CAJ | 학술논문

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-qPCR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。
목적：연구세신뇌대반마어배태발육급운동행위적영향。방법이수정후3h（hpf）반마어배태위실험모형，분별폭로우불동농도（25、50、100、200화400μmol/L）세신뇌용액중，이불가세신뇌위대조조，매격24 h경환1차세신뇌용액，분별재24、48、72화96 hpf등시간점용현미경관찰반마어배태발육형태，기록24 hpf배태자주추동차수，48 hpf심솔、기형솔화사망솔，72화96 hpf부화솔화사망솔，이용RT-qPCR검측96 hpf반마어배태Sepn1기인표체정황，채용Noldus반마어유어행위시빈근종계통평개세신뇌폭로대유어운동행위적영향。결과세신뇌각실험조반마어배태자주추동차수수농도증가이강저，100、200화400μmol/L조여대조조상비유겁현저성통계학차이（P＜0.01）；세신뇌각실험조반마어48 hpf심솔현저강저，여대조조상비균유현저통계학차이（P＜0.01）；72화96 hpf반마어유어출현료척주만곡、심포수종、란황낭수종등기형형태；여대조조상비，100、200화400μmol/L조96 hpf부화솔균하강，구유현저성통계학차이（P＜0.01）；여대조조상비，400μmol/L조사망솔강저，유통계학차이（P＜0.05）；수착세신뇌농도증가，96 hpf반마어유어운동속도화거리명현감소，200화400μmol/L조여대조조상비균유통계학차이（P＜0.05）；동시200화400μmol/L농도조활약도강저，여대조조상비구유현저차이（P＜0.01）；96 hpf시，세신뇌각농도조배태Sepn1기인표체하강，기중100μmol/L조여대조조상비유통계학차이（P＜0.05），이200화400μmol/L조여대조조상비차이경현저（P＜0.01）。결론세신뇌대반마어배태화유어유치기작용，병차영향기운동행위，건의영유인신용세신뇌。
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-qPCR. 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.