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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Autoflex 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。
Objective To analyze the changes of serum proteomics in mice infected with Toxoplasma gondii. Methods Sixteen littermate-matched C57BL / 6J mice were randomly divided into acute infection group (Toxoplasma gondii) and control group (n = 8). Toxoplasma gondii acute infection group were intraperitoneally infected with CF 11 cellulose purified RH Toxoplasma gondii RH strain (1 × 104 / ml) 0.2 ml, the control group were intraperitoneally inoculated 0.2 ml sterile saline. All the mice were sacrificed 4 days after infection, and the serum proteins were captured by WCX magnetic nanoparticles. Serum protein levels were detected by MALDI-TOF MS. And analyzed and compared using Autoflex analysis and Clin ProTools software. Results Compared with the serum protein fingerprints of the mice in the control group, the molecular weight of the Toxoplasma gondii infection group was 13 000, 16 000 Da, , Four kinds of protein expression down. The m / z (m / z) values of the expressed peptides were 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 Respectively 1 866.40, 4 063.71, 8 120.31 and 8 203.83. Conclusion Serum proteome changes significantly in mice infected with Toxoplasma gondii, and WCX nano-magnetic microspheres combined with MALDI-TOF MS can be used to detect specific protein markers in serum of mice infected with Toxoplasma gondii acutely.