Department of Forensic Medicine, ShangHai Medical College of Fudan University, Shanghai 200032, PR China; Shanghai Key Laboratory of Forensic Medicine, Institute of Forensic Sciences, Ministry of Justice, Shanghai 200063, PR China.
Shanghai Key Laboratory of Forensic Medicine, Institute of Forensic Sciences, Ministry of Justice, Shanghai 200063, PR China.
Toxicology. 2012 Mar 11;293(1-3):89-96. doi: 10.1016/j.tox.2012.01.002. Epub 2012 Jan 12.
In order to investigate markers of toxic injury and elucidate the mechanisms underlying methanol intoxication at the protein level, proteomics technology was applied to study variations in retinal protein expression between normal rats and rat models of methanol intoxication.
Seven rats were administered saline and methanol respectively by gavage. After seven days, retinal function was assessed by electroretinography and retinal proteins were extracted and separated by two-dimensional gel electrophoresis. The gels were then stained with AgNO₃ and analyzed with Pdquest software. The differential expression of proteins was analyzed by MALDI-TOF-TOF MS, and related proteins were searched in a protein database.
Twenty-eight spots with significant differences were found, 24 of which were successfully identified. Specifically, there were 14 increased expression proteins, such as aldehyde dehydrogenase, tropomyosin alpha-1 chain, myosin light chain, and crystallin family proteins. There were 10 decreased expression ones, such as glyceraldehyde-3-phosphate dehydrogenase, recoverin, ATP synthase alpha subunit in rats with methanol toxicity.
Retinal function was greatly destroyed upon methanol intoxication. Several key proteins were up- or down-regulated upon induced retinal toxicity, indicating that there are other mechanisms underlying methanol poisoning besides oxidative injury. Together, this data provides insight and knowledge for future studies in this field.
为了从蛋白质水平上研究甲醇中毒的毒性损伤标志物和阐明其作用机制,我们采用蛋白质组学技术研究了正常大鼠和甲醇中毒大鼠模型的视网膜蛋白表达变化。
经灌胃分别给予 7 只大鼠生理盐水和甲醇,7 天后,通过视网膜电图评估视网膜功能,并用二维凝胶电泳提取和分离视网膜蛋白。然后用 AgNO₃染色,用 Pdquest 软件进行分析。采用 MALDI-TOF-TOF MS 分析差异表达蛋白,并在蛋白质数据库中搜索相关蛋白。
发现了 28 个具有显著差异的斑点,其中 24 个成功得到鉴定。具体来说,有 14 种表达上调的蛋白,如醛脱氢酶、原肌球蛋白α-1 链、肌球蛋白轻链和晶体蛋白家族蛋白;有 10 种表达下调的蛋白,如甘油醛-3-磷酸脱氢酶、恢复蛋白和 ATP 合酶α亚基。
甲醇中毒后,视网膜功能受到严重破坏。诱导的视网膜毒性导致一些关键蛋白表达上调或下调,表明甲醇中毒除了氧化损伤之外,还有其他机制。总之,这些数据为该领域的未来研究提供了新的思路和知识。