Proteomic identification of carbonylated proteins in F344 rat hippocampus after 1-bromopropane exposure.
Proteomic identification of carbonylated proteins in F344 rat hippocampus after 1-bromopropane exposure.
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DOI：
10.1016/j.taap.2012.05.021
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发表时间：
2012-08
影响因子：
3.8
通讯作者：
Zhenlie Huang;S. Ichihara;S. Oikawa;Jie Chang;Lingyi Zhang;K. Subramanian;S. Mohideen;G. Ichihara-G.-Ichih
中科院分区：
文献类型：
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作者：
Zhenlie Huang;S. Ichihara;S. Oikawa;Jie Chang;Lingyi Zhang;K. Subramanian;S. Mohideen;G. Ichihara-G.-Ichih
1-溴丙烷（1-BP）在实验动物和人类中都具有神经毒性。先前对大鼠海马的蛋白质组学分析涉及氧化应激中蛋白质表达的改变，表明氧化应激在1-BP诱导的神经毒性中起作用。为了从蛋白质水平了解这种作用，我们将雄性F344大鼠吸入0、400或1000 ppm的1-BP，每天8小时，持续1周或4周，并使用蛋白质羰基测定、二维凝胶电泳（2-DE）、免疫印迹和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-TOF/MS）定量海马蛋白质羰基的变化。海马活性氧和蛋白质羰基显着增加，证明1-BP相关的诱导氧化应激和蛋白质损伤。MALDI-TOF-TOF/MS鉴定了10个具有增加的羰基修饰的个体蛋白质（p<0.05;倍数变化≥1.5）。所鉴定的蛋白质参与多种生物过程，包括糖酵解、ATP产生、酪氨酸催化、GTP结合、鸟嘌呤降解和多巴胺的神经元代谢。磷酸马丙糖异构酶（TPI）活性显著降低，与TPI羰基化呈负相关（p<0.001; r=0.83）。海马和血浆中晚期糖基化终产物（AGE）水平均显著升高，海马AGE与TPI活性呈负相关（p<0.001; r=0.71）。总之，1-BP诱导的大鼠海马神经毒性似乎涉及细胞蛋白的氧化损伤、TPI活性降低和AGEs升高。
1-Bromopropane (1-BP) is neurotoxic in both experimental animals and humans. Previous proteomic analysis of rat hippocampus implicated alteration of protein expression in oxidative stress, suggesting that oxidative stress plays a role in 1-BP-induced neurotoxicity. To understand this role at the protein level, we exposed male F344 rats to 1-BP at 0, 400, or 1000ppm for 8h/day for 1week or 4weeks by inhalation and quantitated changes in hippocampal protein carbonyl using a protein carbonyl assay, two-dimensional gel electrophoresis (2-DE), immunoblotting, and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF-TOF/MS). Hippocampal reactive oxygen species and protein carbonyl were significantly increased, demonstrating 1-BP-associated induction of oxidative stress and protein damage. MALDI-TOF-TOF/MS identified 10 individual proteins with increased carbonyl modification (p<0.05; fold-change≥1.5). The identified proteins were involved in diverse biological processes including glycolysis, ATP production, tyrosine catabolism, GTP binding, guanine degradation, and neuronal metabolism of dopamine. Hippocampal triosephosphate isomerase (TPI) activity was significantly reduced and negatively correlated with TPI carbonylation (p<0.001; r=0.83). Advanced glycation end-product (AGE) levels were significantly elevated both in the hippocampus and plasma, and hippocampal AGEs correlated negatively with TPI activity (p<0.001; r=0.71). In conclusion, 1-BP-induced neurotoxicity in the rat hippocampus seems to involve oxidative damage of cellular proteins, decreased TPI activity, and elevated AGEs.