Yunnan baiyao exerts anti-glioma activity by inducing autophagy-dependent necroptosis

Ethnopharmacological relevance: Yunnan Baiyao (YB), a traditional herbal formulation, has been used for over a century to manage bleeding and enhance blood circulation. Its ingredients are widely recognized for their anti-cancer properties. However, its impact on glioma, the most common primary malignant tumor of the central nervous system, remains unexplored.

Aim of the study: This study aims to investigate the anti-glioma activity of YB in vitro and in vivo, and to elucidate the underlying mechanism of action.

Methods: U-87 MG cells were treated with YB and subjected to cell proliferation assay, colony formation assay, and flow cytometry with Annexin V/PI staining to confirm anti-glioma activity. The induction of Necroptosis and Autophagy was confirmed through live-cell imaging, western blotting, and immunofluorescence analysis. The role of Apoptosis, Necroptosis, Autophagy, and AMPK was validated using specific inhibitors. The in vivo anti-glioma activity of YB was evaluated using subcutaneous and orthotopic xenograft models in nude mice and chemically induced glioma rat models.

Results: YB induced necroptotic rather than apoptotic cell death in glioma U-87 MG cells, as evidenced by increased phosphorylated MLKL levels and plasma membrane disruptions. Rescue experiments further confirmed the role of Necroptosis. Importantly, YB-triggered Necroptosis was found to be dependent on Autophagy induction, which relies on the AMPK signaling pathway. In line with these findings, YB demonstrated significant anti-glioma activity in vivo.

Conclusions: Our study reveals that YB exerts potent anti-glioma effects both in vitro and in vivo through the induction of autophagy-dependent Necroptosis.

Autophagy; Glioma; Necroptosis; Yunnan baiyao.