Arecoline improves neuroinflammation of BV2 cells induced by lipopolysaccharide and its mechanism
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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