ABSTRACT
Objective To study the toxic effect ofNiuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40,200,400,800,1 200 tg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage,exceeding the normal effective dose).While,there was toxicity to embryo development at the concentration greater than 200 tg/mL.The head and eyes of the embryo were smaller,and the pupils were colorless.Zebrafish embryonic heart malformation was observed,heart rate was weak,blood pool congestion happened and back blood flow was not smooth.The abdominal surface of the embryo was poor in transparency,the tail was twisted,the yolk sac elongation structure became thicker,the notochord became thin,there was no normal embryo movement,and the body length became shorter.After the withdrawal,the abnormal phenotype was not recovered,and severity of the abnormal phenotype was concentration-dependent.It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 tg/mL.Its toxic effect occurs during embryonic organogenesis.And it mainly affects the development of embryonic nervous system,cardiovascular system and bone.
ABSTRACT
Objective To study the toxic effect ofNiuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40,200,400,800,1 200 tg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage,exceeding the normal effective dose).While,there was toxicity to embryo development at the concentration greater than 200 tg/mL.The head and eyes of the embryo were smaller,and the pupils were colorless.Zebrafish embryonic heart malformation was observed,heart rate was weak,blood pool congestion happened and back blood flow was not smooth.The abdominal surface of the embryo was poor in transparency,the tail was twisted,the yolk sac elongation structure became thicker,the notochord became thin,there was no normal embryo movement,and the body length became shorter.After the withdrawal,the abnormal phenotype was not recovered,and severity of the abnormal phenotype was concentration-dependent.It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 tg/mL.Its toxic effect occurs during embryonic organogenesis.And it mainly affects the development of embryonic nervous system,cardiovascular system and bone.
ABSTRACT
Objective To investigate the functional role of a soluble secretion factor midkine-a in the process of zebrafish embryonic heart development .Methods Whole-mount in situ hybridization was performed to detect whether midkine-a is expressed in the embryonic heart .In the transgenic embryonic heart of Tg ( pmidkine-a:EGFP) , the expres-sion of EGFP in the heart was monitored .Multiple heat shock treatments were applied to Tg (phsp:midkine-a:EGFP) em-bryos in order to overexpress midkine-a, and the phenotype of the heart was observed .The heterozygous Tg ( phsp:mid-kine-a:EGFP) fish were crossed with homozygous Tg (pcmlc2:dsRed) fish, which specifically expresses RFP in the nucle-oli of embryonic cardiomyocytes to facilitate myocyte number counting .To get Tg ( phsp:midkine-a:EGFP/pcmlc2:dsRed)embryos, and the total number of cardiomyocytes in their heart was counted and compared with the controls when the heat shock induced overexpression of midkine-a.Morphonino knocked out of midkine-a was performed to observe the heart phe-notype.Results Midkine-a was expressed in the zebrafish embryonic heart during development .Overexpression of mid-kine-a led to a smaller heart and reduced total number of cardiomyoctes in a single heart , which might be associated with the smaller heart phenotype .Morphonilo knocked out of midkine-a had no effect upon the heart development .Conclusions Midkine-a impedes zebrafish embryonic heart growth by limiting its cardiomyocyte pool .
ABSTRACT
Aim To study the cardiotoxicity of celastrol to zebrafish embryo.Method 48 h post-fertilization zebrafish embryos were used as model for analysis of heart toxicity and were treated with various concentrations of celastrol.6,12,24 h after treatment morphological and functional changes of embryos hearts were observed.Results Cardiotoxicity was not found in embryos during 24 h treatment with 1 ?mol?L-1 concentration of celastrol.Toxic symptoms of embryos were caused by 2,3,4 ?mol?L-1 concentrations of celastrol with appearance of heart linearization,heart membrane hemorrhage and hemocytes accumulation in cardiac region.Moreover,heart rate decreased significantly with increase of concentration and prolongation of treatment.The EC50(24 h)of decrease of heart rate was about 1.78 ?mol?L-1.Conclusion Celastrol is cardiotoxic to zebrafish embryo.