一种基于神经细胞膜电位荧光探针的河豚毒素毒性检测细胞模型

A cell model for tetrodotoxin toxicity detection based on fluorescent probe of nerve cell membrane potential

目的:本实验建立一种有效的河豚毒素毒性检测细胞模型,为河豚毒素解毒药物高通量筛选奠定基础。方法:用不同浓度膜电位荧光探针DiBAC4(3)在全黑底透96孔细胞培养板中孵育SH-SY5Y人神经母细胞瘤细胞,利用不同浓度藜芦定使细胞去极化,利用荧光酶标仪测定不同浓度河豚毒素对细胞膜电位的影响。结果:以DiBAC4(3)1000nmol/L、藜芦定200μmol/L、河豚毒性100μmol/L作用于SH-SY5Y细胞,观察到河豚毒素使该细胞膜电位变化最显著。结论:通过测量毒素诱导的膜电位变化可以初步预测和量化河豚毒素的毒性,可用于筛选解毒剂。该方法操作简便,并且可以实现高通量自动化筛选。

Objective:This experiment aims to establish an effective cell model for the detection of tetrodotoxin toxicity.It provides a method for high-throughput screening of tetrodotoxin detoxification drugs.Methods:SH-SY5 Y human neuroblastoma cells were cultured with DiBAC4(3)fluorescent probe of membrane potential with different concentrations in a 96-well black cell plate.The cells were depolarized with different concentrations of veratrol.The effects of different concentrations of tetrodotoxin on cell membrane potential were measured by fluorescein microplate.Results:When 1000 nmol/L DiBAC4(3),200μmol/L veratridine,100μmol/L and tetrodotoxin were applied to SH-SY5 Y cells,it was observed that tetrodotoxin caused the most significant changes in the cell membrane potential.Conclusion:The toxicity of tetrodotoxin can be predicted and quantified by measuring the change of toxininduced membrane potential which can be used to screen antidotes.This method is easy to operate and can realize high-throughput automatic screening.