Netrin-1调控AMPK-ULK1通路诱导自噬对周围神经病理性疼痛的作用

神经病理性疼痛（NP）严重影响生活质量。我们发现，轴索导向因子netrin-1受体Unc5H2基因沉默可抑制自噬，降低损伤后施万细胞存活，并加重坐骨神经慢性压迫所致NP，提示netrin-1可能通过自噬参与NP的发生，其机制尚不清楚。已经证实，自噬相关基因ULK1参与Unc5H2的胞内定位，并受AMPK调控，后者可被netrin-1激活并参与一系列炎性反应，影响细胞存活。我们假设netrin-1可通过AMPK-ULK1途径调控自噬，促进神经修复和抑制炎症而改善NP。本研究将建立坐骨神经慢性压迫NP模型，以病毒转染补充netrin-1，同时以siRNA干扰其受体，以及自噬相关因子，明确netrin-1在不同受体介导下对NP的作用及途径，探讨netrin-1对自噬的调控机制；同时，应用抗体中和炎性细胞因子，阐明自噬介导抗炎机制在NP形成中的作用。研究结果对发现治疗新靶点、改善NP有重要意义。

Neuropathic pain (NP) severely affects life quality, and the underlying mechanisms remain unclear. Our preliminary data show for the first time that the knockdown of Unc5H2 (the receptor of netrin-1, an axon growth guidance factor) exaggerated allodynia induced by chronic constrictive injury (CCI) of sciatic nerve in rats. We also found that netrin-1 can rescue the survival of Schwann cells by activating autophagy, and the inhibition of autophagy aggravated NP. It is known that netrin-1 is genetically related to ULK1 (autophagy related gene 1), and can activate AMPK which is an upstream activator of autophagy. Therefore, we raised our hypothesis: netrin-1 ameliorates NP by enhancing neural regeneration and inhibiting inflammation, mediated by AMPK-ULK1 regulated autophagic activation. First, adenovirus-associated virus transfection, siRNA and neutralizing antibodies will be applied in CCI model, neuroplasticity, inflammation, behavior and neuroelectrophysiological tests will be performed, in order to investigate the effects of netrin-1 induced autophagy on NP. Second, Schwann cells will be cultured, different interfering methods will be used to detect the regulating pathway of netrin-1 in the activation of autophagy. The proposed study will reveal the underlying mechanism of netrin-1 and its receptors in the regulation of NP, thus reveal a potential new target for the management of NP.