Network Pharmacology and Experimental Validation Reveal the Potential Antidepressant-Like Effects of Chaihu Shugan San Associated With Glutamate Homeostasis.

Chaihu Shugan San (CHSGS) is a traditional Chinese formula widely used in the treatment of depression, but its antidepressant-like effects remain incompletely understood. This study investigated the effects of CHSGS on stress-induced behavioral abnormalities with a focus on hippocampal glutamate homeostasis. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to identify the chemical constituents of CHSGS, and network pharmacology combined with molecular docking was applied to predict potential targets and signaling pathways. A chronic restraint stress (CRS)-induced mouse model was established to evaluate the antidepressant-like effects of CHSGS through behavioral tests and molecular analyses. UPLC-MS/MS identified flavonoids as the major constituents of CHSGS. Network pharmacology analysis suggested that the AGE-RAGE signaling pathway may be involved in the effects of CHSGS, with several flavonoids, including kaempferol, hesperetin, isorhamnetin, naringenin, and nobiletin, showing potential interactions with key targets. In CRS mice, CHSGS treatment improved behavioral abnormalities, reduced hippocampal glutamate levels, alleviated neuronal Nissl body damage, restored hippocampal GFAP and GLT-1 expression, and inhibited RAGE/NF-κB signaling activation. These findings suggest that CHSGS effectively alleviated CRS-induced behavioral abnormalities and improved hippocampal glutamate homeostasis in mice, which may be associated with the regulation of GLT-1 expression and involvement of the RAGE/NF-κB signaling pathway.