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目的应用蛋白质芯片及表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)技术,筛选硝酸羟胺(HAN)中毒小鼠血清差异蛋白,为研究其中毒损伤机制提供基础资料。方法实验动物为ICR小鼠,雄性,体质量为19～23 g,随机分为对照组(n=8)和染毒组(n=8),对照组小鼠腹腔注射生理盐水,染毒组小鼠腹腔注射HAN(亚致死剂量LD20=158 mg/kg)。染毒24 h后取血,采用弱阳离子蛋白质芯片CM10及SELDI-TOF-MS技术对血清样品进行蛋白质谱分析,得出差异蛋白分子量,用BioMarker Wizard软件进行统计,登录相关数据库搜索分子量接近的血清差异蛋白。结果染毒后,动物出现流涎、紫绀、呼吸困难、肌肉颤动、屏息症状,提示缺氧缺血严重。经蛋白质谱仪分析,获得4种血清差异蛋白,并检索得到相关血清蛋白。结论实验获得了HAN染毒小鼠血清差异蛋白,该差异蛋白有可能参与了HAN中毒损伤的发生和发展过程。
Objective To screen the serum differential proteins of mice poisoned by HAN poisoning by protein chip and surface-enhanced laser desorption / ionization time-of-flight mass spectrometry (SELDI-TOF-MS), and provide the basic information for studying the mechanism of poisoning. Methods The experimental animals were ICR mice, male, weighing 19-23 g. They were randomly divided into control group (n = 8) and exposure group (n = 8). The control mice were injected intraperitoneally with normal saline Mice were intraperitoneally injected with HAN (sublethal LD20 = 158 mg / kg). Blood samples were collected 24 h after infection. Protein profiles of the serum samples were analyzed by using the weak cation protein chip CM10 and SELDI-TOF-MS. The molecular weight of the proteins was calculated and compared with BioMarker Wizard software. The relevant databases were searched for molecular weight close to serum Differential proteins. Results After exposure, the animals developed salivation, cyanosis, dyspnea, muscle flutter and breath-holding symptoms, suggesting severe hypoxia-ischemia. By protein spectrometer analysis, four kinds of serum differential proteins were obtained, and the related serum proteins were retrieved. Conclusions The serum differential proteins of HAN-exposed mice were obtained experimentally, and the differential proteins may be involved in the pathogenesis and progression of HAN poisoning.