P2Y2受体在脊髓损伤大鼠星形胶质细胞介导血管及神经再生与修复中的作用研究

Astrocytes on neural functional recovery after spinal cord injury play an important role. P2Y2 receptors are involved in regulating neurotransmitters, inflammation, astrocyte activation and neural differentiation. While there is no report that P2Y2 receptor mediated angiogenesis and neuronal regeneration in rats with spinal cord injury. In this study, neurobehavioral morphology, molecular biology and metabolomics were used to speculate the mechanism of purinergic receptor P2Y2 in astrocytes mediated neural functional recovery in rats with spinal cord injury. Here, 1H NMR based metabolomic studies was used to analysis the metabolic changes of the injured spinal cords and the related metabolin of P2Y2 receptor. Through immunofluorescence to investigate the role of the P2Y2 receptor on astrocytes activation, activating and inducting endogenous neural stem cells (ENSCs), angiogenesis and neural differentiation after spinal cord injury. The corticospinal tract (CST) fibers were traced by biotinylated dextran amine (BDA) immunofluorescence. We provide a framework for understanding the P2Y2 receptors in astrocytes mediated angiogenesis and neuronal regeneration in rats with spinal cord injury. The findings unveil a new approache for spinal cord injury.

星形角质细胞在脊髓损伤神经功能恢复中起重要作用，P2Y2受体在参与神经活性物质合成与释放、星形胶质细胞活化及神经发育分化等方面均扮演重要角色，但针对P2Y2受体是否介导脊髓损伤后血管、神经再生的相关研究尚无报道。本研究拟应用行为学、分子生物学、代谢组学等方法，观察脊髓损伤后P2Y2受体与神经功能恢复的关系并明确部分机制，通过核磁共振1H谱（1H NMR）分析脊髓损伤区代谢变化，明确P2Y2受体相关代谢物，免疫荧光法观察P2Y2受体对脊髓损伤后星形胶质细胞活化、内源性神经干细胞激活、新生神经元及血管再生的影响，并示踪检测P2Y2受体在皮质脊髓束神经纤维生长中的作用，结果将明确P2Y2受体相关差异代谢物，阐明P2Y2受体在脊髓损伤大鼠星形胶质细胞活化、内源性神经干细胞激活及神经血管再生中的作用，从多层面验证P2Y2受体介导脊髓损伤功能恢复的机制，为临床治疗提供新方法，为新药开发提供新靶点。

脊髓损伤后由于中枢神经固有的再生能力差，神经功能恢复困难，P2Y2受体在参与神经活性物质合成与释放、星形胶质细胞活化及神经发育分化等方面均扮演重要角色，本研究探讨P2Y2受体介导脊髓损伤后血管、神经再生的机制。实验通过应用P2Y2R抑制剂干预大鼠脊髓损伤模型观察其神经保护作用。我们首先建立了大鼠脊髓损伤动物模型，以P2Y2R抑制剂作为干预因素，利用大鼠后肢功能BBB评分法评价大鼠神经功能恢复情况，HE 染色观察脊髓损伤后组织病理改变情况，采用免疫荧光染色观察脊髓损伤后反应性星形胶质细胞活化；应用生物素葡聚糖胺顺行示踪脊髓损伤后皮质脊髓束的神经纤维；免疫荧光染色技术观察损伤后神经干细胞的数量及分布，神经元的分化情况；western blots 检测P2Y2R表达量变化，1H NMR分析了代谢组学变化，初步探讨了P2Y2R抑制剂对大鼠脊髓损伤后神经功能恢复的影响及其可能的作用机制。本研究表明P2Y2R抑制剂可以明显改善大鼠脊髓损伤后神经功能的恢复，脊髓损伤后应用P2Y2R抑制剂能促进神经纤维生长，脊髓损伤后大量的内源性神经干细胞被激活，干预P2Y2R信号通路可诱导内源性神经干细胞部分向神经元方向分化。