针刺促进脊髓可塑性中microRNA124的作用及其对PDXK的调控

Spinal cord injury, as a conmen disease, usually encountered in clinic practice. Acupuncture has been well known as a benefit method for the treatment of SCI, but the underlying mechanism keeps to be elucidated. Our previous study has found PPXK and microRNA 124 were specifically expressed in the spinal cord following transection(SCT) in rats with acupuncture, indicated by proteomics and microarray analysis. This study, therefore, was designed to investigate the role of microRNA 124 in neuroplasticity in SCT rats with acupuncture. Moreover, the level of PDXK was also determined, then the direct regulation and action between microRNA 124 and PDXK was also elucidated. The values of this study is not only useful to support of usage on acupuncture in clinic practice, but also have an important scientific significance and better clinic perspective. It is very interesting and important work with novel creation.

脊髓损伤(SCI)修复机制与治疗研究一直是医学界关注的焦点。针刺能促进损伤脊髓功能部分恢复已在临床得到证实，但分子机制尚不完全清楚。本项目在课题组前期蛋白组学和microRNA芯片发现针刺促进SCI大鼠神经可塑性中吡哆醛激酶（PDXK）和microRNA124表达变化的基础上，拟建立大鼠脊髓横断伤督脉针刺模型，采用基因克隆、RNA干扰、免疫细胞化学、RT-PCR、Western Blot和细胞培养等技术，研究microRNA-124在针刺促进脊髓可塑性中的作用及其对PDXK的调控。项目研究结果不仅有助于进一步推动针刺治疗SCI的临床应用，而且有助于发现影响SCI的新分子和阐明与之相关的微小RNA调控机制，有着重要的科学价值和临床意义，是创新性强的原创性研究工作。

针刺能促进损伤脊髓功能部分恢复已在临床得到证实，但分子机制尚不完全清楚。前期研究中蛋白组学和microRNA 芯片发现针刺促进 SCI 大鼠神经可塑性中吡哆醛激酶 （PDXK） 和 microRNA124 表达变化。本研究探索microRNA-124 及PDXK在针刺促进脊髓可塑性中的作用及microRNA124对 PDXK 的调控。在体外， PDXK过表达可以明显提高新生大鼠皮质神经元的存活率，增加神经元存活数量，而抑制PDXK导致了相反的结果。在体内，制备督脉针刺SD大鼠脊髓横断损伤模型后，PDXK过表达可以明显提高运动功能恢复，对抗胶质细胞和神经元凋亡，而抑制PDXK体内表达导致了负面的结果。同时体外microRNA124添加可以明显提高脊髓神经元的存活率，并对抗缺氧损伤，而加入microRNA124抑制剂导致了相反的结果。在体内，制备督脉针刺SD大鼠脊髓横断损伤模型后，microRNA124仿效剂可以提高运动功能恢复，减少细胞凋亡，而microRNA124抑制剂导致了负面的结果。PDXK 和microRNA124的作用机制与erk，akt和GAP43调节有关。 此外，生物信息学分析表明，PDXK不是microRNA124的靶点。进而，我们用荧光素酶报告基因检测实验发现microRNA339与PDXK的3'UTR位置匹配并以阻止PDXK转录。本研究为未来有关脊髓损伤治疗的临床试验提供了潜在的治疗靶点。