| Environmental hormones are widespread in the environment of our survival, andpose a serious threat to humans and other creatures. The problem of environmentalhormones pollution has become one of the world’s major environmental problems inthe 21st century.In this thesis, we first used HPLC-MS method to make inspections for sevenkinds of representative environmental hormones (estrogen (E1), estradiol (E2),oestriol (E3), ethynyl estradiol (EE2), diethyl stilboestrol (DES), nonylphenol (NP),octylphenol (OP)) in the sewage treatment work’s outflowing water from a universitylcated in eastern of Jinan. We get the conclusion that, in the seven kinds ofrepresentative environmental hormones, only two kinds of environmental hormones(Nonylphenol, Octylphenol) were detected from the water sample, and theirconcentrations were 13.39ng/L and 0.49ng/L, respectively.Based on the above environmental hormone concentrations in the water sample,as well as existing literature on environmental hormones concentrations of differentwater bodys, we determine nonylphenol as the substance of the subsequentexperiments. Then we choose zebrafish as test animal, through acute toxicity, sex ratio,gonadosomatic index to discuss the toxicity of nonylphenol. First, the acute toxicity testwas used to determine 96h LC50 and safe concentration of nonylphenol on male andfemale zebrafish, in concentration. The acute toxicity test results showed that forfemale zebrafish 96h LC50 and safe concentration were 0.902mg/L and 0.0902mg/L,for male zebrafish 96h LC50 and safety concentrations were 0.529mg/L and0.0529mg/L. According to the toxic substance evaluation standard, nonylphenol is poisonousto zebrafish, and male zebrafish are more sensitive to the toxicity of nonylphenol, soin terms of the research on the influence of nonylphenol on zebrafish immune-relatedgene expression, we choose male zebrafish as research animals.To detect the estrogenic effects of nonylphenol on the zebrafish, We exposed zebrafish to solution of different concentrations of nonylphenol for long time. After180 days of exposure to nonylphenol, the results showed that the blank control group,the solvent control group, 20ug/L and 100ug/L of nonylphenol concentrations group,the females ratio were 51.00%, 51.33%, 64.00%, 75.33%, respectively. There was asignificant difference between the two nonylphenol concentrations groups and thecontrol groups (p<0.01). The gonadosomatic index (GSI) of blank control group, solventcontrol group, 20ug/L, 100ug/L nonylphenol concentrations groups were: 4.40%,4.43%, 3.90%, 3.37%, respectively. The gonadosomatic index (GSI) of solvent controlgroup and blank control group were consistent. The gonadosomatic index of malezebrafish of 20ug/L, and 100ug/L nonylphenol concentrations were much lower thanthat of the control group, and there was highly significant difference (p<0.01) betweenthem, the results confirmed that nonylphenol had certain estrogenic effects on thezebrafish.Finally, we used the gene chip technology to detect the effects of differentnonylphenol concentrations on the expression of immune-related genes of thezebrafish. Adult male zebrafish were exposed to 100ug/L solvent control group,20ug/L, and 100ug/L nonylphenol solution group in turn for 7 days. Six malezebrafish were taken out from every group, and RNA was extracted from pooled liversof the six male zebrafish. We used gene chip technology to detect differentlyexpressed genes. The results shows that: compared with the control group, there were1259 genes expressed differently, including 26 immune-related genes, in 20μg/Lnonylphenol concentration group; there were 2024 genes expressed differently,including 21 immune-related genes, in 100μg/L nonylphenol concentration group;there were 718 genes expressed differently, including 12 immune-related genes,insewage treatment group. In both 20μl/L and 100μg/L two nonylphenols concentrationgroup compared with the control results, there were nine immune-related geneexpression expressed differently, eight of those genes expressed down, It proved thatnonylphenol had immune toxicity on zebrafish.In order to detect the accuracy of the gene chip results, we use Real-time PCRtechnology to identify four different expressed genes. The results showed: The vast majority of gene variations were coincide with the gene chip’results , and it confirmedthat the gene chips’results were reliable. |
PDF Full Text Request