目的 探讨槟榔碱对脂多糖（LPS）诱导小鼠小胶质细胞BV2炎症反应的改善作用及机制。方法 取对数生长期的BV2细胞分为空白组和LPS（0.01、0.1、1、10、20 μg/mL）组，CCK-8法检测细胞活力，分光光度法检测一氧化氮（NO）含量。另取细胞分为空白组、模型组、槟榔碱（10、20、40 μmol/L）组。CCK-8法检测细胞活力；分光光度法检测NO含量；酶联免疫吸附法（ELISA）检测上清中肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）、白细胞介素1β（IL-1β）和抗炎因子白细胞介素10（IL-10）水平；实时荧光定量PCR（qPCR）法检测细胞中TNF-α、IL-6、IL-1β、一氧化氮合酶（iNOS）mRNA表达表达；Western blotting法检测Toll样受体4（TLR4）、p-p65、p65、环氧化酶2（COX2）、iNOS、胞内磷脂酰肌醇激酶（PI3K）、蛋白激酶B（Akt）、p-Akt蛋白表达水平。结果 0.01～20 μg/mL LPS诱导对BV2小胶质细胞的细胞活力无显著影响，1 μg/mL LPS诱导显著上调了细胞中NO含量；槟榔碱在10～40 μmol/L内对细胞活力无显著影响，且均可缓解LPS诱导的BV2小胶质细胞损伤。与模型组相比，槟榔碱组可降低细胞内NO含量，下调炎症因子TNF-α、IL-6、IL-1β mRNA相对表达量及其血清水平；抑制TLR4、p-P65、iNOS、COX2、PI3K、p-Akt蛋白表达量。结论 槟榔碱对LPS诱导的BV2小胶质细胞炎症模型具有显著改善作用，其作用机制可能与抑制NO生成、下调iNOS表达、调控TLR4/核因子-κB（NF-κB）、PI3K/Akt信号通路有关。

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-1β 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.

R285;R971

国家重点研发计划（2022YFD1600303）；中国农业科学院农产品加工研究所创新工程（CAAS-ASTIP-2023-IFST）；三亚中国农业科学院国家南繁研究院“南繁专项”（YYLH05）；三亚中国农业科学院国家南繁研究院“南繁专项”（ZDXM2302）