从LncBANCR介导ERK/mTOR调控自噬探讨推拿对SNL大鼠的镇痛效应机制

Neuropathic pain (NPP) is one of the most difficult problems which are hard to break through in pain treatment at present. Previous studies have found that ERK and spinal dorsal horn autophagy are important pathways and nodes of pain conduction and participate in the occurrence and development of NPP，and massage can affect ERK and autophagy, and then achieve the purpose of analgesia,but the relation between them is unclear. Recent studies have also found that ERK / mTOR is an important pathway for autophagy and LncBANCR is involved in ERK regulation. According these studies, we put forward the hypothesis that massage may affect the expression of LncBANCR, regulate the occurrence of autophagy in spinal dorsal horn through ERK/mTOR pathway,thereby inhibiting the occurrence of pain. To verify the hypothesis, we observe the change and expression of autophagy of the dorsal horn of the spinal cord when massage on SNL rats model, and then use PCR, Western blot and immunohistochemistry to detect the effct on ERK, mTOR and autophagy by LncBANCR expression , clarify their correlations and compare with the situation after massage on SNL rats , for clarifing the massage can affect LncBANCR expression then regulate autophagy, clearing molecular mechanism of massage analgesia.

神经病理性疼痛(Neuropathic pain，NPP)是目前疼痛治疗无法突破的难题之一。前期研究分别发现ERK与脊髓背角自噬是疼痛传导的重要通路与节点，参与了NPP的发生发展，而推拿均可以通过影响ERK及自噬，达到镇痛的目的，但它们之间的联系尚不明确。最近研究提示，ERK/mTOR是自噬发生的重要途径，而LncBANCR参与了ERK的调控。据此我们提出假说，推拿可能影响LncBANCR表达，通过ERK/mTOR途径调控脊髓背角自噬，进而抑制疼痛的发生。为验证假说，我们通过SNL大鼠模型，观察推拿对SNL大鼠脊髓背角的自噬表达及变化情况；再运用PCR、Western blot、免疫组化等技术检测LncBANCR表达对ERK/mTOR及自噬的影响，明确其相关性；同时与推拿后大鼠脊髓背角自噬的情况进行比较，以期阐明推拿是通过影响LncBANCR的表达，进而调控自噬达到镇痛的目的。

神经病理性疼痛(neuropathicpain，NPP)，是由躯体感觉系统的损伤或疾病所导致的一种慢性疼痛，是慢性疼痛的主要原因之一，在人群中发病率约8%。有文献报道，LncBANCR可以通过调控自噬及炎症的发生，但LncBANCR与自噬、炎症在NPP疾病中三者关系尚未明确。本实验通过推拿干预SNL模型大鼠，观察推拿对SNL大鼠脊髓背角LncBANCR与自噬相关蛋白LC3、p62表达的影响，探讨推拿对神经病理性疼痛大鼠的作用并分析其作用机理。术前各组大鼠PWTL和PWMT差异无统计学意义；术后干预第1天SNL组、推拿组、假推拿组、假手术组的PWTL和PWMT较正常组显著降低(P＜0.01)；术后干预第3天SNL组、推拿组、假推拿组、假手术组的PWTL和PWMT较正常组显著降低(P＜0.01)，其中推拿组较SNL组PWTL和PWMT显著上升(P＜0.01)；术后干预第7天SNL组、推拿组、假推拿组的PWTL和PWMT较正常组显著降低(P＜0.01)，推拿组PWTL和PWMT高于SNL组 (P＜0.01)。假手术组PWTL和PWMT术后干预第7天逐渐恢复，与正常组比较无显著差异( P＞0.05)。术后干预第7天，SNL组血清IL-1β、脊髓背角LncBANCR、ERK、mTOR、LC3Ⅱ/ LC3Ⅰ和p62蛋白表达水平较正常组均上升(P＜0.01)，与SNL组相比，推拿组可下调血清IL-1β、脊髓背角LncBANCR、ERK、mTOR、LC3Ⅱ/ LC3Ⅰ和p62蛋白表达水平(P＜0.01)。证实推拿可减轻SNL大鼠痛觉过敏反应，其机制是推拿与LncBANCR的表达可能存在相关性，推拿可通过抑制LncBANCR的表达，并通过下调ERK/mTOR通路，下调自噬蛋白LC3和p62蛋白表达，进而抑制自噬过度激活，阻碍炎症因子IL-1β的成熟和分泌，达到阻断NPP传导和维持的作用。