目的 应用高内涵(HCS)技术快速筛选对β淀粉样蛋白(Aβ_25-35)致SH-SY5Y细胞损伤模型有保护作用的中药提取物。方法 选取20种中药饮片,每种药材分别进行20%、40%和95%乙醇提取,以Aβ_25-35损伤的SH-SY5Y细胞为模型,采用Hoechst 33342和PI双染,并利用HCS技术观察60个提取物对模型细胞的保护作用;在此基础上进一步观察活性提取物对模型细胞Caspase-3/7活性的影响,快速筛选对阿尔茨海默病(AD)有潜在治疗作用的中药提取物。结果 HCS筛选发现,在60个提取物中,有17个提取物对模型细胞具有较好的保护作用;进一步研究显示,其中有8个提取物对模型细胞Caspase-3/7活性有较好的抑制作用。并且以刘寄奴40%乙醇部位及丹参20%、40%乙醇部位效果最优。结论 以荧光标记为基础的HCS技术是有效、快速筛选对Aβ_25-35致SH-SY5Y细胞损伤模型有保护作用中药提取物的分析方法,筛选出的中药提取物具有潜在的抗AD活性。

Objective To identify neuroprotective extracts with the protective effects on Aβ_25-35-induced SH-SY5Y cell injury via high content screening (HCS). Methods Hoechst 33342/PI double staining method was used to screen neuroprotective extracts from 60 Chinese materia medica (CMM) extracts. Further more, the effects of neuroprotective extracts on Aβ_25-35-induced changes in the levels of Caspase-3/7 were detected. Results The results showed that 17 extracts had obviously neuroprotective effects. Among the 17 extracts, 8 of them inhibited Aβ_25-35-induced up-regulation of Caspase-3/7. Conclusion HCS is an efficient method to screen neuroprotective extracts with the protective effects of Aβ_25-35-induced SH-SY5Y cell injury. The neuroprotective extracts have potential medicinal value in Alzheimer's disease.

国家科技部重大新药创制项目(2013ZX09402203)