莱菔硫烷对Aβ处理的SH-SY5Y细胞活力、组蛋白乙酰化水平及TrkA表达的影响  

EFFECTS OF SULFORAPHAE ON CELL VIABILITY,HISTONE ACETYLATION LEVELS,AND THE EXPRESSIONS OF TrkAIN SH-SY5Y CELLS EXPOSED TO Aβ

作　　者：施万英[1] 张瑞[2] 马爽[1] 张静竹[2] 李心蕙 安丽[2] 

机构地区：[1]中国医科大学附属第一医院,临床营养科,沈阳110001 [2]中国医科大学公共卫生学院营养与食品卫生学教研室,沈阳110122

出　　处：《营养学报》2016年第6期591-595,共5页Acta Nutrimenta Sinica

基　　金：辽宁省目然科学基金资助项目(No.2014021034);辽宁省教育厅科学研究一般项目(No.L2014289)

摘　　要：目的探讨莱菔硫烷(sulforaphane,SFN)对β淀粉样肽(β-amyloid peptide,Aβ)处理的神经细胞的保护作用及其机制,为阿尔茨海默病(Alzheimer's disease,AD)的防治提供理论依据。方法体外培养人神经母细胞瘤细胞(SH-SY5Y),采用MTT法检测SFN(终浓度为2μmol/L)对Aβ染毒(终浓度分别为10、20、40、80μmol/L)的SH-SY5Y细胞活力的影响,并以Real time PCR、Western blot法分析SFN预处理后的Aβ染毒(终浓度为20μmol/L)细胞TrkA mRNA及其蛋白表达以及组蛋白乙酰化水平(Ace-H3K9、Ace-H4K12)的时相分布变化。结果细胞活力随着Aβ染毒剂量的增加而逐渐降低,其中40、80μmol/L Aβ染毒的细胞活力与对照组比较,差异有统计学意义(P<0.01或P<0.001);SFN预处理后的各组细胞活力未见统计学差异(P>0.05)。与对照组比较,Aβ处理后12、24 h时,细胞TrkA mRNA水平降低,24 h时TrkA蛋白表达水平降低,6、12、24 h时Ace-H3K9、Ace-H4K12水平降低,差异均有统计学意义(P<0.01);与Aβ染毒细胞比较,SFN预处理后的Aβ染毒细胞12、24 h时TrkA mRNA水平升高,24 h时TrkA蛋白表达水平升高,12、24 h时Ace-H3K9以及6、12、24 h时Ace-H4K12水平升高,差异均有统计学意义(P<0.001)。结论 Aβ可使细胞活力下降、TrkA mRNA及其蛋白表达水平降低以及Ace-H3K9、Ace-H4K12水平的升高,SFN对Aβ诱导的上述改变均具有抑制作用。Objective To investigate the effects of sulforaphane （SFN） on SH-SY5Y cells exposed to β -amyloid peptide Aβ, and provide theoretical basis for Alzheimer＇s disease prevention. Methods MTT test was conducted to evaluate the effect of 2 μmol/L SFN pretreatment on cell viability of SH-SY5Y cells exposed to Aβ （final concentrations were 10, 20, 40, 80 μmol/L respectively）. Real time PCR and Western blot were used to detect the relative phase-distribution-expressions of mRNA and protein expressions of TrkA, and Ace-H3K9 and Ace-H4K12 levels in SFN-pretreated SH-SY5Y cells exposed to Aβ （20 μmol/L）. Results Exposure to Aβ decreased the cell viability of SH-SY5Y cells in a dose-response manner, and 40 and 80 μmol/L Aβ significantly decreased the cell viability （P〈0.01 or P〈0.001）. SFN pretreatment did not change the cell viability of SH-SY5Y cells significantly （P〉0.05）. TrkA mRNA transcript levels fell down at 12 h and 24 h, and TrkA protein level dropped at 24 h （P〈0.01）. The levels of Aee-H3K9 and Aee-H4K12 decreased at 6 h, 12 h, and 24 h in cells exposed to Aβ （P〈0.01）. Compared to the cells exposed to AI3, SFN pretreatment up-regulated TrkA mRNA transcript levels at 12 h and 24 h （P〈0.01）, and TrkA protein level at 24 h （P〈0.01）, increased Ace-H3K9 levels at 12 h, 24 h （P〈0.01）, and Ace-H4K12 levels at 6 h, 12 h, 24 h （P〈0.01）. Conclusion When exposed to Aβ, SFN could protect SH-SY5Y cell viability, and maintain the levels of TrkA mRNA and protein expression and decrease Ace-H3K9and Ace-H4K12 levels. [ACTA NUTRIMENTA SINICA, 2016, 38（6）： 591-595]

关 键 词：莱菔硫烷 阿尔茨海默病 Β-淀粉样肽 酪氨酸激酶A 组蛋白乙酰化 

分 类 号：R749.16[医药卫生—神经病学与精神病学]