阿司匹林对Aβ25-35诱导神经元炎性损伤的保护作用研究

Protective effect of aspirin on neuronal inflammatory injury induced by Aβ25-35 in vitro

摘要:

目的 观察阿司匹林对Aβ25-35诱导神经炎性反应中肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）、核因子κB（NF-κB）/p65核蛋白和磷酸化核因子κB抑制蛋白α（pIκB-α）总蛋白表达的影响，探讨NF-κB通路在此过程中的作用。方法 取孕龄17~18 d SD大鼠的海马神经元，将培养至第7天的神经元随机分为4组：（1）Aβ组：加入终浓度为20 μmol/L的Aβ25-35；（2）低剂量阿司匹林组：加入终浓度为50 μmol/L的阿司匹林和20 μmol/L的Aβ25-35；（3）高剂量阿司匹林组：加入终浓度为100 μmol/L的阿司匹林和20 μmol/L的Aβ25-35；（4）空白对照组：加入等量维持培养基。继续培养48 h后，采用双抗体夹心ELISA法测定上清液中TNF-α和IL-1β含量，Western-Blot法检测NF-κB/p65核蛋白和pIκB-α总蛋白表达水平。结果 （1）与对照组比较，Aβ组TNF-α和IL-1β的含量明显升高（P＜0.01），NF-κB/p65核蛋白和pIκB-α总蛋白的表达水平也明显升高（P＜0.01）；（2）与Aβ组比较，低剂量和高剂量阿司匹林均可降低TNF-α和IL-1β的含量（P＜0.05），并减少NF-κB/p65核蛋白和pIκB-α总蛋白表达水平（P＜0.05）；（3）低剂量和高剂量阿司匹林组两组相比差异无统计学意义（P＞0.05）。结论 阿司匹林可能通过抑制NF-κB的激活来减轻Aβ25-35诱导神经元的炎性损伤。

Abstract:

Objective To determine the effect of aspirin on the expression of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), nuclear factor-kappa B (NF-κB)/p65 and total protein phosphorylated inhibitor protein α of nuclear factor κB (pIκB-α) in neuroinflammation induced by β amyloid protein 25-35 (Aβ25-35) in order to investigate the role of NF-κB pathway in this process. Methods Hippocampal neurons were freshly isolated from fetal SD rats at embryonic day 17 to 18 and cultured for 7d. Then the neurons were randomly divided into 4 groups, that is, Aβ treatment group (20 μmol/L Aβ25-35), low dose aspirin group (50 μmol/L aspirin and 20μmol/L Aβ25-35), high dose aspirin group (100μmol/L aspirin and 20μmol/L Aβ25-35), and the blank control group. After 48 hours’ treatment, the levels of IL-1β and TNF-α in the supernatant were measured with ELISA, and the expression level of NF-κB/p65 and pIκB-α with Western blotting. Results Compared with the control group, Aβ treatment remarkably resulted in higher levels of TNF-α and IL-1β (P＜0.01), as well as the expression of NF-κB/p65 and pIκB-α (P＜0.01). Compared with Aβ group, low dose and high dose aspirin groups had lower levels of TNF-α and IL-1β (P＜0.05), as well as the expression of NF-κB/p65 and pIκB-α (P＜0.05). There were no statistical differences between the low and high aspirin groups (P＞0.05). Conclusion Aspirin attenuates neuronal inflammatory injury induced by Aβ25-35 through inhibiting the activation of NF-κB.