Naodesheng decoction regulating vascular function via G-protein-coupled receptors: network analysis and experimental investigations

Abstract

Introduction: Ischemic stroke (IS) is a detrimental neurological disease with limited treatment options. Recanalization of blocked blood vessels and restoring blood supply to ischemic brain tissue are crucial for post-stroke rehabilitation. The decoction Naodesheng (NDS) composed of five Chinese botanical drugs, including Panax notoginseng (Burk.) F. H. Chen, Ligusticum chuanxiong Hort., Carthamus tinctorius L., Pueraria lobata (Willd.) Ohwi, and Crataegus pinnatifida Bge., is a blood-activating and stasis-removing herbal medicine commonly used for the clinical treatment of cerebrovascular diseases in China. However, the material basis of NDS on the effects of blood circulation improvement and vascular tone regulation remains unclear. Methods: A database comprising 777 chemical metabolites of NDS was constructed. Then, the interactions between various herbal metabolites of NDS and five vascular tone modulation G-protein-coupled receptors (GPCRs), including 5-HT1AR, 5-HT1BR, β2-AR, AT1R, and ETBR, were assessed by molecular docking. Using network analysis and vasomotor experiment of the cerebral basilar artery, the potential material basis underlying the vascular regulatory effects of NDS was further explored. Results: The Naodesheng Effective Component Group (NECG) was found to induce relaxation of rat basilar artery rings precontracted using Endothelin-1 (ET-1) and KCl in vitro in a dose-dependent manner. Several metabolites of NDS, including C. tinctorius, C. pinnatifida, and P. notoginseng, were found to be the main plant resources of metabolites with high docking scores. Furthermore, several metabolites in NDS, including formononetin-7-glucoside, hydroxybenzoyl-coumaric anhydride, methoxymecambridine, puerarol, and pyrethrin II, were found to target multiple vascular GPCRs. Metabolites with moderate-to-high binding energy were verified to have good rat basilar artery-relaxing effects, and the maximum artery relaxation effects of all three metabolites, namely, isorhamnetin, kaempferol, and daidzein, were found to exceed 90%. Moreover, metabolites of NDS were found to exert a synergistic effect by interacting with vascular GPCR targets, and these metabolites may contribute to the cerebrovascular regulatory function of NDS. Discussion: The study reports that various metabolites of NDS contribute to its vascular tone regulating effects and demonstrates the multi-component and multi-target characteristics of NDS. Among them, metabolites with moderate-to-high binding scores in NDS may play an important role in regulating vascular function.

Keywords: G-protein-coupled receptor; Naodesheng decoction; material basis; molecular docking; network analysis; vascular function.

FIGURE 1

Flow chart.

FIGURE 2

Vasorelaxant effects of the NECG on the rat endothelium-intact basilar artery preconstricted with ET-1 (10⁻⁸ M) or KCl (60 mM). (A) NECG dose-dependently relaxed ET-1 (10⁻⁸ M)-precontracted basilary artery rings. (B) NECG dose-dependently relaxed KCl (60 mM)-precontracted basilary artery rings. The results are presented as the mean ± SEM (n = 6).

FIGURE 3

Proportion of the herbal metabolites in NDS.

FIGURE 4

CDOCKER scores obtained by molecular docking of the chemical metabolites in NDS with the five vascular GPCR targets. The five vascular GPCR targets are (A) 5-HT1AR, (B) 5-HT1BR, (C) β2-AR, (D) AT1R, and (E) ETBR.

FIGURE 5

(A) Chemical composition (score in the top 50) target interaction network of NDS. (B) Enrichment of the 50 top-scoring metabolites with high docking scores interacting with each of the vascular GPCR targets, which were sorted and calculated according to their herb origin in NDS. (C) Number of top 50 scoring metabolites interacting with multiple vascular GPCR targets.

FIGURE 6

Vasorelaxant effects of isorhamnetin (A), kaempferol (B), and daidzein (C) on the endothelium-intact rat basilar artery precontracted with KCl (60 mM). The results are presented as the mean ± SEM (n = 3).