Single-cell RNA sequencing reveals the link between axonal regeneration and neuropathic pain by modified analysis of a novel peripheral nerve injury rat model

Severe peripheral nerve injury (PNI) often causes significant movement disorders and intractable pain. Therefore, promoting nerve regeneration while avoiding neuropathic pain, a problem that remains unsolved, is key to the clinical treatment of PNI patients. Here, we establish a novel spared nerve crush (SNC) rat model that successfully reproduces axonal regeneration and neuropathic pain after PNI. Subsequently, we obtained single-cell RNA sequencing (scRNA-seq) data from rat directly injured and indirectly injured rat dorsal root ganglion (DRG) neurons at various time points after SNC and found that the PEP1 neuronal subtype in directly injured DRG is of particular interest. Through experimental design, sc-RNA sequence processing (EDSSP) and functional verification, we identified a potential key gene, Adcyap1, that encodes a key molecule linking nerve regeneration and pain after PNI. Our study sheds new light on the intrinsic link between axonal regeneration and neuropathic pain following PNI and provides new molecular targets and ideas for therapeutic intervention.重度周围神经损伤（peripheral nerve injury, PNI）常引发严重的运动功能障碍与难治性疼痛。因此，在促进神经再生的同时规避神经病理性疼痛——这一尚未解决的临床难题，是PNI患者临床治疗的关键所在。本研究构建了一种新型保留神经压榨（spared nerve crush, SNC）大鼠模型，该模型可成功复现PNI后轴突再生与神经病理性疼痛的病理过程。随后，我们在SNC术后不同时间点，分别从直接受损与间接受损的大鼠背根神经节（dorsal root ganglion, DRG）神经元中获取了单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）数据，并发现直接受损DRG中的PEP1神经元亚型具有特殊研究价值。通过实验设计、单细胞RNA测序数据处理（EDSSP）与功能验证，我们鉴定出了潜在关键基因Adcyap1，其编码的关键分子可连接PNI后的神经再生与疼痛通路。本研究为阐明PNI后轴突再生与神经病理性疼痛之间的内在关联提供了新视角，并为治疗干预提供了全新的分子靶点与研究思路。