Integrating UPLC-Q-TOF-MS, network pharmacology, molecular docking, and molecular dynamics simulation to reveal the material basis and mechanism of Xiangshao sanjie oral liquid in treating hyperplasia of mammary glands

Abstract

Xiangshao sanjie oral liquid (XSSJ), a classical traditional Chinese medicine (TCM), has been applied to treat hyperplasia of the mammary gland (HMG) for more than 20 years in China. Whereas its active ingredients and pharmacological mechanisms remained unclear. This paper aims to elucidate the active ingredients in XSSJ and to provide a clear understanding of how these compounds interact with biological pathways and contribute to therapeutic outcomes. This study systematically identified 88 chemical constituents in XSSJ using UPLC-Q-TOF-MS, among which 22 potential active ingredients were screened based on oral bioavailability (OB) and drug-likeness (DL). In a rat HMG model, XSSJ treatment dose-dependently ameliorated mammary hyperplasia, as evidenced by reduced nipple height and diameter, normalized lobular structure, and volume of mammary acinus via histopathological evaluation. The serum levels of IL-1β, IL-6, and TNF-α were significantly decreased compared with the HMG rats. Furthermore, compared with the HMG model group, XSSJ significantly inhibited the levels of p-JNK/JNK, p-ERK/ERK, and p-P38/P38, and decreased the expression of NF-κB, COX-2, and iNOS in mammary gland tissue. Molecular docking and 200 ns molecular dynamics simulations were employed to evaluate stable binding conformations and robust interactions between active ingredients and key targets, and the results confirmed that these ingredient-target complexes exhibited excellent binding properties. These integrated findings demonstrated that the mechanism of XSSJ against HMG was related to MAPK/NF-κB signaling pathway, which could provide valuable experimental evidence and new perspectives to explore the potential mechanisms of XSSJ in treating HMG.