Anti-glioma mechanism of Angelica sinensis and stigmasterol: p53-associated ferroptosis identified via multi-method analysis

Abstract

Although Angelica sinensis (Oliv.) Diels possesses anti-glioma potential, its active compounds and molecular targets and molecular mechanisms are still not fully understood. This study aims to systematically identify the active components of Angelica sinensis involved in glioma treatment, elucidate its molecular mechanisms, and validate these findings through an integrated approach combining network pharmacology and experimental verification. An integrated methodological approach-incorporating network pharmacology, molecular docking techniques, and in vitro experiments-was adopted in the present research. Using network pharmacology, common targets of Angelica sinensis and stigmasterol in glioma were identified and subjected to GO and KEGG enrichment analysis. To evaluate the molecular interaction, docking simulations were performed between active constituents of Angelica sinensis and the core target proteins. Finally, we performed a battery of cellular assays to validate the effects of Angelica sinensis and stigmasterol on proliferation, clonogenicity, migration, invasion, and ferroptosis in U251-MG cells. Network pharmacology predictions indicated that the p53 signaling pathway is a key pathway mediated by stigmasterol. In vitro experiments confirmed that Angelica sinensis and stigmasterol significantly suppressed malignant phenotypes in U251-MG cells. Mechanistically, stigmasterol activated the p53 pathway, upregulating ACSL4 while downregulating GPX4 and SLC7A11 expression, thereby inducing ferroptosis. This study reveals that stigmasterol, an active component of Angelica sinensis, can inhibit the proliferation of glioma cells by activating the p53 signaling pathway to induce ferroptosis. These findings provide a theoretical basis for the development of novel stigmasterol-based therapeutic strategies against glioma.