title: Highly selective screening of the bioactive compounds in Huoxue capsule using immobilized β(2)-adrenoceptor affinity chromatography.
abstract: A highly selective assay was developed for screening compounds that bind to the porcine recombinant β2-adrenoceptor (β2-AR) with affinity chromatography coupled to quadrupole time-of-flight mass spectrometry (Q-TOF-MS). The methodology involved selective screening with immobilized β2-AR, a highly accurate identification via Q-TOF-MS, and a functional evaluation of the screened compounds with a sensitive myograph system. Ferulic acid, hydroxysafflor yellow A (HSYA), and naringin were confirmed to be the bioactive compounds in Huoxue capsule that specifically bound to the β2-AR. These compounds produced a concentration-dependent relaxation of arteries that were contracted by treatment with phenylephrine, and the relaxation caused by these compounds was attenuated in the presence of ICI 118551, a type of β2-AR antagonist. Our data indicate that the use of an immobilized receptor is potentially an alternative method for the rapid screening of bioactive compounds in a complex matrix because of its high specificity. β2-AR affinity chromatography was valuable in focusing attention on the further investigation of ferulic acid, HSYA, and naringin as β2-AR agonists.
authors.affiliation: Department of Pharmacology, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China; Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education/College of Life Science, Northwest University, Xi'an 710069, China.
authors.forename: Shixiang
authors.initials: S
authors.lastname: Wang
authors.affiliation: No. 1 Hospital of Xi'an City, Xi'an 710002, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China. Electronic address: zwj@xjtu.edu.cn.
authors.forename: Weijin
authors.initials: W
authors.lastname: Zang
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education/College of Life Science, Northwest University, Xi'an 710069, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education/College of Life Science, Northwest University, Xi'an 710069, China.
authors.forename: Xinfeng
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Xi
authors.initials: X
authors.lastname: He
authors.affiliation: Shaanxi Provincial People's Hospital, Xi'an 710068, China.
authors.forename: Qinshe
authors.initials: Q
authors.lastname: Liu
authors.affiliation: First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Weiyi
authors.initials: W
authors.lastname: Feng
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education/College of Life Science, Northwest University, Xi'an 710069, China. Electronic address: zhengxh@nwu.edu.cn.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zheng
mesh_terms: D002214:Capsules; D002846:Chromatography, Affinity / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D011412:Propanolamines / Q000737:chemistry; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry*; D012680:Sensitivity and Specificity
chemical_list: D002214:Capsules; D004365:Drugs, Chinese Herbal; D055767:Immobilized Proteins; D011412:Propanolamines; D018343:Receptors, Adrenergic, beta-2; C026777:ICI 118551
keywords: Affinity chromatography; Bioactive compound; High-performance liquid chromatography/mass spectrometry; Huoxue capsule; β(2)-Adrenoceptor