【作者】 王建锋； 毛晓燕； 张超； 卢卫嘉； 陈继军； 秦海燕；

【Author】 WANG Jian-feng,MAO Xiao-yan,ZHANG Chao,LU Wei-jia,CHEN Ji-jun,QIN Hai-yan (The Department of the Fifth Research,Lanzhou Institute of Biological Products,Lanzhou 730046,China)

【机构】 兰州生物制品研究所第五研究室；

【摘要】 目的:以小鼠为动物模型研究E型肉毒毒素中毒主要器官病理变化及血清蛋白变化,为肉毒毒素中毒早期诊断寻找生物标志物及中毒病理学研究奠定基础。方法:腹腔注射清洁级小鼠E型肉毒毒素1个LD⁵⁰,眼眶采集存活小鼠血液,离心分离血清,取心,肝,脾,肺,肾固定在10%的甲醛溶液,做石蜡切片。SDS-PAGE结合2-DE-SDS-PAGE分离血清蛋白,PDQUUest8.0软件对凝胶进行背景消除和点的匹配,离子阱质谱检测差异蛋白点。结果:病理切片可见E型肉毒毒素中毒小鼠主要脏器（心,肝,脾,肺,肾）均出现不同程度病理变化,其中心脏、肝脏病变尤为严重。SDS-PAGE电泳条带显示中毒24 h血清蛋白变化最明显,正常血清和中毒24 h血清2-DE图谱找到20个差异蛋白点,其中14个蛋白点只存在小鼠正常血清中3个只存在肉毒中毒血清中,3个在肉毒中毒呈现低量表达。质谱检测出晶体蛋白家族,它只在正常小鼠血液中表达;急性期蛋白在E型肉毒毒素中毒24 h出现明显变化,其中结合珠蛋白在中毒血清样品低表达,而血清淀粉样蛋白A高表达。结论:小鼠E型肉毒中毒主要器官及血清蛋白均发生明显变化,结合珠蛋白和血清淀粉样蛋白A是中毒早期的活性分子,可作为肉毒中毒早期的生物标志物。更多还原

【Abstract】 Objective:Study on the main pathological changes of organs and serum protein changes in mice suffered from injection botulinum toxin E,to seek biomarker for early diagnosis of botulinum poisoning and lay a foundation for poisoning pathology research.Methods:Clean level mouse was abdominal injected with 1 LD₅₀ botulinum type E toxin.The blood of survived mouse was collected and serum was separated by centrifugation.The heart,liver,spleen,lung,kidney of this mouse was fixed with 10%formaldehyde and paraffin sections was produced.Serum proteins were analyzed by SDS-PAGE and 2-DE-SDS-PAGE.The background of gel was eliminated and the point was matched by PDQUest8.0 software. The differential protein points were identified by ion trap mass spectrometry.Results:Pathological section showed that the major organs-heart,liver,spleen,lung,kidney-of the botulism mouse had different degree pathological changes,among them,heart and liver had striking changes.SDS-PAGE lane display that 24 hours poisoning serum had a dramatic difference compared to other period serum.2-DE patterns of the normal serum and 24 hours poisoning serum had 20 different proteins.There were 14 proteins only existed in normal serum and 3 proteins only existed in poisoning serum. Compared with normal serum,there were 3 proteins emerged low level expression in Poisoning serum.Mass spectrum results showed that crystal proteins family only expressed in normal serum.Acute phase proteins had an obvious change in 24 hours poisoning serum.AP had low-regulation in poisoning serum,but SAA had high level expression in poisoning serum.Conclusion:The serum protein and the main organs changed significantly in botulism type E.AP and SAA are the active molecules in early poisoning,and can be used as biological diagnosis molecular of early poisoning.更多还原