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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。
OBJECTIVE: To screen differentially expressed protein spots related to delayed neurotoxicity (OPIDN) in chicken lumbar spinal cord tissue from three o-tolyl phosphate (TOCP) -induced broiler chickens, and to provide basis for further mass spectrometry analysis. Methods Twenty four adult roman hen chickens were randomly divided into 1 000 mg / kg TOCP group and treated with 40 mg / kg phenylmethyl sulphonyl fluoride (PMSF) for 24 h and then given 1 000 mg / kg TOCP (PMSF intervention group ) And saline-administered control group, 8 rats in each group. The animals were sacrificed on the 5th day after exposure and the total protein was extracted from the lumbar spinal cord. The whole protein profile was obtained by two-dimensional gel electrophoresis. The image analysis software (ImageMaster2D) was used to analyze the silver ion staining. Results According to the characteristics of PMSF, there were significant differences in the TOCP group compared with the control group, 23 points up-regulated and 113 down-regulated points in the POCF intervention group with no significant difference (P> 4 times the increase of 9 points,> 4 times the reduction of 11 points. Conclusion The differential expressions of 9 up-regulated spots and 11 down-regulated spots in chicken lumbar tissues exposed to TOCP may be closely related to their delayed neurotoxicity.