孕期暴露三氯生对秀丽线虫子代运动、生殖的影响

发布时间：2018-07-08 20:33

本文选题：三氯生 + 秀丽线虫　；参考：《新乡医学院》2014年硕士论文

【摘要】：背景：内分泌干扰物(endocrine disrupting chemicals,EDCs),如三氯生(Triclosan,TCS)通过与激素受体结合或影响细胞信号途径等方式,模拟或部分模拟激素,发挥类激素样或抗激素样作用,干扰机体内激素的合成、分泌、转运、结合、代谢及调节等过程,从而影响儿童的生长发育。秀丽线虫作为一种模式动物,具有生命周期短,繁殖速度快,个体小,容易培养且遗传与发育背景清楚等诸多优点,已广泛应用于生物学各领域的研究。秀丽线虫对多种药物、污染物敏感,且行为反应模式稳定、结果可靠,已被用于药物筛选、检测和评价污染。目的：观察三氯生(Triclosan,TCS)对秀丽线虫(Caenorhabditis elegans,c.elegans)子代(F1)的运动能力、后代数目及世代时间、生长发育等生物学指标的影响,为研究TCS毒性效应及其机制提供理论依据。方法：实验设置对照组和4个三氯生暴露组,浓度分别为100nmol/L、1μmol/L、10μmol/L、20μmol/L,从c.elegans L4期开始暴露,暴露时间24h,暴露后转移至无TCS培养板继续培养。观察TCS对c.elegans子代(F1)头部摆动频率、身体弯曲频率、后代数目及世代时间、体长及体宽等生物学指标的影响；采用基因芯片技术分析对照组和TCS20μmol/L差异表达基因及主要作用。各组数据以均数±标准差(x±s)表示,应用SPSS20.0软件包进行统计学分析,多组间均数采用单因素方差分析(ANOVA),以P0.05作为显著性检验标准。结果：1.各浓度TCS水平组c.elegans Fl头部摆动频率较对照组降低(P0.01),ANOVA示,三氯生暴露后,c.elegans Fl头部摆动能力受到显著影响(F=卯.245,P0.01)；各浓度TCS水平组c.elegans F1身体弯曲频率较对照组降低(P0.01),ANOVA示,三氯生暴露后,c.elegans F1身体弯曲能力受到显著影响(F=25.636,P0.01)。 2.与对照组比,暴露在100mol/L、10μmol/L、20μmol/L三氯生中的c.elegans F1个体的后代数目分别降低了4.71%、25.60%、39.45%、58.67%, AN OVA结果示,三氯生暴露后对c.elegans生殖能力产生极显著影响(F=461.593,P0.01)；与对照组相比,三氯生暴露后秀丽线虫的世代时间缩短了2.14%--5.38%(P0.01),并且表现出浓度依赖性表型。ANOVA结果示、三氯生暴对c.elegans F1世代时间产生显著影响(F=27520,P0.01)。 3.与对照组比,暴露在100nmol/L、1μmol/L、101μmol/L、20μmo1/L三氯生中的c.elegans F1L4期幼虫体长及体宽变化不明显(P0.05),无统计学意义。 4.基因芯片结果示：对照组和TCS20μmol/L暴露后组间差异大于2倍,组内差异小的表达差异基因共有113条,其中大部分为与线虫神经发育和调节运动相关的基因；挑选组间差异大于3倍,组内差异小的7个基因进行验证,验证结果与基因芯片相符。结论：1.孕期三氯生暴露对秀丽线虫的子代运动、生殖影响显著,提示三氯生存在生殖毒性及抑制秀丽线虫神经发育。 2.GO分析和Pathway分析的结果说明孕期暴露三氯生后可能是因为基因的差异表达对线虫的运动及生殖能力产生影响。 3.基因芯片结果示差异表达基因较多,由此推测孕期三氯生暴露可能会对秀丽线虫子代产生更广泛的潜在影响。
[Abstract]:Background: endocrine disrupting chemicals (EDCs), such as three chloroplastic (Triclosan, TCS), which mimic or partially mimic hormone like or antisteroid effects by combining with hormone receptors or affecting cell signaling pathways, and interferes with the synthesis, secretion, transport, binding, metabolism, and regulation of hormones in the body. It affects the growth and development of children.
As a model animal, Caenorhabditis elegans has many advantages, such as short life cycle, fast propagation speed, small individual, easy cultivation and clear genetic background, and so on. It has been widely used in various fields of biology. The elegans are sensitive to a variety of drugs and contaminants, and are stable in reaction mode and reliable. Detection and evaluation of pollution.
Objective: To observe the effects of Triclosan (Triclosan (TCS) on the biological indexes of Caenorhabditis elegans (C.elegans) progeny (F1), the number of offspring and generation time, and the growth and development of the offspring (F1), and provide a theoretical basis for the study of the toxicity and mechanism of TCS.
Methods: the experiment set up the control group and 4 three chlorobi exposure groups, the concentration was 100nmol/L, 1 mu mol/L, 10 mu mol/L, 20 mu mol/L, from C.elegans L4 stage to exposure, exposure time 24h, and then transferred to no TCS culture plate to continue culture. The frequency of TCS to C.elegans sub generation (F1), body bending frequency, the number of offspring and generation time were observed. The effect of body length and body width and other biological indexes, the gene chip technique was used to analyze the differentially expressed genes and the main functions of the control group and TCS20 mu mol/L. The data of each group were expressed by the mean number of standard deviation (x + s), and the SPSS20.0 software package was used for statistical analysis. The average number of groups was analyzed by single factor variance analysis (ANOVA), and P0.05 was used as the significance. Test standard.
Results: 1. the head oscillating frequency of C.elegans Fl in the TCS level group was lower than that of the control group (P0.01). ANOVA showed that the head swing ability of C.elegans Fl was significantly affected after three chlorinated exposure (F= sockets.245, P0.01), and the flexural frequency of the body was lower than that of the control group. F1 body flexion ability was significantly affected (F=25.636, P0.01).
2. compared with the control group, the number of descendants of C.elegans F1 individuals exposed to 100mol/L, 10, mol/L, 20 mol/L three chloride decreased by 4.71%, 25.60%, 39.45%, 58.67%, and AN OVA results showed that three chloride exposure had a significant effect on C.elegans reproduction (F=461.593, P0.01); compared with the control group, three chloro was exposed to Caenorhabditis elegans. The generation time shortened 2.14%--5.38% (P0.01) and showed a concentration dependent phenotypic.ANOVA result indicating that three chlorochlorine had a significant effect on the C.elegans F1 generation time (F=27520, P0.01).
3. compared with the control group, the changes of body length and body width of C.elegans F1L4 larvae in 100nmol/L, 1, 101, 101, and 20 mo1/L three chloride were not significant (P0.05). There was no statistical significance.
The results of 4. gene chip showed that the difference between the control group and the TCS20 mu mol/L was more than 2 times, and there were 113 differentially differentially expressed genes in the group. Most of them were genes related to the development of the nematode nerve and the regulation of the movement. The differences between the selected groups were more than 3 times, and 7 genes with small differences in the group were verified, and the results were verified with the gene core. The film coincide.
Conclusion: during the 1. trimester, three chlorinone exposure had a significant effect on the reproduction of the offspring of the nematode Caenorhabditis elegans, suggesting that three chloro had reproductive toxicity and inhibited the development of the Caenorhabditis elegans.
The results of 2.GO analysis and Pathway analysis showed that exposure to three chlorprospan during pregnancy may be due to differential expression of genes affecting the movement and reproductive capacity of nematodes.
3. gene chip results showed that there were more differentially expressed genes. It is speculated that exposure to three chloro chloride during pregnancy may have a wider potential impact on the offspring of the male worm.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R715.3

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