姜黄素抑制β淀粉样蛋白致小胶质瘤细胞神经炎症反应

Curcumin inhibits neuroinflammation mediated by amyloid beta-protein in BV2 cells

目的:采用β-淀粉样蛋白(Aβ_(25-35))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;Aβ_(25-35)模型组:40μmol/L Aβ_(25-35)刺激24 h;Aβ_(25-35)+RAGE受体阻断组:抗RAGE抗体10μmol/L预处理1 h后,加入40μmol/L Aβ_(25-35)刺激24 h;Aβ_(25-35)+Cur治疗组:姜黄素10μmol/L预处理1 h后,加入40μmol/L Aβ_(25-35)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,Aβ_(25-35)模型组、Aβ_(25-35)+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Aβ_(25-35)模型组、Aβ_(25-35)+RAGE受体阻断组相比,Aβ_(25-35)+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Aβ_(25-35)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 boxl （HMGB1） and inflam- matory response of the receptor for advanced glycation end product （RAGE） with inflammation model of Alzheimer＇s disease （AD） selected Aβ25.3：induced BV2 cells for 24 hours later, to further investigate the effects of curcumin on expression of HMGB 1, RAGE, interleukin- 1 β （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 HMGB 1, 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 HMGB 1 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, HMGB 1/IL-1β and TNF-α in supematant significantly declined in group D （P〈0.05）. Conelusion： 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.