Research on protective effect of salvianolic acid B on the mouse model of inflammation in BV2 cells via inhibiting Toll-like receptor 4/nuclear factor κB signaling pathway

ZHU Tong; ZENG Guanghong; ZHOU Jie; CAI Zhibiao

Abstract：Objective To investigate influence and mechanism of salvianolic acid B(Sal B)on neuroinflammatory response of BV-2 microglia in mice induced by MPP+.Methods BV2 cells were divided into blank control group,MPP+group,and low,medium,and high concentration of salvianolic acid B+MPP+treatment group.Cells in the low,medium,and high concentration groups were treated with 10,50,and 100 μM salvianolic acid B for 2 hours,respectively.After that,cells were disposed with lM MPP+for 24 hours.The MPP+group was merely treated with IM MPP+for 24 hours.Flow cytometry and CCK-8 assay were used to detect cellular apoptosis and survival status.Western blot,immunohistochemistry,and immunofluorescence were utilized to detect expression levels of proteins such as toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor κB(NF-κB)p65 in each group of cells.Enzyme-linked immunosorbent assay was applied to measure expression levels of tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β),and interleukin 6(IL-6).Results Compared with the control group,cellular expression levels of proteins including TLR4,MyD88 and p-NF-KB p65,as well as inflammatory factors containing TNF-α,IL-1 β and IL-6 in the MPP+group increased(P＜0.01).In comparision with the MPP+group,cellular expression levels of proteins including TLR4,MyD88 and p-NF-κB p65,as well as inflammatory factors containing TNF-α,IL-1 β and IL-6 in the group treated with low,medium,and high concentrations of salvianolic acid B+MPP+were downregulated in direct proportion to the concentration of salvianolic acid B(P＜0.01).Conclusion In the MPP+-induced mouse inflammatory cell model,salvianolic acid B exerted anti-inflammatory effect by inhibiting activation of the TLR4/NF-κB inflammatory signaling pathway.