Vacuoles are membrane vesicles in eukaryotic cells, the digestive system of cells that break down substances absorbed outside the cell and digest the useless components of the cell itself. Researches on anti-cancer and intractable diseases using vacuoles are being actively conducted. The practical application of this study to animals requires the determination of the biocompatibility of vacuole. In the present study, we evaluated the effects of vacuoles isolated from S. cerevisiae in RAW264.7 cells. This showed a significant increase in the production of nitric oxide produced by macrophage activity. Using Reactive Oxygen Species (ROS) Assay, we identified that ROS is increased in a manner dependent on vacuole concentration. Western blot analysis showed that vacuole concentration-dependently increased protein levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2). Therefore, iNOS expression was stimulated to induce Nitric oxide (NO) production. In addition, pro-inflammatory cytokines levels promoted, such as interleukin 6 and tumor necrosis factor -α. In summary, vacuoles activate the immune response of macrophages by promoting the production of immune-mediated transporters NO, ROS, and pro-inflammatory cytokines.

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