电针通过Nogo-A/NgR与NGF/TrkA的crostalk调节SCI后神经可塑性的机制研究

Spinal cord injury (SCI) in the injured central nervous system, Nogo-A/NgR signaling pathway and P75NTR protein binding inhibition of nerve regeneration,NGF/TrkA signaling pathway and P75NTR protein binding to promote nerve regeneration,Spinal nerves have limited plasticity by the crosstalk of the two signaling pathways.How to improve the crosstalk between the tow so as to promote the neural plasticity after SCI,Is the key to the recovery of neural function after SCI.Our research group from the perspective of Chinese medicine of "flaccidity disease",selection of electro acupuncture on the basis of "Du meridian qi and blood " and "treatment for flaccidity aims at yangming meridiandredge " treatment as a means of optimization of crosstalk between NGF/TrkA and Nogo-A/NgR,copy SCI rat model,Use transmission electron microscopy, western bolt Co-IP, qRT-PCR and other technologies,to observe the expression of related protein and mRNA in SCI rats spinal cord segments injured by using different signal pathway blocking agent and electroacupuncture at different acupoints,Preliminary study of the effect about crosstalk between NGF/TrkA and Nogo-A/NgR on neural plasticity of injured spinal cord in rats,and the effect of Electroacupuncture of "treating the same disease with different governance" in the treatment of SCI.

脊髓损伤(SCI)后受损中枢神经中Nogo-A/NgR信号通路与P75NTR蛋白结合抑制神经再生，NGF/TrkA信号通路与P75NTR蛋白结合促进神经再生，脊髓神经在这两条信号通路的crosstalk下具备有限的神经可塑性。如何改善它们之间的crosstalk从而促进SCI后神经可塑性，是SCI后神经功能恢复的关键。本课题组从中医“痿病”角度出发，选取电针依据“疏通督脉气血”和“治痿独取阳明”治则作为优化NGF/TrkA与Nogo-A/NgR间crosstalk的手段，复制SCI大鼠模型，使用透射电镜、Co-IP、western bolt、qRT-PCR等技术观察运用不同信号通路阻断剂以及使用不同经穴电针干预SCI大鼠受损脊髓节段的相关蛋白及mRNA表达量，初步探讨Nogo-A/NgR和NGF/TrkA的crosstalk对大鼠受损脊髓神经可塑性的影响及电针“同病异治”治疗SCI的机制。

脊髓损伤(SCI) 后对神经可塑性具有抑制作用的 Nogo-A/NgR 信号通路和具有促进作用的NGF/TrkA 信号通路具有 crosstalk。本课题提出“SCI 后神经可塑性受到 Nogo-A/NgR 和 NGF/TrkA 的 crosstalk影响”的假说，通过建立大鼠SCI模型，将120只脊髓损伤大鼠模型随机分为胃经组、督脉组、混合组、对照组4组。观察3d、7d、14d、21d时间节点， 通过神经功能评价，qrt-PCR技术观察Nogo-A/NgR 和 NGF/TrkA信号转导通路相关蛋白GAP-43 和 SYP表达。常规 HE 染色，在光学显微镜下观察脊髓损伤后各时间点组织形态学变化；铀-铅染色后，经透射电镜观察损伤区神经元的超微结构变化。结果①电针刺激通过调控Nogo-A/NgR 和 NGF/TrkA信号通路相关分子蛋白的表达对脊髓损伤大鼠产生治疗作用。②电针刺激可以上调Nogo-A/NgR 和 NGF/TrkA信号通路相关因子基因和相应蛋白表达实现对脊髓损伤的治疗作用。③阻断剂可通过调控Nogo-A/NgR 和 NGF/TrkA信号通路相关分子蛋白的表达对脊髓损伤大鼠产生治疗作用。④阻断剂可以上调Nogo-A/NgR 和 NGF/TrkA信号通路相关因子基因和相应蛋白表达实现对脊髓损伤的治疗作用。实验数据表明：脊髓损伤大鼠下肢神经功能评分提示脊髓损伤后大鼠下肢运动功能存在可逆性的修复现象，可能是中枢神经系统自我修复机制的一部分，同时电针干预对脊髓损伤后大鼠 的下肢运动功能肌力恢复在时间和效果上优于未经干预的脊髓损伤大鼠。证明电针和阻断剂均可改变神经干细胞的微环境，影响一些重要的信号通路的变化，促进脊髓损伤大鼠下肢运动功能的恢复，提高其行为学评分。为脊髓损伤的后续治疗提供了一种新思路，为脊髓损伤后神经再生修复的基础研究及临床转化研究提供参考。