title: Development of fluorescence imaging-based assay for screening cardioprotective compounds from medicinal plants.
abstract: Medicinal plants have been widely recognized as a renewable resource for the discovery of novel leads and drug. In this study, an approach for screening and identification compounds with cardioprotective activity from medicinal plant extracts by cellular-fluorescence imaging technique was developed. It is a cell-based assay for measuring mitochondrial membrane potential changes in H9c2 cardiac muscle cells exposed to H(2)O(2) by using a fluorescence automatic microscopy screening platform. Rhodamine 123 was used as the fluorescent dye to indicate the change of mitochondrial membrane potential. The sensitivity and linear range of the proposed approach were evaluated and validated using vitamin C, an antioxidative compound. The method was applied to screen active components with potent cardioprotective effects from a traditional Chinese formula. The potential cardioprotective components were identified by liquid chromatography coupled with mass spectrometry (LC/MS). Moreover, the utility of the proposed approach was further validated by three compounds (salvianolic acid B, protocatechuic aldehyde, and tanshinone II A) identified from the formula which showed cardioprotective effects in a dose-dependent manner. These applications suggested that the proposed rapid and sensitive screening approach offers an efficient way to discover active components or compounds from medicinal plants.
authors.affiliation: Pharmaceutical Informatics Institute, Zhejiang University, Hangzhou 310058, PR China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiumei
authors.initials: X
authors.lastname: Gao
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Nie
authors.forename: Yingxin
authors.initials: Y
authors.lastname: Yang
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Fan
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002316:Cardiotonic Agents / Q000737:chemistry* / Q000302:isolation & purification*; D002460:Cell Line / Q000166:cytology; D002853:Chromatography, Liquid / Q000379:methods; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000627:therapeutic use; D005453:Fluorescence; D057166:High-Throughput Screening Assays / Q000379:methods*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D013058:Mass Spectrometry / Q000379:methods; D008516:Medicine, Chinese Traditional; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008856:Microscopy, Fluorescence / Q000379:methods; D010946:Plants, Medicinal / Q000737:chemistry*; D051381:Rats; D020112:Rhodamine 123 / Q000032:analysis
chemical_list: D002316:Cardiotonic Agents; D004365:Drugs, Chinese Herbal; C528580:qishen yiqi; D020112:Rhodamine 123; D006861:Hydrogen Peroxide; D001205:Ascorbic Acid