基于斑马鱼幼鱼的抗氧化物筛选模型建立及其应用

Establishment and application of antioxidant screening model based on zebrafish larva

目的 建立一种基于斑马鱼幼鱼的抗氧化物质识别模型,并初步应用于候选物质的筛选。方法 以受精后3 d的斑马鱼幼鱼为受试对象,以12种已知抗氧化物、2种氧化物和4种候选物为受试物,将幼鱼置于含有受试物的养鱼水中处理20 h,根据幼鱼的急性毒性表现确定每种受试物的最大耐受剂量,并以此为最高剂量向下设计多个剂量组处理幼鱼,同时在培养体系中添加活性氧(ROS)指示剂CM-H_(2)DCFDA,20 h后检测处理体系的荧光强度,并计算自由基清除率。结果 12种已知抗氧化物中的11种抗氧化物处理的幼鱼荧光强度降低,表现出清除ROS的作用,2种氧化物增加幼鱼荧光强度,4种候选物中有3种具有清除ROS的作用。结论 该试验体系具有较好的灵敏度。用该体系对实验室的开发产物进行筛选,结果提示其具有一定的应用价值。

Objective To establish a zebrafish larva model for identifying antioxidants and screen candidate substances.Methods Zebrafish larvae with 3 d post fertilization(dpf)were treated with the fish culture water containing 12 known antioxidants,2 oxidants and 4 candidate substances for 20 h. The maximum tolerant concentration(MTC)was determined by the acute toxic effect including death. With the highest dose of MTC,different dosages of tested substance were used to treat 3dpf larvae respectively,and the reactive oxygen species(ROS)indicator CM-H_(2)DCFDA was added to the water at the same time. After 20 h treatment,fluorescent density was detected to calculate ROS clearance rate.Results Eleven tested known antioxidants showed lower fluorescent density in treated zebrafish larvae compared to the control,which suggested their ROS clearance potency. Two oxidants increased the fluorescent density,and three of four candidate substances were found to decrease fluorescent density in zebrafish larvae.Conclusion The study provided a sensitive test system for screening candidate antioxidants. The system was used to screen the products developed in the laboratory,and the results showed that it had certain application value.