Protective Effects of Physalis angulata Leaf Extract on Busulfan-induced Ovarian Dysfunction: In Vivo, Network Pharmacology, and Molecular Dynamics Insights

Cancer is a major cause of mortality worldwide, including in Indonesia. Busulfan, a chemotherapeutic agent for hematopoietic malignancies, is known to impair female reproductive function and induce ovarian failure. Physalis angulata L. contains phytosteroids with the potential to restore hormonal homeostasis. This study evaluated the effects of P. angulata leaf extract on estrous cycles and pregnancy outcomes in female rats exposed to busulfan. Antioxidant activity, phytosteroid, phenolic, and flavonoid contents were analyzed in vitro. In vivo experiments employed a randomized design with five groups: control, busulfan (40 mg/kgBW), P. angulata (300 mg/kgBW for 28 days), sequential busulfan— P. angulata , and simultaneous busulfan— P. angulata . Metabolite profiling was performed using LC-HRMS, while in silico analyses applied network pharmacology, molecular docking, and molecular dynamics simulations. The results showed moderate antioxidant activity attributed to phytosteroids, phenolics, and flavonoids. P. angulata extract improved busulfan-induced estrous cycle and ovarian histology disruption but paradoxically reduced pregnancy rates. Seventy-seven metabolites were identified, with Physalin F acting as an estrogen receptor 1 (ESR1) antagonist and Albanol A as an ESR1 agonist; Albanol A demonstrated superior stability in molecular dynamics simulations. In conclusion, P. angulata leaf extract exerts dual mechanisms: ameliorating reproductive cycle impairment while reducing pregnancy rates in busulfan-treated female rats.