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目的应用高内涵(HCS)技术快速筛选对β淀粉样蛋白(Aβ25-35)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Aβ25-35损伤的SH-SY5Y细胞为模型,采用Hoechst33342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Aβ25-35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 rapidly screen high-content (HCS) extracts of traditional Chinese medicine (TCM) that has a protective effect on SH-SY5Y cell injury induced by β-amyloid protein (Aβ25-35). Methods Twenty Chinese Herbal Medicine Pieces were extracted. Each herb was extracted with 20%, 40% and 95% ethanol respectively. The SH-SY5Y cells damaged by Aβ25-35 were used as the model and Hoechst33342 and PI double staining. Extract on the model cells; on this basis, further observation of the activity of active extract on model cells Caspase-3/7 activity, rapid screening of Alzheimer’s disease (AD) potential therapeutic effect of traditional Chinese medicine extract. Results HCS screening found that among the 60 extracts, 17 extracts had better protective effect on model cells. Further studies showed that 8 of these extracts had a good effect on the activity of Caspase-3/7 Inhibition. And to Liu slaves 40% ethanol and Salvia 20%, 40% ethanol optimal site. Conclusion Fluorescent labeling-based HCS technology is an effective and rapid screening method for the analysis of traditional Chinese medicine extracts that have a protective effect on SH-SY5Y cell injury model induced by Aβ25-35. The selected traditional Chinese medicine extract has potential anti-AD activity.