Polysaccharides from Scrophularia ningpoensis Hemsl. Improve reserpine-induced depression-like behavior by inhibiting HTR2A/HTR2C mediated AKT/GSK3β/β-catenin/CBP/BDNF signaling.

Scrophularia ningpoensis Hemsl. is a traditional Chinese medicine used to regulate blood sugar levels, immunity, etc. We previously isolated polysaccharides from S. ningpoensis Hemsl. (SNPS) and innovatively observed that SNPS exhibit antidepressant properties; however, the underlying mechanism is still unclear. Here, we employed network pharmacology to predict the potential targets and antidepressant mechanism of SNPS. Accordingly, we detected the effects of SNPS on monoamine neurotransmitter synthesis, metabolism, receptor expression and signal transduction in reserpine (RES)-treated mice using ELISA, HPLC-electrochemistry, metabonomics, Golgi-Cox staining and Western blotting. Finally, the mechanism of SNPS on key targets (HTR2A and HTR2C) was verified in vivo and in vitro. Results showed that SNPS ameliorated depression by restoring monoamine neurotransmitter homeostasis and hippocampal neurogenesis. SNPS reversed the depletion of 5-HT, NE and DA by activating the tryptophan (Trp)/5-HT and tyrosine (Tyr)/DA/NE metabolic pathways. SNPS decreased HTR2A and HTR2C contents, leading to the phosphorylation of AKT and GSK3β, followed by increases in β-catenin, CBP and BDNF levels. Mechanistically, SNPS reduced the levels of HTR2A and HTR2C proteins by inhibiting their mRNA transcription, rather than inducing protein degradation. In conclusion, by inhibiting the transcription of HTR2A and HTR2C, SNPS activated the AKT/GSK3β/β-catenin/CBP/BDNF pathway, thereby exerting dose-dependent antidepressant effects.