Lin Xiaoyi, Xu Benhong, Zhang Zaijun, Yang Ying, Liu Gongping, Zhu Feiqi, Ren Xiaohu, Liu Jianjun, Li Shupeng, Huang Xianfeng, Yang Xifei
College of Pharmaceutical Engineering and Life Sciences, Changzhou University No. 21, Gehu Middle Road, Wujin District Changzhou China 213000
Key Laboratory of Modern Toxicology of Shenzhen, Shenzhen Center for Disease Control and Prevention No. 8, Longyuan Road, Nanshan District Shenzhen China 518055
RSC Adv. 2019 Jun 14;9(33):18776-18784. doi: 10.1039/c9ra03054a.
Alzheimer's disease (AD) is a common neurodegenerative disease characterized by pathological processes, including abnormal amyloid deposits and filament tangles, oxidative stress, neuroinflammation, and neurotrophic insufficiency, leading to chronic and prolonged neuronal loss and cognitive deficits. Tetramethylpyrazine (TMP) is one of the main active components of , a traditional Chinese medicine widely used for brain related disease. Here, we synthesized the TMP derivative tetramethylpyrazine dimer (DTMP), and evaluated the potential mechanisms underlying its potential neuroprotective effects using the murine neuron-like cells (N2a) transfected with the human "Swedish" mutant amyloid precursor protein (N2aAPP). ELISA results indicated that DTMP reduced the levels of Aβ and Aβ in N2aAPP. Then through proteomic analysis we identified a total of 208 differentially expressed proteins in N2aAPP cells compared to the wild-type N2a cells (N2aWT), including 144 increased and 64 decreased proteins. 449 differentially expressed proteins were revealed in N2aAPP cells on DTMP treatment with 69 increased and 380 decreased proteins. Bioinformatic analysis suggested that these proteins are enriched in mitochondrial function, the electronic transmission chain, ATP binding, oxidative phosphorylation, GTPase function, the transcriptional translation process, amino acid metabolism, nucleotide binding and others. Given the vital role of mitochondria in the pathogenesis of AD, we selected the electron transport chain pathway-related molecules to further validate these findings. Western-blot analysis demonstrated that DTMP significantly increased the levels of complex I (NDUAA), complex II (SDHB), complex III (UCRI), complex IV (COX5A) and complex V (ATP5A) in N2aAPP cells. The modulation of dysregulated proteins implicated in AD pathogenesis implies the pharmacological mechanisms of DTMP and its potential as a novel therapeutic choice in AD.
阿尔茨海默病（AD）是一种常见的神经退行性疾病，其特征在于病理过程，包括异常的淀粉样蛋白沉积和细丝缠结、氧化应激、神经炎症和神经营养不足，导致慢性和长期的神经元丧失和认知缺陷。川芎嗪（TMP）是一种广泛用于治疗脑部相关疾病的传统中药的主要活性成分之一。在此，我们合成了TMP衍生物川芎嗪二聚体（DTMP），并使用转染了人类“瑞典”突变淀粉样前体蛋白的小鼠神经元样细胞（N2a）评估了其潜在神经保护作用的潜在机制（N2aAPP）。ELISA结果表明，DTMP降低了N2aAPP中Aβ和Aβ的水平。然后通过蛋白质组学分析，我们在N2aAPP细胞中总共鉴定出208种与野生型N2a细胞（N2aWT）相比差异表达的蛋白质，其中144种蛋白质增加，64种蛋白质减少。在用DTMP处理的N2aAPP细胞中发现了449种差异表达的蛋白质，其中69种蛋白质增加，380种蛋白质减少。生物信息学分析表明，这些蛋白质富集在线粒体功能、电子传递链、ATP结合、氧化磷酸化、GTPase功能、转录翻译过程、氨基酸代谢、核苷酸结合等方面。鉴于线粒体在AD发病机制中的重要作用，我们选择电子传递链途径相关分子来进一步验证这些发现。蛋白质印迹分析表明，DTMP显著增加了N2aAPP细胞中复合物I（NDUAA）、复合物II（SDHB）、复合物III（UCRI）、复合物IV（COX5A）和复合物V（ATP5A）的水平。对AD发病机制中失调蛋白质的调节暗示了DTMP的药理机制及其作为AD新型治疗选择的潜力。
