title: Tracking antiangiogenic components from Glycyrrhiza uralensis Fisch. based on zebrafish assays using high-speed countercurrent chromatography.
abstract: Natural products are some of the most important sources of lead compounds for drug discovery. The advanced isolation technique of lead compounds of natural origin using therapeutically relevant bioassays is capable of enhancing work efficiency from complex multiconstituent extracts. In the present study, a bioassay-guided isolation strategy combined with bioactivity screening was used to identify novel angiogenesis inhibitors from licorice (Glycyrrhiza uralensis Fisch.) based on the zebrafish model and rapid preparative separation by high-speed countercurrent chromatography. Zebrafish embryos at 24 h postfertilization were chosen as the angiogenesis inhibition model for bioactivity screening. A solvent system (n-hexane-ethyl acetate-methanol-water) with different ratios was optimized and applied in the high-speed countercurrent chromatography separation of two fractions, Fr5 and Fr6, from the ethyl acetate extract of licorice. Blood circulation and vascular outgrowth in intersegmental vessels were found to be simultaneously inhibited by isoliquiritigenin and isolicoflavonol in a dose-dependent manner. Thus, these two compounds were identified and considered as active inhibitors against angiogenesis. These experimental results indicate that zebrafish bioassays combined with high-speed countercurrent chromatography may provide an alternative pathway for the rapid isolation of bioactive natural products.
authors.affiliation: School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
authors.forename: Liwen
authors.initials: L
authors.lastname: Han
authors.forename: Yanqiang
authors.initials: Y
authors.lastname: Yuan
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.forename: Qiuxia
authors.initials: Q
authors.lastname: He
authors.forename: Yubo
authors.initials: Y
authors.lastname: Li
authors.forename: Xiqiang
authors.initials: X
authors.lastname: Chen
authors.forename: Xiuhe
authors.initials: X
authors.lastname: Liu
authors.forename: Kechun
authors.initials: K
authors.lastname: Liu
mesh_terms: D020533:Angiogenesis Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology*; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D003377:Countercurrent Distribution / Q000379:methods*; D029882:Glycyrrhiza uralensis / Q000737:chemistry*; D010936:Plant Extracts / Q000302:isolation & purification* / Q000494:pharmacology*; D015027:Zebrafish / Q000254:growth & development*
chemical_list: D020533:Angiogenesis Inhibitors; D010936:Plant Extracts