基于VAMP7介导的细胞自噬探讨突触结构重塑对神经病理性疼痛的影响

突触结构重塑是导致神经病理性疼痛反复发作的重要病理基础，其具体机制尚未阐明，并可能影响疼痛长期疗效。研究表明自噬与许多疾病组织结构重塑密切相关，可能是极有前景的治疗靶点；VAMP7是调控自噬体膜融合关键蛋白SNAREs成员，我们前期证明VAMP7在疼痛大鼠脊髓表达下调，并伴突触结构重塑，而上调其表达能激活脊髓内自噬并改善疼痛症状，据此我们提出假说：“VAMP7介导的细胞自噬参与突触结构重塑，在神经病理疼痛发生和迁延不愈中起着关键作用”。本项目拟采用GFP-LC3转基因大鼠和原代脊髓神经元，分别建立疼痛动物模型和细胞剌激模型，应用自噬抑制剂和激动剂、基因沉默和过表达等手段干预自噬及VAMP7表达，通过观察自噬荧光、自噬蛋白表达、突触结构及疼痛行为等指标，探讨VAMP7介导的自噬在突触结构重塑及疼痛中的作用。本项目有望揭示VAMP7介导的自噬在神经病理性疼痛中的机制，为新镇痛靶点确立提供依据。

The synaptic structural remodeling was one of important pathological fundament for neuropathic pain. However, the detailed pathogenesis of synaptic structural remodeling in neuropathic pain is still unclear. Autophagy was supposed to play an important role during such remodeling process. Meanwhile, it was suggested neuron autophagy is closely related with the synaptic structural remodeling. But it is not clear the relationship between autophagy and tissue structural remodeling in neuropathic pain. VAMP7 is an important member in SNAREs protein family, and played an important role in the membrane fusion during autophagy. Meanwhile, VAMP7 is essential for neuronal morphogenesis. Besides, our bioinformatics and molecular biology analyses demonstrated the decreased expression of VAMP7 in neuropathic pain, and the up-regulation of VAMP7 expression could relieve pain syndrome by activating autophagy process. Therefore, we speculate that autophagy process mediated by the VAMP7 may be related with the synaptic structural remodeling in neuropathic pain. In this study we plan to discuss whether the VAMP7 expression can influence the synaptic structural remodeling and neuropathic pain by regulation the autophagy process on both cell (spinal cord neurons) and animal (SNL model on GFP-LC3 transgenic rats) levels. Besides, we plan to verify the difference on treatment effect for neuropathic pain between the up-regulation of the VAMP7 expression and the application of gabapentin. This study would provide new theoretical basis for the research of neuropathic pain mechanism and treatment.