Investigation On The Anti-depression Mechanism Of Fufang Chaigui Fang Supercritical CO₂ Extraction

Ethnopharmacology:Depression is a common emotional and mental disorder, which characters long-lasting depressed emotion. The morbidity of depression is as high as 15-20%, which seizes more and more concerns of the medical world and public health. Because of the complex pathogenesis of depression, although some new antidepressants are developed, the need for elucidating the mechanisms of depression and antidepressants is more and more urgent. Taking the side effects and the lagging effects into consideration, researchers are paying more attention to developing new antidepressants.Our research group has done much research on the anti-depression component of Xiaoyaosan, the famous formula in traditional Chinese medicine. Fufang Chaigui Fang was composed after two herbal drugs removed from the formula, after which the extraction method was confirmed to be supercritical fluid CO2 extraction [1]. The supercritical fluid CO2 extraction component (FFCGF) was obtained. This project will discuss the mechanism of anti-depression effect of FFCGF.Objective:Based on the previous results, this project investigated the anti-depression mechanisms from the serum 1H NMR metabonomics, cAMP-PKA-CREB-BDNF signaling pathway and inflammatory cytokines on the hippocumpus and serum of CUMS rats.Methods:The anti-depression effect and the mechanisms of Fufang Chaigui Fang were investigated through serum 1H NMR metabonomics analysis and cAMP signal pathway on CUMS rat hippocampus and serum. The concentration of cAMP signaling pathway factors and inflammatory cytokines were measured using ELISA and mRNA level of PKA, BDNF, TrkB and CREB were measured using Real Time-qRT-PCR. The serum H NMR metabonomics was conducted using principle component analysis (PCA).Results:1. The principle component analysis (PCA) scores plot of serum 1H NMR metabonomics analysis showed that the trajectories of CUMS group was separated from the control group, suggesting the CUMS model was duplicated successfully; after Fufang Chaigui Fang administration, the trajectories showed the tendency of coming back to the control group, demonstrating the anti-depression effect of Fufang Chaigui. The PCA loading plots illuminated that the concentrations of isoleucine, choline and TMAO were decreased and the levels of lactate, threonine and β-glucose were increased compared with those of control group (P< 0.05 or P< 0.01). After drug administration, those levels were corrected to those of control group (P< 0.05 or P< 0.01), vertified the anti-depression effect of Fufang Chaigui Fang.2. Compared with the control group, the cAMP, PKA, p-CREB and BDNF levels were decreased significantly (P< 0.01); The results of Real Time-qRT-PCR demonstrated that the levels of PKA, BDNF, TrkB and CREB mRNA in the hippocampus of CUMS group rats decreased compared with those of control group.3. Compared with the control group, the inflammatory cytokines (such as IL-1, IL-6, PGE2 and COX-2) showed significantly increase (P < 0.01), after Fufang Chaigui Fang administration, the levels decreased significantly (P< 0.05 or P< 0.01), suggesting the anti-depression effect of Fufang Chaigui Fang were conducted through the regulation of inflammatory cytokines.Conclusion:1. The anti-depression mechanism of Fufang Chaigui Fang might be relevant with the normalization of energy metabolism and amino acid metabolism.2. The anti-depression mechanism of Fufang Chaigui Fang might be relevant with cAMP-PKA-CREB-BDNF signaling pathway.3. The anti-depression mechanism of Fufang Chaigui Fang might be relevant with the regulation with inflammatory cytokines.