Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE-RAGE-Related Signaling.

Modified Huanglian Wendan Decoction (HLWDD) is used for polycystic ovary syndrome with insulin resistance (PCOS-IR), but its biological basis remains incompletely defined. This study integrated network pharmacology, molecular docking, 100-ns molecular dynamics (MD), batch-specific chemical characterization, and in vivo evaluation in a letrozole/high-fat diet rat model. HPLC fingerprinting showed high batch reproducibility, while UHPLC-HRMS/MS identified 997 candidate library assignments, including 118 higher-confidence features. Network analysis identified 136 candidate compounds and 532 overlapping PCOS-related targets, prioritizing AGE-RAGE signaling. Representative docked complexes remained stable during MD and showed negative MM/PBSA binding free energies. In vivo, HLWDD improved estrous-cycle recovery and ovarian morphology, reduced fasting insulin, HOMA-IR, reproductive hormones, and inflammatory markers, and modulated ovarian AGE, RAGE, STAT3, and AKT1 expression. High-dose HLWDD also reduced p-STAT3/STAT3 and increased p-AKT/AKT1 relative to the Model group. These findings support an association between HLWDD treatment and AGE-RAGE/STAT3/AKT-related signaling without establishing causal pathway mediation.