Bupleurum falcatum L. root extract suppresses norepinephrine-induced M2 macrophage polarization by regulating IL-4/STAT6 axis.

Chronic psychological stress promotes cancer progression by activating tumor-associated macrophages (TAMs), particularly through polarization toward the pro-tumorigenic M2 phenotype. However, therapeutic strategies targeting stress-induced macrophage polarization remain limited. This study investigated whether the ethanolic extract of Bupleurum falcatum L. root (EBF) can modulate chronic stress-driven TAM polarization. To simulate a stress-associated microenvironment, norepinephrine (NE) was used to treat 4 T1 breast cancer cells. Conditioned media from NE-treated cells (NE CM) significantly upregulated M2 macrophage markers and STAT6 phosphorylation in RAW 264.7 cells; however, these effects were markedly attenuated by EBF. Consequently, EBF inhibited M2 macrophage-induced cancer cell migration. Notably, EBF treatment modulated the tumor cell secretome, as CM from 4 T1 cells co-treated with NE and EBF failed to induce M2 polarization. Network pharmacology analysis identified interleukin (IL)-4 as a key mediator-upregulated by NE and suppressed by EBF-among the overlapping genes shared by cancer metastasis, BF, and M2 macrophage-associated gene sets. Furthermore, saikosaponins A, C, and D consistently contributed to these regulatory effects. Collectively, our findings demonstrate that EBF inhibits NE-driven M2 polarization and subsequent macrophage-mediated cancer progression, suggesting its potential as a therapeutic agent to modulate the tumor microenvironment under chronic stress conditions.