Identify the Therapeutic Role and Potential Mechanism of α-Cyperone in diminished ovarian reserve Based on Network Pharmacology, Molecular docking, Lip-MS and Experimental Validation

Research hypothesis: α-Cyperone regulates the hormone metabolism/steroideogenesis pathway by directly binding to the key proteins of ovarian granulosa cells (KGN), thereby alleviating DOR. In vitro, limited proteolysis–mass spectrometry (LiP–MS) has provided a way to observe changes in both protein abundances and structures on a proteome-wide scale. We extracted whole-cell protein samples from CTX-induced KGN cells, divided the samples into two aliquots, and one sample was incubated with AC for 24h. Two samples were treated with proteinase K (PK), and standard sample preparation was performed on two duplicate samples. All peptides were subjected to liquid chromatography-tandem MS (LC-MS/MS) in data independent acquisition (DIA) mode for DIA analysis .