Genipin, A Gut Microbiota‐Derived Metabolite of Xihuang Pill, Suppresses Triple‐Negative Breast Cancer Angiogenesis by Inhibiting HIF ‐1α/ HSPG2 Axis

Xihuang Pill (XHP) has already been demonstrated with multitarget potential in the treatment of breast cancer (BC), especially for triple-negative breast cancer (TNBC). However, its antiangiogenic mechanisms remain unclear yet. This study aims to demonstrate that Genipin, a key component of XHP, can suppress angiogenesis and to delineate its underlying molecular mechanisms. We adopted a network pharmacology approach to screen gut microbiota-derived metabolites and their potential as therapeutic targets for BC. In vitro and in vivo experiments were used to evaluate the impacts of Genipin on the proliferation, migration, and angiogenesis of TNBC cells. RNA sequencing, western blot, and other functional assays were also employed to reveal the HIF-1α/HSPG2 axis as potential biological mechanisms. Genipin was identified as the sole shared compound between gut microbial metabolites and XHP ingredients. We found that Genipin inhibited TNBC cell proliferation and migration, suppressed endothelial cell motility, and reduced microvessel density, eventually leading to alleviation of tumor hypoxia. These effects were mediated through downregulation of the HIF-1α/HSPG2 signaling axis. Knockdown of HSPG2 mimicked Genipin's antiangiogenic and antimigratory effects, which could not be reversed by dimethyloxalylglycine (DMOG). In vivo, Genipin was also found to significantly suppress tumor growth and angiogenesis in both mouse and zebrafish models. Overall, Genipin, a gut microbiota-activated metabolite and a key component of XHP, suppresses TNBC angiogenesis by targeting the HIF-1α/HSPG2 pathway. These findings provide mechanistic insights into the therapeutic potential of XHP and highlight Genipin as a promising candidate for antiangiogenic therapy in TNBC.