This study aims to study the effective substance and mechanism of Ziziphi Spinosae Semen extract in the treatment of
insomnia based on serum metabolomics and network pharmacology. The rat
insomnia model induced by
p-chlorophenylalanine(PCPA) was established. After
oral administration of Ziziphi Spinosae Semen extract, the general morphological observation,
pentobarbital sodium-induced sleep test, and histopathological evaluation were carried out. The potential
biomarkers of the extract in the treatment of
insomnia were screened by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS) combined with multivariate analysis, and the related metabolic pathways were further analyzed. The "component-target-pathway" network was constructed by ultra-high performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry(UHPLC-Q-Exactive-MS/MS) combined with network pharmacology to explore the effective substances and mechanism of Ziziphi Spinosae Semen in the treatment of
insomnia. The results of
pentobarbital sodium-induced sleep test and histopathological evaluation(
hematoxylin and
eosin staining) showed that Ziziphi Spinosae Semen extract had good theraputic effect on
insomnia. A total of 21 endogenous
biomarkers of Ziziphi Spinosae Semen extract in the treatment of
insomnia were screened out by serum metabolomics, and the metabolic pathways of
phenylalanine,
tyrosine and
tryptophan biosynthesis,
phenylalanine metabolism, and
nicotinate and
nicotinamide metabolism were obtained. A total of 34 chemical constituents were identified by UHPLC-Q-Exactive-MS/MS, including 24
flavonoids, 2
triterpenoid saponins, 4
alkaloids, 2
triterpenoid acids, and 2
fatty acids. The network pharmacological analysis showed that Ziziphi Spinosae Semen mainly acted on target
proteins such as
dopamine D2 receptor(DRD2),
5-hydroxytryptamine receptor 1 A(HTR1 A), and alpha-2 A
adrenergic receptor(ADRA2 A) in the treatment of
insomnia. It was closely related to neuroactive
ligand-receptor interaction, serotonergic synapse, and calcium signaling pathway.
Magnoflorine, N-
nornuciferine, caaverine,
oleic acid,
palmitic acid,
coclaurine,
betulinic acid, and
ceanothic acid in Ziziphi Spinosae Semen may be potential effective compounds in the treatment of
insomnia. This study revealed that Ziziphi Spinosae Semen extract treated
insomnia through multiple metabolic pathways and the overall correction of metabolic disorder profile in a multi-component, multi-target, and multi-channel manner. Briefly, this study lays a foundation for further research on the mechanism of Ziziphi Spinosae Semen in treating
insomnia and provides support for the development of innovative Chinese drugs for the treatment of
insomnia.