摘要
目的用斑马鱼模型对BCG-CpG-DNA进行安全性评价。方法以斑马鱼胚胎为实验材料,将BCG-CpG-DNA设置4个浓度组(0.15、1.5、15、75 mg/L),暴露斑马鱼,以胚胎培养液暴露为空白对照组,以三甲基氯化锡(TMT)和全氟辛烷磺酸盐(PFOS)暴露为阳性对照组,胚胎期6 hp(f受精后6 h)脱膜,8 hpf进行暴露毒性实验,研究其对斑马鱼发育、遗传、免疫以及行为的毒性影响。每组做3个重复,试验重复3次。结果未观察到BCG-CpG-DNA各浓度组的斑马鱼胚胎明显的畸形和孵化死亡情况,其自主运动、触摸运动、行为检测、免疫强度检测与空白对照组相比,差异均无统计学意义(P>0.05)。TMT对照组暴露对斑马鱼生长、发育、免疫及其神经等均有不同程度的影响,导致心胞肿大,对光暗刺激反应敏感,相对荧光强度明显增强,嗜中性粒细胞数量增多,对炎症的敏感性增强,与空白对照组相比,差异有统计学意义(P<0.05)。结论 BCG-CpG-DNA暴露对斑马鱼生长、发育、免疫及其神经均无明显的急性毒性作用,在斑马鱼的胚胎暴露中具有较好的安全性。
Objective To evaluate the safety of BCG-CpG-DNA by using zebrafish model.Methods The embryos of zebrafish were exposed to BCG-CpG-DNA at concentrations of 0.15,1.5,15 and 75 mg / L respectively,using those exposed to culture liquid as blank control,and those exposed to TMT and PFOS as positive controls.Membrane shedding test was performed 6 h,while exposure toxicity test 8 h post fertilization to investigate the toxic effect on development,genetics,immune status and behavior of zebrafish.Each test was performed for 3 times in triplicate.Results No obvious monstrosities or deaths during incubation were observed in various test groups,while the autonomous movement,touch movement,behavior and immune level showed no significant difference with those in blank control group（P 0.05）.However,TMT exposure showed effects at various degrees on the growth,development,immune status and nerve of zebrafish,causing enlargement of vesicula cardiacum,sensitivity to light and dark variation as well as increased relative fluorescence intensity,increased expression of neutrophil and enhanced sensitivity to inflammation,which showed significant difference with those in blank control group（P 0.05）.Conclusion BCG-CpG-DNA exposure showed no obviously acute toxic effect on the growth,development,immune status and nerve of zebrafish,indicating high safety.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第12期1618-1622,共5页
Chinese Journal of Biologicals
基金
"十二五"国家重大科技专项(2012ZX10004701-003)
"十一五"国家高技术研究发展计划(863计划)(2006AA02A240)
