去水卫矛醇应用于斑马鱼胚胎的安全性评价

目的:利用斑马鱼胚胎评价抗肿瘤药物去水卫矛醇(DAG)的安全性。方法:以受精后6 h(6 hpf)发育正常的斑马鱼胚胎作为实验对象,分别暴露于A,B两个不同实验组的DAG溶液中,A组:98.00,110.00,125.00,141.00,160.00 mg·L~(-1)的低质量浓度剂量组,B组:661.00,871.00,1 148.00,1 514.00,1 995.00 mg·L~(-1)的高质量浓度剂量组,两组均设空白组,每隔24 h更换一次DAG溶液。观察药物处理至不同时间点时斑马鱼胚胎的发育情况,计算半数致死浓度(LC50),半数致畸效应浓度(EC50),治疗指数(TI)和安全指数(SI)。结果:B组斑马鱼胚胎在48 hpf时心率随着DAG浓度增大而下降,48,72,96 hpf孵化率随着观察时间点不同呈现不同趋势,与空白组比较有统计学差异(P0.01)。斑马鱼胚胎的畸形率和死亡率均随着给药浓度增大和给药时间的延长而增加(P0.01)。72 hpf的LC50为1 851.33 mg·L~(-1),EC50为184.50 mg·L~(-1),TI为10.03 mg·L~(-1),SI为1.59 mg·L~(-1);96 hpf的LC50为1 071.96 mg·L~(-1),EC50为74.76 mg·L~(-1),TI为14.34 mg·L~(-1),SI为2.94 mg·L~(-1);6 dpf的LC50为133.12 mg·L~(-1)。结论:DAG的浓度与斑马鱼胚胎发育的毒性存在量效关系;DAG作用时间长短与斑马鱼胚胎发育毒性也存在明显的剂量效应。DAG安全性评价指数相对较高。

Safety Evaluation of Dianhydrogalactitol to Zebrafish Embryos

Objective: To evaluate the safety of dianhydrogalactitol (DAG) in zebrafish embryos. Method: The 6 hours post fertilization (hpf), normally developed zebrafish embryos were exposed to DAG solution in two different experimental groups (A and B). Group A was a low-dose concentration group:98.00, 110.00, 125.00, 141.00, 160.00 mg·L^-1, while group B was a high-dose concentration group:661.00, 871.00, 1 148.00, 1 514.00, and 1 995.00 mg·L^-1. The corresponding blank controls were set for both groups. The DAG solution was refreshed every 24 h. Microscope was used to observe embryos development at different treating periods. Finally, the median lethal concentration (LC₅₀), median effect concentration (EC₅₀), therapeutic index (TI) and safety index (SI) were calculated. Result: Zebrafish heart rate of group B at 48 hpf was decreased with the increase of concentration of DAG solution. In addition, 48, 72, 96 hpf hatchabilities showed different trends at different observation time points (48, 72, 96 hpf). Difference was statistically significant compared with controls in both groups (P<0.01). Deformity rate and mortality rate of zebrafish embryos were increased with the increasing concentration of DAG solution and extended medication time (P<0.01). The LC₅₀ was 1 851.33 mg·L^-1, EC₅₀ of 184.50 mg·L^-1, TI of 10.03 mg·L^-1, and SI of 1.59 mg·L^-1at 72 hpf. LC₅₀ at 96 hpf was 1 071.96 mg·L^-1, EC₅₀ of 74.76 mg·L^-1, TI of 14.34 mg·L^-1, and SI of 2.94 mg·L^-1. LC₅₀ at 6 dpf was 133.12 mg·L^-1. Conclusion: There is a significant dose-dependent relationship between the concentration of DAG solution and the toxicity in zebrafish embryos development. There is also a significant dose-dependent relationship between the exposure time and the toxicity. In conclusion DAG's safety evaluation index is relatively high.