摘要
目的探究氧化钕对斑马鱼早期发育的影响,揭示纳米氧化钕心脏毒性机制。方法将状态良好的胚胎按40个/孔置于6孔板中,分别暴露于终浓度为0(对照)、50、100、200、400、800 mg/L纳米氧化钕悬液,并轻轻摇匀,于28℃孵育箱中培养,每两天换液一次,连续染毒5 d。观察斑马鱼胚胎死亡、畸形情况,实时监控斑马鱼胚胎心率变化情况,原位杂交染色观察心脏肌球蛋白轻链2(cardiac myosin light chain2,cmlc2)基因的表达情况。通过TUNEL染色方法检测细胞凋亡情况。结果与对照组相比,各浓度纳米氧化钕染毒组斑马鱼胚胎的死亡率和畸形率均较高,差异具有统计学意义(P<0.05);且随着纳米氧化钕染毒浓度的升高,斑马鱼胚胎的死亡率及畸形率呈上升趋势。200 mg/L纳米氧化钕染毒组斑马鱼胚胎的心率降低,差异有统计学意义(P<0.05)。原位杂交结果显示:200 mg/L纳米氧化钕染毒组病理性损害较100 mg/L染毒组和对照组更明显,同时,200 mg/L纳米氧化钕染毒组cmlc2基因表达量也高于对照组和100 mg/L纳米氧化钕染毒组。TUNEL染色结果表明,100、200 mg/L纳米氧化钕染毒组斑马鱼胚胎TUNEL染色阳性心肌细胞数均较高,差异有统计学意义(P<0.05)。结论纳米氧化钕染毒可能是通过上调斑马鱼胚胎心脏发育标记基因cmlc2,导致心肌细胞凋亡,从而发挥心脏毒性作用。
Objective To explore the effects of nano-neodymium oxide on the early development of zebrafish,and further to reveal the mechanism of cardiac toxicity.Methods The well-developed zebrafish embryos were placed in the 6-well plates,40 embryos in each well,and exposed to the nano-neodymium oxide suspension of the 0(the control group),50,100,200,400 and800 mg/L respectively,at 28℃incubator for five consecutive days.The suspension was shaken slightly and changed every two days.Then the death and malformation of zebrafish embryos were observed,and the change of heart rate was observed with real time monitoring.Subsequently,the expression of cmlc2 gene was determined by using in situ hybridization and the apoptosis level was detected by TUNEL staining.Results Compared with the control group,the mortality and malformation rate of zebrafish embryos in nano-neodymium oxide-exposed groups were significantly higher(P<0.05)with a dose dependent manner,the mortality and malformation rate of zebrafish embryos increased with the increase of nano-neodymium oxide exposure doses.The heart rate of zebrafish embryos in the 200 mg/L nano-neodymium oxide group decreased significantly(P<0.05).The pathological changes were more obvious in the 200 mg/L group compared with the control and the 100 mg/L group accordance to the results of in situ hybridization,and the cmlc2 gene expression was higher than that of the control group and the 100 mg/L group.The number of TUNEL-positive cardiomyocytes(red dotted distribution)in zebrafish embryos exposed to 100 and 200 mg/L nano-neodymium oxide was significantly higher(P<0.05).Conclusion Nano-neodymium oxide may present heart toxic effect through up-regulation of zebrafish embryo heart development marker gene cmlc2 and the related myocardial cell apoptosis.
作者
陈昱
舒凡
许迎春
郭宣娇
安慧荣
赵立东
巴德仁贵
谢伟
邵国
吴刚
贾小娥
CHEN Yu;SHU Fan;XU Ying-chun;GUO Xuan-jiao;AN Hui-rong;ZHAO Li-dong;BADE Ren-gui;XIE Wei;SHAO Guo;WU Gang;JIA Xiao-e(Biomedicine Research Center,Neuroscience Institute,Baotou Medical College,Baotou,Inner Mongolia 014040,China;不详)
出处
《环境与健康杂志》
CAS
北大核心
2020年第2期122-125,188,共5页
Journal of Environment and Health
基金
国家自然科学基金(81660204,81160341,81901918)
包头医学院科学研究基金(BYJJ-YF 201610)
内蒙古自治区青年科技英才(NJYT-17-B32)
内蒙古自治区草原英才青年创新创业人才一层次(Q2017047)
内蒙古自治区研究生创新项目(201610127)
关键词
纳米氧化钕
斑马鱼
心率
心脏肌球蛋白轻链2
Nano-neodymium oxide
Zebrafish
Heart rate
Cardiac myosin light chain2
作者简介
陈昱(1993-),男,硕士研究生,从事神经与发育研究;通讯作者:贾小娥,E-mail:evangeline2004@163.com;通讯作者:吴刚,E-mail:wugang0525@163.com
