Tandem mass tag-based proteomic analysis reveals the treatment mechanism of Bushen Huoxue Formula on psychological stress-induced premature ovarian insufficiency

Ethnopharmacological relevance: Bushen Huoxue formula (BSHXF) is a Chinese herbal prescription composed of eleven herbs widely used to treat psychological stress-induced premature ovarian insufficiency (POI) in clinical. However, the underlying mechanism is still unclear. Aim of the study: The purpose of this study was to clarify the underlying molecular mechanisms of BSHXF in the treatment of psychological stress-induced POI. Materials and Methods: The rat model was induced by corticosterone (CORT, 40mg/kg). Drugs were administered to rats once daily for 21 days. The serum E-2, FSH and AMH levels were examined by enzyme-linked immunosorbent assays. Tandem mass tag-based proteomic analysis was used to identify differentially expressed proteins. Western blot was used to verify the results of proteomic. Results: Our results indicate that BSHXF can improve ovarian disfunction. The levels of serum FSH were signally enhanced in model group compared to control group. As respected, BSHXF treatment for 3 weeks led to the decreased FSH levels than the model group. The concentrations of AMH showed an obvious decrease in the model group and were increased by BSHXF treatment. Moreover, the size and number of follicles in the BSHXF groups were similar to those in the control group. The proteomic screened out that Np4 and Angptl4 were simultaneously enriched by GO and KEGG, thus these two proteins were chosen for further study. Conclusions: These findings revealed that BSHXF might regulate the expression of Np4 and Angptl4 to improve psychological stress-induced POI.