基于“AR-NOX-ROS”信号解析灯盏细辛有效组分保护缺血性脑中风的作用机制研究

The reactive oxygen species-induced peroxidation is an important mechanism for the development of ischemic stroke. Our previous study found that the expression of aldose reductase (AR) and NADPH oxidase (NOX) up-regulated after cerebral ischemic stroke, AR and NOX has a good therapeutic effect on cerebral ischemia stroke, and the mechanism of treatment is related with reactive oxygen species-induced peroxidation, so we speculate that AR may be the upstream signal molecules regulating NOX. Based on "clinical efficacy of traditional Chinese medicine and its essential components → brain protection plus anti-oxidative stress→anti-cerebral ischemic stroke mechanism of chinese medicine herb”, we proposed that "AR-NOX-ROS" Signaling Mechanism of Protective of Isolated Essential Components of Erigeron breviscapus on Ischemia Stroke. In the transient MCAO model, using knockout in vivo , the mechanism of AR-NOX-ROS in cerebral ischemic injury was investigated by molecular biology method, which will compared with that effected by essential components of Erigeron breviscapus and ultimately form a safe and effective, clear composition, quality control, a clear mechanism of the new effective ingredients compatibility compound. This is of great value to improve the pathophysiology of cerebral ischemic injury and to further expand the application of active ingredients in traditional Chinese medicine.

活性氧引起的过氧化反应是缺血性脑中风发生的重要机制，本项目组前期发现脑缺血中风后醛糖还原酶（AR）及NADPH氧化酶（NOX）表达上调，抑制AR及NOX对脑缺血具有良好治疗效果，且发现其治疗效果与对抗脑缺血后的过氧化及自由基损伤有关，我们推测AR可能是调控NOX的上游信号分子，本课题组依据 “临床确有疗效的中药及其制剂→脑神经保护＋抗氧化损伤→中药脑靶向神经保护药效物质基础”的研究思路，提出基于“AR-NOX-ROS”信号研究灯盏细辛的有效组分抗脑缺血中风的作用机制，应用基因敲除鼠建立脑缺血中风模型，通过分子生物学方法深入探讨脑缺血损伤后AR-NOX-ROS的调控机制，以及与灯盏细辛的有效组分干预作用进行比较，最终形成一个安全有效、成分清楚、质量可控、机制明确的新型有效成分配伍，完善脑缺血损伤的的病理生理学机制，开发缺血性中风治疗新靶点的有效药物。

活性氧引起的过氧化反应是缺血性脑中风发生的重要机制，本项目组前期发现脑缺血中风后醛糖还原酶（AR）及NADPH氧化酶2（NOX2）表达上调，抑制AR及NOX2对脑缺血具有良好治疗效果，且发现其治疗效果与对抗脑缺血后的过氧化及自由基损伤有关，因此提出了灯盏细辛有效组分保护脑缺血中风的AR-NOX-ROS作用机制研究，研究结果显示脑缺血损伤后，AR及NOX1、NOX2、NOX4的mRNA及蛋白表达均上调，灯盏细辛有效组分灯盏花乙素改善脑缺血后神经功能评分及减少了脑梗塞面积，同时ELISA检测3-NT、4-HNE、8-OHDG、NT-3、PAPR1和ROS表达显著减少，具有抗氧化作用，而且下调了AR及NOX1、NOX2、NOX4的mRNA及蛋白水平。研究进一步应用AR基因敲除小鼠建立脑缺血中风模型探讨AR与NOX之间是否存在直接的调控机制，实验结果显示AR基因敲除小鼠缺血中风后NOX1、NOX2、NOX4的mRNA及蛋白水平均下调，同时AR基因敲除能保护脑缺血损伤，及对抗脑缺血引起的过氧化损伤，灯盏花乙素在对抗脑缺血损伤中可能发挥AR抑制剂的功效，本研究结果明确了灯盏花乙素抗脑缺血损伤的AR-NOX-ROS作用机制。