中性红摄取实验预测化学品急性毒性研究  

Study on neutral red uptake cytotoxicity assay for acute toxicity prediction on chemicals

作　　者：曾丽海[1] 黄振烈[1] 杨杏芬[2] 葛怡琛[1] 殷霄[1] 赖关朝[1] 

机构地区：[1]广东省职业病防治院,广东省职业病防治重点实验室,广东广州510300 [2]广东省疾病预防控制中心,511430

出　　处：《中国职业医学》2014年第6期632-638,共7页China Occupational Medicine

基　　金：广东省职业病防治重点实验室(2012A061400007);广东省化学中毒与核辐射突发事件医学救援应急技术研究中心(2012A032100002);广东省医学科研基金(B2013020)

摘　　要：目的评估BALB/c小鼠胚胎成纤维细胞中性红摄取(3T3 NRU)实验预测化学品急性毒性的能力及其替代经典急性经口毒性实验的可行性。方法第1阶段从细胞毒性登记库(RC)中选取11种化学品进行暴露24.0、48.0、72.0 h的3T3 NRU实验,将所得半抑制浓度(IC50)与RC中大鼠急性毒性半数致死剂量(LD50)建立回归模型。第2阶段选取15种化学品进行暴露24.0、48.0、72.0 h的3T3 NRU实验,将所得IC50值根据RC回归模型计算LD50预测值,对受试化学品进行急性毒性分级,与经口急性毒性实验LD50值的急性毒性分级结果比较。结果第1阶段实验结果显示:暴露24.0、48.0、72.0 h的3T3 NRU实验IC50值均可与RC提供的LD50建立在RC回归模型可信区间内的回归模型[暴露24.0 h组:lg(LD50)=0.470×lg(IC50)+0.582,决定系数(R2)=0.937,F=132.805,P<0.05;暴露48.0 h组:lg(LD50)=0.475×lg(IC50)+0.638,R2=0.925,F=111.107,P<0.05;暴露72.0 h组:lg(LD50)=0.480×lg(IC50)+0.639,R2=0.933,F=125.514,P<0.05],3T3 NRU实验可应用RC回归模型预测急性毒性的LD50值。第2阶段实验结果显示:暴露24.0、48.0和72.0 h的3T3 NRU实验LD50预测值与经口急性毒性实验LD50值的急性毒性分级一致率分别为60.0%(9/15)、66.7%(10/15)和60.0%(9/15);3T3 NRU实验判断急性毒性为5级的化学品,其急性经口毒性实验的分级均为5级。结论 3T3 NRU实验适合于对低毒的化学品进行检测,可用RC回归模型预测3T3 NRU实验的急性毒性LD50值;对于预测毒性分级为4级及以下者,3T3 NRU实验的LD50预测值可作为经口急性毒性实验始剂量。Objective To evaluate the toxicity predictive ability of 3T3 neutral red uptake （NRU） cytotoxicity assay on chemicals and to discuss the possibility of 3T3 NRU as an alternative test for acute toxicity of chemicals. Methods At stage 1, 11 chemicals from registry of cytotoxicity （ RC ） were examined by 3T3 NRU cytotoxicity assay for 24.0, 48.0, and 72.0 hours, and regressive model was established between the half inhibitive concentration （ IC50 ） from 3T3 NRU cytotoxicity assay and median lethal dose （LD50） from RC by regressive analysis. At stage 2, 15 chemicals were detected by 3T3 NRU cytotoxicity assay for 24. 0, 48.0, and 72.0 hours. The regression model of RC was used to estimate the predicted value of LD50 with IC50 from 3T3 NRU cytotoxicity assay and the toxicity was classified. The results of toxicity classification from predicted value of LD50 were compared with classification from acute toxicity assay. Results The stage 1 results showed that IC50 from 3T3 NRU cytotoxicity assay had positive linear correlation with LD50 from RC [ 24. 0 hours, lg（LD50） =0. 470×lg（IC50） ＋0. 582, R^2 =0. 937, F = 132. 805, P 〈0. 05; 48.0 hours, lg（LD50 ） =0. 475×lg（ IC50 ） ＋ 0.638, R2 =0.925, F=111.107, P〈0.05; 72.0 hours： lg（LD50） =0.480 ×lg（lC50） ＋0.639, R2 =0.933, F= 125. 514, P〈0. 05 ], and the regression lines obtained from 3T3 NRU cytotoxicity assay were within the acceptance interval of RC model. LD50 of 3T3 NRU cytotoxicity assay could be predicted with the regression model of RC. The stage 2 results showed that the classification consistency between the results of 3T3 NRU eytotoxicity assay and acute oral toxicity assay were 60. 0% （24. 0 hours, 9/15） , 66.7% （48. 0 hours, 10/15 ） and 60. 0% （72. 0 hours, 9/15） respectively. The chemicals which were judged to be level 5 classified by 3T3 NRU assay were in the same class classified by acute oral toxicity assay. Conclusion 3T3 NRU eytotoxicity assay may be suitable to predict the chemicals with low

关 键 词：中性红摄取实验 急性毒性 细胞毒性实验 替代方法 化学品 

分 类 号：R114[医药卫生—卫生毒理学] R13[医药卫生—公共卫生与预防医学]