Baicalin alleviates Mycoplasma gallisepticum-induced oxidative stress and inflammation via modulating NLRP3 inflammasome-autophagy pathway.

Baicalin is a well-known flavonoid compound, possess therapeutic potential against inflammatory diseases. Previous studies reported that Mycoplasma gallisepticum (MG) induced inflammatory response and immune dysregulation inside the host body. However, the underlying molecular mechanisms of baicalin against MG-infected chicken-like macrophages (HD11 cells) are still illusive. Oxidant status and total reactive oxygen species (ROS) were detected by ELISA assays and flow cytometry respectively. Mitochondrial membrane potential (ΔΨM) was evaluated by immunofluorescence microscopy. Transmission electron microscopy was used for ultrastructural analysis. The hallmarks of inflammation and autophagy were determined by western blotting. Oxidative stress and reactive oxygen species (ROS) were significantly enhanced in the MG-infected HD11 cells. MG infection caused disruption in the mitochondrial membrane potential (ΔΨM) compared to the control conditions. Meanwhile, baicalin treatment reduced MG-induced reactive oxygen species (ROS), oxidative stress and alleviated the disruption in ΔΨM. The activities of inflammatory markers were significantly enhanced in the MG-infected HD11 cells. Increased protein expressions of TLR-2-NF-κB pathway, NLRP3-inflammasome and autophagy-related proteins were detected in the MG-infected HD11 cells. Besides, baicalin treatment significantly reduced the protein expressions of TLR-2-NF-κB pathway and NLRP3 inflammasome. While, the autophagy-related proteins were significantly enhanced with baicalin treatment in a dose-dependent manner in the MG-infected HD11 cells. The results showed that baicalin prevented HD11 cells from MG-induced oxidative stress and inflammation via the opposite modulation of TLR-2-NF-κB-mediated NLRP3-inflammasome pathway and autophagy, and baicalin could be a promising candidate for the prevention of inflammatory effects caused by MG-infection in macrophages.