Objective To investigate the effects of arecoline on lipopolysaccharide(LPS)induced BV2 inflammatory response in mouse microglia and its mechanisms.Method BV2 cells were divided into blank group and LPS group(0.01,0.1,1,10,20 μg/mL).The cell viability was detected by CCK-8 method,and the content of NO was detected by spectrophotometry.Cells were divided into blank group,model group and arecoline group(10,20,40 μmol/L).Cell viability was detected by CCK-8 method.NO content was detected by spectrophotometry.The levels of TNF-α,IL-6,IL-1β,and anti-inflammatory factor IL-10 in supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative PCR(qPCR)was used to detect the mRNA expression of TNF-α,IL-6,IL-1β,iNOS in cells.The expression levels of TLR4,p-P65,P65,COX2,iNOS,PI3K,Akt,and p-Akt were detected by Western blotting.Result In the concentration range(0.01—20 μg/mL),LPS induction had NO significant effect on the cell viability of BV2 microglia,and 1 μg/mL LPS induction significantly increased the content of NO in the cells.Arecaline had no significant effect on cell viability in the range of 10 to 40 μmol/L,and could alleviate LPS-induced damage of BV2 microglia.Compared with the model group,arecoline group could decrease the intracellular NO content,down-regulate the mRNA expression of inflammatory factors TNF-α,IL-6,IL-1 β and their serum levels.The expression levels of TLR4,p-P65,iNOS,COX2,PI3K and p-Akt were inhibited.Conclusion Arecarecine can significantly improve the LPS-induced inflammation model of BV2 microglia,and its mechanism may be related to inhibiting NO production,down-regulating the expression of iNOS,and regulating the TLR4/NF-κB and PI3K/Akt signaling pathways.
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