Chen Binglin, Guo Jiabao, Gong Chan, Zhu Chenchen, Wu Yang, Wang Shengbo, Zheng Yili, Lu Haixia
Department of Neurobiology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, No. 76 Yanta West Road, Yanta District, Xi'an, 710061, China.
The Second Clinical Medical College, Xuzhou Medical University, Xuzhou, China.
Sci Rep. 2025 Jan 18;15(1):2391. doi: 10.1038/s41598-025-86661-0.
Neuropathic pain (NP) is a complex and prevalent chronic pain condition that affects millions of individuals worldwide. Previous studies have shown that prior exercise protects against NP caused by nerve injury. However, the underlying mechanisms of this protective effect remain to be uncovered. Therefore, the purpose of this study is to investigate how prior exercise affects protein expression in NP model rats and thus gain comprehensive insights into the molecular mechanisms involved. To achieve this objective, 6-week-old male Sprague-Dawley rats were randomly assigned into three groups, named as chronic constriction injury (CCI) of the sciatic nerve, CCI with prior 6-week swimming training (CCI_Ex), and sham operated (Sham). The CCI_Ex group underwent 6 weeks of swimming training before CCI surgery, while the CCI and sham groups had no intervention. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were used as the main observation indicators to evaluate the behavioral changes associated with pain. Tissues from the spinal dorsal horn of the rats in the three groups were collected at 4 weeks after operation. LC-MS/MS proteomic analysis based on the label-free approach was used to detect protein profiles, and volcano plots, Venn diagrams, and clustering heatmaps were used to identify differentially expressed proteins (DEPs). Gene Ontology (GO) annotations, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and protein-protein interaction networks were employed to explore the biological importance of DEPs. At 14, 21, and 28 days following CCI, CCI rats with prior exercise showed a significant increase in the MWT and TWL of the injured lateral hind paw compared with those without exercise. A total of 122 proteins with significant changes in abundance were detected after CCI surgery, and 55 proteins were detected in the comparison between the CCI_Ex and CCI groups. GO and KEGG enrichment analysis revealed that oxygen transport capacity and the complement and coagulation cascades may be the critical mechanism by which prior exercise protects against NP. Serpina1, DHX9, and Alb are the key proteins in this process and warrant further attention, as confirmed by the results of Western blot analysis. In conclusion, this study provides new evidence that active physical activity can accelerate the relief of hyperalgesia after NP. Proteomic analyses revealed the potential target proteins and pathways for this process, offering valuable data resources and new insights into the pathogenesis and therapeutic targets of NP.
神经病理性疼痛（NP）是一种复杂且普遍的慢性疼痛病症，影响着全球数百万人。先前的研究表明，预先进行运动可预防由神经损伤引起的NP。然而，这种保护作用的潜在机制仍有待揭示。因此，本研究的目的是探究预先运动如何影响NP模型大鼠的蛋白质表达，从而全面深入了解其中涉及的分子机制。为实现这一目标，将6周龄雄性Sprague-Dawley大鼠随机分为三组，分别命名为坐骨神经慢性缩窄损伤（CCI）组、预先进行6周游泳训练后的CCI组（CCI_Ex）和假手术组（Sham）。CCI_Ex组在CCI手术前进行6周的游泳训练，而CCI组和假手术组未进行干预。采用机械缩足阈值（MWT）和热缩足潜伏期（TWL）作为主要观察指标，评估与疼痛相关的行为变化。术后4周收集三组大鼠脊髓背角组织。基于无标记方法的液相色谱-串联质谱（LC-MS/MS）蛋白质组学分析用于检测蛋白质谱，并使用火山图、韦恩图和聚类热图来鉴定差异表达蛋白（DEP）。利用基因本体论（GO）注释、京都基因与基因组百科全书（KEGG）通路以及蛋白质-蛋白质相互作用网络来探究DEP的生物学重要性。在CCI后的第14、21和28天，与未运动的大鼠相比，预先运动的CCI大鼠受伤侧后爪的MWT和TWL显著增加。CCI手术后共检测到122种丰度有显著变化的蛋白质，在CCI_Ex组和CCI组的比较中检测到55种蛋白质。GO和KEGG富集分析表明，氧运输能力以及补体和凝血级联反应可能是预先运动预防NP的关键机制。丝氨酸蛋白酶抑制剂A1（Serpina1）、解旋酶9（DHX9）和白蛋白（Alb）是这一过程中的关键蛋白质，蛋白质印迹分析结果证实了这一点，值得进一步关注。总之, 本研究提供了新的证据，表明积极的体育活动可以加速NP后痛觉过敏的缓解。蛋白质组学分析揭示了这一过程的潜在靶蛋白和途径，为NP的发病机制和治疗靶点提供了有价值的数据资源和新的见解。
