Comparative study on microassays for screening acetylcholinesterase inhibitors
- VernacularTitle:乙酰胆碱酯酶抑制剂微量筛选模型的比较研究
- Author:
Qianyun SUN
;
Fumei YANG
- Publication Type:Journal Article
- Keywords:
acetylcholinesterase;
acetylcholinesterase inhibitor;
screening model;
Alzheimers disease
- From:
Chinese Pharmacological Bulletin
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To provide practical microassays for screening acetylcholinesterase (AChE) inhibitors in drug discovery.Methods The optimal conditions of assaying the activity of AChEs from electric eel,rat brain homogenate and cobra venom were determined in 96-well plates under physiological pH value and temperature by orthogonal matrix method.The concentrations of AChE,substrate and DTNB,and reaction time were optimized.After the effects of sample solvent (DMSO) used in the assay and stopping reagent on enzyme activity were assessed,the assay conditions were finally selected,and 492 kinds of extracts from Guizhou ethno-drugs were screened.Results Practical microassays for screening AChE inhibitors were successfully constituted by using AChEs mentioned above.The data analysis of screening results revealed that electric eel AChE possessed a high sentivity to inhibitors,and cobra venom AChE shared high similarity with rat brain homogenate in positive results.Conclusion Microassays constituted in this work possessed advantages of being easy,rapid,reliable,cost saving and flexible.AChE from electric eel was especially suitable for screening AChE inhibitors from extracts,and AChE from cobra venom was more suitable to be used in screening AChE inhibitors from large numbers of compounds.
