三邻甲苯磷酸酯暴露鸡大脑总蛋白双向电泳图谱差异表达分析

Differential expression analysis of total proteins in the cerebrum of hens exposed to TOCP by two-dimensional electrophoresis

目的:筛选三邻甲苯磷酸酯(tri-ortho-cresyl phosphate,TOCP)暴露鸡大脑总蛋白中的差异表达蛋白,为蛋白质谱分析提供依据。方法:成年罗曼鹤母鸡12只随机分成TOCP染毒组和对照组,染毒组按1000mg/kg经口1次给予TOCP,对照组给予生理盐水。染毒后24d处死动物,冷环境下分离大脑,匀浆后高速离心提取总蛋白。通过双向凝胶电泳获得完整的全蛋白质图谱,运用图像分析软件(Image Master2D)对银离子染色的电泳图谱进行分析。结果:在鸡大脑总蛋白中,对照组和染毒组分别检测到1332和1008个蛋白点,能相互匹配的占77.78%。与对照组比较,染毒组的蛋白表达谱中差异表达蛋白有262个,其中蛋白差异表达达到4倍以上的蛋白点数有43个。结论:TOCP暴露可导致鸡大脑多种蛋白差异表达,其中一些蛋白的差异表达可能与TOCP诱发迟发性神经毒性有关。

OBJECTIVE：To screen the differential expression of proteins in the cerebrum of hens exposed to tri-ortho-cresyl phosphate（TOCP）so as to provide evidence for mass spectroscopy.METHODS：Twelve Roman hens were randomly divided to 2 groups,with six in each group.The hens in experiment group were treated with a single oral dosage of 1000 mg/kg TOCP,the hens in control group received the same volume of sodium chloride.On the 24th day after TOCP administration,hens were sacrificed.Their cerebrum were dissected and homogenized in ice bath.Total proteins of the cerebral tissues were separated by isoelectric focusing as the first dimension and SDS-PAGE as the second dimension.The two-dimensional electrophoresis（2-DE）maps were visualized after silver staining and analyzed by ImageMaster2D software.RESULTS：Among the total proteins from the control group and the experiment group,1 332 and 1 008 protein spots,respectively,were detected.Comparison analysis showed that 77.78% of the total protein spots were matched between the two groups.Compared with control group,262 differentially expressed protein spots were detected in experiment group and 43 protein spots showed 4 fold change.CONCLUSION：Proteomic analysis could identify the differentially expressed proteins in the two groups.And some of these protein spots may contribute to the occurrence and development of TOCP-induced delayed neurotoxicity.