三邻甲苯磷酸酯暴露鸡脊髓神经中Stathmin的差异表达

The differential expression of Stathmin in the spinal cord tissue of hens exposed to tri-o-cresylphosphate

目的从三邻甲苯磷酸酯（TOCP）暴露鸡的脊髓组织中筛选可能与调控微管解聚作用相关的差异表达蛋白，为探讨有机磷化合物诱发的迟发性神经毒性（OPIDN）作用机制提供靶蛋白依据。方法42只罗曼鹤母鸡随机分成1000mg／kgTOCP组、给予40mg／ks苯甲基磺酰氟（PMSF）后再给予1000ms／ksTOCP的干预组和生理盐水对照组，每组14只。染毒第5和20天，每组分别处死4只动物，低温环境下分离腰髓，提取总蛋白。用双向电泳和质谱分析技术，筛选和鉴定可能与调控微管解聚作用相关的差异表达蛋白。结果染毒第5天，TOCP暴露组部分鸡出现OPIDN症状，并随时间的推移其症状逐渐加重。双向电泳和质谱分析结果表明，在染毒早期与对照组和PMSF前干预组比较，TOCP组鸡腰髓组织中微管解聚蛋白Stathmin表达分别下调3．4和2．8倍，而PMSF前干预组与对照组之间，Stathmin的表达无明显差异。结论TOCP暴露诱导鸡脊髓神经组织Stathmin表达明显下调，而且该蛋白表达下调与微管的大量聚集及其OPIDN发生的机制可能有关。

Objective To screen the differently expressed proteins related to regulating the depolymerization of microtubules in the spinal cord of hens exposed to tri-o-cresyl phosphate （TOCP） and to provide target protein evidence for exploring the mechanisms of the delayed neurotoxicology （OPIDN） induced by organophosphorus compounds （OPs）. Methods Forty two Roman hens were randomly divided into three groups, i.e. TOCP group treated with 1000 mg/kg TOCP; intervention group treated with 40 mg / kg phenylmethanesulfonyl fluoride （PMSF） before 1000 mg/kg TOCP treatment and control group treated with tap water. Four hens in each group were sacrificed on the 5th and 20th days after exposure, respectively. Spinal cords were separated and ho- mogenated at low temperature, and the total proteins were extracted. The OPIDN symptoms observed and record- ed in the remaining 6 hens in each group. The differently expressed proteins related to regulating the depolymer- ization of microtubules were screen by two-dimensional electrophoresis and mass spectroscopy （MS）. Results The OPIDN symptoms appeared on the 5th day after exposure in TOCP group, which were gradually serious with time. The results by two-dimensional electrophoresis and MS showed that the Stathmin expression was downreg- ulated 3.4 times and 2.8 times in TOCP group, respectively, as compared with the control and PMSF intervention groups. However, there was no significant difference of the Stathmin expression between control group and PMSF intervention group. Conclusion The Stathmin expression in the spinal cord tissues of hens exposed to TOCP significantly downregulated. Moreover, the downregulated Stathmin expression may be related to excess poly- merization of microtubules and the mechanism of OPIDN.