Sorbinil和Zopolrestat对酵母工程菌中人醛糖还原酶活性的抑制作用

Inhibition of Human Aldose Reductase Activity in Engineering Yeast Strain by Sorbinil and Zopolrestat

目的利用人醛糖还原酶基因(AR)及其与绿色荧光蛋白基因(GFP)融合的酵母细胞表达模型,选用两种经典的AR抑制剂(ARI)-Sorbinil和Zopolrestat,初步观察模型在药物筛选方面的功能。方法复苏菌种并培养,在细胞培养液中加入ARI,观察细胞生长、GFP表达及荧光强度、AR蛋白表达及AR活性。结果 GFP荧光表达及AR蛋白的表达不受影响,但AR活性可被Zopolrestat抑制。结论 GFP标记的AR基因的酵母细胞模型可用于ARI的初步筛选,易重复,费用经济。Zopolrestat对酵母工程菌中表达的AR活性呈现出良好的抑制效应,可考虑作为该模型进行ARI筛选时的对照药物。

Objective To evaluate the inhibition effect of two classical aldose reductase inhibitors （ARIs）, Sorbinil and Zopolrestat on a potential ARI screening platform. Methods Aldose reductase （AR） and AR - GFP fusion genes were inserted into the yeast Saccharomyces cerevisiae cells, respectively. Sorbinil and Zopolrestat were added into the culture medium of the engineering XAR and XAG stains. The cell growth, GFP fluorescent signal, the target protein expression levels and AR activities were observed. Results The GFP fluorescent signal and protein level of AR was not inhibited by the ARIs. A relatively lower activity of AR was observed in Zopolrestat - treated group. Conclusion The yeast expression stain of AR tagged with GFP demonstrated a potential application in ARI screening. Zopolrestat showed a better inhibitory effect. Therefore it may be considered as the control drug for screening ARIs.