姜黄素减弱Aβ_(25-35)致大鼠原代小胶质细胞的神经炎症反应  

Curcumin reduces neuroinflammation stimulated by Aβ_(25-35) in primary rat microglial cells

作　　者：刘绪华[1] 王孝庆[1] 王中苏[1] 赵航[1] 钱小伟[1] 曹红[1] 李军[1] 

机构地区：[1]温州医科大学附属第二医院麻醉科,浙江温州325027

出　　处：《中国病理生理杂志》2016年第9期1635-1641,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81271204);浙江省科技厅公益项目(No.2016C37098)

摘　　要：目的:研究姜黄素(Cur)对β-淀粉样蛋白(Aβ)刺激下大鼠原代小胶质细胞活力及高迁移率族蛋白1(HMGB1)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)表达的影响。方法:取新生SD大鼠大脑皮层行混合胶质细胞原代培养,摇床振摇法分离小胶质细胞并行Iba-1免疫细胞化学鉴定。在培养的小胶质细胞中加入Aβ_(25-35)作用24 h后,观察细胞形态并用CCK-8实验确定造模浓度及Cur的治疗浓度。将大鼠原代小胶质细胞分为5组:正常组、Aβ_(25-35)模型组、Cur组、Aβ_(25-35)+Cur治疗组、Aβ_(25-35)+DMSO组;用Western blot法检测细胞内HMGB1、晚期糖基化终末产物受体(RAGE)、NF-κB的表达情况,取上清液用ELISA检测HMGB1、IL-1β、TNF-α的表达。结果:Iba-1的阳性率在95%以上,培养的大鼠原代小胶质细胞可用于实验。Western blot实验结果示Aβ_(25-35)活化诱导后,细胞内的HMGB1、RAGE和NF-κB表达明显升高(P<0.05),加入姜黄素后细胞内的HMGB1、RAGE和NF-κB表达明显减少(P<0.05)。ELISA结果示Aβ_(25-35)活化诱导后,细胞上清液中HMGB1、IL-1β、TNF-α明显升高(P<0.05),加入姜黄素后上清液中HMGB1、IL-1β、TNF-α明显下降(P<0.05)。结论:姜黄素可明显抑制Aβ_(25-35)刺激下大鼠原代小胶质细胞的神经炎症反应。AIM： To investigate the effects of curcumin（ Cur） on the expression of High mobility group box 1protein（ HMGB1）,interleukin-1β（ IL-1β）,tumor necrosis factor-α（ TNF-α） in amyloid-β（ Aβ）-induced primary rat microglial cells. METHODS： Microglia were derived from the cerebral cortices of postnatal rat brains. The cells were identified by immunocytochemistry using mouse anti rat Iba-1 monoclonal antibody. A cell model using primary rat microglial cells incubated with Aβ25-35 as an inflammation model of Alzheimer＇s disease（ AD） was set up. The morphological characters of primary rat microglial cells were observed. The concentration of Aβ25-35 and the treatment concentration of curcumin were selected by CCK-8 assay. Cultured primary rat microglial cells were divided into 5 groups： normal cell group,Aβ25-35 group,Cur group,Aβ25-35＋ Cur group and Aβ25-35＋ DMSO group. The expression of HMGB1,NF-κB,and receptor for advanced glycation end products（ RAGE） was detected by Western blot. The levels of HMGB1,IL-1β,and TNF-α in the culture supernatant were measured by ELISA. RESULTS： The purity of primary microglias determined by Iba-1 immunofluorescence was more than 95%. The protein levels of HMGB1,RAGE and NF-κB were significantly increased after Aβ25-35 stimulation. After treatment with Cur,the protein levels of HMGB1,RAGE and NF-κB were significantly decreased（ P〈0. 05）. The levels of HMGB1,IL-1β and TNF-α in the supernatant were significantly increased after Aβ25-35 stimulation. Cur significantly decreased the level of HMGB1,IL-1β and TNF-α in the supernatant. CONCLUSION： Curcumin significantly inhibits neuroinflammation stimulated by Aβ25-35 in primary rat microglial cells.

关 键 词：阿尔茨海默病 姜黄素 Β-淀粉样蛋白 小胶质细胞 高迁移率族蛋白1 

分 类 号：R363.2[医药卫生—病理学] R749.16[医药卫生—基础医学]