基于斑马鱼模型的抗结核药物肝脏毒性比较研究

Comparative study on liver toxicity of anti-tuberculosis drugs based on the zebrafish model

目的以肝脏绿色荧光转基因斑马鱼Tg(L-FABP:EGFP)为模型,研究一线抗结核药物异烟肼、吡嗪酰胺和乙胺丁醇对斑马鱼肝脏的影响以及比较其肝毒性大小。方法用不同浓度的异烟肼、吡嗪酰胺和乙胺丁醇分别处理发育至72h的斑马鱼,于加药后24、48和72hpe(hour post-exposure)观察斑马鱼的死亡率、畸形率和肝脏形态变化情况,荧光显微镜下观察药物对斑马鱼肝脏荧光面积和荧光强度的影响。结果随着给药浓度的增加和给药时间的延长,异烟肼、吡嗪酰胺和乙胺丁醇导致斑马鱼死亡率、畸形率升高。在72hpe,与空白对照组相比,1、2.5和5mmol/L吡嗪酰胺组斑马鱼肝脏荧光面积显著下降,5mmol/L异烟肼组斑马鱼肝脏荧光面积显著下降,10、20和30mmol/L乙胺丁醇组斑马鱼肝脏荧光面积显著下降。5mmol/L浓度下,异烟肼导致斑马鱼肝脏荧光面积降低程度高于吡嗪酰胺。结论异烟肼、吡嗪酰胺和乙胺丁醇均有肝脏毒性,乙胺丁醇肝毒性最小。1和2.5mmol/L浓度下吡嗪酰胺导致斑马鱼发生肝损伤程度强于异烟肼,但5mmol/L下异烟肼肝脏损伤程度强于吡嗪酰胺,其有关机制需进一步研究。

Objective To research hepatotoxicity of the first-line anti-tuberculosis drugs(isoniazid,pyrazinamide,and ethambutol)using the transgenic zebrafish line Tg(L-FABP:EGFP).Methods Thirty zebrafish larvae were treated with different concentrations of isoniazid,pyrimidine,and ethambutol.The mortality,malformation rate,and liver morphology changes in zebrafish larvae were measured at 24,48 and 72hpe(hour post-exposure).The effects of the drugs on the fluorescence area and intensity of the zebrafish liver were observed using fluorescence microscope.Results Isoniazid,pyrazinamide and ethambutol led to the increase of mortality and malformation rate of zebrafish with the increase of drug concentration and duration.At 72hpe,the liver area of zebrafish in the 1,2.5 and 5mmol/L pyrazinamide group,5mmol/L isoniazid group and 10,20 and 30mmol/L ethambutol group significantly decreased compared with the control group.Isoniazid reduced the liver area of zebrafish more serious than pyrazinamide at the concentration of 5mmol/L.Conclusion Isoniazid,pyrazinamide,and ethambutol had liver toxicity,and ethambutol had the least hepatotoxicity.At 1 and 2.5mmol/L concentration,pyrazinamide caused higher liver damage than isoniazid,but isoniazid was more toxic than pyrazinamide at 5mmol/L.The relevant mechanism needs further study.