姜黄素抑制β淀粉样蛋白致小胶质瘤细胞神经炎症反应

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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：40 μmol/L Aβ25-35刺激24 h；Aβ25-35+RAGE受体阻断组：抗RAGE抗体10 μmol/L预处理1 h后，加入40 μmol/L Aβ25-35刺激24 h；Aβ25-35+Cur治疗组：姜黄素10 μmol/L预处理1 h后，加入40 μmol/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。

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关键词 ：阿尔茨海默病；姜黄素；淀粉样&beta, 蛋白；BV2细胞；高迁移率族蛋白质类；糖基化终末产物受体

Abstract：Objective: To explore the relationship between high mobility group box1 (HMGB1) and inflammatory response of the receptor for advanced glycation end product (RAGE) with inflammation model of Alzheimer’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/L Aβ25-35, 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/L Aβ25-35, 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: Curcumin may reduce Aβ25-35-induced neuroinﬂammation in BV2 cells through inhibiting HMGB1 expression/extracellular released and inhibition NF-κB pathway, partly correlated with RAGE expression down-regulatd.

Key words： Alzheimer’s disease curcumin amyloid beta-protein BV2 cells high mobility group proteins receptor for advanced glycation end product

收稿日期: 2016-03-31

基金资助:国家自然科学基金资助项目（81271204）；浙江省科技厅公益项目（2016C37098）

通讯作者: 李军，主任医师，教授，硕士生导师，Email：lijun0068@163.com

作者简介: 张旭彤（1974-），男，浙江温州人，副主任医师，博士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2017/V47/I2/85

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