目的 筛选具有抗氧化及神经系统保护活性的淫羊藿Epimedii Folium黄酮类化合物（flavonoids extract from E. Folium，FEE），为缺血性脑卒中药物的开发奠定实验基础。方法 以1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基清除能力与铁离子还原抗氧化能力（ferric reducing ability of plasma，FRAP）为评价指标，筛选FEE中具有抗氧化活性的化合物，并以此为研究对象，建立强氧化剂过氧化氢诱导的SH-SY5Y细胞氧化应激损伤模型和PC12细胞氧糖剥离损伤模型评估其神经细胞保护活性，并观察其对急性脑缺血再灌注（middle cerebral artery occlusion，MCAO）大鼠模型的保护作用。结果 淫羊藿素、淫羊藿次苷I、金丝桃苷、槲皮苷、β-脱水淫羊藿素具有较高的DPPH自由基清除能力和总抗氧化能力（P＜0.01）；淫羊藿素、金丝桃苷和槲皮苷可显著抑制SH-SY5Y细胞氧化应激损伤，保护缺糖缺氧损伤的PC-12细胞，表现为细胞活力升高（P＜0.01）、活性氧水平降低（P＜0.05、0.01）、SH-SY5Y细胞丙二醛产生减少（P＜0.01）且胞内超氧化物歧化酶水平升高（P＜0.05、0.01）；淫羊藿素、金丝桃苷、槲皮苷可改善缺血再灌注大鼠脑组织抗氧化能力、调节神经功能、减少脑梗死面积（P＜0.05、0.01），具有神经保护作用。结论 淫羊藿素、金丝桃苷、槲皮苷具有良好的体外抗氧化及神经系统保护活性，对MCAO大鼠具有保护作用。

Objective To screen effective compounds with antioxidant and neuroprotective activities of flavonoids extract from Yinyanghuo (Epimedii Flolium, FEE) for laying the experimental foundation for the development of drugs with ischemic stroke. Methods The study used 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability and ferric reducing ability of plasma (FRAP) as indicators to screen compounds with high antioxidant activities from FEE. Based on the results, the oxidative stress model induced by hydrogen peroxide in SH-SY5Y cells and oxygen-glucose deprivation model in PC-12 cells were established to evaluate the neuroprotective activities. The acute middle cerebral artery occlusion (MCAO) model was used to verify their protective effects on rats.Results Antioxidant assay showed that icaritin, icariside I, hyperoside, quercitrin, β-anhydroicaritin had high DPPH radical scavenging ability and total antioxidant capacity (P < 0.01). Evaluation of neuroprotective activity showed that icaritin, hyperoside and quercitrin could significantly inhibit oxidative stress injury in SH-SY5Y cells and improve oxygen-glucose deprivation in PC12 cells, as demonstrated by increased cell viability (P < 0.01), decreased levels of reactive oxygen species (P < 0.05, 0.01), decreased production of malondialdehyd (P < 0.01) and increased levels of superoxide dismutase (P < 0.05, 0.01) in SH-SY5Y cells. Effects of cerebral ischemia-reperfusion showed that icaritin, hyperoside and quercitrin could improve the anti-oxidation ability of brain tissue, regulate nerve function and reduce the area of cerebral infarction (P < 0.05, 0.01), showing a good neuroprotective effects. Conclusion Icaritin, hyperoside and quercitrin had antioxidant and neuroprotective activities, exhibiting protective effects on rats with MCAO.

R285.5

国家中医药管理局“岐黄学者”中医药领军人才项目