Wang Xin, Xu Mei, Frank Jacqueline A, Ke Zun-Ji, Luo Jia
Department of Pharmacology and Nutritional Sciences, University of Kentucky College of Medicine, Lexington, KY 40536, United States.
Department of Biochemistry, Shanghai University of Traditional Chinese Medicine, Shanghai, China 201203.
Toxicol Appl Pharmacol. 2017 Apr 1;320:26-31. doi: 10.1016/j.taap.2017.02.009. Epub 2017 Feb 11.
Thiamine (vitamin B1) deficiency (TD) plays a major role in the etiology of Wernicke's encephalopathy (WE) which is a severe neurological disorder. TD induces selective neuronal cell death, neuroinflammation, endoplasmic reticulum (ER) stress and oxidative stress in the brain which are commonly observed in many aging-related neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD) and progressive supranuclear palsy (PSP). However, the underlying cellular and molecular mechanisms remain unclear. The progress in this line of research is hindered due to the lack of appropriate in vitro models. The neurons derived for the human induced pluripotent stem cells (hiPSCs) provide a relevant and powerful tool for the research in pharmaceutical and environmental neurotoxicity. In this study, we for the first time used human induced pluripotent stem cells (hiPSCs)-derived neurons (iCell neurons) to investigate the mechanisms of TD-induced neurodegeneration. We showed that TD caused a concentration- and duration-dependent death of iCell neurons. TD induced ER stress which was evident by the increase in ER stress markers, such as GRP78, XBP-1, CHOP, ATF-6, phosphorylated eIF2α, and cleaved caspase-12. TD also triggered oxidative stress which was shown by the increase in the expression 2,4-dinitrophenyl (DNP) and 4-hydroxynonenal (HNE). ER stress inhibitors (STF-083010 and salubrinal) and antioxidant N-acetyl cysteine (NAC) were effective in alleviating TD-induced death of iCell neurons, supporting the involvement of ER stress and oxidative stress. It establishes that the iCell neurons are a novel tool to investigate cellular and molecular mechanisms for TD-induced neurodegeneration.
硫胺素(维生素B1)缺乏症(TD)在韦尼克脑病(WE)的病因中起主要作用,韦尼克脑病是一种严重的神经疾病。TD会在大脑中引发选择性神经元细胞死亡、神经炎症、内质网(ER)应激和氧化应激,这些现象在许多与衰老相关的神经退行性疾病中普遍存在,如阿尔茨海默病(AD)、帕金森病(PD)、亨廷顿病(HD)和进行性核上性麻痹(PSP)。然而,其潜在的细胞和分子机制仍不清楚。由于缺乏合适的体外模型,这一研究领域的进展受到阻碍。源自人类诱导多能干细胞(hiPSC)的神经元为药物和环境神经毒性研究提供了一种相关且强大的工具。在本研究中,我们首次使用人类诱导多能干细胞(hiPSC)衍生的神经元(iCell神经元)来研究TD诱导神经退行性变的机制。我们发现TD导致iCell神经元出现浓度和时间依赖性死亡。TD诱导了ER应激,这可通过ER应激标志物如GRP78、XBP-1、CHOP、ATF-6、磷酸化eIF2α和裂解的caspase-12的增加得以体现。TD还引发了氧化应激,这表现为2,4-二硝基苯(DNP)和4-羟基壬烯醛(HNE)表达的增加。ER应激抑制剂(STF-083010和salubrinal)和抗氧化剂N-乙酰半胱氨酸(NAC)可有效减轻TD诱导的iCell神经元死亡,这支持了ER应激和氧化应激的参与。这表明iCell神经元是研究TD诱导神经退行性变的细胞和分子机制的一种新工具。