DIA-based proteomic analysis reveals Scutellaria barbata D. Don Exosome-like nanovesicles promote ferroptosis in HCC...

This study systematically characterizes ferroptosis-mediated antitumor mechanisms of exosome-like nanovesicles derived from Scutellaria barbata D. Don (SBENs) in hepatocellular carcinoma (HCC). SBENs were isolated by ultracentrifugation combined with sucrose gradient centrifugation and characterized by NTA, TEM, and DIA-based proteomic analysis. Ferroptosis induction was assessed in HepG2 cells and subcutaneous xenograft models through biochemical assays, qPCR, Western blot, and histopathological examination. SBENs were isolated using ultracentrifugation and sucrose gradient centrifugation. NTA and TEM revealed subspherical SBENs with an average diameter of 134.1 nm. Proteomic analysis identified modulation of ferroptosis-related proteins. Cellular assays showed SBENs suppressed HepG2 proliferation, migration, invasion, and ferroptosis. The ferroptosis effect induced by SBENs in HepG2 cells is marked by mitochondrial depolarization, elevated Fe²⁺, MDA, ROS, depleted GSH, and reduced SLC7A11/GPX4 expression. Mitochondrial dysfunction involved downregulated ND1, CYTB, COX1 and TFAM suppression, mechanistically associated with Nrf2/SLC7A11/GPX4 pathway inhibition. In xenografts, SBENs attenuated tumor growth, reduced Ki-67 expression, and induced ferroptosis without altering serum ALT, AST, BUN, CRE levels or causing histological damage. SBENs induced HepG2 ferroptosis via mitochondrial dysfunction and Nrf2/SLC7A11/GPX4 pathway inhibition, effectively triggering HCC ferroptosis in subcutaneous tumor xenografts with safety profiles.