细辛脑对斑马鱼胚胎发育及运动行为的影响  

Influence of asarone on zebrafish embryonic development and behavior

作　　者：尚艳楠 蒲韵竹[2] 李小满[2,4] 陈怡君[3] 刘萍[3] 薛江东[1] 李前[2] 黄春倩[2] 马德慧[1] 赵宝全[2] 

机构地区：[1]内蒙古民族大学,通辽028000 [2]抗药物与毒理学国家重点实验室,军事医学科学院毒物药物研究所,北京100850 [3]解放军总医院海南分院药剂科,三亚572013 [4]中国科学院近代物理研究所,兰州730000

出　　处：《国际药学研究杂志》2015年第2期176-182,共7页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金资助项目(81173572)

摘　　要：目的研究细辛脑对斑马鱼胚胎发育及运动行为的影响。方法以受精后3h(hpf)斑马鱼胚胎为实验模型,分别暴露于不同浓度(25、50、100、200和400μmol/L)细辛脑溶液中,以不加细辛脑为对照组,每隔24 h更换1次细辛脑溶液,分别在24、48、72和96 hpf等时间点用显微镜观察班马鱼胚胎发育形态,记录24 hpf胚胎自主抽动次数,48 hpf心率、畸形率和死亡率,72和96 hpf孵化率和死亡率,利用RT-q PCR检测96 hpf斑马鱼胚胎Sepn1基因表达情况,采用Noldus斑马鱼幼鱼行为视频跟踪系统评价细辛脑暴露对幼鱼运动行为的影响。结果细辛脑各实验组斑马鱼胚胎自主抽动次数随浓度增加而降低,100、200和400μmol/L组与对照组相比有极显著性统计学差异(P<0.01);细辛脑各实验组斑马鱼48 hpf心率显著降低,与对照组相比均有显著统计学差异(P<0.01);72和96 hpf斑马鱼幼鱼出现了脊柱弯曲、心包水肿、卵黄囊水肿等畸形形态;与对照组相比,100、200和400μmol/L组96 hpf孵化率均下降,具有显著性统计学差异(P<0.01);与对照组相比,400μmol/L组死亡率降低,有统计学差异(P<0.05);随着细辛脑浓度增加,96 hpf斑马鱼幼鱼运动速度和距离明显减小,200和400μmol/L组与对照组相比均有统计学差异(P<0.05);同时200和400μmol/L浓度组活跃度降低,与对照组相比具有显著差异(P<0.01);96 hpf时,细辛脑各浓度组胚胎Sepn1基因表达下降,其中100μmol/L组与对照组相比有统计学差异(P<0.05),而200和400μmol/L组与对照组相比差异更显著(P<0.01)。结论细辛脑对斑马鱼胚胎和幼鱼有致畸作用,并且影响其运动行为,建议婴幼儿慎用细辛脑。Objective To investigate the effect of asarone on the zebrafish embryonic development and behavior. Methods The 3 hours post fertilization（hpf）zebrafish embryos were exposed to different concentrations（0,25,50,100,200 and 400 μmol/L）of asarone solution. The asarone solution was replaced each 24 h. Microscope was used to observe embryos morphology at 24,48,72 and 96 hpf,respectively. Spontaneous movements at 24 hpf,heart rate at 48 hpf,hatch rate,deformity rate and mortality rate were evaluated.The sepn1 gene expression of zebrafish in 96 hpf was detected by RT-q PCR. With Noldus tracking system the behaviors of zebrafish larvae exposed to asarone were recorded. Results After zebrafish embryos were exposed to the different concentrations of asarone,their spontaneous movements were decreased. Compared with the control group,the 100,200 and 400 μmol/L groups were significantly decreased（P〈0.01）. There was a significant difference（P〈0.01） in 48 hpf heart rate between all asarone groups and control group. Spine curvature,pericardial edema,yolk sac edema and other deformity were observed at 72 hpf and 96 hpf.Compared with the control group,the 100,200 and 400 μmol/L groups showed significanly decreased hatch rate（P〈0.01）. There was a significant difference（P 0.05） in mortality rate between the 400 μmol/L group and control group. With concentrations of asarone increasing, the speed and distance of movement of 200 and 400 μmol/L group were significantly reduced compared to the control group（P〈0.05）. There was a significant difference between the 200,400 μmol/L groups and the control group in activity（P〈0.01）.At 96 hpf,the expression of sepn1 gene in arone groups were decreased compared to control group, especially the 100 μmol/L group（P〈0.05）,and the 200 and 400 μmol/L groups（P〈0.01）. Conclusion Asarone had teratogenic effect on zebrafish embryos and larvae,and inhibitory effect on larvae behavior.We should be careful to use asarone in infant medication.

关 键 词：细辛脑 斑马鱼 胚胎发育 婴幼儿用药 

分 类 号：R285.5[医药卫生—中药学]