title: Screening bioactive compounds from Ligusticum chuanxiong by high density immobilized human umbilical vein endothelial cells.
abstract: High throughput screening methodologies play a very important role in screening bioactive compounds from complex media. In this work, a new strategy for attaching cells onto amino microspheres using human umbilical vein endothelial cells (HUVECs) as a probe was developed. The immobilization depended on the specific affinity between integrin on the cells and the RGD peptide, which was coated on poly[oligo (ethylene glycol) methacrylate] by atom transfer radical polymerization. Validated application of the stationary phase was performed in the analysis of Ligusticum chuanxiong extraction by high performance affinity chromatography-mass spectrometry. Three compounds were screened as the bioactive compounds of Ligusticum chuanxiong. Two of them were identified as 3-butyl-hexahydroisobenzofuran-1(3H)-one and tetramethylpyrazine (TMP), whereas the other one remains indistinct. The association constant of vascular endothelial growth factor (VEGF) and TMP binding to VEGF receptor (VEGFR) on HUVECs were calculated to be (1.04 ± 0.08) × 10(11) M(-1) and (9.84 ± 1.11) × 10(8) M(-1) by zonal elution. Molecular docking showed that one hydrogen bond was formed between N atom of TMP and 3-N atom of imidazole group in histidine(223) of VEGFR. Both zonal elution and molecular docking indicated that TMP and VEGF bind to the same site of VEGFR on HUVECs. It is possible to become a promising tool for high throughput screening of the bioactive compounds binding to HUVECs through broad application of the stationary phase.
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, 710069, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Guangxin
authors.initials: G
authors.lastname: Liu
authors.forename: Huanmei
authors.initials: H
authors.lastname: Sun
authors.forename: Liujiao
authors.initials: L
authors.lastname: Bian
authors.forename: Xinfeng
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zheng
mesh_terms: D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D057166:High-Throughput Screening Assays; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D030004:Ligusticum / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D062105:Molecular Docking Simulation; D010936:Plant Extracts / Q000378:metabolism / Q000494:pharmacology*; D011719:Pyrazines / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor / Q000378:metabolism; D013499:Surface Properties; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D010936:Plant Extracts; D011719:Pyrazines; D042461:Vascular Endothelial Growth Factor A; D040262:Receptors, Vascular Endothelial Growth Factor; C017953:tetramethylpyrazine