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弓形虫急性感染小鼠血清蛋白质质谱分析的初步研究
引用本文:周永华,黄玉政,陶永辉,王玠,许永良,高琪.弓形虫急性感染小鼠血清蛋白质质谱分析的初步研究[J].中国血吸虫病防治杂志,2012,24(3):311-314.
作者姓名:周永华  黄玉政  陶永辉  王玠  许永良  高琪
作者单位:江苏省寄生虫病防治研究所、 卫生部寄生虫病预防与控制技术重点实验室、 江苏省寄生虫分子生物学重点实验室、 江苏省寄生虫病学重点学科 (无锡 214064)
摘    要:目的 分析弓形虫急性感染小鼠血清蛋白质组学变化。方法 16只同窝配对的C57BL/6J小鼠, 随机分成弓形虫急性感染组和对照组, 每组8只。弓形虫急性感染组每鼠腹腔感染经CF?11纤维素纯化的RH株弓形虫速殖子 (1×104 /ml) 0.2 ml, 对照组每鼠腹腔接种0.2 ml 灭菌生理盐水。感染后4 d, 处死全部小鼠, 应用弱阳离子 (WCX) 纳米磁性微球捕获小鼠血清蛋白, 采用基质辅助激光解析电离飞行时间质谱 (MALDI?TOF?MS) 技术检测两组小鼠血清蛋白质变化, 并采用Au? toflex analysis 与Clin ProTools 软件进行分析与比较。结果 在分子量为1 000~16 000 Da区段, 与对照组小鼠血清蛋白指纹图谱比较, 弓形虫急性感染小鼠血清蛋白质谱有13种蛋白表达差异有统计学意义, 其中9种蛋白质表达上调, 4种蛋白质表达下调。表达上调的多肽质荷比 (m/z值)分别为1 932.76、 1 976.85、 2 090.53、 5 004.5、 5 776.01、 5 803.05、 5 847.99、 5 877.51和7 501.58, 表达下调的多肽m/z值分别为1 866.40、 4 063.71、 8 120.31和8 203.83。结论 弓形虫急性感染小鼠血清蛋白质组发生显著改变, 采用WCX纳米磁性微球联用MALDI?TOF?MS技术可检出弓形虫急性感染小鼠血清中的特异性蛋白标志物。

关 键 词:刚地弓形虫    血清蛋白质    比较蛋白质组学    质谱分析    弱阳离子纳米磁性微球    小鼠  

Preliminary study on serum proteomics in mice with acute Toxoplasma gondii infection by using functionalized magnetic beads and MALDI-TOF-MS technique
ZHOU Yong-hua , HUANG Yu-zheng , TAO Yong-hui , WANG Jie , XU Yong-liang , GAO Qi.Preliminary study on serum proteomics in mice with acute Toxoplasma gondii infection by using functionalized magnetic beads and MALDI-TOF-MS technique[J].Chinese Journal of Schistosomiasis Control,2012,24(3):311-314.
Authors:ZHOU Yong-hua  HUANG Yu-zheng  TAO Yong-hui  WANG Jie  XU Yong-liang  GAO Qi
Institution:Jiangsu Institute of Parasitic Diseases|Key Laboratory on Technology for Parasitic Disease Prevention and Control|Ministry of Health|Jiangsu Provincial Key Laboratory on Molecular Biology of Parasites|Jiangsu Provincial Key Subject on Parasitic Diseases| Wuxi 214064|China
Abstract:Objective To detect and analyze the serum protein biomarkers in mice with acute Toxoplasma gondii infection.Methods The serum samples from 8 C57BL/6J mice with acute Toxoplasma gondii infection and 8 normal healthy paired mice were prepared with WCX magnetic beads,and then analyzed on PBSⅡ-C mass spectrometer reader.The protein spectra of the serum samples were normalized by the Ciphergen Protein Chip software.The peak labeling was performed by the Biomarker Wizard software.The specific protein biomarkers were screened by the Biomarker Pattern software to construct a diagnostic model for acute Toxoplasma gondii infection.Results A total of 13 distinguished proteomic peaks were detected.Nine peaks were of up-regulated expressions including m/z values of 1 932.76,1 976.85,2 090.53,5 004.5,5 776.01,5 803.05,5 847.99,5 877.51 and 7 501.58,respectively;and four peaks were of down-regulated expressions including m/z values of 1 866.40,4 063.71,8 120.31 and 8 203.83,respectively.Conclusion The potential protein biomarkers for acute Toxoplasma gondii infection are discovered in mouse serum by MALDI-TOF-MS combined with WCX magnetic beads.
Keywords:Toxoplasma gondii  Serum protein  Comparative proteomics  Mass chromatographic analysis  Weak cationic exchange magnetic beads  Mouse
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