PGN对Aβ1-42寡聚体促进BV2细胞分泌IL-1β的影响

摘要

Objective To investigate the effect of pro-inflammatory medium, peptidoglycan (PGN), on the secretion of incerleukin-1β (IL-1β) in amyloid protein β 1-42 (Aβ1-42) oligomer activated BV2 cells. Methods BV2 cells were cultivated and replaced microglia by cell passage method. BV2 cells were firstly separated into four groups and PGN (20 μg/mL), Aβ1-42 oligomer (0.5 μmol/L). PGN (20 μg/mL) +Aβ1-42 oligomer (0.5 μmol/L), and PGN (20 μg/mL) +SB202190, were given respectively. Then, the PGN treated BV2 cells were further divided into 5 groups and treated with different doses of PGN (0, 5, 10, 20, 40 μg/mL, respectively). A|31-42 oligomers were prepared according to Klein WL (2002). The concentration of 1L-1β in BV2 cell culture fluid was determined by ELISA. Results IL-1β secretion in BV2 cells could be activated by PGN. Aβ1-42 oligomers and PGN + Aβ1-42 oligomers, and the IL-1β peak was at the 24 h, 12 h and 24 h, separately. IL-1β concentration was higher in PGN+Aβ1-42 oligomers group in comparison with PGN and Aβ1-42 oligomers groups (P<0. 05, P<0. 05, separately). Compared to control group, the secretion of IL-1β activated by PGN increased significantly ( P < 0. 01 ). Dose-dependent relationship existed between the concentration of PGN and the 1L-1β secretion level in BV2 cells CP<0. 05). SB202190, mitogen-activated protein kinase (MAPK) inhibitor, decreased the secretion of IL-1β in BV2 cells (P<0. 01). Conclusions PGN can activate the secretion of IL-1β in BV2 cells, and also promote IL-lf) secretion in Aβ stimulated BV2 cells. P38 MAPK inhibitor could decrease IL-1β secretion, suggesting p38MAPK may participate in the process of IL-1β secretion in BV2 cells.%目的 探讨前炎介质肽聚糖(peptidoglycan,PGN)激活BV2细胞内吞β淀粉样蛋白(amyloid protein β,Aβ)1-42寡聚体后,对白细胞介素-1β(IL-1β)分泌的影响及其机制.方法 采用细胞株传代法培养BV2细胞替代小胶质细胞,按Klein方法制备Aβ1-42寡聚体.将BV2细胞分为PGN(20 μg/mL)组,A,β1-42寡聚体(0.5 μmol/L)组、PGN(20 μg/mL)+ Aβ1-42寡聚体(0.5 μmol/L)组,比较BV2细胞分泌IL-1β水平；将BV2细胞子PGN(20 μg/mL)及PGN+SB202190,比较两组IL-1β分泌水平；将不同浓度的PGN(0、5、10、20、40 μg/mL)加入BV2细胞中培养,分别检测培养液中IL-1β的水平.采用ELISA方法测定BV2细胞培养液中IL-1β的水平.结果 PGN、Aβ1-42寡聚体、PGN+ Aβ1-42寡聚体均可激活BV2细胞分泌IL-1β,分泌IL-1β高峰分别为孵育后24 h、12h、24 h;孵育后6、12、24 h同时间相比,PGN+ Aβ1-42寡聚体组BV2细胞分泌IL-1β水平均较PGN组和Aβ1-42寡聚体组明显增多(均P＜0.05)；PGN激活BV2细胞分泌IL-1β的水平与PGN浓度有剂量依赖关系(P＜0.05)；丝裂原活化蛋白激酶(MAPK)抑制剂SB202190使BV2细胞分泌IL-1β量明显减少(P＜0.01).结论 PGN可激活BV2细胞分泌IL-1β,且促进Aβ刺激BV2细胞分泌IL-1β增多,p38 MAPK抑制剂可抑制IL-1β分泌,推测p38 MAPK可能参与BV2细胞分泌IL-1β的过程.