原代神经元及PC12细胞用于药物体外急性毒性评价的实验研究  

Experimental Study on Primary Rat Cortical Neurons and PC12 cells Used for the Evaluation of Acute Cytotoxicity

作　　者：巫玮[1] 周捷[1] 潘雪刁[1] 潘琴[1] 石磊[1] 臧林泉[1] 

机构地区：[1]广东药学院药科学院新药筛选中心／药理学教研室,广东广州510006

出　　处：《医学临床研究》2009年第3期369-373,共5页Journal of Clinical Research

基　　金：国家自然科学基金资助（No.30572177）;教育部重点基金资助（No.208105）

摘　　要：【目的】研究原代神经元及PC12细胞用以评价药物或化合物的急性毒性，以判断是否可以替代动物实验，为建立稳定的体外高通量筛选化合物和药物的急性毒性评价体系奠定基础。【方法】原代分离培养大鼠皮质神经元，传代培养大鼠类神经元细胞——PC12细胞，取17种药物和化合物，分别稀释不同浓度，与上述两种细胞分别孵育24h，与空白对照组比较，以MTT法检测药物／化合物对细胞的抑制率，据此量效曲线计算出IC50，与大鼠口服LD50进行相关性分析。【结果】体外神经元的IC50与体内LD50相关性为：R^2=0．7253，PC12细胞毒性检测结果的IC50与动物体内急性毒性LD50相关性为：R^2=0．6441，二者均有明显相关性。【结论】利用体外分离培养的原代神经元以及PC12细胞，在短时间（24h）内可以评价药物或化合物的急性毒性，并可以预测化合物对人及动物的毒性，基本可以替代或减少动物实验。[Objective]To study the primary culture of isolated neonate rat neurons and the passage culture of PC12 cells so as to evaluate acute cytotoxicity of medicines or compounds, and decide whether it can replace animal experiment, and provide the proof for establishing the high throughput screening method of acute toxicity of medicines or compounds. [Methods]Primary rat cortical neurons and PC12 cells were studied in culture to evaluate their poten- tial to the screening for cytotoxicity. Confluent monolayers were incubated in the absence or presence of increasing concentrations of test chemicals for 24 h, and the MTT assay was used to assess toxicity. Inhibitory concentrations （IC50） were extrapolated from concentration-effect curves after linear regression analysis. Seventeen chemicals were tested with each cell line and rat half lethal dose （LD50） were compared. [Results]IC50 of neurons in vitro was significantly correlated to LD50 of neurons in vivo （R^2= 0. 7253）. IC50 by cytotoxicity testing of PC12 was also signifi cantly correlated to LD50 of acute cytotoxicity in animal bodies （R^2=0. 6441）. [Conclusion] Primary neurons and PC12 cells may be used to evaluate the acute cytotoxicity of medicines and compounds in a short time（24h）, and can predict the compounds toxicity to human and animals. Therefore, it can replace or decrease animal experiments.

关 键 词：细胞毒性 免疫 神经元 PC12细胞 

分 类 号：R-332[医药卫生]