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???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
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dc.contributor.advisor | 李妮鍾 | |
dc.contributor.author | Ping-Shang Wu | en |
dc.contributor.author | 吳蘋珊 | zh_TW |
dc.date.accessioned | 2021-06-17T01:31:45Z | - |
dc.date.available | 2017-09-12 | |
dc.date.copyright | 2017-09-12 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-08-03 | |
dc.identifier.citation | 1. Ufnal, M., A. Zadlo, and R. Ostaszewski, TMAO: A small molecule of great expectations. Nutrition, 2015. 31(11-12): p. 1317-23.
2. Treberg, J.R., et al., The accumulation of methylamine counteracting solutes in elasmobranchs with differing levels of urea: a comparison of marine and freshwater species. J Exp Biol, 2006. 209(Pt 5): p. 860-70. 3. Mackay, R.J., et al., Trimethylaminuria: causes and diagnosis of a socially distressing condition. Clin Biochem Rev, 2011. 32(1): p. 33-43. 4. Randrianarisoa, E., et al., Relationship of Serum Trimethylamine N-Oxide (TMAO) Levels with early Atherosclerosis in Humans. Sci Rep, 2016. 6: p. 26745. 5. Koeth, R.A., et al., Intestinal microbiota metabolism of L-carnitine, a nutrient in red meat, promotes atherosclerosis. Nat Med, 2013. 19(5): p. 576-85. 6. Wang, Z., et al., Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature, 2011. 472(7341): p. 57-63. 7. Wu, L. and B.H. Juurlink, Increased methylglyoxal and oxidative stress in hypertensive rat vascular smooth muscle cells. Hypertension, 2002. 39(3): p. 809-814. 8. Ufnal, M., A. Zadlo, and R. Ostaszewski, TMAO: A small molecule of great expectations. Nutrition, 2015. 31(11): p. 1317-1323. 9. Carvajal, K. and R. Moreno-Sánchez, Heart Metabolic Disturbances in Cardiovascular Diseases. Archives of Medical Research, 2003. 34(2): p. 89-99. 10. Ascunce, R.R., A.C. Nayar, and C.K. Phoon, Cardiac magnetic resonance findings in a case of carnitine deficiency. Texas Heart Institute Journal, 2013. 40(1): p. 104. 11. Adachi, T., et al., Decreased serum carnitine is independently correlated with increased tissue accumulation levels of advanced glycation end products in haemodialysis patients. Nephrology (Carlton), 2012. 17(8): p. 689-94. 12. Stanley, C.A., M.J. Bennett, and E. Mayatepek, Disorders of mitochondrial fatty acid oxidation and related metabolic pathways, in Inborn metabolic diseases. 2006, Springer. p. 175-190. 13. Buyse, J., et al., Dietary L-carnitine supplementation enhances the lipopolysaccharide-induced acute phase protein response in broiler chickens. Vet Immunol Immunopathol, 2007. 118(1-2): p. 154-9. 14. Longo, N., C. Amat di San Filippo, and M. Pasquali, Disorders of carnitine transport and the carnitine cycle. Am J Med Genet C Semin Med Genet, 2006. 142C(2): p. 77-85. 15. Hwu, W.-L., et al., Deficiency of the carnitine transporter (OCTN2) with partial N-acetylglutamate synthase (NAGS) deficiency. Journal of inherited metabolic disease, 2007. 30(5): p. 816. 16. Tseng, C.-H., Mortality and causes of death in a national sample of diabetic patients in Taiwan. Diabetes care, 2004. 27(7): p. 1605-1609. 17. 行政院衛生福利部 (2017)。105 年台灣地區主要死亡原因統計。2017年06月19日, 取自 http://www.mohw.gov.tw/cht/Ministry/。 18. Criqui, M.H., et al., Progression of peripheral arterial disease predicts cardiovascular disease morbidity and mortality. Journal of the American College of Cardiology, 2008. 52(21): p. 1736-1742. 19. Tang, W.H., et al., Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N Engl J Med, 2013. 368(17): p. 1575-84. 20. Bánhegyi, G., et al., Endoplasmic reticulum stress. Annals of the New York Academy of Sciences, 2007. 1113(1): p. 58-71. 21. Woltjer, R.L., et al., Effects of chemical chaperones on oxidative stress and detergent-insoluble species formation following conditional expression of amyloid precursor protein carboxy-terminal fragment. Neurobiol Dis, 2007. 25(2): p. 427-37. 22. Vrbanac, J.J. and S.H. Zeisel, The measurement of dimethylamine, trimethylamine, and trimethylamine N-oxide using capillary gas chromatography-mass spectrometry. Analytical biochemistry, 1990. 187(2): p. 234-239. 23. Chaferpericas, C., R. Herraezhernandez, and P. Campinsfalco, Liquid chromatographic determination of trimethylamine in water. Journal of Chromatography A, 2004. 1023(1): p. 27-31. 24. Abeling, N., et al., Diagnosis of a new case of trimethylaminuria using direct proton NMR spectroscopy of urine. Journal of inherited metabolic disease, 1995. 18(2): p. 182-184. 25. Bain, M.A., et al., Quantifying trimethylamine and trimethylamine-N-oxide in human plasma: interference from endogenous quaternary ammonium compounds. Anal Biochem, 2004. 334(2): p. 403-5. 26. Arthur, C.L. and J. Pawliszyn, Solid phase microextraction with thermal desorption using fused silica optical fibers. Analytical chemistry, 1990. 62(19): p. 2145-2148. 27. Kadar, H., et al., A multiplexed targeted assay for high-throughput quantitative analysis of serum methylamines by ultra performance liquid chromatography coupled to high resolution mass spectrometry. Arch Biochem Biophys, 2016. 597: p. 12-20. 28. Miller, M.J., et al., Chronic oral l-carnitine supplementation drives marked plasma TMAO elevations in patients with organic acidemias despite dietary meat restrictions. 2016. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/67426 | - |
dc.description.abstract | 背景
氧化三甲胺,又稱三甲胺氧化物( Trimethylamine N-oxide、TMAO )為腸道菌群代謝的產物,當富含有膽鹼(choline)及肉鹼(carnitine)等食物進入腸道會被腸內道細菌代謝成三甲胺(Trimethylamine、TMA),三甲胺會再經由肝臟的酵素(FMO3)代謝成氧化三甲胺。以往對氧化三甲胺的認知為體內的代謝物,在體中是無毒性的小分子,對人體也無不好的影響,然而,近年來有研究指出,氧化三甲胺會造成動脈粥狀硬化(Atherosclerosis),增加心血管疾病的機率。雖然也有一些研究顯示氧化三甲胺反而是細胞遭遇壓力下的保護機轉,所以血液中TMAO上升對人體的確切影響仍然未明。 肉鹼(L-carnitine)是先天代謝異常患者如有機酸血症(organic academia)、原發性肉鹼缺乏症(primary carnitine deficiency)以及脂肪酸代謝異常(fatty acid oxidation defect)所需要使用的藥物,藉以維持體內代謝的穩定。因為有研究指出肉鹼的代謝物氧化三甲胺會造成動脈粥狀硬(Atherosclerosis),這引起了此肉鹼的使用在先天代謝異常患者的疑慮,到底補充肉鹼對於這類患者心血管的影響如何?目前並沒有解答。 因為氧化三甲胺在人體極為微量,以往用生化儀器檢測不易,近年來因為質譜儀(mass spectrometry, MS)的發展才有許多人體的微量物質能被檢測,目前氣相層析串聯質譜儀(gas chromatography-tandem mass spectrometry,GC-MS)、液相層析串聯質譜儀(liquid chromatography-tandem mass spectrometry, LC-MS/MS)用在人體微量分子的檢測。目前只有鮮少的醫療院所有在檢測氧化三甲胺,我們欲運用質譜儀(MS)去開發一個簡易、快速、精準的氧化三甲胺檢測技術。 因此我們想探討長期使用高劑量口服肉鹼患者其血液中氧化三甲胺的變化,藉以了解這種長期使用高劑量口服肉鹼治療對先天性代謝異常病人的影響。尤其是先天代謝異常患者照護越來越進步的年代,許多以前無法存活的患者都可以長大,我們希望能及早知道目前肉鹼治療對病人可能的影響,並擬定治療措施。 方法 我們收集長期服用肉鹼的先天代謝異常患者(包括有機酸血症與脂肪酸代謝異常)病友16人、與不需服用肉鹼的先天代謝異常患者10人及年齡性別相符的健康人8人作為對照組參加此研究,我們將採10 mL周邊血液,並利用氣相質譜儀及液相質譜儀,分析其血中三甲胺及氧化三甲胺的含量。 結果 結果顯示本分析方法操作簡單且快速,對氧化三甲胺的檢量線線性 (濃度範圍1至500 ng mL-1、R2 > 0.990)、靈敏度(最低可定量濃度為1 ng mL-1) 及最低可偵測濃度為1 ng mL-1,且有良好的精密度(CV < 9.67 % )及準確度(82.84 % - 111.39 % )。長期服用肉鹼的先天代謝異常患者、不需服用肉鹼的先天代謝異常患者與健康人的血中氧化三甲胺濃度分別為2779.67 ng mL-1、464.31 ng mL-1、71.52 ng mL-1。利用此方法我們發現有服用肉鹼的病人血中氧化三甲胺濃度會比沒有服用肉鹼的人高 ( p =0.016 )。 結論 藉由此研究,我們建立了檢測血漿氧化三甲胺的方法,並確認了肉鹼與氧化三甲胺的關聯性,做為日後治療方向擬定的參考。 | zh_TW |
dc.description.abstract | Background
Trimethylamine N-oxide (TMAO) is a gut-derived metabolite which is generated by bacterial conversion of phosphatidylcholine, choline, betaine and carnitine. Carnitine is abundant in red meat. It is subsequently converted to gaseous trimethylamine. For this reason, TMAO has been proposed to constitute a link between diet and cardiovascular disease(CVD). High TMAO concentrations may increase atherosclerosis by suppressing reverse cholesterol transport (RCT) and bile acid synthesis, therefore, it has been suggested that chronic L-carnitine supplementation might be harmful and may accelerate atherosclerosis. On the other hand, carnitine supplementation is an important treatment in certain types of inborn error of metabolism (IEM). In addition to organic acidemias, fatty acid oxidation defect is another group of the IEM and needs long-term carnitine supplementation. The current treatment for the disease is carnitine supplementation to keep intracellular carnitine concentration. To evaluate the correlation between atherosclerosis and cardiovascular events and carnitine supplementation, the study focused on the correlation between long-term oral carnitine supplementation (more than 1 year) and the TMAO in blood. This study has focused on developing a simple, rapid, and cost-effective quantitative assay for TMAO in plasma. We used the gas chromatographic -mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) and in the purpose of comparing which one can detect TMAO more sensitively and specifically. Material and method We collected plasma samples from 16 IEM patients who need long-term carnitine supplementations, 11 IEM patients with normal acylcarnitine profiles and 8 healthy subjects as control. We collected 10 ml peripheral blood and analyze their Trimethylamine (TMA) and TMAO by using GC-MS and LC-MS. Result We have developed an easy and rapid LC-MS assay for quantification of TMAO. The calibration linear range is 0.5 ng mL-1 to 500 ng mL-1. The LOD is 0.5 ng mL-1. The intra- and inter-day precision (CV< 10%) and high accuracy (< 20% error) were observed using this method. The TMAO level in patient with carnitine supplementation, patient without carnitine supplementation and healthy control is 2779.67 ng mL-1、464.31 ng mL-1、71.52 ng mL-1, respectively. Furthermore, the plasma samples collected in our clinical study has demonstrated that the supplement of carnitine resulted in relative high TMAO in plasma. Conclusion Via this study, we established the simple method to detect TMAO, and also established the correlation between oral carnitine supplementation and TMAO. This will help future treatment strategies formulation in IEM patients. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T01:31:45Z (GMT). No. of bitstreams: 1 ntu-106-P04448011-1.pdf: 1875308 bytes, checksum: 0b4d19f2d9714b78fd2209ef5a26e2e9 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 誌謝 i
中文摘要 ii 英文摘要 iv Contents vi List of Figures viii List of Tables x Terminology xi Chapter 1. Introduction 1 1.1The Basis of TMAO 1 1.2 Correlation of cardiovascular disease and TMAO 3 1.3 Motivation 5 Chapter 2. Materials and Methods 6 2.1 Instrument 6 2.1.1 GC-MS 6 2.1.2 LC-MS 7 2.2 Extraction 8 2.3 Chemical and reagents 9 2.3.1 For GC-MS Method 9 2.3.2 For LC-MS Method 9 2.4 Preparation of Standards, Calibration and QC 10 2.4.1 GC-MS 10 2.4.2 LC-MS 10 2.5 Sample preparation 12 2.5.1 GC-MS 12 2.5.2 LC-MS 13 2.6 Method validation 13 2.7 Participant recruitment 14 Chapter 3. Result 15 3.1 Condition for GC-MS 15 3.2 Identification of the TMAO by LC-MS 16 3.2.1 Accuracy and precision 17 3.2.2 Recovery and carryover 17 3.2.3 The storage and stability 18 3.3 Distribution of TMAO in normal subjects 19 3.4 Distribution of TMAO levels in patients with long-term carnitine supplementation 19 Chapter 4. Discussions 21 4.1 Method validation 21 4.1.1 Calibration and linearity 21 4.1.2 Accuracy and precision 22 4.1.3 Recovery and carryover 22 4.1.4 The storage and stability 23 4.2 Detection of TMAO in plasma 25 4.3 Patients with long-term carnitine supplementation 26 4.4 The application of TMAO measurement in clinical chemistry 27 4.5 Limitation of the study 29 Chapter 5. Conclusions 30 Reference 31 | |
dc.language.iso | zh-TW | |
dc.title | 探討長期服用carnitine病人血中Trimethylamine N-oxide的濃度 | zh_TW |
dc.title | Plasma Trimethylamine N-oxide level
in patients with long-term carnitine supplementation | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 陳珮珊 | |
dc.contributor.oralexamcommittee | 胡務亮,簡潁秀 | |
dc.subject.keyword | 氧化三甲胺,三甲胺,動脈粥狀硬化,肉利用氣相質譜儀及鹼,先天代謝異常,液相層析質譜儀,氣相層析質譜儀, | zh_TW |
dc.subject.keyword | Trimethylamine N-oxide (TMAO),Trimethylamine (TMA),Gas Chromatography-Tandem Mass Spectrometry (GC-MS),Liquid chromatography-tandem mass spectrometry(LC-MS),atherosclerosis,inborn error of metabolism (IEM), | en |
dc.relation.page | 60 | |
dc.identifier.doi | 10.6342/NTU201702388 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-08-03 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
Appears in Collections: | 分子醫學研究所 |
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