三邻甲苯基磷酸酯对人成神经瘤SH-SY5Y细胞分化的影响

Effect of Tri-ortho-cresyl Phosphate on the Differentiation of Human Neuroblastoma SH-SY5Y Cells

目的观察三邻甲苯基磷酸酯(Tri-ortho-cresyl phosphate,TOCP)对人成神经瘤SH-SY5Y细胞形态分化的影响,为更全面的了解TOCP的毒性提供依据。方法选用人成神经瘤细胞SH-SY5Y为细胞模型,以全反式维甲酸(ATRA)为诱导分化工具药物,在诱导中以及诱导后以不同浓度TOCP作用,动态观察SH-SY5Y细胞形态学变化,台盼蓝拒染法检测死亡细胞百分比。结果 ATRA对SH-SY5Y细胞作用7 d可导致细胞出现长出较长突起的分化特征,在诱导分化过程中及诱导分化后以不同剂量TOCP作用于细胞,各剂量组与对照组相比,无论是突起长度还是分化细胞比例差异均有统计学意义(P<0.05),其中,0.6 mmol/L组抑制效果最强。结论 TOCP可抑制SH-SY5Y细胞的分化,并呈现一定时间和剂量依赖关系。

Objective To study the effect of tri-ortho-cresyl phosphate（TOCP） on differentiation of human neuroblastoma SH-SY5Y cells,and to provide the evidence for comprehensively understanding the toxicity of TOCP.Methods SH-SY5Y cells were induced to differentiate by 10 mol/L all-trans retinoic acid（ATRA） for 7 days.During or after cell differentiating,SH-SY5Y cells were incubated with TOCP in different concentrations（0,0.2,0.4,and 0.6 mmol/L）.The cells were fixed in methanol and stained with Coomassie Blue R250.Percent of differentiated cells was counted and length of the longest neurite on each differentiated cell was measured under inverted phase-contrast microscopy.Trypan blue exclusion was used to detect the antiproliferative effect on SH-SY5Y cells.Results ATRA could induce the morphological differentiation of axon-like growth of SH-SY5Y cells after 7 days of treatment.Not only during cell differentiating but also after cell differentiating,there were statistically significant differences in length of neurite and percents of differentiated cells between each TOCP treatment group and control group（P0.05）.The inhibition on differentiation of 0.6 mmol/L TOCP group was the most efficacious.Conclusions TOCP could affect retinoic acid-induced neuronal differentiation of SH-SY5Y cell and cause the inhibition of axon-like growth in a time and dose dependent manner.