Mechanisms of Baishao (Radix Paeoniae Alba) and Gancao (Radix Glycyrrhizae) on major depressive disorder: network pharmacology and in vivo validation

doi:10.19852/j.cnki.jtcm.2025.05.013

Abstract

Abstract:

OBJECTIVE: To elucidate the potential molecular mechanisms of Baishao (Radix Paeoniae Alba) (APR) and Gancao (Radix Glycyrrhizae) (GR) in the treatment of major depressive disorder (MDD).

METHODS: Based on the network pharmacology strategy, the therapeutic targets of APR-GR for MDD are predicted, differentially expressed genes from the Integrated Gene Expression database for MDD patients. Topological networks are constructed, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways are enriched, their pharmacological potential molecular mechanisms are discussed, and molecular docking analysis is performed to further motivate compositional and target interactions. Finally, the CUMS mouse model is used for validation.

RESULTS: Based on the pharmacological network analysis, 17 candidate genes were identified, including muscarinic acetylcholine receptor M1(CHRM1), muscarinic acetylcholine receptor M2 (CHRM2), β2-adrenergic receptor (ADRB2), adrenergic α1A receptor (ADRA1A) and 5-hydroxytryptamine transfer protein (SLC6A4), etc. which are primarily involved in reactive oxygen species metabolism, neural response, oxidative stress response and other biological processes. Further analysis revealed that these targets are closely related to Ca²⁺, cyclic adenosine monophosphate, etc., and exhibit optimal binding sites after molecular docking. Finally, in vivo experiments were performed and it was found that APR-GR significantly improved depression-like behavior and hippocampal impairment in mouse models, increasing brain levels of 5-hydroxytryptamine, dopamine and norepinephrine and decreasing serum levels of corticotropin releasing hormone, corticosterone and adreno cortico tropic hormone, while upregulating the expression of CHRM1, CHRM2 and ADRA1A in the hippocampus and downregulating the expression of SLC6A4 and ADRB2.

CNCLUSION: This research sheds light on the potential molecular mechanism of APR-GR to improve MDD.

Key words: depressive disorder, major, animal experimentation, molecular docking simulation, network pharmacology, Baishao (Radix Paeoniae Alba), Gancao (Radix Glycyrrhizae)

PEI Ke, LI Yong, LIN Zhe, LYU Guangfu. Mechanisms of Baishao (Radix Paeoniae Alba) and Gancao (Radix Glycyrrhizae) on major depressive disorder: network pharmacology and in vivo validation[J]. Journal of Traditional Chinese Medicine, 2025, 45(5): 1067-1077.

Figures/Tables 5

Figure 1 Network pharmacological results A: volcanic map of the GEO differential gene in patients with MDD; B: venny diagram of APR-GR against MDD; C: network topology results; D: drug-ingredient-target-disease network diagram; F: GO enrichment analysis scatter diagram; G: relationship between pathways and proteins. MDD: Major depressive disorder; GEO: Gene Expression Omnibus; GO: Gene Ontology; APR-GR: Baishao (Radix Paeoniae Alba) and Gancao (Radix Glycyrrhizae).

Figure 2 Molecular docking pattern results A: docking model of mairin with ADRA1A; B: docking model of mairin with ADRA1B; C: docking model of mairin with ADRB2; D: docking model of mairin with CHRM1; E: docking model of mairin with CHRM2. F: docking model of mairin with SLC6A4. CHRM1: muscarinic acetylcholine receptor M1; CHRM2: muscarinic acetylcholine receptor M2; ADRA1A: adrenergic a1A receptor; ADRB2: β-2 adrenergic receptor; SLC6A4: serotonin transporter; ADRA1B: adrenergic a1B receptor.

Table 1 Molecular docking results of the main active ingredient with the core target

Target	Sitosterol	Mairin (KJ/mol)
ADRA1A	－12.53	－11.93
SLC6A4	－11.24	－10.94
ADRB2	－10.87	－10.07
ADRA1B	－9.37	－9.84
CHRM1	－8.56	－8.88
CHRM2	－7.17	－8.97

Figure 3 Behavioral test results and hippocampal morphology results A: effect of APR-GR on sugar water preference; B: OFT movement trajectory diagram; B1: control group; B2: model group; B3: APR-GR group; C: OFT results; C1: effect of APR-GR on center residence time; C2: effect of APR-GR on corner residence time; C3: effect of APR-GR ontotal travel distance; D: MWM, TST and FST experimental results. D1: effect of APR-GR in TST; D2: effect of APR-GR in FST; D3: effect of APR-GR incubation period in MWM; D4: effect of APR-GR movement in MWM; E: MWM movement trajectory diagram; E1: control group; E2: model group; E3: APR-GR group; F: mouse hippocampus pathological sections (× 200); F1: control group; B2: model group; B3: APR-GR group; control group: drink and eat freely + the same volume of normal saline was given by gavage, model group: chronic unpredictable mild stress, APR-GR: model group + APR-GR (1.5 g·kg-1·d-1). TST: tail suspension test; FST: forced swimming test; MWM: morris water maze; OFT: open field test; APR-GR: Baishao (Radix Paeoniae Alba) and Gan Cao (Radix Glycyrrhizae). Data are presented as mean ± standard deviation (n = 10) and analyzed using the t-test statistical method; aP < 0.01, compared with the control group; bP < 0.01, compared with the model group.

Figure 4 biochemical indicator results and the expression of related proteins in brain A: effects on biochemical indicators in brain. A1: effects of APR-GR on the levels of 5-HT in brain; A2: effects of APR-GR on the levels of DA in brain; A3: effects of APR-GR on the levels of NE in brain; B: effects on biochemical indicators in serum. B1: effect of APR-GR in serum CRH levels; B2: effect of APR-GR in serum ACTH levels; B3: effect of APR-GR in serum CORT levels; C: Western blot result. C1: correlated protein band diagram; C2: effect of APR-GR on ADRA1A expression; C3: effect of APR-GR on SLC6A expression; C4: effect of APR-GR on CHRM1 expression; C5: effect of APR-GR on ADRB2 expression; C5: effect of APR-GR on CHRM2 expression; control group: drink and eat freely + the same volume of normal saline was given by gavage, model group: chronic unpredictable mild stress, APR-GR: model group + APR-GR (1.5 g·kg-1·d-1); ACTH: adreno cortico tropic hormone, CORT: corticosterone; CRH: corticotropin releasing hormone; 5-HT: 5-hydroxytryptamine; DA: dopamine; NE: norepinephrine; CHRM1: muscarinic acetylcholine receptor M1; CHRM2: muscarinic acetylcholine receptor M2; ADRA1A: adrenergic a1A receptor; ADRB2: β-2 adrenergic receptor; SLC6A4: serotonin transporter; APR-GR: Baishao (Radix Paeoniae Alba) and Gan Cao (Radix Glycyrrhizae). Data are presented as mean ± standard deviation (n = 10) and analyzed using the t-test statistical method. aP ≤ 0.01, compared with the control group; bP ≤ 0.01, compared with the model group.

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