Urine Metabolomics Reveals the Therapeutic Effects and Compatibility Rules of Shaofuzhuyu Decoction in Endometriosis of Cold Coagulation and Blood Stasis

Ethnopharmacological relevance: Shaofuzhuyu Decoction (SFZYD) is a classical formula for treating endometriosis of cold coagulation and blood stasis (ECB). The clinical efficacy is definite, but the formula compatibility rules are yet to be clarified.Aim of the study: This study aims to find ECB-related biomarkers and explore the therapeutic effects and compatibility rules of SFZYD in ECB rats.Materials and methods: This study used autologous transplantation combined with ice-water immersion to establish the ECB model. Metabolomics and multivariate statistical analysis were used to identify the biomarkers and related pathways of ECB in urine samples of rats. Based on the traditional Chinese medicine (TCM) theory of "Monarch-Minister-Adjuvant-Courier", the therapeutic effects of SFZYD on ECB rats and its compatibility were evaluated by combining biochemical indicators (CA125, IL-6) and the calculation of the biomarker callback intensity (the ratio of the absolute biomarker peak area change between the treatment and model groups to the biomarker peak area in the model group). The "biomarker-target-biological pathway" network was constructed using network pharmacology analysis.Results: This study successfully established the ECB rat model. The results of biochemical indicators showed that the therapeutic effects on ECB were ranked as SFZYD > Mon > Min > Adj. A total of 18 biomarkers for ECB in 7 pathways were identified. Among the identified ECB-related biomarkers, SFZYD, Mon, Min, and Adj were able to return the levels of 14, 9, 10, and 6 biomarkers, respectively. The results of the biomarker callback intensity showed that SFZYD > Mon > Min > Adj. The Mon group could independently return Phenol glucuronide (HMDB0060014), which was associated with 17 ECB-related protein targets and distributed in 76 biological pathways. The Min group could independently return Hippuric acid (HMDB0000714) and Isonicotinylglycine (HMDB0041912), which were associated with 20 ECB-related protein targets and distributed in 55 biological pathways. Only the combination of the Mon, Min, and Adj groups could return 15-Hydroxynorandrostene-3,17-dione glucuronide (HMDB0010353), Mycophenolic acid-O-acyl-glucuronide (HMDB0060491), and 3,3-Dimethylglutaric acid (HMDB0002441), which was associated with 48 ECB-related protein targets and distributed in 249 biological pathways.Conclusions: SFZYD could significantly improve the biochemical indicators and metabolic abnormalities of ECB. The biomarkers returned by Monarch drugs were closely related to hormone regulation, and those returned by Minister drugs were closely related to the inhibition of endometriosis pain. This study showed that metabolomics is a powerful tool to reveal the formula compatibility rules.