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研究生: 王瀞嬴
Wang, Jing-Ying
論文名稱: 利用iTRAQ化學標定方法進行小鼠肝臟中塑膠微粒毒性之差異蛋白質體學分析
Differential proteomic analysis of microplastic toxicity in mouse liver by iTRAQ technology
指導教授: 陳頌方
Chen, Sung-Fang
口試委員: 陳怡婷
Chen, Yi-Ting
陳翰民
Chen, Han-Min
陳頌方
Chen, Sung-Fang
口試日期: 2023/07/28
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2023
畢業學年度: 111
語文別: 中文
論文頁數: 65
中文關鍵詞: 塑膠微粒iTRAQ差異蛋白質體學
英文關鍵詞: Microplastics, iTRAQ, Differential proteomics
研究方法: 實驗設計法
DOI URL: http://doi.org/10.6345/NTNU202301580
論文種類: 學術論文
相關次數: 點閱:90下載:0
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  • 謝誌 i 摘要 ii Abstract iii 目錄 iv 表目錄 vii 圖目錄 viii 第一章 緒論 1 1-1 前言 1 1-2 環境中的塑膠微粒 2 1-2-1 塑膠微粒對生物體的影響 2 1-3 蛋白質體學 5 1-3-1 蛋白質身分鑑定 5 1-3-2 差異蛋白質體學 7 1-4 同重元素相對與絕對定量(iTRAQ) 9 1-5 質譜儀技術 12 1-5-1 質譜分析系統 12 1-5-2 電灑游離 13 1-5-3 四極桿飛行時間質譜儀 14 1-6 液相層析質譜 15 1-6-1 液相多維分離技術 15 1-7 研究動機 17 第二章 實驗材料與方法 18 2-1 實驗儀器與設備 18 2-2 BALB/c小鼠試驗 19 2-3 蛋白質樣品製備 20 2-4 樣品濃縮純化 20 2-5 蛋白質濃度測定 21 2-6 蛋白質水解與iTRAQ化學標定 22 2-7 第一維分離策略 23 2-8 自製C18離心管柱去鹽 25 2-9 奈升級超高效能液相層析電灑串聯式質譜儀(NanoLC-ESI- MS/MS) 26 2-9-1 奈升級超高效能液相層析儀 26 2-9-2 四極桿飛行時間質譜儀 28 2-10 數據分析 29 第三章 結果與討論 30 3-1 小鼠試驗重複劑量毒性評估 30 3-1-1 體重變化 30 3-1-2 血清生化分析 31 3-1-3 組織病理切片 34 3-2 蛋白質樣品濃度測定 35 3-3 第一維液相分離—鹼性逆向層析 36 3-4 基於iTRAQ標定的相對定量分析 38 3-4-1 蛋白質身分鑑定及相對定量 38 3-4-2 統計分析 39 3-5 生物資訊分析 47 3-5-1 Gene Ontology分析 47 3-5-2 Metascape分析 50 3-5-3 差異蛋白質功能討論 52 第四章 結論與未來展望 57 參考文獻 58

    1. Williams, A. T.; Rangel-Buitrago, N., The past, present, and future of plastic pollution. Marine Pollution Bulletin 2022, 176, 113429.
    2. UNEP, U. N. E. A. o. t. U. N. E. P., End Plastic Pollution: Towards an International Legally Binding Instrument. United Nations Environment Programme: 2022.
    3. Cole, M.; Lindeque, P.; Halsband, C.; Galloway, T. S., Microplastics as contaminants in the marine environment: A review. Marine Pollution Bulletin 2011, 62 (12), 2588-2597.
    4. Boucher, J.; Friot, D., Primary microplastics in the oceans: a global evaluation of sources. Iucn Gland, Switzerland: 2017; Vol. 10.
    5. Dodson, G. Z.; Shotorban, A. K.; Hatcher, P. G.; Waggoner, D. C.; Ghosal, S.; Noffke, N., Microplastic fragment and fiber contamination of beach sediments from selected sites in Virginia and North Carolina, USA. Marine Pollution Bulletin 2020, 151, 110869.
    6. Schmidt, C.; Krauth, T.; Wagner, S., Export of Plastic Debris by Rivers into the Sea. Environmental Science & Technology 2017, 51 (21), 12246-12253.
    7. Kanhai, L. D. K.; Gårdfeldt, K.; Lyashevska, O.; Hassellöv, M.; Thompson, R. C.; O'Connor, I., Microplastics in sub-surface waters of the Arctic Central Basin. Marine Pollution Bulletin 2018, 130, 8-18.
    8. Law, K. L.; Morét-Ferguson, S. E.; Goodwin, D. S.; Zettler, E. R.; DeForce, E.; Kukulka, T.; Proskurowski, G., Distribution of Surface Plastic Debris in the Eastern Pacific Ocean from an 11-Year Data Set. Environmental Science & Technology 2014, 48 (9), 4732-4738.
    9. Udovicki, B.; Andjelkovic, M.; Cirkovic-Velickovic, T.; Rajkovic, A., Microplastics in food: scoping review on health effects, occurrence, and human exposure. International Journal of Food Contamination 2022, 9 (1), 7.
    10. Schymanski, D.; Goldbeck, C.; Humpf, H.-U.; Fürst, P., Analysis of microplastics in water by micro-Raman spectroscopy: Release of plastic particles from different packaging into mineral water. Water Research 2018, 129, 154-162.
    11. Setälä, O.; Fleming-Lehtinen, V.; Lehtiniemi, M., Ingestion and transfer of microplastics in the planktonic food web. Environmental Pollution 2014, 185, 77-83.
    12. Wang, C.; Zhao, J.; Xing, B., Environmental source, fate, and toxicity of microplastics. Journal of Hazardous Materials 2021, 407, 124357.
    13. Atugoda, T.; Vithanage, M.; Wijesekara, H.; Bolan, N.; Sarmah, A. K.; Bank, M. S.; You, S.; Ok, Y. S., Interactions between microplastics, pharmaceuticals and personal care products: Implications for vector transport. Environment International 2021, 149, 106367.
    14. Xu, S.; Ma, J.; Ji, R.; Pan, K.; Miao, A.-J., Microplastics in aquatic environments: Occurrence, accumulation, and biological effects. Science of The Total Environment 2020, 703, 134699.
    15. Kim, J.-H.; Yu, Y.-B.; Choi, J.-H., Toxic effects on bioaccumulation, hematological parameters, oxidative stress, immune responses and neurotoxicity in fish exposed to microplastics: A review. Journal of Hazardous Materials 2021, 413, 125423.
    16. Shiu, R.-F.; Vazquez, C. I.; Chiang, C.-Y.; Chiu, M.-H.; Chen, C.-S.; Ni, C.-W.; Gong, G.-C.; Quigg, A.; Santschi, P. H.; Chin, W.-C., Nano- and microplastics trigger secretion of protein-rich extracellular polymeric substances from phytoplankton. Science of The Total Environment 2020, 748, 141469.
    17. Porter, A.; Lyons, B. P.; Galloway, T. S.; Lewis, C., Role of Marine Snows in Microplastic Fate and Bioavailability. Environmental Science & Technology 2018, 52 (12), 7111-7119.
    18. Zhang, Q.; Xu, E. G.; Li, J.; Chen, Q.; Ma, L.; Zeng, E. Y.; Shi, H., A Review of Microplastics in Table Salt, Drinking Water, and Air: Direct Human Exposure. Environmental Science & Technology 2020, 54 (7), 3740-3751.
    19. Senathirajah, K.; Attwood, S.; Bhagwat, G.; Carbery, M.; Wilson, S.; Palanisami, T., Estimation of the mass of microplastics ingested – A pivotal first step towards human health risk assessment. Journal of Hazardous Materials 2021, 404, 124004.
    20. Ragusa, A.; Svelato, A.; Santacroce, C.; Catalano, P.; Notarstefano, V.; Carnevali, O.; Papa, F.; Rongioletti, M. C. A.; Baiocco, F.; Draghi, S.; D'Amore, E.; Rinaldo, D.; Matta, M.; Giorgini, E., Plasticenta: First evidence of microplastics in human placenta. Environment International 2021, 146, 106274.
    21. Leslie, H. A.; van Velzen, M. J. M.; Brandsma, S. H.; Vethaak, A. D.; Garcia-Vallejo, J. J.; Lamoree, M. H., Discovery and quantification of plastic particle pollution in human blood. Environment International 2022, 163, 107199.
    22. Prata, J. C.; da Costa, J. P.; Lopes, I.; Duarte, A. C.; Rocha-Santos, T., Environmental exposure to microplastics: An overview on possible human health effects. Science of The Total Environment 2020, 702, 134455.
    23. Deng, Y.; Zhang, Y.; Lemos, B.; Ren, H., Tissue accumulation of microplastics in mice and biomarker responses suggest widespread health risks of exposure. Scientific Reports 2017, 7 (1), 46687.
    24. Lu, L.; Wan, Z.; Luo, T.; Fu, Z.; Jin, Y., Polystyrene microplastics induce gut microbiota dysbiosis and hepatic lipid metabolism disorder in mice. Science of The Total Environment 2018, 631-632, 449-458.
    25. Jin, Y.; Lu, L.; Tu, W.; Luo, T.; Fu, Z., Impacts of polystyrene microplastic on the gut barrier, microbiota and metabolism of mice. Science of The Total Environment 2019, 649, 308-317.
    26. Zheng, H.; Wang, J.; Wei, X.; Chang, L.; Liu, S., Proinflammatory properties and lipid disturbance of polystyrene microplastics in the livers of mice with acute colitis. Science of The Total Environment 2021, 750, 143085.
    27. Luo, T.; Wang, C.; Pan, Z.; Jin, C.; Fu, Z.; Jin, Y., Maternal Polystyrene Microplastic Exposure during Gestation and Lactation Altered Metabolic Homeostasis in the Dams and Their F1 and F2 Offspring. Environmental Science & Technology 2019, 53 (18), 10978-10992.
    28. Wright, S. L.; Kelly, F. J., Plastic and Human Health: A Micro Issue? Environmental Science & Technology 2017, 51 (12), 6634-6647.
    29. Prata, J. C., Airborne microplastics: Consequences to human health? Environmental Pollution 2018, 234, 115-126.
    30. Yang, Y.-F.; Chen, C.-Y.; Lu, T.-H.; Liao, C.-M., Toxicity-based toxicokinetic/toxicodynamic assessment for bioaccumulation of polystyrene microplastics in mice. Journal of Hazardous Materials 2019, 366, 703-713.
    31. Huang, H.; Wei, F.; Qiu, S.; Xing, B.; Hou, J., Polystyrene microplastics trigger adiposity in mice by remodeling gut microbiota and boosting fatty acid synthesis. Science of The Total Environment 2023, 890, 164297.
    32. Wilkins, M. R., Proteome research: new frontiers in functional genomics. Springer Science & Business Media: 1997.
    33. Wilkins, M. R.; Sanchez, J.-C.; Gooley, A. A.; Appel, R. D.; Humphery-Smith, I.; Hochstrasser, D. F.; Williams, K. L. J. B.; reviews, g. e., Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it. 1996, 13 (1), 19-50.
    34. Edman, P.; Högfeldt, E.; Sillén, L. G.; Kinell, P.-O. J. A. c. s., Method for determination of the amino acid sequence in peptides. 1950, 4 (7), 283-293.
    35. Han, X.; Aslanian, A.; Yates, J. R., Mass spectrometry for proteomics. Current Opinion in Chemical Biology 2008, 12 (5), 483-490.
    36. Krause, E.; Wenschuh, H.; Jungblut, P. R., The Dominance of Arginine-Containing Peptides in MALDI-Derived Tryptic Mass Fingerprints of Proteins. Analytical Chemistry 1999, 71 (19), 4160-4165.
    37. Thiede, B.; Höhenwarter, W.; Krah, A.; Mattow, J.; Schmid, M.; Schmidt, F.; Jungblut, P. R., Peptide mass fingerprinting. Methods 2005, 35 (3), 237-247.
    38. Buncherd, H.; Roseboom, W.; de Koning, L. J.; de Koster, C. G.; de Jong, L., A gas phase cleavage reaction of cross-linked peptides for protein complex topology studies by peptide fragment fingerprinting from large sequence database. Journal of Proteomics 2014, 108, 65-77.
    39. Domon, B.; Broder, S., Implications of New Proteomics Strategies for Biology and Medicine. Journal of Proteome Research 2004, 3 (2), 253-260.
    40. Aggarwal, K.; Choe, L. H.; Lee, K. H., Shotgun proteomics using the iTRAQ isobaric tags. Briefings in Functional Genomics 2006, 5 (2), 112-120.
    41. Thompson, A.; Schäfer, J.; Kuhn, K.; Kienle, S.; Schwarz, J.; Schmidt, G.; Neumann, T.; Hamon, C., Tandem Mass Tags:  A Novel Quantification Strategy for Comparative Analysis of Complex Protein Mixtures by MS/MS. Analytical Chemistry 2003, 75 (8), 1895-1904.
    42. Ludwig, C.; Gillet, L.; Rosenberger, G.; Amon, S.; Collins, B. C.; Aebersold, R., Data-independent acquisition-based SWATH-MS for quantitative proteomics: a tutorial. Molecular Systems Biology 2018, 14 (8), e8126.
    43. Ross, P. L.; Huang, Y. N.; Marchese, J. N.; Williamson, B.; Parker, K.; Hattan, S.; Khainovski, N.; Pillai, S.; Dey, S.; Daniels, S.; Purkayastha, S.; Juhasz, P.; Martin, S.; Bartlet-Jones, M.; He, F.; Jacobson, A.; Pappin, D. J., Multiplexed Protein Quantitation in Saccharomyces cerevisiae Using Amine-reactive Isobaric Tagging Reagents*. Molecular & Cellular Proteomics 2004, 3 (12), 1154-1169.
    44. Wiese, S.; Reidegeld, K. A.; Meyer, H. E.; Warscheid, B., Protein labeling by iTRAQ: A new tool for quantitative mass spectrometry in proteome research. PROTEOMICS 2007, 7 (3), 340-350.
    45. Yamashita, M.; Fenn, J. B., Electrospray ion source. Another variation on the free-jet theme. The Journal of Physical Chemistry 1984, 88 (20), 4451-4459.
    46. Delahunty, C.; Yates Iii, J. R., Protein identification using 2D-LC-MS/MS. Methods 2005, 35 (3), 248-255.
    47. Li, D.; Dück, R.; Schmitz, O. J., The advantage of mixed-mode separation in the first dimension of comprehensive two-dimensional liquid-chromatography. Journal of Chromatography A 2014, 1358, 128-135.
    48. Yang, F.; Shen, Y.; Camp, D. G.; Smith, R. D., High-pH reversed-phase chromatography with fraction concatenation for 2D proteomic analysis. Expert Review of Proteomics 2012, 9 (2), 129-134.
    49. Kishimoto, Y.; Hiraiwa, M.; O'Brien, J. S., Saposins: structure, function, distribution, and molecular genetics. Journal of Lipid Research 1992, 33 (9), 1255-1267.
    50. Meyer, R. C.; Giddens, M. M.; Coleman, B. M.; Hall, R. A., The protective role of prosaposin and its receptors in the nervous system. Brain Research 2014, 1585, 1-12.
    51. Männistö, P. T.; Kaakkola, S. J. P. r., Catechol-O-methyltransferase (COMT): biochemistry, molecular biology, pharmacology, and clinical efficacy of the new selective COMT inhibitors. 1999, 51 (4), 593-628.
    52. Lo, V.; Erickson, B.; Thomason-Hughes, M.; Ko, K. W. S.; Dolinsky, V. W.; Nelson, R.; Lehner, R., Arylacetamide deacetylase attenuates fatty-acid-induced triacylglycerol accumulation in rat hepatoma cells. Journal of Lipid Research 2010, 51 (2), 368-377.
    53. Trickett, J. I.; Patel, D. D.; Knight, B. L.; Saggerson, E. D.; Gibbons, G. F.; Pease, R. J., Characterization of the Rodent Genes for Arylacetamide Deacetylase, a Putative Microsomal Lipase, and Evidence for Transcriptional Regulation * 210. Journal of Biological Chemistry 2001, 276 (43), 39522-39532.
    54. Grenier, J.; Tomkiewicz, C.; Trousson, A.; Rajkowski, K. M.; Schumacher, M.; Massaad, C., Identification by microarray analysis of aspartate aminotransferase and glutamine synthetase as glucocorticoid target genes in a mouse Schwann cell line. The Journal of Steroid Biochemistry and Molecular Biology 2005, 97 (4), 342-352.
    55. Tordjman, J.; Leroyer, S.; Chauvet, G.; Quette, J.; Chauvet, C.; Tomkiewicz, C.; Chapron, C.; Barouki, R.; Forest, C.; Aggerbeck, M.; Antoine, B., Cytosolic Aspartate Aminotransferase, a New Partner in Adipocyte Glyceroneogenesis and an Atypical Target of Thiazolidinedione *. Journal of Biological Chemistry 2007, 282 (32), 23591-23602.
    56. Palmieri, F., The mitochondrial transporter family (SLC25): physiological and pathological implications. Pflügers Archiv 2004, 447 (5), 689-709.
    57. An, Y. A.; Chen, S.; Deng, Y.; Wang, Z. V.; Funcke, J.-B.; Shah, M.; Shan, B.; Gordillo, R.; Yoshino, J.; Klein, S.; Kusminski, C. M.; Scherer, P. E., The mitochondrial dicarboxylate carrier prevents hepatic lipotoxicity by inhibiting white adipocyte lipolysis. Journal of Hepatology 2021, 75 (2), 387-399.
    58. Murugan, M.; Fedele, D.; Millner, D.; Alharfoush, E.; Vegunta, G.; Boison, D., Adenosine kinase: An epigenetic modulator in development and disease. Neurochemistry International 2021, 147, 105054.
    59. Pereira, C. V.; Peralta, S.; Arguello, T.; Bacman, S. R.; Diaz, F.; Moraes, C. T., Myopathy reversion in mice after restauration of mitochondrial complex I. EMBO Molecular Medicine 2020, 12 (2), e10674.
    60. Tiranti, V.; Viscomi, C.; Hildebrandt, T.; Di Meo, I.; Mineri, R.; Tiveron, C.; D Levitt, M.; Prelle, A.; Fagiolari, G.; Rimoldi, M.; Zeviani, M., Loss of ETHE1, a mitochondrial dioxygenase, causes fatal sulfide toxicity in ethylmalonic encephalopathy. Nature Medicine 2009, 15 (2), 200-205.

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