Effects of extracellular vesicles of Clostridium butyricum on migration, PANoptosis, and macrophage polarization in lipopolysaccharide-induced RAW264.7 macrophages

Extracellular vesicles (EVs) are nano-sized membrane-bound vesicles released by cells, playing a crucial role in intercellular communication and exchange between cells and their hosts. EVs from probiotics serve as important carriers, enabling probiotics to interact with host cells and exert their beneficial effects. Clostridium butyricum-derived EVs have been shown to exhibit a high yield, contain minimal toxic substances, and possess the ability to modulate cytokine expression in macrophages. This study aimed to investigate the effects of C. butyricum-derived EVs on cytokine expression, migration, PANoptosis, and polarization in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. EVs were isolated from C. butyricum and characterized using transmission electron microscopy and nanoparticle tracking analysis. Quantitative PCR (q-PCR) was employed to assess cytokine expression in RAW264.7 macrophages, while the transwell assay was used to evaluate their migration capacity. Flow cytometry and q-PCR were further utilized to examine the PANoptosis and polarization of the macrophages. The results demonstrated that C. butyricum-derived EVs could promote the expression of proinflammatory cytokines, such as IL-6, IL-12, and TNF-α (p