观察时间和换液频率对斑马鱼评价模型的影响
|
摘要
目的通过设置不同的观察时间和换液频率,研究两者对斑马鱼模型的影响,优化评价模型。方法分别进行斑马鱼胚胎给药5 d,每天换液、实验7 d,每天换液及给药5 d不换液实验,检测3种实验方法中各组斑马鱼的死亡率、自由活动、形态发育和骨骼发育情况,并对3种实验方法的结果进行比较。结果较给药5 d每天换液实验,延长观察时间,使斑马鱼死亡率升高、畸形程度增加、骨骼矿化呈现出一定的剂量趋势,适当延长观察时间,可使药物的毒性作用表达更加充分;不换液实验中,斑马鱼死亡率略低、畸形程度减轻,不换液给药一定程度上降低了药物的毒性作用。结论定期更换药液,保证暴露浓度和适当延长观察时间,可使斑马鱼药物毒性评价模型的结果更加准确、可靠。
Aim To study the effects of different observation time and liquid exchange frequency on zebrafish model and optimize the evaluation model. Methods De-membrane zebrafish embryos of 6 h fertilization were exposed for 5 days and changed solution for 4/5 every 24 hours. The mortality rate was measured during the experiment, and the autonomous activity, morphological development and bone development of young fish were detected at the end of the experiment. Meanwhile, the experiment was carried out for 7 days too, that was, after 5 days of exposure, the observation time was prolonged until the 7 th day. No liquid exchange test was no change of solution during 5 days of exposure. The other test methods were the same, and the results of the three test methods were compared. Results Compared with the daily liquid exchange test for 5 days, prolonged observation time increased the mortality and the degree of deformity, showing a certain dose-effect of bone mineralization, and decreased the teratogenic dose of the positive drug all-trans retinoic acid(ATRA). Proper prolongation of observation time could make the expression of drug toxicity more fully. In the non-exchange test, the mortality of zebrafish was slightly lower, the degree of deformity was reduced, and the teratogenic dose of the positive drug ATRA increased. The toxicity of the drug was reduced to a certain extent. Conclusion The results of drug toxicity evaluation model of zebrafish can be more accurate and reliable if the exposure concentration of drug is maintained by regular change of liquid and proper prolongation of observation time.
Aim To study the effects of different observation time and liquid exchange frequency on zebrafish model and optimize the evaluation model. Methods De-membrane zebrafish embryos of 6 h fertilization were exposed for 5 days and changed solution for 4/5 every 24 hours. The mortality rate was measured during the experiment, and the autonomous activity, morphological development and bone development of young fish were detected at the end of the experiment. Meanwhile, the experiment was carried out for 7 days too, that was, after 5 days of exposure, the observation time was prolonged until the 7 th day. No liquid exchange test was no change of solution during 5 days of exposure. The other test methods were the same, and the results of the three test methods were compared. Results Compared with the daily liquid exchange test for 5 days, prolonged observation time increased the mortality and the degree of deformity, showing a certain dose-effect of bone mineralization, and decreased the teratogenic dose of the positive drug all-trans retinoic acid(ATRA). Proper prolongation of observation time could make the expression of drug toxicity more fully. In the non-exchange test, the mortality of zebrafish was slightly lower, the degree of deformity was reduced, and the teratogenic dose of the positive drug ATRA increased. The toxicity of the drug was reduced to a certain extent. Conclusion The results of drug toxicity evaluation model of zebrafish can be more accurate and reliable if the exposure concentration of drug is maintained by regular change of liquid and proper prolongation of observation time.
引文
[1] 许冰洁, 张立将, 李春启, 等. 斑马鱼胚胎评价5种药物的发育毒性与模型验证[J]. 中国药理学通报, 2016,32(1):74-9.[1] Xu B J, Zhang L J, Li C Q, et al. Model validation and evaluation of developmental toxicity of five drugs using zebrafish embryos[J]. Chin Pharmacol Bull, 2016,32(1):74-9.
[2] 吴玉琼, 陈 莹, 胡永乐, 等. 四种新型农药对斑马鱼胚胎发育的毒性效应[J]. 生物技术通报, 2017,33(6):155-61.[2] Wu Y Q, Chen Y, Hu Y L, et al. Toxic effects of four currently-used pesticides on zebrafish embryonic development[J]. Biotechnol Bull, 2017,33(6):155-61.
[3] Huang D, Li H, He Q, et al. Developmental toxicity of diethylnitrosamine in zebrafish embryos/ Juveniles related to excessive oxidative stress[J]. Water Air Soil Pollut, 2018, 229(3):81-91.
[4] 陈 颖, 王 茉, 宋 捷, 等. 补骨脂水煎液和醇提物对斑马鱼骨骼发育的影响及其毒性作用[J].中国药理学与毒理学杂志, 2017,31(6):661-9.[4] Chen Y, Wang M, Song J, et al. Effect of extract ethanol and water decoction of Psoralea corylifolia L. on bone development and their toxicities in zebrafish[J] . Chin J Pharm Toxicol, 2017,31(6): 661-9.
[5] 陈锡强, 王荣春, 王希敏, 等. 斑马鱼皮肤损伤模型的建立及分子机制[J]. 中国药理学通报, 2018, 34(3): 433-7.[5] Chen X Q, Wang R C, Wang X M, et al. Model establishment of skin damage and molecular mechanism on transgenic zebrafish embryos[J]. Chin Pharmacol Bull, 2018, 34(3): 433-7.
[6] Brannen K C, Panzica-Kelly J M, Danberry T L, et al. Development of a zebrafish embryo teratogenicity assay and quantitative prediction model[J]. Birth Defects Res B Dev Reprod Toxicol, 2010, 89(1): 66-77.
[2] 吴玉琼, 陈 莹, 胡永乐, 等. 四种新型农药对斑马鱼胚胎发育的毒性效应[J]. 生物技术通报, 2017,33(6):155-61.[2] Wu Y Q, Chen Y, Hu Y L, et al. Toxic effects of four currently-used pesticides on zebrafish embryonic development[J]. Biotechnol Bull, 2017,33(6):155-61.
[3] Huang D, Li H, He Q, et al. Developmental toxicity of diethylnitrosamine in zebrafish embryos/ Juveniles related to excessive oxidative stress[J]. Water Air Soil Pollut, 2018, 229(3):81-91.
[4] 陈 颖, 王 茉, 宋 捷, 等. 补骨脂水煎液和醇提物对斑马鱼骨骼发育的影响及其毒性作用[J].中国药理学与毒理学杂志, 2017,31(6):661-9.[4] Chen Y, Wang M, Song J, et al. Effect of extract ethanol and water decoction of Psoralea corylifolia L. on bone development and their toxicities in zebrafish[J] . Chin J Pharm Toxicol, 2017,31(6): 661-9.
[5] 陈锡强, 王荣春, 王希敏, 等. 斑马鱼皮肤损伤模型的建立及分子机制[J]. 中国药理学通报, 2018, 34(3): 433-7.[5] Chen X Q, Wang R C, Wang X M, et al. Model establishment of skin damage and molecular mechanism on transgenic zebrafish embryos[J]. Chin Pharmacol Bull, 2018, 34(3): 433-7.
[6] Brannen K C, Panzica-Kelly J M, Danberry T L, et al. Development of a zebrafish embryo teratogenicity assay and quantitative prediction model[J]. Birth Defects Res B Dev Reprod Toxicol, 2010, 89(1): 66-77.