姜黄素抑制β淀粉样蛋白致小胶质瘤细胞神经炎症反应

摘要

目的:采用β-淀粉样蛋白(Aβ25-35)致小鼠小胶质瘤细胞(BV2细胞)炎症建立阿尔茨海默病(AD)模型,观察高迁移率族蛋白1(HMGB1)与糖基化终末产物受体(RAGE)介导炎症反应的关系,并探讨姜黄素(Cur)对BV2细胞活力、HMGB1、RAGE、IL-1β、TNF-α、NF-κB表达的影响.方法:将对数生长期的BV2细胞分为4组.对照组:不做任何处理;Aβ25-35模型组:40μmol/L Aβ25-35刺激24 h;Aβ25-35+RAGE受体阻断组:抗RAGE抗体10μmol/L预处理1 h后,加入40μmol/L Aβ25-35刺激24 h;Aβ25-35+Cur治疗组:姜黄素10μmol/L预处理1 h后,加入40μmol/L Aβ25-35刺激24 h.孵育24 h后行细胞形态学观察,CCK8检测细胞活性,Western blot法检测细胞HMGB1、NF-κB、RAGE蛋白的表达情况,ELISA法检测上清液HMGB1、IL-1β、TNF-α的含量.结果:与对照组相比,Aβ25-35模型组、Aβ25-35+RAGE受体阻断组细胞活力明显下降,胞内HMGB1表达明显升高(P<0.05),总NF-κB表达增加(P<0.05),上清液中IL-1β和TNF-α含量升高(P<0.05).与Aβ25-35模型组、Aβ25-35+RAGE受体阻断组相比,Aβ25-35+Cur治疗组细胞内HMGB1、RAGE、NF-κB表达明显下降(P<0.05),上清液中HMGB1、IL-1β、TNF-α含量明显下降(P<0.05).结论:姜黄素可减轻Aβ25-35引起的BV2细胞炎症反应,其机制与抑制HMGB1表达及核外释放、抑制NF-κB通路有关,与RAGE表达下调部分相关.%Objective: To explore the relationship between high mobility group box1 (HMGB1) and inflam-matory response of the receptor for advanced glycation end product (RAGE) with inflammation model of Alzheim-er's disease (AD) selected Aβ25-35-induced BV2 cells for 24 hours later, to further investigate the effects of curcumin on expression of HMGB1, RAGE, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) in Aβ25-35-induced BV2 cells. Methods: Cultured BV2 cells in logarithmic growth phase were divided into 4 groups: control group (group A, non-treament), model Aβ25-35 group (group B, 40 μmol/LAβ25-35, 24 h), Aβ25-35+anti-RAGE antibody group (group C, 10 μmol/L anti-RAGE antibody 1 h before+40 μmol/L Aβ25-35, 24 h) and Aβ25-35+curcumin treatment group (group D, 8 μmol/L curcumin 1 h before+40 μmol/LAβ25-35, 24 h). The morphological character of BV2 cells was observed 24 hours later and cells viability was examined by CCK8, the level expression of HMGB1, NF-κB, RAGE in cells were detected by western blotting. The level secretion of HMGB1, IL-1β, TNF-α were detected by ELISA 24 hours later. Results: Compared with group A, the cell viability in group B and C were significantly declined and the level of HMGB1 protein expression in cells was significantly increased (P<0.05), the expression of total NF-κB were significantly increased (P<0.05), IL-1β and TNF-α in supernatant were significantly increased (P<0.05). Compared with group B and C, the cell viability, the level of HMGB1 and NF-κB protein expression in cells significantly declined, HMGB1/IL-1β and TNF-α in supernatant significantly declined in group D (P<0.05). Conclusion: Cur-cumin may reduce Aβ25-35-induced neuroinflammation in BV2 cells through inhibiting HMGB1 expression/extra-cellular released and inhibition NF-κB pathway, partly correlated with RAGE expression down-regulatd.