蜂胶分级提取物的神经保护功效及成分研究

The Neuroprotective Effect and the Effective Components of Ethanol Extracts of Propolis

研究蜂胶中具有神经保护功效的活性成分,对N2a细胞进行氧糖剥夺(OGD)模拟体内缺血再灌注模型。采用CCK-8法检测细胞存活率并以其作为活性筛选指标,以吉姆萨染色法观测细胞形态学指标,对蜂胶分级醇提物进行抗脑缺血再灌注损伤的活性研究。结合HPLC进行成分分析。结果表明:蜂胶分级醇提物的最大作用质量浓度不超过20μg/m L,以ODG4h复糖复氧24 h为最佳模型条件,对蜂胶分级醇提物进行活性筛选,蜂胶70%醇溶物活性最高,5,10μg/m L和15μg/m L剂量组细胞存活率和细胞形态较模型组细胞均显示有极显著差异,且神经保护活性存在剂量依赖关系。HPLC分析表明,70%醇溶物中含量较高的黄酮类成分为白杨素和松属素,可作为评估蜂胶神经保护功效活性的指标。

To confirm the neuroprotective effective components in ethanol extracts of propolis, a model of N2 a cells injured by OGD was established. Cell survival rate measured by CCK-8 assay was considered as the screening criterion.GIMSA dying assay was used to observe morphology changes of N2 a cells. HPLC were used as detection and analysis method. The result showed that the action concentration was no more than 20 μg/mL. The appropriate time of OGD that could induce N2 a cells injury was 4 hours. 70% ethanol extract group showed the highest protective effect that the cell survival rate and the morphology of N2 a cells of 5 μg/mL group, 10 μg/mL group and 15 μg/mL group was significantly higher than the model group. The result of HPLC qualitative analysis of 70% ethanol extract suggested that major constituents were chrysin, pinocembrin, which can be used as evaluation index of the neuroprotective effect of propolis.