基于酵母的高灵敏度抗朊病毒药物筛选模型建立及应用

Construction and application of a high sensitive anti-prion drug screening model based on yeast

建立一种高灵敏度抗朊病毒药物筛选模型研究传统中药的抗朊病毒作用。通过比较五种酵母朊病毒[PSI+]菌株在表型、生长特性和对抗朊病毒化合物敏感性方面的差异,优选出高药物敏感性酵母朊病毒[PSI+]菌株,并引入半变性琼脂糖凝胶电泳技术建立了一种高灵敏度抗朊病毒药物筛选模型;利用该模型研究了20种中药水提物治愈朊病毒的活性,发现淫羊藿和两面针水提物浓度为0.1 g.mL-1时,对酵母朊病毒[PSI+]的5 d治愈率分别为12.5%±3.5%和56.2%±9.1%,蛋白水平实验表明淫羊藿和两面针水提物具有治愈酵母朊病毒[PSI+]作用。

A high sensitive yeast-based, anti-prion drug screening model was developed to study anti-prion effects of traditional Chinese medicines. By comparing five yeast prion[PSI^＋] strains in growth characteristic, phenotype and sensitivity to anti-prion compounds, an optimized [PSI^＋] strain with high sensitivity to drugs was determined and a semi-denaturing detergent-agarose gel electrophoresis （SDD-AGE） technique was introduced to establish a high sensitive anti-prion drug screening model. Using this improved model, effects of aqueous extracts from 20 kinds of traditional Chinese medicines on the curing yeast prion [PSI^＋] were investigated. The results showed that the curing rates of [ PSI＋ ] cells at a 5-day treatment with the aqueous extracts from Epimedium brevicornum and Zanthoxylum nitidum at concentration of 0. 1 g . mL^-1 were 12.5% _＋ 3.5% and 56. 2% _＋ 9. 1% , respectively. Protein experiments showed that the aqueous extracts from Epimedium brevicornum and Zanthoxylum nitidum were able to cure yeast prion [PSI^＋]