黄芪甲苷配伍川芎嗪靶向NETosis治疗缺血性脑卒中的机制研究

中性粒细胞胞外诱捕网（NETs）是导致缺血性脑卒中静脉溶栓抵抗及再灌后炎症损伤的关键因素，血小板是NETs形成（NETosis）的诱导剂。临床常用治疗脑卒中药物川芎嗪能显著抗血小板、改善微循环，但对急性期炎症疗效欠佳限制了其广泛应用；黄芪甲苷为补气圣药黄芪的主要有效成分，能明显抑制缺血再灌注炎症损伤。分子对接提示黄芪甲苷与Gal-3、川芎嗪和整合素α-IIb结合，进一步的预实验表明二者配伍抗脑卒中作用优于单用，且显著抑制NETs形成。鉴此，提出“黄芪甲苷靶向中性粒细胞，川芎嗪锚定血小板，二者配伍共同抑制NETosis，减轻脑缺血再灌损伤”的假说。本项目拟通过MCAO大鼠及多种细胞模型，评价黄芪甲苷配伍川芎嗪干预脑卒中及NETosis的作用；并通过细胞热转变分析等技术筛选作用靶点，进而用基因干扰等手段对关键靶点反向验证。相关结果为脑卒中治疗提供新策略，也为阐释芪芎药对配伍机制提供思路。

Neutrophil extracellular traps (NETs) are the key factors leading to venous thrombolysis resistance and inflammatory cascade damage after reperfusion in ischemic stroke. Platelets are inducers of NETosis. Tetramethylpyrazine, the commonly used drug for treating ischemic stroke, significantly inhibits platelet function and improves microcirculation, but its unsatisfactory efficacy on acute phase inflammation limits its widespread use. Astragalus, the famous Qi-invigorating medicine, is of the highest frequency use in Chinese herbal formula to treat stroke. Its main active ingredient, Astragaloside IV, has a significant effect on ischemia-reperfusion inflammation, and the result of molecular docking suggests that Astragaloside IV combines with Gal-3, Tetramethylpyrazine anchors integrin α-IIb. Further preliminary experiments show that the combination of the two can significantly inhibit the formation of NETs. In view of this, we proposed the hypothesis that "Astragaloside IV targets neutrophils and Tetramethylpyrazine anchors platelets, the combination of the two inhibits NETosis and furthermore reduces cerebral ischemia-reperfusion injury." This project intends to evaluate the effect of Astragaloside IV combined with Tetramethylpyrazine on stroke and NETosis through MCAO rats and various cell models. Cellular thermal shift assay combined with other technology are used to screen biological target, and then reverse verification of key genes will be conducted. The relevant results will provide a new strategy for the treatment of stroke, and a reference for explaining the compatibility mechanism of Qixiong pair.