Aifu Nuangong Plaster alleviates primary dysmenorrhea in rats by inhibiting PI3K-AKT pathway activity

Primary dysmenorrhea (PD) is a widespread gynecological condition causing severe menstrual pain, often driven by chronic uterine inflammation and excessive prostaglandin activity. Conventional oral treatments for traditional Chinese medicine (TCM) formulations like Aifu Nuangong Wan (AFNGW) face challenges such as first-pass metabolism and gastrointestinal side effects, limiting their efficacy and patient compliance. This study addresses these limitations by developing a novel transdermal plaster, Aifu Nuangong Plaster (AFNGP), to deliver active compounds more effectively and clarify its underlying molecular mechanisms against PD.

Researchers developed Aifu Nuangong Plaster (AFNGP), a transdermal formulation derived from Aifu Nuangong Wan. They used UPLC-Q-Exactive Orbitrap-MS to identify 82 compounds in the AFNGP extract and 19 prototype components absorbed into serum. A Deep-PK model predicted transdermal properties. A PD rat model was induced with estradiol benzoate and oxytocin to evaluate AFNGP's therapeutic effects, administered in a dose-dependent manner. Network pharmacology, RNA-seq analysis, and Western blotting were employed to elucidate the functional mechanisms, focusing on key protein expression.

Pharmacodynamic assessments confirmed that AFNGP significantly improved symptoms in the PD rat model. Treated rats exhibited reduced writhing responses and alleviated uterine histopathological damage. AFNGP also decreased levels of PGF₂α and IL-6 in both serum and uterine tissues in a dose-dependent manner. Integrated network pharmacology and RNA-seq analysis strongly indicated that AFNGP's therapeutic effects on PD are primarily mediated through the PI3K-AKT signaling pathway. This was further validated by Western blotting experiments. > AFNGP consistently inhibited the expression of key proteins within the PI3K-AKT signaling pathway, confirming its mechanistic role in alleviating primary dysmenorrhea.