A Dynamic Hormone-Cycling in Vitro Model for Phase-Specific Pharmacological Evaluation in Adenomyosis
DOI:
https://doi.org/10.54097/ns5y4426Keywords:
Adenomyosis, Estrogen Receptor, co-culture model, Menstrual Cycle Regulation, Herbal Medicine.Abstract
Adenomyosis is an estrogen-dependent uterine disorder characterized by inflammation, abnormal estrogen receptor signaling, and aberrant smooth-muscle activation and proliferation, all of which fluctuate across different phases of the menstrual cycle. However, conventional in vitro models lack the ability to simulate dynamic hormonal cycling, limiting the study of phase-specific mechanisms and therapeutic responses. This study established a high estradiol-primed human endometrial stromal cell and human uterine smooth muscle cell co-cultured adenomyosis-like model (H-CoC) and applied controlled estradiol/progesterone ratios to simulate proliferative, luteal, and menstrual phases. Tanshinone IIA and Paeonol were applied as a combined herbal intervention. Markers such as COX-2, PGE2, and estrogen receptor (ER- /ER- ) expression were measured. The model successfully reproduced key features of adenomyosis, including elevated COX-2 and PGE2 levels and phase-dependent alterations in ER- and ER- expressions. Herbal treatment exhibited distinct phase-specific regulatory effects. The strongest activity was observed in the proliferative phase and the menstrual phase. A shift toward ER-β predominance was observed during the proliferative phase, whereas both ER-α and ER-β were significantly suppressed during the menstrual phase, consistent with reduced COX-2, PGE2, and Ki-67 levels, reflecting coordinated anti-inflammatory and anti-proliferative effects. This study establishes a physiologically relevant platform that captures adenomyosis- and menstrual cycle-related pathogenetic changes, and highlights the phase-dependent nature of herbal responsiveness under different hormonal contexts. This system provides a foundation for future in vivo validation and patient-derived organoid studies, supporting cycle-informed evaluation of non-hormonal therapeutic candidates.
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