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目的建立基于高分化PC12细胞模型的神经毒性物质高内涵筛选方法,并对该方法进行评价。方法应用高内涵筛选技术定量检测11种药物对高分化PC12细胞的细胞数量、线粒体质量、细胞核面积、细胞膜通透性、Mn SOD水平、γH2A.X水平、细胞面积、细胞圆度、细胞宽度、细胞长度和细胞宽长比等参数的影响。结果安眠酮、多虑平和地西泮等在抑制细胞存活的同时,引起线粒体质量改变,膜通透性增大,Mn SOD水平、γH2A.X水平升高,细胞长度减小、圆度增大、面积减小等;苯巴比妥、异戊巴比妥能够促进细胞存活或促进细胞神经突起生长。结论与文献已有数据进行比较,神经毒性检测正确率高,多参数表征的一般毒性较单一参数表征的一般毒性检测正确率要高。可使用高分化PC12细胞作为高内涵筛选的神经毒性评价模型,以细胞数量、线粒体质量、Mn SOD水平和γH2A.X水平作为细胞毒性指标,以细胞宽长比作为神经毒性指标,对神经毒性物质进行有效筛选。
Objective To establish a high-content screening method for neurotoxic substances based on a well-differentiated PC12 cell model and evaluate the method. Methods The high content screening technique was used to quantitatively detect the cell number, mitochondrial mass, nucleus area, cell membrane permeability, Mn SOD level, γH2A.X level, cell area, cell roundness, cell width, Cell length and cell length ratio and other parameters. Results Both methamphetamine, doxepin and diazepam could inhibit the cell survival while inducing the change of mitochondrial mass, membrane permeability, Mn SOD level, γH2A.X level, cell length decreasing and roundness increasing , Area reduction, etc .; phenobarbital, amobarbital can promote cell survival or promote cell neurite growth. Conclusions Compared with the existing data in the literature, neurotoxicity detection accuracy is high, the general toxicity of multi-parameter characterization is higher than the general toxicity detection of single parameter characterization. Well-differentiated PC12 cells can be used as a neurotoxicity evaluation model for high content screening. Taking the cell number, mitochondrial mass, Mn SOD level and γH2A.X level as cytotoxicity indexes and the cell aspect ratio as a neurotoxicity index, neurotoxic substances Effective screening.