Abstract / Summary
Abstract Diminished ovarian reserve (DOR) is a common reproductive disorder that severely impairs female fertility and lacks effective treatments. Protecting granulosa cells (GCs) has emerged as a promising therapeutic strategy. Yijing decoction (YJD), a classical Chinese herbal formula, exhibits clinical efficacy against DOR; however, its mechanisms are unclear. This study aimed to evaluate the effects of YJD on ovarian reserve function and explore the potential molecular mechanisms underlying these effects. Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to identify the constituents of YJD and its serum-absorbed prototypes. Network pharmacology and molecular docking were then performed to predict therapeutic targets and validate component-target interactions. A DOR mouse model was established by intraperitoneal injection of cyclophosphamide, followed by oral administration of YJD or estradiol valerate (positive control) for 8 weeks. Ovarian function was evaluated by estrous cycle monitoring, hormone profiling, and histopathological analysis. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining was used to assess apoptosis in ovarian tissues. In isolated GCs, apoptosis, reactive oxygen species (ROS) levels, mitochondrial membrane potential, mitochondrial ultrastructure, and adenosine triphosphate levels were measured using flow cytometry, transmission electron microscopy, and commercial kits, respectively. Finally, the predicted signaling pathways were validated by immunofluorescence, reverse transcription quantitative polymerase chain reaction, and Western blotting. Thirty-two compounds were identified in the YJD-medicated serum. Network pharmacology analysis revealed 319 intersecting targets between YJD and DOR, with 22 core hubs (for example, HIF1A, PPARGC1A, and BCL2) mainly enriched in oxidative stress, ROS scavenging, the intrinsic apoptosis pathway, and HIF-1 signaling. Based on these findings, we selected PGC-1α, SIRT3, and HIF-1α for experimental validation. In cyclophosphamide-induced DOR mice, YJD treatment augmented ovarian weight and gonadosomatic index. It effectively restored estrous cycle regularity, significantly attenuated serum follicle-stimulating hormone and luteinizing hormone levels, and markedly elevated E 2 and anti-Müllerian hormone concentrations, thereby mitigating follicular atresia. Furthermore, YJD preserved mitochondrial homeostasis, reduced ROS accumulation, and suppressed GC apoptosis. Mechanistically, YJD upregulated PGC-1α and SIRT3 expression while downregulating HIF-1α in GCs. YJD enhances mitochondrial function and inhibits apoptosis in GCs, thereby maintaining ovarian reserve function. These effects may be associated with changes in the PGC-1α/SIRT3/HIF-1α signaling pathway. Further functional studies are required to validate the causal role of this signaling pathway.