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硫化氢中毒机理尚未在蛋白质层面阐明,寻找血液中的蛋白质标识物在临床及法医学上具有重要意义。近年发展起来的液相色谱-质谱技术的定量蛋白质组学(iTRAQ/TMT)技术为筛选生物标识物提供了新的有力工具。采用蛋白质组学方法研究了硫化氢中毒的大鼠模型的结果。大鼠暴露于150mg/m~3浓度受控的硫化氢环境中,时间分别为0、2和4h。收集大鼠血液后,分离血浆并用蛋白质浓缩试剂盒(ProteoMiner)去除高丰度蛋白质,并通过串联质谱标签标记技术和纳流液相色谱-串联质谱(Nano HPLC MS/MS)进行定量蛋白分析。实验采取两组平行重复以确认鉴定及定量结果。在2次重复实验中,共有520个蛋白被鉴定并进行后续生物信息学分析。结果表明:蛋白质组学技术是在蛋白质水平上研究并揭示硫化氢中毒的机理。
The mechanism of hydrogen sulfide poisoning has not been elucidated at the protein level. It is of clinical and forensic importance to search for protein markers in the blood. The quantitative proteomics (iTRAQ / TMT) technology developed in recent years by liquid chromatography-mass spectrometry has provided a new and powerful tool for screening biomarkers. Proteomics was used to study the results of a rat model of hydrogen sulfide poisoning. Rats were exposed to a controlled hydrogen sulfide environment at 150 mg / m3 for 0, 2 and 4 h, respectively. After collecting rat blood, plasma was separated and high-abundance proteins were removed using a Proteominer and quantified by tandem mass spectrometry and Nano-HPLC-MS / MS. Two parallel replicates of the experiment were used to confirm the identification and quantitation results. In two replicates, a total of 520 proteins were identified and subjected to subsequent bioinformatic analysis. The results show that: Proteomics is the study of the protein level and reveals the mechanism of hydrogen sulfide poisoning.