影响斑马鱼幼鱼心脏发育和运动行为的囊胚期暴露二甲亚砜浓度探讨  

Effects of concentrations of dimethyl sulfoxide on zebrafish larvae′s heart development and motor behavior during blastocyst stage

作　　者：山花 贺亚南 冯元州 何嘉玲 朝宝 刘彩萍 王永安 董武 张东威[1] 赵宝全 SHAN Hua;HE Ya-nan;FENG Yuan-zhou;HE Jia-ling;CHAO Bao;LIU Cai-ping;WANG Yong-an;DONG Wu;ZHANG Dong-wei;ZHAO Bao-quan(Clinical Medical College,Inner Mongolia University for Nationalities,Tongliao 028000,China;State Key Laboratory of Toxicology and Medical Countermeasures,Institute of Toxicology and Pharmacology,Academy of Military Medical Sciences,Beijing 100850,China;National Research Institute for Family Planning,Beijing 100081,China)

机构地区：[1]内蒙古民族大学临床医学院,内蒙古通辽028000 [2]军事科学院军事医学研究院毒物药物研究所,抗毒药物与毒理学国家重点实验室,北京100850 [3]国家卫生健康委科学技术研究所,北京100081

出　　处：《中国药理学与毒理学杂志》2022年第7期495-501,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(82060816)。

摘　　要：目的探讨影响斑马鱼幼鱼心脏发育和运动行为的二甲亚砜(DMSO)浓度。方法受精后4 h斑马鱼囊胚暴露含终浓度(V/V)为0.15%,0.30%,0.45%和0.60%DMSO的E3孵化液中,持续6 d。用Noldus斑马鱼幼鱼行为学分析系统,于暴露2,3和4 d,记录胚胎和幼鱼心率;暴露6 d进行行为学实验,检测幼鱼游动时间、游动速度、游动距离和游动频次;暴露2,4和6 d,RT-PCR检测心脏发育相关基因心房特异性肌球蛋白重链(amhc)和心室特异性肌球蛋白重链(vmhc)基因及运动相关基因谷氨酸脱羧酶1b(gad1b),γ氨基丁酸A型受体亚基α1(gabra1)和γ氨基丁酸转氨酶(abat)mRNA表达水平。结果与正常对照组相比,0.3%~0.60%DMSO对胚胎(暴露2 d)心率无影响;幼鱼(暴露4 d)心率显著降低(P<0.05,P<0.01)。行为学实验结果显示,与正常对照组相比,暴露6 d,0.3%~0.60%DMSO组幼鱼游动时间、游动速度、游动距离和游动频次显著下降(P<0.05)。RT-PCR结果显示,与正常对照组相比,暴露2 d,仅0.60%DMSO组胚胎vmhc mRNA表达水平显著降低(P<0.05),DMSO 0.15%~0.60%组gad1b mRNA表达水平均显著降低(P<0.01),仅0.45%DMSO组gabra1 mRNA表达水平显著降低(P<0.05),0.15%DMSO组abat mRNA表达水平显著上调(P<0.01),而DMSO 0.45%和0.60%组显著下调(P<0.01);暴露4 d,0.15%~0.60%DMSO组gad1b,gabra1和abat mRNA表达水平均显著下降(P<0.01);暴露6 d,0.15%~0.60%DMSO组gad1b mRNA表达水平均显著下降(P<0.01),abat mRNA表达水平显著上调(P<0.01),0.60%DMSO组gabra1 mRNA表达水平显著上调(P<0.01)。结论DMSO浓度≥0.3%,持续暴露6 d,对斑马鱼心率、运动行为、心脏发育相关基因和运动相关基因表达有影响。OBJECTIVE To explore the effects of different concentrations of dimethyl sulfoxide(DMSO)on the heart development and motor behavior of zebrafish juveniles.METHODS Zebrafish blastocysts at 4 h after fertilization were exposed to E3 incubation medium containing final concentrations(V/V)of 0.15%,0.30%,0.45%and 0.60%DMSO for 6 d.Using the Noldus behavioral analysis system of zebrafish larvae,heart rates of embryos and larvae were recorded on the 2nd,3rd and 4th days of exposure while behavioral experiments were performed on the 6th day of exposure to detect the duration,speed and distance and frequency of swimming of the larvae.On the 2nd,4th and 6th days of exposure,RT-PCR was used to detect the expression levels of cardiac development-related genes-atrial-specific myosin heavy chain(amhc),and ventricle-specific myosin heavy chain(vmhc),and those of exercise-related genes-glutamate decarboxylase 1b(gad1b),γ-aminobutyric acid type A receptor subunitα1(gabra1)andγ-aminobutyric acid transaminase(abat)mRNA.RESULTS Compared with the normal control group,0.3%-0.60%DMSO had no effect on the heart rate of embryos(after exposure of 2 d),but juveniles(after exposure of 4 d)had significantly-decreased heart rate(P<0.05,P<0.01).The results of behavioral experiments showed that compared with the normal control group,the duration,speed,distance and frequency of swimming among the juvenile fish exposed to 0.3%-0.60%DMSO for 6 d decreased significantly(P<0.05).RT-PCR results showed that compared with the normal control group,only the 0.60%DMSO group had a significant decrease in the expression level of vmhc mRNA(P<0.05),while the 0.15%-0.60%DMSO group had a significant decrease in the expression level of gad1b m RNA after 2 d of exposure(P<0.01).Only the 0.45%DMSO group had a significantly decreased expression level of gabra1 mRNA(P<0.05).The expression level of abat mRNA in the 0.15%DMSO group was significantly increased(P<0.01),but was significantly down-regulated in the 0.45%and 0.60%DMSO groups(P<0.01).After 4 d of exposure,

关 键 词：二甲亚砜 斑马鱼 心脏 运动行为 

分 类 号：R965[医药卫生—药理学]