Effects of organophosphorus compounds on ATP production and mitochondrial integrity in cultured cells
Effects of organophosphorus compounds on ATP production and mitochondrial integrity in cultured cells
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DOI：
10.1007/bf03036450
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发表时间：
2005-01-01
影响因子：
3.7
通讯作者：
Ehrich, M
中科院分区：
文献类型：
--
作者：
Massicotte, C;Knight, K;Ehrich, M
最近的研究在体内和体外表明，线粒体功能障碍后，暴露于有机磷（OP）酯。由于线粒体ATP产生对于细胞完整性是重要的，因此在人神经元细胞系（SH-SY 5 Y神经母细胞瘤细胞）和从鸡胚分离的初级背根神经节（DRG）细胞中检查了在OP神经毒物存在下的ATP产生，并随后培养以实现轴突的成熟。选择这些细胞培养系统来评价与暴露于OP化合物相关的线粒体呼吸链的毒性效应，OP化合物会引起和不会引起OP诱导的迟发性神经病（OPIDN），这是一种神经毒性酯酶（NTE）抑制之前的疾病。浓度和时间响应的研究是在神经母细胞瘤细胞暴露于苯基水杨苷磷酸盐（PSP）和mipafox，这两种化合物很容易诱导迟发性神经病变的母鸡，或对氧磷，这不会。苯甲基磺酰氟（PMSF）被列为NTE的非神经性抑制剂。用1 μ M PSP、mipafox或对氧磷处理9日龄鸡胚DRG的纯化神经元培养物12小时。生物发光法测定ATP产生的原位评价表明PSP处理的神经母细胞瘤细胞和鸡DRG神经元中的ATP浓度均降低。Mipafox可降低DRG细胞ATP生成，但对SH-SY 5 Y细胞无影响。这种低能状态存在于线粒体呼吸链的几个水平，包括复合物I、II、III和IV，尽管复合物I受到的影响最严重。对氧磷和PMSF在所有复合物中均无效，并且有效时需要比PSP所需的浓度更高的浓度。结果表明，线粒体是OP化合物的重要早期靶点，暴露导致ATP产生耗尽。神经元，而不是雪旺氏细胞线粒体在DRG中暴露于PSP和mipafox后的靶向验证了在这些细胞中的神经特异性染料，四甲基罗丹明的损失。在从DRG分离的暴露于对氧磷的神经元中或在用任何测试化合物处理的许旺细胞中未观察到这种损失。
Recent studies in vivo and in vitro suggested that mitochondrial dysfunction follows exposure to organophosphorus (OP) esters. As mitochondrial ATP production is important for cellular integrity, ATP production in the presence of OP neurotoxicants was examined in a human neuronal cell line (SH-SY5Y neuroblastoma cells) and primary dorsal root ganglia (DRG) cells isolated from chick embryos and subsequently cultured to achieve maturation with axons. These cell culture systems were chosen to evaluate toxic effects on the mitochondrial respiratory chain associated with exposure to OP compounds that do and do not cause OP-induced delayed neuropathy (OPIDN), a disorder preceded by inhibition of neurotoxic esterase (NTE). Concentration- and time-response studies were done in neuroblastoma cells exposed to phenyl saligenin phosphate (PSP) and mipafox, both compounds that readily induce delayed neuropathy in hens, or paraoxon, which does not. Phenylmethylsulfonyl fluoride (PMSF) was included as a non-neuropathic inhibitor of NTE. Purified neuronal cultures from 9 day-old chick embryo DRG were treated for 12 h with 1 mu M PSP, mipafox, or paraoxon. In situ evaluation of ATP production measured by bioluminescence assay demonstrated decreased ATP concentrations both in neuroblastoma cells and chick DRG neurons treated with PSP. Mipafox decreased ATP production in DRG but not in SH-SY5Y cells. This low energy state was present at several levels of the mitochondrial respiratory chain, including Complexes I, II, III, and IV, although Complex I was the most severely affected. Paraoxon and PMSF were not effective at all complexes, and, when effective, required higher concentrations than needed for PSP. Results suggest that mitochondria are an important early target for OP compounds, with exposure resulting in depletion of ATP production. The targeting of neuronal, rather than Schwann cell mitochondria in DRG following exposure to PSP and mipafox was verified by loss of the mitochondrial-specific dye, tetramethylrhodamine, in these cells. No such loss was seen in paraoxon exposed neurons isolated from DRG or in Schwann cells treated with any of the test compounds.