β淀粉样蛋白25-35对BV2细胞TREM2/NF-κB信号通路的影响

Effect of Aβ25-35 on TREM2/NF-κB Signaling Pathway in BV2 Microglia

目的:探讨不同浓度β-淀粉样蛋白25-35(Aβ25-35)对小胶质细胞髓样细胞触发受体2(TREM2)/核转录因子-κB(NF-κB)信号通路的影响。方法:将BV2细胞随机分为5组,分别加入0、5、10、20、40μmol/L的Aβ25-35,作用24、48 h,镜下观察细胞形态,CCK-8法测定细胞活性,酶联免疫吸附实验(ELISA)测定肿瘤坏死因子-α(TNF-α)表达,实时荧光反转录-聚合酶链反应(RT-PCR)测定NF-κBp65、TREM2 mRNA表达。结果:显微镜下观察干预48 h后Aβ25-35≥10μmol/L细胞成片死亡,干预24 h后,CCK-8检测显示,不同浓度Aβ25-35干预BV2细胞的存活率均>70%,ELISA和RT-PCR检测显示,Aβ25-3520μmol/L和40μmol/L组促炎因子TNF-α和TREM2 mRNA的表达均明显升高,Aβ25-3520μmol/L的NF-κB p65 mRNA表达明显升高,与对照组相比差异有统计学意义(P<0.05)。结论:Aβ25-35浓度20μmol/L作用24 h,能够激活小胶质细胞发生炎症反应,可能与TREM2/NF-κB信号通路的激活有关。

Objective:To investigate the effects of different concentrations of beta-amyloid peptide 25-35(Aβ25-35)on triggering receptor expressed on myeloidcells 2(TREM2)/nuclear factor-κB(NF-κB)signaling pathway in microglia.Methods:BV2 cells were randomly divided into 5 groups,treated with 0,5,10,20,and 40μmol/L Aβ25-35 for 24 h and 48 h.Cell morphology was observed under microscope,cell proliferation activity was determined by CCK-8 method,TNF-αexpression was determined by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of NF-κB p65 and TREM2 were determined by reverse transcription-polymerase chain reaction(RT-PCR).Results:Under the microscope,the cells died in slices after induction by Aβ25-35≥10μmol/L for 48 h.After 24 hours of intervention,the survival rate of BV2 cells treated with different concentrations of Aβ25-35 was>70%.Compared with the control group,the expression of pro-inflammatory cytokines TNF-αand TREM2 mRNA significantly increased in Aβ25-3520,40μmol/L,and the expression of NF-κB p65 mRNA significantly increased in Aβ25-3520μmol/L,the difference was statistically significant(P<0.05).Conclusion:The concentration of Aβ25-35 at 20μmol/L for 24 h can activate microglia to induce inflammation,which may be related to the activation of TREM2/NF-κB signaling pathway.