Study Of Toxic Effect And Effector Screening Of Ethanol On Zebrafish Embryo Development

Ethanol is a major component of alcoholic beverages and also been abused widely.Excessive drinking can cause alcoholism,alcohol dependence and many diseases.It is also a common source of ethanol teratogenicity,maternal fetal alcohol syndrome can lead to alcohol consumption during pregnancy,the symptoms are head and facial abnormalities,developmental delay,movement coordination,learning disabilities,it is a great burden to society and family.Many researches have been done on the mechanism of ethanol toxicity,among which ethanol induced developmental abnormalities is a research hotspot.This paper aims to study toxic mechanism of ethanol on zebrafish embryo development,and analysis oxidative damage,DNA methylation and carcinogenesis induced by ethanol.To establish an abnormal development model of zebrafish embryo after ethanol exposure,observe mortality rate,hatching rate and malformation rate of embryos after exposure.To analysis enzyme activity and mRNA expression level related to oxidative stress,and detect oxidative stress effect in zebrafish embryos.To screen and identify specific proteins after the ethanol exposure,and clarify the relationship between protein expression and DNA methylation after ethanol exposure by using epigenetic and proteomics methods.The main results of this study are as follows:1 Ethanol had toxic effect on zebrafish embryos and in a dose dependent manner.LD50-96 h of ethanol on zebrafish embryos was 0.333mol/L.Ethanol inhibited significantly zebrafish embryo development,can cause development delay and malformation,there were a dose-effect relation.Main malformation type were head deformity,heart malformation,and trunk deformity etc.2 Ethanol exposure could cause oxidative damage in embryonic cells,SOD and CAT activity were increased significantly;induced abnormal expression of oxidative stress related genes(cat,sod1)in zebrafish embryos,showed that ethanol could cause doxidative damage on zebrafish embryos.Ethanol exposure induced abnormal expression of DNA methylation related genes(DNMT1,MeCP2 and tet3),endocrine related genes(KISS2 and sox9a)and embryo development related genes(sox2)in zebrafish embryos.The difference was statistically significant(p<0.05).3 It was found that several proteins such as hnRNPab,eef1b2 and Pentraxin were up and down regulated significantly by two-dimensional fluorescence differential gel electrophoresis and mass spectrometry.6 Ethanol could inhibit expression of hnRNPab protein in zebrafish embryos,and there was a dose-response relationship.In conclusion,ethanol had toxic effects on zebrafish embryo development,including lethal and teratogenic effects.Ethanol could cause doxidative damage and DNA methylation aberration during zebrafish embryo development.Ethanol could interfere early embryonic development;caused abnormal development of multiple biosystems;and suggesteda certain reproductive toxicity.Protein hnRNPab could be a toxic effector of ethanol on zebrafish embryo development.