Anas barbariae 200K Improves Phagocytosis, Modulates Cell Stiffness and Oxidative Stress in Microglial Cells In Vitro

Background: In clinical practice, Anas barbariae 200K, a homeopathic medicine, is traditionally used for influenza-like illnesses. During viral infection, the innate immunity response, which is the first line of defense, involves phagocytosis and oxidative stress. We investigated the effects of Anas barbariae 200K on phagocytosis, cell stiffness, and reactive oxygen species production using microglial cells, a subpopulation of macrophages specific to the central nervous system, often used to study the inflammatory processes, oxidative stress, and phagocytosis generated during influenza-like episodes. Methods: To measure phagocytosis, the resting microglial cells were incubated with pHRodo BioParticles conjugated to E. coli, and the engulfment of the beads by the cells was measured using fluorescence microscopy. The production of total intracellular and mitochondrial reactive oxygen species (ROS) was measured with Cell-ROX or MitoSOX probe in inflamed cells using fluorescence microscopy. To study cell stiffness, the resting or inflamed cells were incubated with Anas barbariae 200K, and stiffness was assessed using atomic force microscopy. Results: The results showed that Anas barbariae 200K significantly enhanced phagocytosis by 1.5-fold compared with the vehicle in resting cells. In inflamed microglial cells, Anas barbariae 200K significantly decreased ROS production by 40% (total intracellular ROS) and by 67% (mitochondrial ROS) compared with vehicle. Moreover, Anas barbariae 200K rapidly increased cell stiffness in resting cells by 41% compared with vehicle, while in inflamed cells, cell stiffness was decreased by 21% with Anas barbariae 200K compared with vehicle. Anas barbariae 200K caused a reorganisation of filamentous actin in inflamed cells, with marked relocation of actin at the cell extremities. Conclusion: These results demonstrate an effect of Anas barbariae 200K at the cellular level, impacting cell elasticity, mitochondria, and phagocytosis-stimulating activity. The effect of Anas barbariae 200K on cell stiffness might modulate phagocytosis and actin reorganisation.