MST1-JNK通路调控线粒体自噬对周围神经病理性疼痛的作用

线粒体功能异常影响神经病理性疼痛（NP）的形成和慢性化，线粒体自噬与之密切相关。我们发现基因沉默哺乳动物Ste20样激酶1（Mammalian ste20-like kinase 1, MST1）保护坐骨神经慢性压迫所致NP，并可与调控线粒体自噬有关，其机制尚不清除。已经证实MST1磷酸化激活JNK，后者诱导线粒体自噬，并介导MST1作为活性氧感受器的作用；MST1受到长链非编码RNA MNX1-AS1转录后抑制。我们假设：MST1受MNX1-AS1调控，籍JNK诱导线粒体自噬异常和氧化失衡，进而促进NP的形成和慢性化。本研究将建立坐骨神经慢性压迫NP模型，选择MST1基因敲除及野生型动物，应用体内基因沉默/过表达及药物处理策略，探讨MST1对NP的作用和调控途径；并建立施万细胞糖氧剥夺模型，阐明MST1调控线粒体自噬的环节及对线粒体功能的作用。本研究有助于阐明NP的机制和发现治疗新靶点。

The induction and chronification of neuropathic pain are affected by mitochondrial dysfunction which can be resulted from abnormal mitophagy. Our preliminary data showed for the first time that the knockdown of mammalian ste20-like kinase 1 (MST1) protected the animals from nerve injury-induced allodynia, which could be related to the effect of MST1 on mitophagy. However, the underlying mechanisms remain unclear. It is reported that MST 1 activates c-jun N-terminal kinase（JNK）by phosphorylation, and the activated JNK can induce mitophagy and mediate the downstream effect of MST1 in oxidative stress. In addition, MST 1 is methylated and thus inactivated via Lnc RNA MNX1-AS1. Therefore, we put forward the hypothesis: MST1, controlled by MNX1-AS1, augments the induction and chronification of peripheral neuropathic pain by regulating mitophagy and oxidative stress via JNK pathway. In this study, the mouse model of chronic constrictive injury in the unilateral sciatic nerve will be established, both MST1 knockout and wild type mice will be used, siRNA and adnovirus-associated virus transfection will be applied, and behavior and neuroelectrophysiological tests will be performed. Mitophagy, reactive oxgen species, apoptosis and neuroplasticity will be detected in the injured nerve, dorsal root ganglia and posterior horn. This part is to investigate the effect of MST1 on peripheral neuropathic pain and herein the role of its regulation in mitophagy. In addition, Schwann cells will be cultured, different interfered methods will be used, mitophagy and mitochondrial function will be detected. This part is to clarify the cellular functional consequence of MST1 in mitophagic regulation in peripheral nerves as well as the underlying mechanism. The proposed study will reveal the roles of MST1 in the regulation of peripheral neruopathic pain, thus suggest a potential new therapeutic target.