中草药醛糖还原酶抑制剂筛选模型的建立

Establishment of Screening Model for Aldose Reductase Inhibitor(ARI) from Chinese Herbal Medicine

目的:通过观察中药知母对醛糖还原酶活性的抑制作用,建立一种适用于筛选中草药醛糖还原酶抑制剂的模型。方法:大鼠12只随机分成3组:空白对照组、知母组、阳性药物组,分别给予生理盐水、知母水提液、依帕司他水溶液灌胃。采用“通法”给药方案制备含药血清,部分血清经过丙酮预处理。将知母水提液和含药(知母)血清分别加入体外建立的以DL-甘油醛为底物、NADPH为辅酶的醛糖还原酶活性测定体系。结果:丙酮处理过的空白血清较未处理的空白血清对酶活性的影响更小;知母水提液对醛糖还原酶抑制率达99%,含药(知母)血清对醛糖还原酶抑制率达67%。结论:运用血清药理学实验方法建立的筛选模型能有效、可靠的进行中草药醛糖还原酶抑制剂的筛选。

Objective：To set up a screening model of aldose reductase inhibitor （ARI） from Chinese herbal medicine by observing the inhibitory effect of anemarrhenae on aldose reductase（AR）. Methods：12 rats were randomly divided into 3 groups： anemarrhenae group,positive drug group and negative control group, They were orally administrated with anemarrhenae extration,epalrestat and NS, respectively. The rats were then sacrificed by collecting blood from the abdominal aorta one hour later after the last intragastric administration. Some blood serum was pre - disposed with acetone. Then Anemarrhenae serum and Anemarrhenae extraction were add to AR reaction system that used DL - glyceraldehyde as substrate and NADPH as coenzyme. The AR activity was detected by microplate scanning spectrophotometer. Results：The blank serum pre-deposed with acetone had less impact on AR than blank serum; Anemarrhenae extraction and anemarrhenae serum both had great inhibitory effect on AR, the inhibitory rates were 99% and 67% ,respectively. Conclusion：The new screening model can efficiently and credibly search for the ARI.