应用鸡胚尿囊绒膜锥虫蓝吸收法评价化学物质的眼刺激性

Chorioallantoic membrane trypan blue test predicts the potential human eye irritation induced by various chemicals

背景:传统多采用兔眼刺激实验(Draize实验)评价化学物质眼刺激性,即将化学物涂进兔眼睛后观察其变化,这种方法费时,费力,且给动物带来很大痛苦。各国实验室都在寻找动物替代实验的方法。目的:采用鸡胚尿囊绒膜-锥虫蓝吸收实验作为兔眼刺激实验替代方法评价化学物眼刺激性的可行性。设计、时间及地点:观察实验,于2008-05/09在中国检验检疫科学研究院毒理学实验室完成。材料:无特定病原体级受精鸡卵,9日龄,购于北京梅里亚通实验动物技术有限公司。方法:将1g/L的锥虫蓝溶液滴加在受化学物质刺激损伤的鸡胚尿囊绒膜上,通过计算鸡胚尿囊绒膜受损伤后吸收锥虫蓝的量,半定量测定化学物质对鸡胚尿囊绒膜的刺激性,并将所得结果与体内兔眼刺激实验评分结果进行比对,作相关性分析。同时预测了5种常见有机溶剂的眼刺激性。主要观察指标:鸡胚尿囊绒膜受刺激损伤后吸收锥虫蓝的量。结果:鸡胚尿囊绒膜-锥虫蓝吸收法与体内兔眼刺激实验评分结果相关性很好(R2=0.9259)。5种常见有机溶剂未稀释时,甘油无眼刺激性,丙酮、乙醇、二甲基亚砜、甲醇均有较高刺激性。当稀释为1%后,丙酮、乙醇、二甲基亚砜无眼刺激性,但甲醇刺激性仍然较高。结论:鸡胚尿囊绒膜-锥虫蓝吸收法可以代替兔眼刺激试验对化学物质眼刺激性进行评价。

BACKGROUND： Draize test has been used to predict eye potential irritation of chemicals, limiting by time-consuming, strenuous, and causing great pain for animals. At present, studies based on alternative models to replace this assay are in development. OBJECTIVE： To use the hen＇egg test-chorioallantoic membrane trypan blue （CAM-TB） as an alternative method of Draize eye irritation test to test the chemical eye irritation. DESIGN, TIME AND SETTING： The alternative experiment was performed at the Laboratory of Toxicology, Chinese Academy of Inspection and Quarantine, from May to September 2008. MATERIALS： SPF hatching eggs of 9-day old was bought from MeriaI-Vital Laboratory Animal Technology Co., Ltd. METHODS： Using the chorioallantoic membrane trypan blue test, trypan blue solution were placed onto the CAM which injured by chemicals, the irritation potential of chemicals to CAM were calculated using the amount of trypan blue （nmol） absorbed by the CAM. The results were compared with Draize eye irritation test. Moreover eye irritations of 5 common solutions were forecasted by the CAM-TBS test. MAIN OUTCOME MEASURES： The amount of trypan blue （nmot） absorbed by the CAM. RESULTS： The correlation coefficient between CAM-TB score and Draize eye irritation was 0.925 9. Glycerol had no eye irritation potential, while acet, alcohol, methanol and DMSO had. When they were diluted to 1%, the eye irritation of acet, alcoho and DMSO were little, while methanol had eye irritation still. CONCLUSION： CAM-TBS can be used as an alternative method of Draize eye irritation to test chemical eye irritation.