论文总字数：23986字

摘 要

背景：

现今空气中煤烟PM_2.5的含量非常高，已经严重影响了环境质量，对人体健康也造成了极大伤害。但到目前为止，对于PM_2.5体内致毒机理并不十分清楚。本课题组近年来致力于纳米材料的毒性和毒理学研究，对煤烟PM_2.5在模式生物秀丽线虫中的毒性作用进行了初步研究，并发表了两篇相关SCI论文。为了进一步在RNA 水平上研究解析煤烟PM_2.5致秀丽线虫体内毒性作用机制，因此开展本项目的研究。

秀丽线虫的发育与遗传背景是相对研究清楚的模式动物，其繁殖速度快，实验周期短，易于实验室培养。因此，课题组利用秀丽线虫分析体系，进一步探讨煤烟PM_2.5潜在的负面影响和相应的作用机理。

方法：

利用新一代测序技术确定在煤烟PM_2.5暴露的秀丽线虫体内异常表达的 miRNA，并研究miRNA对煤烟PM_2.5致秀丽线虫体内毒性的调控作用及作用机制。

结果：

1.筛选出9个在煤烟PM_2.5暴露的秀丽线虫体内异常表达miRNA。其中mir-231在煤烟PM_2.5致秀丽线虫体内毒性和转运中起到重要作用。mir-231突变体对煤烟PM_2.5毒性具有抗性作用。

2. Smk-1可能是mir-231调节煤烟PM_2.5致秀丽线虫体内毒性和转运的候选靶基因，smk-1突变体对煤烟PM_2.5毒性敏感。

3.Mir-231—smk-1双突变实验，可最终证明mir-231与smk-1具有靶向关系。

结论：

将可能帮助我们获得新的评价与检测PM_2.5毒性的分子评价终点，并有助于进一步制定有效的策略预防与抑制PM_2.5对于生物体的毒害。

关键词：煤烟PM_2.5 mir-231 smk-1 靶向关系 毒性评价

abstract

Background:

Today's air PM_2.5 (Coal combustion related fine particulate matter) content is very high, has seriously affected the environmental quality, has also caused great harm to the human health. However, for the PM_2.5 body toxic mechanism is not very clear. In recent years, the research group has devoted itself to the toxicity and toxicology research of nanomaterials, and studied the toxic effects of PM_2.5 in the model nematode, and published two related SCI papers. In order to find out the mechanism of toxicity in the analysis of the toxic effects of PM_2.5 in the RNA level, the research of this project was carried out.

The development and genetic background of the C. elegans is relatively clear. As a model animal, its breeding speed is quick, the experimental period is short, easy to laboratory culture. Therefore, the research group used the C. elegans analysis system to further explore the potential negative effects of PM_2.5 and the corresponding mechanism of action.

Method:

The miRNAs were screened by using a new generation of sequencing techniques to study the abnormal expression of miRNAs in the beautiful nematodes exposed to soot PM_2.5, and the effects of miRNA on the toxicity of soot PM_2.5 to nematodes were studied.

Result:

     1. Abnormal expression of miRNAs in 9 healthy nematodes exposed to PM_2.5. Among them, mir-231 plays an important role in the toxicity and transport of soot PM_2.5. The mir-231 mutant has a resistance to PM_2.5 toxicity.

     2. Smk-1 may be a candidate target gene for the toxicity and transport of smk-1, which is regulated by mir-231, and smk-1 mutant is sensitive to soot PM_2.5 toxicity.

    3.Mir-231-smk-1 double mutagenesis experiments, it can be concluded that mir-231 has a targeted relationship with smk-1.

Conclusion:

Will help us to obtain new evaluation and detection of PM_2.5 toxicity at the molecular evaluation end point and help to further develop effective strategies to prevent and inhibit PM_2.5 toxicity to organisms.

Key words:

PM_2.5 mir-231 smk-1 targeting relationship toxicity evaluation

目 录

摘 要 2

abstract 3

第一章 microRNA的研究进展 7

1.1 microRNA的发现和作用机制 7

1.1.1 microRNA的发现 7

1.1.2 microRNA的加工产生和作用机理 7

1.2 microRNA参与线虫的发育和衰老 7

1.3 microRNA调控线虫衰老的作用机制 8

1.3.1 microRNA调控胰岛素/IGF-1信号通路 8

1.3.2 microRNA调控TOR信号通路 9

1.3.3 microRNA调控自噬 10

第二章 基于转录组测序全基因筛选秀丽线虫中煤烟PM_2.5毒性相关的miRNA 12

2.1 材料与方法 12

2.1.1 实验材料 12

2.1.2 实验方法 13

2.2 实验结果 14

2.2.1 全基因组内筛选鉴定miRNA 14

2.2.2 筛选PM_2.5暴露后差异表达的miRNA 14

2.2.3 miRNA突变体的毒性效应 16

第三章 mir-231调控PM_2.5致秀丽线虫体内毒性调控作用的分子机制 19

3.1 mir-231 突变体中的毒性效应 19

3.2 预测mir-231的靶基因 20

3.3 smk-1 突变体毒性效应 21

3.4 smk-1与mir-231突变体运动行为的分析 22

3.5 PM_2.5暴露对秀丽线虫肠道通透性的影响 22

3.6 mir-231与smk-1靶向关系的确立 23

3.7 smk-1在调节煤烟PM_2.5毒性的下游调控基因 24

第四章 总结讨论 26

参考文献 28

致 谢 30

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