本研究的主要目的為建立以底泥與人類細胞株為檢測樣本來評估環境累積毒性對人類健康的影響。工業化的發展加劇了對環境的迫害，而底泥是所有環境污染物的最終儲存庫，因此底泥是評估環境污染物經由生物累積對人類健康影響很好的樣本。新竹科學園區是世界著名的半導體製造工業園區，客雅溪流經新竹科學園區和新竹市區。因半導體產業所衍生的工業廢棄物具有引發癌症、出生缺陷與其他嚴重疾病等，因此客雅溪的水質評估值得被關注。雖然過去有許多研究表明魚類等底棲生物的退化、生長與底泥的污染有關聯，但探討底泥汙染對人類健康的影響知之甚少。因此，本研究以客雅溪三個不同都市發展程度區域的底泥樣本探討工業發展衍生環境污染物對多種不同人類細胞株的影響。採樣點分別：上游為未開發區，中游為生活/工業廢水排放口，下游為生活/工業廢水累積區。我們的研究結果顯示，客雅溪下游的底泥萃取物比中、上游的底泥萃取物更具細胞毒性。下游底泥萃取物顯著增加了所有細胞中的活性氧物質的表現。而氧化壓力的累積易誘發改變細胞的基本行為能力，如細胞活性、細胞貼附能力與細胞爬行能力。根據MTT細胞存活率試驗結果顯示下游底泥萃取物顯著降低了腦、口腔、肺、乳腺、肝、胰腺、子宮頸、前列腺和結腸直腸細胞的活性。此外，與其他流域相比，下游底泥萃取物抑制大多數細胞的貼附能力與傷口癒合能力。肝臟是環境毒素的最終目標，越來越多的流行病學統計指出，肝臟代謝綜合症的成因與暴露於環境污染有很大的關聯。因此，本研究試圖證實底泥污染物對肝損傷的影響。我們的研究結果表明，下游底泥萃取物比其他區域更具肝細胞毒性。同時，下游底泥萃取物顯著增加肝細胞的活性氧類總量，並引起線粒體功能喪失，導致細胞凋亡，更進而造成血清中天門冬胺酸轉胺酶（GOT）/丙胺酸轉胺酶（GPT）蛋白表現增加。此外，在本研究中，我們還發現下游底泥萃取會通過抑制胰島素接受器受質-1 (IRS-1)/ 蛋白激酶B (AKT)/ 肝醣合成酶激酶3 (GSK3β)，進而影響肝醣合成酶(GYS)活性，以減少肝醣合成。本研究是首次以底泥為樣本探討環境累積毒性對人體健康的影響。我們誠摯地希望本研究能為環境汙染與大眾健康提供新的科學依據，並以全球視角應對公共衛生的挑戰。

The main objective of this study was to establish a human cell-based platform to assess the effects of sediment toxicity on human cell lines. Sediments are long-term repositories for environmental pollutants via Environmental cycles, sediment pollution also has increased with industrialization. Sediment is a good sample to evaluate the effect of environmental pollutants on human health via bioaccumulation. The Hsinchu Science Park is one of the most prominent semiconductor manufacturing centers in the world, and the Ke-Ya River flows through Hsinchu Science Park and the Hsinchu urban district. Because semiconductor wastes potentially contribute to higher-than-normal rates of cancers, birth defects, and serious diseases, the quality assessment of the Ke-Ya River has prompted widespread concerns. While previous studies have shown an association between the degradation of fish populations and sediment pollutants, very little is known about the issues on human health. Herein, the effects of sediment from three sediment-sampling sites of the Ke-Ya River on 11 different human cell lines were directly evaluated. The upstream represents the undeveloped zone, the middle-stream represents the household/industrial wastewater zone, and the downstream represents the accumulation zone. Our results indicated that the sediment pollution of the downstream Ke-Ya River was more cytotoxic than that of the middle stream and upstream. Downstream sediment extract (DSE) significantly increased reactive oxygen species (ROS) levels across all cell types. Accordingly, oxidative stress can trigger redox-sensitive pathways and alter essential biological processes such as cell viability, cell adhesion, and cell motility. Importantly, the MTT assay indicated that DSE significantly decreased the viability of brain, oral, lung, breast, liver, pancreatic, cervical, prostate, and colorectal cells. Furthermore, the adhesive ability and wound healing ability of most cells were greatly reduced in the presence of DSE compared to other conditions. Liver is the final target of environmental toxins; more and more epidemiological studies had shown that liver metabolic syndrome including the impairment of hepatic glycogen synthesis have relentlessly risen by exposure to environmental pollutants. Therefore, this study also sought to confirm the effects of sediment pollutants on liver damage. Our results indicated that the industrial wastewater sediment (downstream) was more cytotoxic than other zones. DSE significantly increased reactive oxygen species (ROS) levels, caused mitochondrial dysfunction, lead to cell apoptosis, and result in glutamic oxaloacetic transaminase (GOT)/ Glutamic pyruvic transaminase (GPT) protein released. Additionally, to elucidate the underlying mechanism by sediment pollutants perturb hepatic glycogen synthesis, we investigated the effect of o different sediment samples from pollution situations on glycogen synthesis in liver cell lines. In this study, we found that downstream sediment extract induced more severe impairments in liver cells, and disturber glycogen synthesis than in other conditions; these include decreased hepatic glycogen synthesis via inhibition and IRS-1/AKT/glycogen synthase kinase3β (GSK3β) mediated glycogen synthase (GYS) inactivation. This study shows the results of the first analyses completed on the sediment cytotoxicity in human cells, and stimulated ROS levels are crucial for cellular life, also provides the first in vitro evidence for sediment accumulated toxicity in disturbance of liver glycogen synthesis and caused hepatic cell damage. We sincerely hope that our study provides a scientific basis for further investigations with a global perspective on public health challenges.

中文摘要 I
Abstract III
Acknowledgement V
Table of Contents VII
List of Figures and Tables IX
Abbreviations XI
Chapter 1 Introduction 1
1.1 Environmental pollution and human health 1
1.2 Sediment quality and urban-industrial pollution 3
1.3 Sediment-associated pollutants and cytotoxicity 5
1.4 Environmental pollutants and liver damage 6
Chapter 2 Materials and Methods 7
2.1 Sediment sampling locations and extraction 7
2.2 Sediment physical and chemical property analysis 9
2.3 Sediment pollutants analysis 10
2.4 Cell lines and cell culture 11
2.5 MTT cell viability assay 12
2.6 Reactive oxygen species (ROS) detection 13
2.7 Cell adhesion assay 13
2.8 Wound healing assay 14
2.9 JC-10 assay 14
2.10 Apoptosis assay 14
2.11 Glycogen colorimetric assay 15
2.12 Immunoblotting analysis 15
2.13 GOT/GPT detection 16
2.14 Statistical analysis 16
Chapter 3 Results 17
3.1 Sediment physical & chemical characteristics 17
3.2 Sediment pollution composition 19
3.3 Sediment extract toxicity and cell viability 21
3.4 Sediment extract effect on ROS production 22
3.5 Sediment extract effect on cell adhesion ability 24
3.6 Sediment extract effect on cell wound healing 25
3.7 The cytotoxic effect of sediment extract on liver cell lines 29
3.8 The impact of sediment extract on ROS production 32
3.9 The impact of sediment extract on mitochondrial membrane potential 34
3.10 The impact of sediment extract on GOT/GPT protein release 36
3.11 The impact of sediment extract on glycogenesis and glycogen production 37
Chapter 4 Discussion 41
Chapter 5 Concussion 48
Chapter 6 Reference 50
Appendix 55
The graphic abstract of previous studies 56
Publications list 58
Frist author full text 61

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