Zhao Xiu-Lan, Zhu Zhen-Ping, Zhang Tian-Liang, Zhang Cui-Li, Yu Li-Hua, Xie Ke-Qin
Institute of Toxicology, Shandong University, 44 Wenhua Xi Road, Jinan 250012, China.
Toxicol Lett. 2004 Sep 10;152(2):139-47. doi: 10.1016/j.toxlet.2004.04.012.
Tri-ortho-cresyl phosphate (TOCP) is an organophosphorus ester. It is capable of producing organophosphorus ester induced delayed neurotoxicity (OPIDN) in human being and sensitive animals, which is characterized by ataxia that progresses to paralysis after 1-3 weeks following exposure to some organophosphorus ester. In present study, 18 adult hens were divided randomly into three groups, i.e. two experimental groups and control group (n = 6 each group). All hens were 10 months old and weighted 1.5-2.0 kg. The hens in two experimental groups were treated with TOCP by gavage at single dosages of 375 and 750 mg/kg respectively. TOCP was dissolved in corn oil and administered at 0.65 ml/kg. Six control hens received an equivalent volume of corn oil by gavage. All hens were sacrificed after 21 days of treatment and the sciatic nerves were dissected, homogenized and used for the determination of cytoskeletal proteins by western blotting. The levels of neurofilament (NF) subunits were decreased both in supernatant and pellet of sciatic nerves, and the most noticeable decrease in levels of NF subunits protein was observed in neurofilament medium (NF-M). Compared to the control hens, neurofilament heavy (NF-H) level decreased by 36 and 38% (P < 0.01) in the pellet and by 27 and 26% (P < 0.05) in the supernatant of sciatic nerves of hens treated with 375 and 750 mg/kg TOCP respectively. The reduction of NF-M were 36 and 68% (P < 0.01) in pellet, 50 and 67% (P < 0.01) in supernatant at 375 and 750 mg/kg dosage respectively. The neurofilament light (NF-L) lessened slightly, but the relative percentage of integrated optical density (IOD) was no significant alteration when compared to the control hens. There were significant decreases in levels of alpha-tubulin, beta-tubulin in pellet and alpha-tubulin, beta-tubulin, beta-actin in supernatant of sciatic nerves in TOCP-treated hens. Thus, the decreases of cytoskeletal proteins suggested the possible involvement of them in delayed neurotoxicity.
磷酸三邻甲苯酯(TOCP)是一种有机磷酸酯。它能够在人类和敏感动物中引发有机磷酸酯诱导的迟发性神经毒性(OPIDN),其特征为共济失调,在接触某些有机磷酸酯后1至3周会发展为麻痹。在本研究中,将18只成年母鸡随机分为三组,即两个实验组和对照组(每组n = 6)。所有母鸡均为10月龄,体重1.5至2.0千克。两个实验组的母鸡分别以375和750毫克/千克的单次剂量经口灌胃给予TOCP。TOCP溶解于玉米油中,以0.65毫升/千克的剂量给药。六只对照母鸡经口灌胃给予等量的玉米油。治疗21天后处死所有母鸡,解剖坐骨神经,匀浆并用于通过蛋白质印迹法测定细胞骨架蛋白。坐骨神经的上清液和沉淀中神经丝(NF)亚基的水平均降低,并且在神经丝中间丝(NF-M)中观察到NF亚基蛋白水平下降最为明显。与对照母鸡相比,在分别用375和750毫克/千克TOCP处理的母鸡的坐骨神经沉淀中,神经丝重链(NF-H)水平分别下降了36%和38%(P < 0.),在上清液中分别下降了27%和26%(P < 0.05)。在375和750毫克/千克剂量下,沉淀中NF-M的减少分别为36%和68%(P < 0.01),上清液中分别为50%和67%(P < 0.01)。神经丝轻链(NF-L)略有减少,但与对照母鸡相比,积分光密度(IOD)的相对百分比无显著变化。在TOCP处理的母鸡的坐骨神经沉淀中,α-微管蛋白、β-微管蛋白水平显著降低,在上清液中,α-微管蛋白、β-微管蛋白、β-肌动蛋白水平显著降低。因此,细胞骨架蛋白的减少表明它们可能参与了迟发性神经毒性。