Aucubin Inhibit LPS/IFN-γ-induced BV2 Cell Inflammation Through Regulating NF-κB Pathways

Microglia are important immune cells in the central nervous system.Activated microglia promote transcription,translation and secrete them outsidethe of various pro-inflammatory cytokines,chemokines and enzymes,and it acting on other microglia and neurons,further causing microglia activated.Neuronal damage and death well induces neuroinflammation and damages the central nervous system,which is one of the triggers of common neurodegenerative diseases,for example Alzheimer’s disease and Parkinson’s disease.Inhibition of microglia activation is one of the important directions for the treatment of neurodegenerative diseases development.The discovery of drugs that can effectively inhibit microglia activation plays an important role in the treatment and slowing of neurodegenerative diseases.Aucubin(AU)is a natural active substance extracted from Chinese herbal medicine.It is widely found in Eucommia ulmoides,Plantain,Codonopsis pilosulae and Radix astragali.In this study,we used LPS/IFN-g to induce the activation of BV2 cells,and the effect of AU on the activated BV2 cells and its mechanism.The research contents and results of this topic are as follows:(1)Preparation of BV2 cell model and AU cytotoxicity analysis: MTT assay was used to find that AU had no obvious cytotoxicity.The drug concentration of the final preparation of the BV2 cell inflammatory model was determined to be 0.1 mg / mL LPS + 10 U / mL IFN-g.(2)Effect of AU on BV2 cell inflammation model: by measuring cell viability,nitric oxide release and nitric oxide synthase activity,interleukin-1,interleukin-6,tumor necrosis factor-a in cell culture supernatant And its mRNA level in the cell.The results showed that AU can inhibit the transcription and secretion of interleukin-1,interleukin-6 and tumor necrosis factor-a genes in BV2 cells,and inhibit the activity of various nitric oxide synthase,and release of nitric oxide..(3)Mechanism of inhibition of BV2 cell activation by AU: By Western blot assay,AU can effectively inhibit the phosphorylation of NF-κB p65 protein and transfer to the nucleus in BV2 cells to inhibit the activation of NF-κB signaling pathway..For the determination of the specific target of activation of NF-κB signaling pathway by AU,in this experiment weused molecular docking,it was found that the possible target of AU was interleukin-1receptorIn conclusion,the results of this experiment provide an in vitro experimental basis for the treatment of neurological diseases caused by microglia activation though AU,and provide a potential therapeutic drug for the treatment of neuroinflammatory diseases.