Integration of metabolomics and bioinformatics to study the central neuromodulatory effects of Jiawei Erzhi formula in ovariectomized rats

Postmenopausal osteoporosis (PMOP) is a metabolic bone disease caused by a decrease in estrogen production triggered by the decline of ovarian function, leading to an imbalance in bone homeostasis. Jiawei Erzhi Formula (JWEZF), which is a classic formula, has been shown to improve bone microstructure and bone metabolism in PMOP rats after dosing. Additionally, the clinical application shows that the formula can alleviate anxiety, insomnia, and other symptoms in patients. The present study focused on investigating the mechanism of the effect of JWEZF on the central nervous system (CNS) of PMOP rats. Firstly, hematoxylin-eosin staining and biochemical indicator examination were used to determine the efficacy of JWEZF. The non-targeted metabolomics was used to investigate the metabolic profiles of the spinal cord and brain tissue of PMOP rats, and the metabolite-response-enzyme-gene network was built by combining the results of bioinformatics analysis, followed by a western blot to validate the key signaling pathways. JWEZF corrected neuronal organization disorder in PMOP rat brain tissue and regulated neuropeptide levels in serum and bone tissue. JWEZF carries out its key regulatory actions through amino acid metabolism, according to the metabolomics study of the spinal cord and brain. JWEZF affects the 5-hydroxytryptamine (5-HT) pathway in tryptophan metabolism mainly through the regulation of TPH2 and MAOA proteins based on joint bioinformatics analysis and Western blot evaluations. JWEZF regulates PMOP via the 5-HT pathway in the spinal cord and brain, which provides the foundation for the treatment of PMOP through the central nervous system.