Exploring the Mechanism of Danggui Shaoyao San in Intervention of Uterine Inflammation Based on Network Pharmacology and Experimental Validation

OBJECTIVE To explore the therapeutic effect and mechanism of Danggui Shaoyao San(DGSYS) on Escherichia coli(E.coli) - induced uterine inflammation in mice.

METHODS Network pharmacology were used to predict the potential active ingredients and targets of DGSYS in treating uterine inflammation, and the potential biological processes and related signaling pathways of DGSYS in treating uterine inflammation were obtained through GO and KEGG enrichment analysis. The efficacy and mechanism of DGSYS in treating uterine inflammation were explored by constructing an E.coli-induced mouse model.

RESULTS The network pharmacology analysis revealed that DGSYS contains 54 potential active compounds and interacts with 99 related targets, while uterine inflammation is associated with 1302 targets, leading to a total of 42 shared genes. Through PPI network analysis, 15 key targets were identified for the treatment of uterine inflammation using DGSYS, including IL6, TNF, AKT1, etc. Further GO and KEGG enrichment analyses indicated that DGSYS may be involved in biological processes such as lipopolysaccharide response, inflammatory response, and NF-κB signaling pathways. Specifically, the TNF and Toll-like receptor signaling pathways played crucial roles in treating uterine inflammation. Animal experimental results showed that after modeling with bacterial solution, the uterus of the model group mice exhibited significant congestion, swelling, endometrial inflammatory infiltration accompanied by congestion and bleeding, disordered arrangement, necrosis, and shedding of local endometrial epithelial cells. Additionally, there was decreased expression of mucin MUC1 and lower mRNA and protein levels of Occludin-1, Claudin-1, and ZO-1 in the uterine tissue compared with the control group. The pro-inflammatory factors IL-6 and TNF-α, as well as IL-1β mRNA expression levels increased, while the anti-inflammatory factor IL-10 mRNA expression level decreased. Moreover, the protein expression levels of inflammatory factors IL-6, TNF-α, IL-18, and IL-1β all increased. After drug intervention, these changes in the model group mice were alleviated, with reduced uterine inflammatory infiltration, maintained mucosal barrier homeostasis, and regulated secretion of inflammatory factors. Notably, the middle and high dose groups of DGSYS demonstrated better efficacy in treating mouse uterine inflammation.

CONCLUSION  Through network pharmacology prediction and experimental validation, it has been demonstrated that DGSYS can protect the expression of endometrial mucin and tight junction proteins in the mouse uterus. This protection subsequently mitigates the overactivation of the NF-κB/NLRP3 signaling pathway, thereby alleviating the uterine inflammatory response in mice.