Perfusion enzyme affinity selection system based on hollow fiber for screening active compounds from total saponins of Kudingcha
10.16438/j.0513-4870.2019-0917
- VernacularTitle:基于中空纤维吸附靶酶的流体灌注系统用于苦丁茶总皂苷活性成分筛选
- Author:
Yu HOU
1
;
Jiao ZHENG
2
;
Peng-fei TU
1
;
Xiao-ni AI
1
;
Yong JIANG
1
Author Information
1. State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100191, China
2. Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
- Publication Type:Research Article
- Keywords:
perfusion;
hollow fiber;
total saponins of Kudingcha;
enzyme-target screening;
lipase;
atherosclerosis
- From:
Acta Pharmaceutica Sinica
2020;55(3):495-500
- CountryChina
- Language:Chinese
-
Abstract:
Atherosclerosis is one of the causes of many cardiovascular diseases. Lipid metabolism disorder is an important risk factor for atherosclerosis. Lipase-targeted screening may help discovery of hypolipidemic and anti-atherosclerotic drugs. Traditional methods for enzyme-based screening exhibit drawbacks of tedious operation steps, reduced enzyme activity, slow mass transfer, as well as high false positive rate. In this paper, an integrated perfusion enzyme affinity selection system based on hollow fiber was constructed to screen hypolipidemic and anti-atherosclerotic compounds from traditional Chinese medicines, and the total saponins of Kudingcha were taken as a case. First, we built a hollow fiber based perfusion system and optimized the methodology for enzyme affinity selection. Then, two active compounds of kudinosides A and C were identified as potential lipase inhibitors from the total saponins of Kudingcha by the proposed system. Last, the activity of kudinosides A and C was verified by lipase inhibitory assay and formation of foam cell model induced by low density lipoprotein aggregates, exhibiting the reliability of the system. This platform shows the advantages of integration, fast mass transfer, simple operation, low cost, as well as improved throughput and efficiency, which is especially suitable for rapid screening active components from traditional Chinese medicine.
