Huang Xianfeng, Wang Jing, Chen Xiao, Liu Pan, Wang Shujin, Song Fangchen, Zhang Zaijun, Zhu Feiqi, Huang Xinfeng, Liu Jianjun, Song Guoqiang, Spencer Peter S, Yang Xifei
College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
Key Laboratory of Modern Toxicology of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen, China.
Front Pharmacol. 2018 Apr 4;9:199. doi: 10.3389/fphar.2018.00199. eCollection 2018.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder that has proved refractory to drug treatment. Given evidence of neuroprotection in animal models of ischemic stroke, we assessed the prenylflavonoid xanthohumol from the Common Hop ( L.) for therapeutic potential in murine neuroblastoma N2a cells stably expressing human Swedish mutant amyloid precursor protein (N2a/APP), a well-characterized cellular model of AD. The ELISA and Western-blot analysis revealed that xanthohumol (Xn) inhibited Aβ accumulation and APP processing, and that Xn ameliorated tau hyperphosphorylation via PP2A, GSK3β pathways in N2a/APP cells. The amelioration of tau hyperphosphorylation by Xn was also validated on HEK293/Tau cells, another cell line with tau hyperphosphorylation. Proteomic analysis (2D-DIGE-coupled MS) revealed a total of 30 differentially expressed lysate proteins in N2a/APP vs. wild-type (WT) N2a cells (N2a/WT), and a total of 21 differentially expressed proteins in lysates of N2a/APP cells in the presence or absence of Xn. Generally, these 51 differential proteins could be classified into seven main categories according to their functions, including: endoplasmic reticulum (ER) stress-associated proteins; oxidative stress-associated proteins; proteasome-associated proteins; ATPase and metabolism-associated proteins; cytoskeleton-associated proteins; molecular chaperones-associated proteins, and others. We used Western-blot analysis to validate Xn-associated changes of some key proteins in several biological/pathogenic processes. Taken together, we show that Xn reduces AD-related changes in stably transfected N2a/APP cells. The underlying mechanisms involve modulation of multiple pathogenic pathways, including those involved in ER stress, oxidative stress, proteasome molecular systems, and the neuronal cytoskeleton. These results suggest Xn may have potential for the treatment of AD and/or neuropathologically related neurodegenerative diseases.
阿尔茨海默病（AD）是一种进行性神经退行性疾病，已证明对药物治疗具有抗性。鉴于在缺血性中风动物模型中有神经保护的证据，我们评估了来自啤酒花（Humulus lupulus L.）的异戊烯基黄酮黄腐酚在稳定表达人类瑞典突变淀粉样前体蛋白（N2a/APP）的小鼠神经母细胞瘤N2a细胞中的治疗潜力，N2a/APP是一种特征明确的AD细胞模型。酶联免疫吸附测定（ELISA）和蛋白质免疫印迹分析表明，黄腐酚（Xn）抑制β淀粉样蛋白（Aβ）积累和淀粉样前体蛋白（APP）加工，并且Xn通过蛋白磷酸酶2A（PP2A）、糖原合成酶激酶3β（GSK3β）途径改善N2a/APP细胞中的tau蛋白过度磷酸化。Xn对tau蛋白过度磷酸化的改善作用在HEK293/Tau细胞（另一种存在tau蛋白过度磷酸化的细胞系）中也得到了验证。蛋白质组学分析（二维差异凝胶电泳耦合质谱法）显示，与野生型（WT）N2a细胞（N2a/WT）相比，N2a/APP细胞中有总共30种差异表达的裂解物蛋白，在有或没有Xn的情况下，N2a/APP细胞裂解物中有总共21种差异表达的蛋白。一般来说，这51种差异蛋白根据其功能可分为七个主要类别，包括：内质网（ER）应激相关蛋白；氧化应激相关蛋白；蛋白酶体相关蛋白；ATP酶和代谢相关蛋白；细胞骨架相关蛋白；分子伴侣相关蛋白，以及其他蛋白。我们使用蛋白质免疫印迹分析来验证Xn在几个生物学/致病过程中引起的一些关键蛋白的变化。综上所述，我们表明Xn减少了稳定转染的N2a/APP细胞中与AD相关的变化。潜在机制涉及多种致病途径的调节，包括那些参与ER应激、氧化应激、蛋白酶体分子系统和神经元细胞骨架的途径。这些结果表明Xn可能具有治疗AD和/或神经病理学相关神经退行性疾病的潜力。
