山楂叶总黄酮调控微环境促进脊髓损伤后神经再生的分子机制研究

Spinal cord injury leads to high rate of disability. Regular clinical medications have proved to be ineffective, and there is an urgent need for development of effective drugs for the treatment of acute spinal cord injuries. The research group found in previous study that the flavonoids in hawthorn leaves, the main pharmacological constituents in region-featured drug Guang hawthorn in Guangxi Province, could effectively protect damaged nerves and initiate nerve regeneration.No relative reports had been published in China or abroad. We hypothesized that the flavonoids in hawthorn leaves could regulate the microenvironment after spinal cord injury through NF-κB/Wnt-β-catenin dual signaling pathway and promote nerve regeneration. Therefore, based on the NF-κB/Wnt-β-catenin dual signaling pathway and metabolomics, we will apply Axillary Reverse Mapping, RNA interference, Dual Luciferase Reporter Gene Technology, Gas Chromatography-Mass Spectrometer and other experimental technology.From aspects including animals, cells, genes, proteins, and metabonomics, we will systematically study the mechanism of how flavonoids in hawthorn leaves modulates the expression of inflammatory factors and neurotrophic factors, and the process of neuronal autophagy after spinal cord injury.Explaining the molecular mechanism of promoting nerve regeneration after spinal cord injury, it can provide new therapeutic strategies for clinical treatment of spinal cord injury.

脊髓损伤导致高致残率，目前临床常规用药被证明效果不明确，研发有效治疗急性脊髓损伤的药物迫在眉睫。课题组前期研究发现，广西道地药材广山楂叶的主要药理成分山楂叶总黄酮能有效保护受损脊髓并启动神经再生，国内外未见相关报道。我们推测山楂叶总黄酮通过靶向NF-κB/Wnt-β-catenin双信号通路调控脊髓损伤后微环境的改变，促进神经功能的恢复。为此，我们将基于NF-κB/Wnt-β-catenin双信号通路和代谢组学，采用反向示踪技术、RNA干扰、双荧光素酶报告基因技术、气相色谱-质谱联用技术等实验技术，从动物、细胞、基因、蛋白质、代谢组学等多方面进行实验设计，系统、深入地研究山楂叶总黄酮调控脊髓损伤后炎性因子、神经营养因子的表达和神经元自噬的过程，阐述其促进脊髓损伤后神经再生的分子机制，为临床治疗脊髓损伤提供新的策略。

项目的背景：脊髓损伤（Spinal Cord Injury, SCI）导致高致残率，目前临床常规用药被证明效果不明确，研发有效治疗急性脊髓损伤的药物迫在眉睫。课题组前期研究发现，广西道地药材广山楂叶的主要药理成分山楂叶总黄酮（Total Flavonoids of Hawthorn Leaves, TFHL）能有效保护受损脊髓并启动神经再生，国内外未见相关报道。因此，本课题组拟在前期研究有所发现的基础上系统深入的探索TFHL对SCI后损伤脊髓局部微环境及神经再生的影响及其作用机制。 .主要研究内容：1.探究TFHL在SCI后保留残存的脊髓功能、促进损伤脊髓结构及运动功能重建的作用；2.探究TFHL通过抑制炎症因子、增强自噬和促进神经营养因子的分泌改变脊髓受损神经周围的微环境，促进SCI恢复的机制；3.探究TFHL调控NF-κB通路抑制炎症因子、增强自噬抑制SCG10和调控Wnt/β-catenin通路增加神经营养因子分泌治疗SCI的分子机制；4.基于Q-Orbitrap高分辨液质联用技术分析TFHL的成分；5.基于1H NMR代谢组学方法探讨TFHL治疗脊髓损伤大鼠的机制。.重要结果：TFHL通过调控TLR4/NF-κB信号通路改变炎症因子表达，增强脊髓神经元自噬相关蛋白表达，调控星形胶质细胞Wnt/β-catenin通路增加神经营养因子分泌改善SCI后微环境，进而保护损伤脊髓组织结构，减少神经元凋亡，促进神经元轴突再生和运动功能恢复。.科学意义：通过本课题的研究，阐明了广西道地药材广山楂叶的主要药理成分山楂叶总黄酮对SCI后神经微环境及损伤神经元再生的影响，深入解析其分子机制，为SCI及神经再生的治疗提供新的策略，为广山楂叶这一广西少数民族特色药物的有效开发利用提供科学的理论依据。