姜黄素抑制BV2细胞RAGE/NF-κB通路发挥对多巴胺能神经细胞的保护作用  

The protection on doparminergic nerve cells exerted by Curcumin via inhibiting RAGE/NF-κB pathway of BV2 cells

作　　者：李梦楠[1] 马振凯[1] 白宏英[1] LI Mengnan;MA Zhenkai;BAI Hongying(Department of Neurology,The Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州大学第二附属医院,河南郑州450014

出　　处：《中风与神经疾病杂志》2017年第12期1072-1075,共4页Journal of Apoplexy and Nervous Diseases

基　　金：河南省基础与前沿技术研究计划项目(No.142300410461)

摘　　要：目的探讨小胶质细胞在多巴胺能神经细胞损伤中所起的作用以及姜黄素通过抑制小胶质细胞的炎症反应发挥保护作用。方法选用鼠小胶质细胞系BV2细胞株和神经元SH-SY5Y细胞作为研究对象,利用鱼藤酮(Rotenone,RO)建立细胞损伤模型,10 nmol/L RO处理BV2细胞6 h后再经不同浓度(1、5、10μmol/L)姜黄素(Curcumin,Cur)处理4 h,收集BV2细胞上清液作为条件培养基处理SH-SY5Y细胞,并设置空白对照组。采用四甲基偶氮唑盐法(MTT法)检测两种细胞活力,Western blot检测细胞BV2细胞内RAGE/NF-κB表达,ELISA法测定BV2细胞培养上清液细胞因子含量。结果 1~10μmol/L的Cur并未影响BV2细胞的增殖活力;与空白对照组相比,RO组RAGE/NF-κB的蛋白表达明显升高(P<0.01),与RO组相比,Cur处理组RAGE/NF-κB的蛋白表达量显著减少(P<0.05);与空白对照组相比,RO组炎症因子的分泌量明显升高(P<0.01),与RO组相比,Cur处理组随着浓度的升高炎症因子的分泌量逐渐减少(P<0.05);受条件培养基处理的SH-SY5Y细胞,与对照组相比,RO组细胞活力明显减弱(P<0.01),但Cur处理组的细胞活力较RO组升高,差异具有统计学意义(P<0.05)。结论 Cur通过抑制小胶质细胞反应,从而保护多巴胺能神经细胞。ObjectiveTo explore the effects of microglia on the impairment of dopaminergic cells and the mechanisms of Cur’s protective effects on them by inhibitting microglia inflammatory response.MethodsSetting rat BV2microglial cell line and SH SY5Y cells as the subjects.The cellular model of PD was established by RO.Then BV2cells were stimulated by Cur of different dose(1,5,10μmol/L)for4h following the preincubation of RO(10nmol/L)for6h,and the blank control group was established.Besides,SH SY5Y cells were treated with the medium from Cur pretreated BV2cells which were conditioned with RO.Then cell viability was assessed with MTT;the protein expression of RAGE/NFκB was detected by Western blotting;cytokines was measured by ELISA.Results1~10μmol/L Cur did not affect proliferation activity of BV2cells;compared with the blank control group,RAGE/NFκB protein in RO group expression increased significantly(P<0.01),compared with RO group,RAGE/NFκB in Cur treatment group significantly reduced(P<0.05);similarly,compared with the blank control group,the production of inflammatory markers in RO group increased significantly(P<0.01),compared with RO group,concentration of inflammatory factor in Cur treatment group gradually reduced(P<0.05);cell viability of SH SY5Y cells treated with conditioned medium reduced obviously compared with control group(P<0.01),but cell vitality in Cur treatment group was much higher than RO group(P<0.05).ConclusionCur protect dopaminergic neurons by inhibiting the microglia reaction.

关 键 词：小胶质细胞(BV2) 条件培养基 SH SY5Y细胞 姜黄素 鱼藤酮 

分 类 号：R742.5[医药卫生—神经病学与精神病学]