Investigation of the pharmacodynamic material basis and mechanism of Gexia Zhuyu Decoction in the treatment of primary dysmenorrhea with qi stagnation and blood stasis based on Chinmedomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Gexia Zhuyu Decoction (GXZYD), a classic traditional Chinese medicine prescription, is widely used for the treatment of primary dysmenorrhea with qi stagnation and blood stasis (QBPD). However, its pharmacodynamic material basis remains unclear.

AIM OF THE STUDY: This study aimed to assess the therapeutic effects of GXZYD on QBPD, and to elucidate its core pharmacodynamic material basis and targets using the Chinmedomics strategy.

MATERIALS AND

METHODS

The therapeutic effects of GXZYD were evaluated in a rat model of QBPD established by unpredictable stimuli combined with estrogen and progesterone intervention. Metabolic biomarkers in the QBPD model rats were analyzed using metabonomics, while the blood-absorbed constituents of GXZYD were characterized by serum pharmacochemistry. Correlation analysis, network analysis and molecular docking were performed to screen the key pharmacodynamic substances, metabolic biomarkers, and targets. In addition, the regulatory effects of these components on the identified targets were investigated through in vitro experiments.

RESULTS

GXZYD significantly prolonged writhing latency, reduced writhing times, and improved the levels of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α) and calcium signaling pathway-related proteins in QBPD model rats. Metabonomics revealed that 32 metabolic biomarkers were differentially regulated by various doses of GXZYD. Serum pharmacochemical analysis characterized 184 compounds derived from GXZYD that were absorbed into the blood. Correlation analysis pinpointed 15 pharmacodynamic substances associated with both the metabolic biomarkers and pharmacodynamic indicators. Among them, paeoniflorin, hesperidin, neohesperidin, luteolin, and daidzein were identified as the key active components. In vitro experiments demonstrated that these key components exert their therapeutic effects by modulating the protein expression of cytochrome P450 19A1 (CYP19A1), estrogen receptor 1 (ESR1), and matrix metalloproteinase 2 (MMP2) in human endometrial epithelial cells.

CONCLUSIONS

This study clarified the pathogenesis of QBPD and the pharmacodynamic basis of GXZYD. The findings indicate that the key pharmacodynamic substances may alleviate QBPD by targeting CYP19A1, ESR1, and MMP2 in endometrial epithelial cells, thereby regulating the release of prostaglandins. These results provide a theoretical foundation for the modern interpretation and clinical application of this classical formula.