PMSF对TOCP暴露鸡腰髓组织蛋白表达谱影响
Effect of phenylmethyl sulfonylfluoride(PMSF) on total differential expression proteins in the spinal cord tissue of TOCP-treated hens
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摘要: 目的 从三邻甲苯磷酸酯(TOCP)暴露鸡的腰髓组织中筛选迟发性神经毒性(OPIDN)相关差异表达蛋白点,为进一步质谱分析提供依据。方法 成年罗曼鹤母鸡24只,随机分成1 000 mg/kg TOCP组,先给40 mg/kg苯甲基磺酰氟(PMSF)后24 h再投1 000 mg/kg TOCP组(PMSF干预组)和给予生理盐水的对照组,每组8只。染毒后5 d天处死动物,分离腰髓后提取总蛋白;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(ImageMaster2D)对银离子染色的电泳图谱进行分析。结果 根据PMSF的作用特点,在TOCP组中选择与对照组比较差异有统计学意义,与PMSF干预组差异无统计学意义的蛋白匹配点,共获得上调点23个,下调点113个,其中>4倍的上调点9个,>4倍的下调点11个。结论 TOCP暴露鸡腰髓组织中9个上调点和11个下调点的差异表达蛋白与其迟发性神经毒性可能密切相关。Abstract: Objective To screen differential expression proteins in the lumbar spinal cord tissue of triorthocresyl phosphate(TOCP)-treated hens and to provide evidence for mass spectroscopy.Methods Twenty-four adult Roman hens were randomly divided into three groups and 8 hens in each group.The hens in TOCP group were treated with a single oral dosage of 1 000 mg/kg TOCP.The hens in the PMSF intervention group first were given 40 mg/kg PMSF and then treated with 1 000 mg/kg TOCP 24 hours later.The control group received the same volume of sodium chloride.On the 5th day after the exposure,the hens were sacrificed and their lumbar spinal cord tissue were dissected and homogenized in ice bath.Total proteins of the lumbar spinal cord tissue were separated by isoelectric focusing as the first dimension and sodium dodecyl sulfate-ployacrylanide gel electrophoresis(SDS-PAGE)as the second dimension.The two-dimensional electrophoresis maps
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