摘要
目的研究诱发迟发性神经病的氟丙胺磷对人成神经细胞瘤SK-N-SH细胞分化的影响及其作用机理。方法MTT法测定细胞增殖,全反式维A酸诱导细胞分化,显微测量有机磷酸酯对细胞分化的影响,比色法测定NTE活力和Western印迹分析总的和磷酸化的神经纤丝重链蛋白以及肌动蛋白的表达。结果50μmol·L-1对氧磷和氟丙胺磷对细胞增殖没有影响;此浓度下对氧磷不抑制细胞的分化,而氟丙胺磷抑制细胞分化;氟丙胺磷抑制神经病靶标酯酶,而对氧磷对其不抑制;氟丙胺磷抑制总的和磷酸化的神经纤维丝重链蛋白的表达,而对氧磷对两种状态的神经纤维丝重链蛋白没有明显的作用。对氧磷和氟丙胺磷对肌动蛋白的表达都无影响。结论氟丙胺磷对人成神经细胞瘤细胞分化的抑制是通过对总的和磷酸化的神经纤维丝重链蛋白表达的抑制实现的,并伴随着神经病靶标酯酶的抑制。
AIM To investigate the effect of mipafox that induced delayed neuropathy on neural differentiation in human neuroblastoma SK-N-SH cells and elucidate its mechanism. METHODS Inhibitory effects of mipafox （50 μmol·L^-1 ） and paraoxon（50μmol·L^-1 ） on cell proliferation were evaluated by MTT method; cell differentiation was induced by all trans-retinoic acid and the effect of organophosphates on neural differentiation was measured by microscopy technique; activity of neuropathy target esterase （NTE） was assayed by photometry; the expression levels of total and phosphorylated neurofilament heavy chain, actin were detected by Western blotting. RESULTS Mipafox and paraoxon had no effect on cell proliferation. Mipafox, but not paraoxon, significantly decreased neurite length in the retinoic acid-induced differentiation of human neuroblastoma SK-N-SH cells, compared to cells treated with vehicle. Activity of NTE was significantly inhibited by mipafox in differentiated cells, in contrast, paraoxon did not affect NTE activity. Western blot analyses revealed that the level of total neurofilament heavy chain protein （NF-H） was decreased to （23. 75 ±3.30） % of control cells and decreased NF-H phosphorylation was also observed following mipafox treatment. Neither mipafox nor paraoxon had significant effect on the levels of actin. CONCLUSION Mipafox inhibited the neural differentiation in human neuroblastoma SK-N-SH cell via the decrease in total NF-H and NF- H phosphorylation levels, which was associated with the inhibition of NTE activity.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2006年第6期494-499,共6页
Chinese Journal of Pharmacology and Toxicology
基金
重庆市自然科学基金资助项目(CSTC2005BB5072)~~
关键词
杀虫药
有机磷
成神经细胞瘤
神经病
靶标酯酶
insecticides, organophosphate
neuroblastoma
neuropathy target esterase
