乙酰葛根素对Aβ_(25-35)诱导的BV-2小胶质细胞NF-κBp65和iNOS表达的影响  

Effect of acetylpuerarin on the expression of NF-κBp65 and iNOS in BV-2 microglia cells induced by Aβ_(25-35)

作　　者：蔡巧英[1] 孟庆慧[1] 李梅[1] 赵荣艳[1] 

机构地区：[1]潍坊医学院护理学院,潍坊261053

出　　处：《神经解剖学杂志》2016年第3期397-402,共6页Chinese Journal of Neuroanatomy

基　　金：山东省自然科学基金(ZR2013HL069);潍坊医学院科技创新研究基金(K1301017)

摘　　要：目的:研究乙酰葛根素对β-淀粉样蛋白诱导的小鼠BV-2小胶质细胞核转录因子-κBp65和诱导型一氧化氮合酶表达的影响。方法:体外培养小鼠BV-2小胶质细胞,用凝聚态β-淀粉样蛋白(amyloid-β25-35,Aβ_(25-35))诱导小胶质细胞活化建立阿尔茨海默病炎症细胞模型。实验细胞随机分为6组:空白对照组,Aβ_(25-35)组,Aβ_(25-35)+乙酰葛根素(浓度分别为:0.1,0.4,1.6μmol/L)剂量组和含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)抑制剂(Z-DEVD-fmk)组。在倒置相差显微镜下观察细胞形态的变化;硝酸还原酶法检测乙酰葛根素对一氧化氮(nitric oxide,NO)的影响;利用Western Blot分析乙酰葛根素对NF-κBp65和i NOS蛋白表达的影响;通过Realtime PCR分析乙酰葛根素对NF-κBp65和i NOS基因表达的影响。结果:Aβ_(25-35)诱导后小胶质细胞由静息状态转变为阿米巴样,乙酰葛根素可减轻细胞发生的形态改变。乙酰葛根素呈剂量依赖性的抑制炎性因子NO的产生,抑制NF-κBp65和i NOS的表达。结论:乙酰葛根素可以改善BV-2小胶质细胞的形态变化,减轻BV-2小胶质细胞的炎性反应,其机制可能与其抑制NF-κB信号通路的激活有关,且具有剂量依赖性。Objective： To study the effects of acetylpuerarin on the expression of nuclear transcription factor-kappa Bp65（ NF-κBp65） and inducible nitric oxide synthase（ i NOS） in mouse BV-2 microglia cells induced by amyloid-β25-35（ Aβ_（25-35））. Methods： The mouse BV-2 microglia cells were cultured and activated by Aβ_（25-35） in condensed state as Alzheimer＇s disease inflammatory cell model in vitro and were randomly divided into six groups： control group,Aβ_（25-35） group,Aβ_（25-35）＋ acetylpuerarin（ 0. 1,0. 4 or 1. 6 μmol / L） groups and caspase-3 inhibitor group. The changes of BV-2microglia cellular morphology were observed under inverted phase contrast microscope; the effect of acetylpuerarin on nitric oxide（ NO） was detected by nitrate reductase method; the protein expression of NF-κBp65 and i NOS were examined by Western Blot; the mRNA expression of NF-κBp65 and i NOS were measured by Real-time PCR. Results： After stimulated by Aβ_（25-35）,the cells changed from resting state into ameboid and acetylpuerarin mitigated the changes. Acetylpuerarin inhibited the production of inflammatory factor NO as well as the expression of NF-κBp65 and i NOS in a dose-dependent manner. Conclusion： Acetylpuerarin can improve the morphological changes of BV-2 microglia cells and inhibit the inflammatory response in BV-2 microglia cells. The mechanism may be related to its suppresion of the activation of NF-κBsignal pathway,which occurred in a dose-dependent manner.

关 键 词：阿尔茨海默病 乙酰葛根素 核转录因子-κBp65 诱导型一氧化氮合酶 BV-2小胶质细胞 

分 类 号：R749.16[医药卫生—神经病学与精神病学]