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

Abstract

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.