Amelioration of PTSD-like behaviors in mice by the aqueous extract of Ziziphi Spinosae Semen via inhibition of GPR50 and regulation of synaptic plasticity

doi:10.12092/j.issn.1009-2501.2026.08.005

Abstract

Abstract:

AIM: To evaluate the effects of the aqueous extract of Ziziphi Spinosae Semen (ZSS) on single prolonged stress (SPS)-induced post-traumatic stress disorder (PTSD)-like behaviors in mice, and to explore its potential mechanism from the perspective of hippocampal G protein-coupled receptor 50 (GPR50) and synaptic plasticity. METHODS: The UPLC-UV method was employed to determine the content of the main components, saponin A and saponin B, in the aqueous extract of ZSS. C57BL/6 mice were randomly divided into Control group, SPS group, SPS+ZSS-L, M, H groups and SPS+PRX group. Behavioral experiments were used to evaluate PTSD-like behaviors in SPS mice. Molecular docking was employed to evaluate the stability of the binding between the active ingredients in aqueous extract of ZSS and GPR50. Western blot combined with immunofluorescence were applied to detect GPR50, PSD95, GluN2A, and GluN2B protein expression in mouse hippocampal tissue. Bioinformatics analysis was used to predict key targets for ZSS treatment of PTSD. RESULTS: This study conducted qualitative and quantitative analyses of the main active ingredients saponin A and B in the aqueous extract of ZSS. Compared with the Control group, mice in the SPS group exhibited PTSD-like behaviors such as anxiety, depression, and difficulty in fear memory extinction (P<0.05), with elevated expression of GPR50 protein in the hippocampus and abnormal expression of synaptic plasticity-related proteins PSD95, GluN2A, and GluN2B. The aqueous extract of ZSS significantly improved PTSD behaviors in SPS mice (P<0.05) and inhibited GPR50 expression (P<0.05). A total of 209 potential targets of the aqueous extract of ZSS for treating PTSD were screened. These targets were enriched in pathways associated with functions such as inhibiting impaired synaptic plasticity and exerting antioxidant properties. Molecular docking demonstrated that ZSS saponin A and saponin B, bind stably to GPR50. CONCLUSION: The aqueous extract of ZSS can ameliorate SPS-induced PTSD-like behaviors in mice, and its mechanism may be associated with the inhibition of GPR50 to alleviate synaptic plasticity impairment.

Key words: post-traumatic stress disorder, G protein-coupled receptor 50, Ziziphi Spinosae Semen, synaptic plasticity

CLC Number:

R749
R965.2

Zixuan GAO, Cunbao HE, Shaojie YANG, Bin WANG, Guoqi ZHU, Hui CHENG, Lixia CHEN. Amelioration of PTSD-like behaviors in mice by the aqueous extract of Ziziphi Spinosae Semen via inhibition of GPR50 and regulation of synaptic plasticity[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(8): 1053-1063.

Figures/Tables 8

Fig.1 HPLC chromatograms of Ziziphi Spinosae Semen (ZSS) solution and standard solution A: jujuboside A; B: the aqueous extract of ZSS; C: jujuboside B.

Fig.2 The aqueous extract of Ziziphi Spinosae Semen (ZSS) improved anxiety-like behaviors in SPS mice ($ \overline{\boldsymbol{x}} $±s, n=8) A: schematic representation of the time point of administration of the ZSS; B-F: time in central area, distance in central area, center entries, total distance and mean speed of OFT; G: percentage of time in open arms in EPMT; H: movement traces of OFT. bP<0.05, compared with the control group; eP<0.05, compared with SPS group.

Fig.3 The aqueous extract of Ziziphi Spinosae Semen (ZSS) promoted the extinction of fear memory in SPS mice ($ \overline{\boldsymbol{x}} $±s, n=8) A: freezing time in the re-exposure phase; B: freezing time in the extinction phase. bP<0.05, compared with the control group; eP<0.05, compared with SPS group.

Fig.4 Potential targets and enrichment analysis of Ziziphi Spinosae Semen (ZSS) in treatment of PTSD A: active component targets and disease targets Venn diagram; B: PPI network.

Fig.5 GO and KEGG enrichment analysis of potential targets of Ziziphi Spinosae Semen (ZSS) treatment for PTSD A-C: GO analysis; D: KEGG analysis; E: drug-component-target network.

Fig.6 Visualization of molecules docking between main components of Ziziphi Spinosae Semen (ZSS) and key targets. A: jujuboside A-GPR50; B: jujuboside B-GPR50.

Fig.7 The aqueous extract of Ziziphi Spinosae Semen (ZSS) inhibited the expression of GPR50 protein in the hippocampus ($ \overline{\boldsymbol{x}} $±s, n=6) A: representative blots; B: quantitative data of GPR50 (n=3); C-H: schematic immunofluorescence of GPR50 in the CA1, CA3 and DG region of mouse hippocampus and mean fluorescence intensity of GPR50 (Nucleus: blue; GPR50: red) (n=3). bP<0.05, compared with the control group; eP<0.05, compared with SPS group.

Fig.8 Effects of the aqueous extract of Ziziphi Spinosae Semen (ZSS) on the expression levels of PSD95, GluN2A and GluN2B proteins in mouse hippocampus ($ \overline{\boldsymbol{x}} $±s, n=3) A: representative blot and quantitative analysis of PSD95; B: representative blot and quantitative analysis of GluN2A; C: representative blot and quantitative analysis of GluN2B; bP<0.05, compared with the control group; eP<0.05, compared with SPS group.

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