Jieyu I formula and its primary component quercetin alleviate anxiety by modulating neuroregeneration and inflammation via the calcium signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Jieyu I formula(JY-Ⅰ) was modified from SiNi San, and its anti-inflammatory and neuroprotective effects were investigated in preliminary studies. However, its potential mechanism of action in interfering with anxiety is unclear.

OBJECTIVE

The aim of this study was to investigate the therapeutic effect of JY-I on anxiety and to elucidate its mechanism of action.

METHODS

Network pharmacology was utilized to identify the key molecular targets and mechanisms underlying the anxiolytic effects of JY-I. A mouse chronic restraint stress (CRS) model was used to evaluate the anxiety-improving effects of JY-I. Mood improvement was assessed by behavioral tests (OFT and EPM, etc.) and histological analyses (including HE staining and Nissl staining). In the in vitro experiments, the pharmacodynamic effects of JY-I were assessed by the CORT-induced stress model of HT-22 cells, and on the other hand, cells were intervened with the calcium signaling antagonist, KN-62, to observe the possible mechanisms of JY-I intervention. The calcium ion signaling pathway function was examined by Western blot and qPCR.

RESULTS

A total of 74 active compounds, 857 potential therapeutic targets and 1714 anxiety-related target genes were obtained. The number of common targets between JY-I and anxiety was 285. KEGG pathway analysis showed that the calcium signaling pathway was closely related to anxiety, which may be a potential mechanism of the anxiolytic activity of JY-I. Network pharmacological analysis showed that the active ingredients included quercetin, kaempferol and lignocerol, and molecular docking analysis showed that the main components of JY-I had good binding ability with CaMK2 and CAM. Animal experiments showed that JY-I improved body weight, reduced anxiety-like behavior and neuroinflammation, and promoted nerve regeneration in CRS mice. In vitro experiments further showed that both JY-I-containing serum and its active ingredient quercetin prevented CORT-induced HT-22 cell damage and increased cell viability, and JY-I activated the calcium signaling pathway (CaMK2/CAM/CREB) in vivo and in vitro, and the therapeutic effects of quercetin were similar to those of the drug-containing serum.The CaMK2 inhibitor KN-62 reversed the therapeutic effects of both drug-containing serum and the therapeutic effect of quercetin.

CONCLUSIONS

JY-I ameliorates CRS-induced anxiety in mice by a mechanism that may be related to the modulation of the calcium signaling pathway.