Background:
Chaihu-Shugan-San is a classical prescription to treat depression. According to the
traditional Chinese medicine (TCM) principle, the 2 decomposed recipes in
Chaihu-Shugan-San exert synergistic effects, including Shu Gan (stagnated Gan-Qi dispersion) and Rou Gan (Gan nourishment to alleviate
pain). However, the specific mechanism of
Chaihu-Shugan-San on depression and its compatibility rule remain to be explored. Objective: We aimed to explore the anti-depression mechanisms and analyze the advantage of TCM compatibility of
Chaihu-Shugan-San. Methods: The chronic unpredictable mild stress (CUMS) rat model was established.
Antidepressant effects were evaluated by
sucrose preference test (SPT), and forced swimming test (FST). Tandem Mass Tag (TMT)-based quantitative proteomics of the hippocampus was used to obtain differentially expressed
proteins (DEPs). Bioinformatics analysis including Gene Ontology (GO), pathway enrichment, and
protein-
protein interaction (PPI) networks was utilized to study the DEPs connections. At last, the achieved key targets were verified by western blotting. Results:
Chaihu-Shugan-San increased
weight gain and food intake, as well as exhibited better
therapeutic effects including enhanced
sucrose preference and extended immobility time when compared with its decomposed recipes. Proteomics showed
Chaihu-Shugan-San, Shu Gan, and Rou Gan regulated 110, 12, and 407 DEPs, respectively. Compared with Shu Gan or Rou Gan alone, the expression of 22
proteins was additionally changed by
Chaihu-Shugan-San treatment, whereas the expression of 323
proteins whose expression was changed by Shu Gan or Rou Gan alone were not changed by
Chaihu-Shugan-San treatment. Bioinformatics analysis demonstrated that
Chaihu-Shugan-San affected
neurotransmitter's release and transmission cycle (e.g., γ-
aminobutyric acid (
GABA),
glutamate,
serotonin,
norepinephrine,
dopamine, and
acetylcholine).
GABA release pathway is also targeted by the 22 DEPs. Unexpectedly, only 2 pathways were enriched by the 323 DEPs: Metabolism and Cellular responses to external stimuli. Lastly, the expression of Gad2,
Vamp2, and Pde2a was verified by western blotting. Conclusions:
Chaihu-Shugan-San treats depression via multiple targets and pathways, which may include regulations of 110 DEPs and some
neurotransmitter's transmission cycle. Compared with Shu Gan and Rou Gan, the 22
Chaihu-Shugan-San advanced
proteins and the affected
GABA pathway may be the advantages of
Chaihu-Shugan-San compatibility. This research offers data and theory support for the clinical application of
Chaihu-Shugan-San.