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.

Initial Cell Adhesion onto a Phospholipid Polymer Brush Surface Modified with a Terminal Cell Adhesion Peptide

“…Hence, the poly­(MPC) brush surface functionalized with cell adhesion molecules at the grafted water-soluble poly­(MPC) chain termini allowed us to study the interaction between the immobilized cell adhesion molecules and the corresponding cell membrane proteins. Analysis of the cell adhesion behavior using several kinds of polymer brush surfaces having cell adhesion molecules has been conducted. − However, cell adhesion molecules were immobilized at the functional groups at the side chains in the monomer unit in almost all reports. Such an immobilization method considerably impairs the structural clarity of the polymer brush surfaces.…”

Two‐dimensional chromatography in screening of bioactive components from natural products

“…Li et al used human umbilical vein endothelial cells (HUVECs) as a probe to adhere cells to amino microspheres. 85 Wang et al prepared an affinity chromatography column containing immobilised β 2 -adrenoceptors using a diazonium salt reaction. 86 Su et al prepared a DNA affinity chromatography column by forming a phosphoramidate bond between a phosphate group at one end of a calf thymus DNA (ct-DNA) molecule and an amino group on aminopropyl silica stationary phase.…”

“…Li et al . used human umbilical vein endothelial cells (HUVECs) as a probe to adhere cells to amino microspheres 85 . Wang et al .…”

Preparation and Application of Modified VEGFR-2 Cell Membrane Chromatographic Separation System

“…[14] and Ligusticum chuanxiong [21]. Screening of active components from traditional Chinese herb Aconitum carmichaeli Debx.…”

“…The VEGFR-CMC system for screening bioactive compounds from Ligusticum chuanxiong was prepared by attaching cells onto amino microspheres using HUVECs as a probe [21]. The high stability of this cell membrane-based stationary phase was due to the high affinity between integrin on the cells and RGD peptide that was coated onto the poly[oligo (ethylene glycol) methacrylate] material [21].…”