[资源分享] 生物组织质谱成像方法的建立及其在微波辐射后大鼠海马组织的蛋白组分析中的应用

生物组织质谱成像方法的建立及其在微波辐射后大鼠海马组织的蛋白组分析中的应用
  刘念;刘锋;许彬;高亚兵;李向红;魏开华;张学敏;杨松成
  　　通过探讨影响质谱成像实验的主要因素,建立了优化的基于基质辅助激光解吸电离飞行时间质谱的生物组织质谱成像方法。优化的实验条件为:基质为SA;V(乙腈)∶V(水)=1∶1为溶剂,基质浓度20g/L。质谱分析条件:线性正离子模式,激光能量范围为30%~50%,照射200次,采样点间距200μm。将本方法用于高功率微波辐射后大鼠海马的蛋白组分析,获得了正常和微波辐射后大鼠海马组织质谱图像。对照组每张切片获得5902张质谱图,320张组分质谱图像;模型组每张切片获得5993张质谱图,346张质谱图像。自编了数据分析软件计算Hits值分布,Hits值的高低与组分的分布以及量有关系。统计分析表明,差异分子199个,其中上调194个,下调5个。这些差异分子的发现对于明确微波辐射致伤效应和致伤机理的研究具有重要意义。
【作者单位】：国家生物医学分析中心 北京100850(刘念;刘锋;许彬;魏开华;张学敏;杨松成);军事医学科学院放射医学研究所 北京100850(高亚兵);解放军总医院 北京100853(李向红)
【关键词】：质谱成像;微波辐射;基质辅助激光解吸电离飞行时间质谱;海马;组织切片;蛋白质组;生物标志物
【基金】：国家自然科学基金(No.20475062)资助项目
【分类号】：Q503;Q42
【DOI】：CNKI:SUN:FXHX.0.2008-04-003
【正文快照】：
　　1引言生物组织质谱成像(imaging mass spectrometry,IMS)通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)或串联飞行时间质谱(TOF/TOF-MS)直接扫描覆盖有基质的生物组织样品,获得组织切片中化合物的分布[1]。该技术无需将组织中的多肽、蛋白质等分子提取出来,无需对目标药
Establishment of Imaging Mass Spectrometry of Biological Tissues and Its Application on the Proteome Analysis of Microwave Radiated Rat Hippocampus
  Liu Nian1;Liu Feng1;Xu Bin1;Gao Ya-Bing2;Li Xiang-Hong3;Wei Kai-Hua1;Zhang Xue-Min1;Yang Song-Cheng1 1(National Center of Biomedical Analysis;Beijing 100850)2(Institute of Radiation Medicine;Academy of Military Medical Sciences;B eijing 100850)3(Chinese People's Liberation Army General Hospital;Beijing 100853)
  　Imaging mass spectrometry based on MALDI-TOF-M S for biological tissues was established.The key experimental factors were opti mized as follows:20 g/L of sinapic acid as matrix,V(ACN)∶ V(H2O)=1∶1 as matrix solvent;MALDI-TOF-MS was operated at positive linear scan mode,30%-50% of laser power range,200 laser shots per scan and 200 μm as the laser moving step.This method was used to th e proteome analysis of SD-rat brain hippocampus after microwave radiation.5902 spectra and 320 MS images per slice for normal group were acquired,599 3 spectra and 346 MS images per slice for microwave radiation group were acqui red.Software was developed to calculate the Hits values,which reflected the component distribution and abundance.By statistic analysis of normal species an d radiation group,total 199 differentiate molecules were found,194 molecules w ere up regulated.These molecules might be important for investigating the injur y mechanism of hippocampus microwave radiation.


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