217. eCollection 2023.
Sciatica (neuropathic pain [NP]) is a common disease characterized by pain from radiation along the sciatic nerve. The aim of this study was to study the genes associated with chronic systolic injury of sciatic nerve (SCN-CCI) in rats by RNA-Seq technique, and to explore their potential as therapeutic targets.
Sciatic nerve rat model was obtained by ligation of sciatic nerve and divided into two groups: SCN-CCI group and Sham group. Behavioral assessments were performed to evaluate pain sensitivity, following which their spinal cord dorsal horn were resected and RNA sequencing was conducted to identify differentially expressed genes (DEGs). Bioinformatics and functional enrichment analysis was performed to identify promising DEGs and their related biological processes and pathways associated with SCN-CCI. PPI network analysis and hub gene identification were conducted. QRT-PCR, western blot, ELISA, and immunofluorescence staining were performed on rat models to validate the expression of these hub genes and investigate related proteins and inflammatory markers.
The SCN-CCI rat model was successfully obtained, exhibiting increased pain sensitivity compared to the Sham group, as indicated by decreased mechanical allodynia thresholds, thermal latencies, and increased paw withdrawals. RNA-Seq analysis identified 117 DEGs in the SCN-CCI rat model, involved in various biological processes and pathways related to sciatica. PPI network analysis revealed hub genes, including Ly6g6e, which exhibited significant differential expression. QRT-PCR and Western blot analysis confirmed the expression patterns of these hub genes. Pain behavior assessment demonstrated reduced pain thresholds and increased paw flinching responses in the SCN-CCI group. Furthermore, the SCN-CCI group showed upregulated expression of Ly6g6e, increased protein levels of Ly6g6e, CGRP, and NGF, as well as elevated levels of IL-1β, MCP-1, and IL-6, and microglial cell activation in the spinal dorsal horn. ELISA results confirmed the increased levels of IL-1β, MCP-1, and IL-6 in the spinal dorsal horn.
These comprehensive findings provide valuable insights into the SCN-CCI rat model, DEGs associated with sciatica, hub genes (Ly6g6e as promising targets), pain behavior changes and molecular alterations.
坐骨神经痛（神经性疼痛[NP]）是一种常见疾病，其特征是沿坐骨神经放射痛。本研究旨在通过RNA测序技术研究与大鼠坐骨神经慢性收缩损伤（SCN-CCI）相关的基因，并探索其作为治疗靶点的潜力。
通过结扎坐骨神经获得大鼠坐骨神经模型，并分为两组：SCN-CCI组和假手术组。进行行为评估以评价疼痛敏感性，随后切除其脊髓背角并进行RNA测序以鉴定差异表达基因（DEG）。进行生物信息学和功能富集分析以鉴定有前景的DEG及其与SCN-CCI相关的相关生物学过程和途径。进行蛋白质-蛋白质相互作用（PPI）网络分析和枢纽基因鉴定。对大鼠模型进行实时定量聚合酶链反应（QRT-PCR）、蛋白质免疫印迹法（western blot）、酶联免疫吸附测定（ELISA）和免疫荧光染色，以验证这些枢纽基因的表达并研究相关蛋白质和炎症标志物。
成功获得SCN-CCI大鼠模型，与假手术组相比，其疼痛敏感性增加，表现为机械性异常性疼痛阈值降低、热潜伏期缩短和爪部退缩增加。RNA测序分析在SCN-CCI大鼠模型中鉴定出117个DEG，涉及与坐骨神经痛相关的各种生物学过程和途径。PPI网络分析揭示了枢纽基因，包括Ly6g6e，其表现出显著差异表达。QRT-PCR和蛋白质免疫印迹法分析证实了这些枢纽基因的表达模式。疼痛行为评估表明SCN-CCI组疼痛阈值降低，爪部退缩反应增加。此外，SCN-CCI组显示Ly6g6e表达上调，Ly6g6e、降钙素基因相关肽（CGRP）和神经生长因子（NGF）蛋白水平增加，以及脊髓背角中白细胞介素-1β（IL-1β）、单核细胞趋化蛋白-1（MCP-1）和白细胞介素-6（IL-6）水平升高，以及小胶质细胞活化。ELISA结果证实脊髓背角中IL-1β、MCP-1和IL-6水平升高。
这些综合研究结果为SCN-CCI大鼠模型、与坐骨神经痛相关的DEG、枢纽基因（Ly6g6e作为有前景的靶点）、疼痛行为变化和分子改变提供了有价值的见解。
