Proteomic analysis of hippocampal proteins in acrylamide-exposed Wistar rats
Proteomic analysis of hippocampal proteins in acrylamide-exposed Wistar rats
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DOI：
10.1007/s00204-019-02484-9
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发表时间：
2019-05
影响因子：
6.1
通讯作者：
Daichi Nagashima;Lingyi Zhang;Y. Kitamura;S. Ichihara;Eri Watanabe;Cai Zong;Y. Yamano;T. Sakurai;S. Oikawa;G. Ichihara
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文献类型：
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作者：
Daichi Nagashima;Lingyi Zhang;Y. Kitamura;S. Ichihara;Eri Watanabe;Cai Zong;Y. Yamano;T. Sakurai;S. Oikawa;G. Ichihara
丙烯酰胺已在工业上使用，也存在于某些高温烹饪的食物中。先前的报告描述了丙烯酰胺相关的人类中毒，表现为共济失调、记忆障碍和/或幻觉。本研究的目的是通过分析丙烯酰胺暴露大鼠海马中各种蛋白质的表达水平来表征丙烯酰胺神经毒性的分子机制。雄性Wistar大鼠灌胃给予丙烯酰胺0、2和20 mg/kg 1周或0、0.2、2和20 mg/kg 5周。实验结束后，取海马组织，提取蛋白质，进行二维差异凝胶电泳结合基质辅助激光解吸电离飞行时间/飞行时间质谱（MALDI-TOF/TOF/MS）分析。MALDI-TOF/TOF/MS在1周暴露组中鉴定出2种蛋白质的显著变化，在5周暴露组中鉴定出22种蛋白质的显著变化。20 mg/kg丙烯酰胺染毒5周后，大鼠海马中9种蛋白表达上调，13种蛋白表达下调。基于生物过程GO的PANTHER过度表达检验显示，注释到烟酰胺核苷酸代谢过程、辅酶生物合成过程、丙酮酸代谢过程和碳水化合物代谢过程的蛋白质显著过度表达。该测试还显示，对于分子功能的GO，注释为肌酐激酶活性的蛋白质以及对于细胞组分的GO，注释为髓鞘、细胞质部分和细胞体的蛋白质中存在显著的过度表达。与先前对暴露于1-溴丙烷的大鼠海马蛋白质进行的蛋白质组学研究相比，发现磷酸丙糖异构酶、线粒体肌酸激酶U型、肌酸激酶β型和蛋白酶体亚基α 1型是受丙烯酰胺和1-溴丙烷影响的蛋白质，这表明软亲电体具有共同的神经毒性机制。
Acrylamide has been used industrially and also found in certain foods cooked at high temperatures. Previous reports described acrylamide-related human intoxication who presented with ataxia, memory impairment, and/or illusion. The aim of this study was to characterize the molecular mechanisms of neurotoxicity of acrylamide by analyzing the expression levels of various proteins in the hippocampus of rats exposed to acrylamide. Male Wistar rats were administered acrylamide by gavage at 0, 2, and 20 mg/kg for 1 week or 0, 0.2, 2, and 20 mg/kg for 5 weeks. At the end of the experiment, the hippocampus was dissected out and proteins were extracted for two-dimensional difference gel electrophoresis combined with matrix-assisted laser-desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF/MS). MALDI-TOF/TOF/MS identified significant changes in two proteins in the 1-week and 22 proteins in the 5-week exposure groups. These changes were up-regulation in 9 and down-regulation in 13 proteins in the hippocampus of rats exposed to acrylamide at 20 mg/kg for 5 weeks. PANTHER overrepresentation test based on the GO of biological process showed significant overrepresentation in proteins annotated to nicotinamide nucleotide metabolic process, coenzyme biosynthetic process, pyruvate metabolic process, and carbohydrate metabolic process. The test also showed significant overrepresentation in proteins annotated to creatinine kinase activity for the GO of molecular function as well as myelin sheath, cytoplasmic part, and cell body for the GO of cellular component. Comparison with a previous proteomic study on hippocampal proteins in rats exposed to 1-bromopropane identified triosephosphate isomerase, mitochondrial creatine kinase U-type, creatine kinase β-type and proteasome subunit α type-1 as proteins affected by exposure to acrylamide and 1-bromopropane, suggesting a common mechanism of neurotoxicity for soft electrophiles.