目的 探明艾片抗脑缺血再灌注损伤的作用及其机制。方法 借助PharmMapper等数据库获得艾片（左旋龙脑）抗脑缺血再灌注损伤（cerebral ischemia-reperfusion injury，CIRI）的潜在靶点并进行基因本体（gene ontology，GO）功能富集分析和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析。分别设置假手术组、模型组、溶剂模型（1%聚山梨酯-80）组、阳性药（尼莫地平）组及艾片高、中、低剂量（0.20、0.10、0.05 g/kg）组，采用预防和治疗给药的方式对线栓法制备的CIRI模型大鼠进行干预，Zea Longa评分法对不同时间点（再灌注4、22.5、46.5、70.5 h）模型大鼠神经功能进行评价；2，3，5-氯化三苯基四氮唑（2，3，5-triphenyltetrazoliumchloride，TTC）染色法观察大鼠脑梗死率；蛋白质印迹（Western blotting）和免疫组织化学法对预测的通路进行初步验证。结果 网络药理学预测显示，艾片抗CIRI的潜在靶点有63个，可能与调控生物过程及叉头转录因子O亚族（forkhead transcription factor of the O class，FOXO）信号通路、磷脂酰肌醇-3激酶（phosphatidylinositol-3-kinases，PI3K）/蛋白激酶B（protein kinase B，Akt）信号通路、雌激素信号通路等有关。实验结果显示，艾片0.20、0.10 g/kg组能极显著降低模型大鼠脑梗死率，显著升高模型大鼠缺血侧皮层中磷酸化叉头转录因子O亚族3a（phosphorylated-forkhead box O3a，p-FOXO3a）蛋白表达量和p-Akt/Akt和p-FOXO3a/FOXO3a值，极显著降低细胞死亡调解因子（Bcl-2 interacting mediator of cell death，Bim）和半胱氨酸天冬氨酸蛋白酶-3（cysteinyl aspartate specific proteinase 3，Caspase-3）蛋白表达量，艾片0.20 g/kg组还能不同程度改善模型大鼠各时间点神经功能损伤。结论 艾片可能是通过调节Akt/FOXO3a/Bim信号通路改善CIRI，发挥抗细胞凋亡作用。

Objective To investigate the role and possible mechanism of different doses of L-borneolum prevention plus treatment in cerebral ischemia-reperfusion injury (CIRI). Methods With the help of databases such as PharmMapper, the potential targets of L-borneolum against CIRI was obtained, then the gene ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were processed. The chosen rats were randomly divided into seven groups (sham group, model group, vehicle (1% tween) group, positive drug (nimodipine) group, and high, medium and low doses of L-borneolum (0.20 g/kg, 0.10 g/kg, 0.05 g/kg) groups. The rat model of CIRI was established by suture method and administrated in the manner of pretreatment plus treatment. Zea Longa scoring was used to detect neurobehavioral changes at various time (reperfusion 4, 22.5, 46.5, and 70.5 h) of CIRI rats. Cerebral infarction volume was detected by 2,3,5-triphenyltetrazoliumchloride (TTC) staining. In addition, Western blotting and immunohistochemistry analysis were performed to verify the predicted pathway. Results Network pharmacology predicted that there were 63 potential targets of L-borneolum against CIRI, these targets were enriched in the forkhead transcription factor of the O class (FOXO) signaling pathway, phosphatidylinositol-3-kinases (PI3K)/protein kinase B (Akt) signaling pathway, and estrogen signaling pathway, etc. Experimental results showed that following L-borneolum (0.20 g/kg and 0.10 g/kg) treatment, the cerebral infarct sizes in CIRI rats were significantly attenuated, the expression of phosphorylated-forkhead box O3a (p-FOXO3a) and the ratio of p-Akt/Akt and p-FOXO3a/FOXO3a was increased, and Bcl-2 interacting mediator of cell death (Bim) and cysteinyl aspartate specific proteinase 3 (Caspase-3) protein expression was decreased in ischemic cortex. In addition, L-borneolum group (0.20 g/kg) can also improve the nerve function impairments of CIRI rats at various time points. Conclusion Neuroprotective and anti-apoptotic effect of L-borneolum against CIRI is mediated via the Akt/FOXO3a/Bim signaling pathway.

R285

国家自然科学基金面上项目（81873023）；国家自然科学基金面上项目（81473371）；成都中医药大学中药“性-效-用”理论与实践创新团队（CXTD2018004）；成都中医药大学西南特色中药资源重点实验室开放研究基金资助项目（2020XSGG025）