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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 study the neuroprotective effect of propolis on the N2a cells in vitro and in vivo by OGD. Cell viability was detected by CCK-8 assay and cell viability was measured by Giemsa staining. The morphological characteristics of the ethanol extract were evaluated by anti-ischemic reperfusion injury. Combined with HPLC analysis of ingredients. The results showed that the maximum concentration of propolis grade ethanol extract did not exceed 20μg / m L, and ODG4h complexed oxygen was the best model for 24 h. The cell viability and cell morphology at the doses of 5, 10μg / mL and 15μg / mL were significantly different from those in the model group, and the neuroprotective activity was dose-dependent. HPLC analysis showed that the higher content of flavonoids in 70% alcohol solubles was chrysin and pine hormone, which could be used as an index to evaluate the neuroprotective activity of propolis.