Mechanisms of Suanzaoren (Ziziphi Spinosae Semen) and its processed products in treating insomnia: an integrated study based on network pharmacology and metabolomics

doi:10.19852/j.cnki.jtcm.2025.06.010

Abstract

Abstract:

OBJECTIVE: To elucidate the potential mechanisms and identify bioactive components of Suanzaoren (Ziziphi Spinosae Semen) (ZSS) and it’s processed products for insomnia treatment.

METHODS: The principal components of ZSS and its processed products were analyzed using Ultra-Performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The effects of ZSS on insomnia were assessed in para-chlorophenylalanine (PCPA)-induced rats. Additionally, to investigate the mechanisms of action in insomnia, serum and hippocampal non-targeted metabolomics were performed alongside network pharmacology and molecular docking.

RESULTS: UPLC-Q-TOF-MS was used to analyze the main components of ZSS and its processed products. In the insomnia rat model induced by PCPA, both ZSS and its processed products demonstrated therapeutic effects, with the processed products showing particularly significant effects, significantly increasing the levels of the Gamma-Aminobutyric Acid and melatonin. Non-targeted metabolomics identified 23 and 10 potential differential metabolites in serum and hippocampus, respectively, involving pathways such as lipid metabolism and amino acid metabolism. Network pharmacology identified three core targets, mainly associated with pathways such as Transient Receptor Potential channel regulation, Advanced Glycation End-products-Receptor for Advanced Glycation End-products signaling pathway, and neuroactive ligand-receptor interaction. Molecular docking confirmed that the main active components of ZSS can stably bind to these core targets, thereby exerting an effect in improving insomnia.

CONCLUSIONS: Combining in vivo experiments, metabolomics, and network pharmacology, we have identified potential pharmacological mechanisms by which ZSS and its derivatives mitigate insomnia. These findings provide a theoretical foundation for the further development and utilization of ZSS, supporting the innovation of new therapeutic agents for insomnia treatment.

Key words: Suanzaoren (Ziziphi Spinosae Semen), insomnia, metabolomics, network pharmacology

ZHANG Yuan, CHENG Shizan, HUA Yue, SHI Ji, SU Guoming, ZHANG Chao, LIAN Jing, LIU Pengpeng, JIA Tianzhu. Mechanisms of Suanzaoren (Ziziphi Spinosae Semen) and its processed products in treating insomnia: an integrated study based on network pharmacology and metabolomics[J]. Journal of Traditional Chinese Medicine, 2025, 45(6): 1307-1316.

Figures/Tables 8

Table 1 Exercise time (s, x - ?± s)

Group	n	Horizontal movement	Vertical movement	Movement time
NC	6	67.5±5.7^a	18.2±2.6^a	125.1±6.0^a
MC	6	96.8±3.8	31.8±2.3	175.43±8.5
PC	6	70.0±5.3^b	22.0±2.6^a	135.29±4.7^a
ZSS	6	79.5±4.6^a	27.3±5.1	158.00±7.2^a
FZSS	6	77.0±6.5^a	27.0±2.6	157.43±6.5^a
SZSS	6	69.7±4.3^a	21.0±3.2^a	138.14±9.7^a

Table 2 Body weight changes ( x - ?± s)

Group	n	0 d	3 d	6 d	9 d	12 d	15 d	17 d
NC	6	261.6±1.7	268.0±3.7^b	271.5±3.2^b	272.2±3.1^b	273.3±2.1^b	281.8±2.0^b	286.3±0.6^b
MC	6	262.5±2.1	239.8±2.3	226.9±3.1	235.7±3.9	245.0±5.1	251.0±6.2	260.8±7.9
PC	6	263.8±3.9	249.2±4.3^a	256.2±6.7^b	265.6±6.9^b	277.5±9.9^b	287.9±6.1^b	297.4±8.5^b
ZSS	6	256.2±4.1	246.3±6.1	242.2±3.9^b	250.8±5.1^b	263.9±10.5^b	275.3±12.3^b	282.9±11.3^b
FZSS	6	259.8±6.0	246.8±4.3	252.3±6.9^b	260.6±4.2^b	271.6±6.9^b	280.2±9.2^b	293.2±15.5^b
SZSS	6	259.1±4.9	241.6±7.0	249.8±2.8^b	256.9±3.4^b	268.2±4.8^b	276.1±6.3^b	285.8±4.5^b

Table 3 Neurogenic factor levels in the serum of the rats ( x - ?± s)

Group	n	5-HT (ng/mL)	GABA (μmol/L)	ACH (ng/mL)	DA (pg/mL)	MT(ng/L)
NC	6	17.4±0.6	9.0±1.0	70.0±3.6^b	1592.3±56.6^b	71.0±5.0^b
MC	6	16.7±0.5	8.6±0.7	86.0±3.3	3073.9±424.6	44.0±3.1
PC	6	17.8±0.8^a	9.4±1.0	68.7±1.7	2648.8±244.0^b	68.9±4.9^b
ZSS	6	18.6±1.1^b	9.3±1.2	75.3±3.8	2924.6±510.7^a	50.2±4.8
FZSS	6	18.4±0.6^a	11.4±1.1^b	79.1±5.1	2927.4±229.0	60.4±2.4^b
SZSS	6	18.7±0.8^b	11.8±1.8^b	75.9±13.2^b	1859.0±545.1^a	71.7±4.2^b

Figure 1 The degree of brain tissue damage in each group of rats HE staining images (× 100, scale bar = 50 μm) of hippocampus. A: NC group, B: MC group, C: PC group, D: ZSS group, E: FZSS group, F: SZSS group. NC group: treated only with physiological saline; MC group: treated only with PCPA 400 mg/kg; PC group: treated with 400 mg/kg PCPA + 0.52 mg/kg PC; ZSS group: treated only with 400 mg/kg PCPA + 3 g/kg ZSS; FZSS group: treated with 400 mg/kg PCPA + 3 g/kg FZSS; SZSS group: treated with 400 mg/kg PCPA + 3 g/kg SZSS. ZSS: Suanzaoren (Ziziphi Spinosae Semen); MC: model control; PC: positive control; FZSS: frying Suanzaoren (Ziziphi Spinosae Semen); SZSS: steaming Suanzaoren (Ziziphi Spinosae Semen); PCPA: para-chlorophenylalanine.

Figure 2 Immunohistochemistry analysis of rats A: expression of GABA in the hippocampal; B: expression of 5-HT in the hippocampal; A1, B1: NC group; A2, B2: MC group; A3, B3: PC group; A4, B4: ZSS group; A5, B5: FZSS group; A6, B6: SZSS group. NC group: treated only with physiological saline; MC group: treated only with PCPA 400 mg/kg; PC group: treated with 400 mg/kg PCPA + 0.52 mg/kg PC; ZSS group: treated only with 400 mg/kg PCPA + 3 g/kg ZSS; FZSS group: treated with 400 mg/kg PCPA + 3 g/kg FZSS; SZSS group: treated with 400 mg/kg PCPA + 3 g/kg SZSS. ZSS: Suanzaoren (Ziziphi Spinosae Semen); MC: model control; PC: positive control; FZSS: frying Suanzaoren (Ziziphi Spinosae Semen); SZSS: steaming Suanzaoren (Ziziphi Spinosae Semen); PCPA: para-chlorophenylalanine; GABA: gamma-aminobutyric acid; 5-HT: 5-hydroxytryptamine.

Table 4 mRNA expression levels in rats ( x ? -± s)

Item	NC (n = 6)	MC (n = 6)	PC (n = 6)	ZSS (n = 6)	FZSS (n = 6)	SZSS (n = 6)
CHRNA7	1.04±0.13^a	2.37±0.21	1.71±0.16^b	2.16±0.10^a	1.21±0.09^a	1.07±0.08^a
DRD2	1.05±0.24^a	3.52±0.70	1.48±0.51^a	1.73±0.19^a	1.66±0.18^a	1.05±0.04^a

Figure 3 The metabolic network diagram involved in the improvement of insomnia by Suanzaoren (Ziziphi Spinosae Semen) TCA: tricarboxylic acid cycle.

Figure 4 Network pharmacological analysis results A: network topology analysis of the main components of ZSS and shared targets; B: PPI analysis of shared targets. ZSS: Suanzaoren (Ziziphi Spinosae Semen); PPI: protein protein interaction.

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