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研究生:池孟修
研究生(外文):Meng-Hsiu Chih
論文名稱(外文):The Culprit of Gout: Triggering Factors and Formation of Monosodium Urate Monohydrate
指導教授:李度李度引用關係
指導教授(外文):Tu Lee
學位類別:碩士
校院名稱:國立中央大學
系所名稱:化學工程與材料工程學系
學門:工程學門
學類:化學工程學類
論文出版年:2015
畢業學年度:103
語文別:英文
論文頁數:88
中文關鍵詞:痛風尿酸單鈉鹽玻尿酸尿酸
外文關鍵詞:goutmonosodium urate monohydratehyaluronateuric acid dihydrate
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痛風是一種常見尿酸鈉鹽結晶引發的關節炎。據估計全世界至少有2.5%的人深受痛風所苦。高尿酸血症為現今醫學公認引發痛風的主因,但是只有少部分高尿酸血症患者會出現痛風的症狀。而且,目前對於尿酸鈉鹽於人體的形成機制尚未明朗。因此,本篇論文將有四大研究目標,以試圖解開痛風形成之謎。第一,觀察尿酸鈉鹽結晶於不同鈉離子溶液中的形成與成長。第二,研究尿酸晶體在仿生溶液中轉化成尿酸鈉鹽之過程,並且找出pH值與痛風形成之關聯性。第三,瞭解玻尿酸與體內常見陽離子之反應,以及其對於尿酸鈉鹽結晶之影響。第四,檢測20種人體常見胺基酸對尿酸鈉鹽結晶的影響。
依據實驗結果與相關文獻參照,本論文提出八項重要發現:(1) 在人體條件下,只需要調整溶液的鈉離子濃度即可製備出不同形貌的尿酸鈉鹽結晶分別為球晶、海膽狀結晶與蝴蝶結狀團聚;(2)尿酸鈉鹽結晶之成長演進,依序從球晶、海膽狀結晶到最後形成蝴蝶結狀團聚;(3)球晶為所有尿酸鹽晶型的前驅型態;(4)藉由玻尿酸、鈉離子與鈣離子溶液,我們觀察到全新形貌的尿酸鈉鹽;(5)我們藉由降低溶液的pH值促使尿酸結晶沉澱,並且觀察尿酸結晶於仿生液中轉化為尿酸鈉鹽之過程;(6)尿酸的轉化可以解釋醫學上對於痛風相關發現,我們進而提出痛風於人體真正的發病機制;(7)我們提出玻尿酸鏈、鈣離子與尿酸離子會形成複合物藉此抑制尿酸鈉鹽之形成。而且可以說明為何只有少數的高尿酸血症患者會罹患痛風;和(8)某些特定的氨基酸能夠抑制尿酸鈉鹽的形成。

Gout is the most widespread crystal-induced inflammatory arthritis by monosodium urate monohydrate (MSUM). It was estimated at least 2.5% people suffering from it all over the world. Hyperuricemia is essential for the MSUM formation, but only some of patients are able to develop gout. Therefore, there are 4 aims in the research to reveal the mystery of MSUM formation in vivo: First, to produce beachballs and other MSUM morphologies with various Na+ ions level which will unravel the pathogenesis of gout. Second, to observe the conversion from uric acid to MSUM in the simulated body fluid system. Third, to explore the synergistic effect of hyaluronate, Na+ ion and Ca2+ ion in MSUM crystallization. Forth, to find the influence of the common 20 kinds of amino acid in human body on MSUM crystallization. According to the experimental results and reference, we proposed 8 significant findings in this study: various morphologies of MSUM: (1) beachball, spherulite and bow-like aggregate could be prepared at different Na+ ion levels at 37C, (2) morphological transformation from beachball to spherulite to bow-like aggregate can be achieved, (3) beachball is a disordered, first-formed precursor for all MSUM morphologies, (4) a new type of MSUM fishtail morphology was observed in the hyaluronate, Na+ and Ca2+ ion containing solutions, (5) the dilemma of MSUM deposition at thermodynamically unfavorable low pH condition was solved by the newly discovered kinetic pathway for the phase conversion of uric acid dihydrate (UAD) to MSUM, (6) the UAD-to-MSUM pathway can also be used to explain the effects brought about by the inflammatory response and acidosis, and why MSUM deposition in cartilage and connective tissue, (7) the question of why only a fraction of hyperuricemic patients has gout was answered by the solubilization of urate through the complex formation with hyaluronate chains and Ca2+ ions, and (8) some amino acids can influence the size of MSUM crystal.
摘要 i
Abstract ii
Acknowledgments iv
Table of Contents v
List of Figures vii
List of Tables x
Chapter 1 1
Introduction 1
1.1 Gout and the Discovery of Monosodium Urate Monohydrate (MSUM) 1
1.2 Introduction of MSUM 5
1.3 Aims of the Research 9
1.4 References 14
Chapter 2 20
Experimental Methods 20
2.1 Materials 20
2.2 Instrumentation 25
2.2.1 Polarizing Optical Microscopy (POM) 25
2.2.2 Scanning Electron Microscopy (SEM) 25
2.2.3 Powder X-ray Diffraction (PXRD) 26
2.2.4 Fourier Transform Infrared Spectroscopy (FT-IR) 26
2.2.5 Thermogravimetric Analysis (TGA) 27
2.2.6 Ultraviolet and Visible (UV/Vis) Spectroscopy 27
2.3 Experimental Methods 28
2.3.1 Morphological Studies of MSUM under Various Na+ Ion Levels 28
2.3.2 pH Effect and Conversion from UAD to MSUM 29
2.3.3 Synergistic Effect of Ca2+, K+ and Na+ Ions and Hyaluronate Chains on MSUM Crystallization 31
2.3.4 The Influence of Various Amino Acids on MSUM Crystallization 33
2.4 References 35
Chapter 3 36
Results & Discussion 36
3.1 Morphological Studies of MSUM under Various Na+ Ion Levels 36
3.2 pH Effect and Conversion of MSUM 43
3.2.1 Crystallization of Uric Acid Dihydrate (UAD) and Uric Acid Anhydrate (UAA) 43
3.2.2 Conversion from UAD to MSUM 45
3.2.3 Conversion from UAA to MSUM 50
3.3 Synergistic Effect of Ca2+, K+ and Na+ Ions and Hyaluronate Chains on MSUM Crystallization 53
3.4 The Influence of Various Amino Acids on MSUM Crystallization 60
3.5 References 66
Chapter 4 69
Conclusions & Future Works 69
4.1 Conclusions 69
4.2 Future Work 71
4.3 References 72

Chapter 1
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Schubert, G.; Reck, G.; Jancke, H.; Kraus, W.; Patzelt, C. Uric Acid Monohydratea New Urinary Calculus Phase. Urol. Res. 2005, 33(3), 231-238.
Zellelow, A. Z.; Kim, K. H.; Sours, R. E.; Swift, J. A. Solid-State Dehydrate of Uric Acid Dihydrate. Cryst. Growth Des. 2010,10(1), 418-425.
Narayana Kalkura, S.; Vaidyan, V. K.; Kanakavel, M.; Ramasamy, P. Crystallization of Uric Acid. J. Cryst. Growth 1993, 132(3-4), 617-620.
Grases, F.; Villacampa, A. I.; Costa-Bauzá, A.; Sӧhnel, O. Uric Acid Calculi Scanning Mircosc. 1999, 13(2-3), 223-234.
Presores, J. B.; Swift, J. A. Solution-mediated Phase Transformation of Uric Acid Dihydrate. CrystEngComm 2014, 16(31), 7278-7284.
Sheehan, J. K.; Arundel, C.; Phelps, C. F. Effect of the Cations Sodium, Potassium and Calcium on the Interaction of Hyaluronate Chains: a Light Scattering and Viscometric Study. Int. J. Biol. Macromol. 1983, 5(4), 222-228.
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Chapter 2
Calvert, P. D.; Fiddis, R. W.; Vlachos, N. Crystal Growth of Monosodium Urate Monohydrate. Colloids Surf. 1985, 14(1), 97-107.
Finlyason, B.; Smith, A. Stability of First Dissociable Proton of Uric Acid J. Chem. Eng. Data 1974, 19(1), 94-97.
Perrin, C. M.; Dobish, M. A.; Keuren E. V.; Swift, J. A. Monosodium Urate Monohydrate Crystallization. CrystEngComm 2011, 13(4), 1111-1117.
Presores, J. B.; Swift, J. A. Solution-mediated Phase Transformation of Uric Acid Dihydrate. CrystEngComm 2014, 16(31), 7278-7284.
Oyane, A.; Kim, H. M.; Furuya, T.; Kokubo, T.; Miyazaki, T.; Nakamura, T. Preparation and Assessment of Revised Simulated Body Fluids. J. Biomed. Res. A. 2003, 65(2), 188-195.
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Grases, F.; Villacampa, A. I.; Costa-Bauzá, A.; Sӧhnel, O. Uric Acid Calculi Scanning Mircosc. 1999, 13(2-3), 223-234.
Tak, H. K.; Wilcox, W. R.; Cooper, S. M. Crystallization of Monosodium Urate and Calcium Urate at 37 oC. J. Colloid Interface Sci. 1980, 77(1), 195-201.
Strachan, R. K.; Smith, P.; Gardner, D. L. Hyaluronate in Rheumatology and Orthopaedics: Is There a Role? Ann. Rheum. Dis. 1990, 49(11), 949-952.

Chapter 3
McCarty, D. J.; Hollander, J. L. Identification of Urate Crystals in Gouty Synovial Fluid. Ann. Intern. Med. 1961, 54(3), 452-460.
McGill, N. W.; Hayes, A. Dieppe, P. A. Morphological Evidence for Biological Control of Urate Crystal Formation in Vivo and in Vitro. Scand. J. Rheumatol. 1992, 21(5), 215-219.
Kalkura, S. N.; Girija, E. K.; Kanakavel, M.; Ramasamy, P. In-Vitro Crystallization of Spherulites of Monosodium Urate Monohydrate. J. Mater. Sci. – Mater. Med. 1995, 6(10), 577-580.
Parekh, B. B.; Vasant, S. R.; Tank, K. P.; Raut A.; Vaidya, A. D. B.; Joshi, M. J. In Vitro Growth and Inhibition Sturdies of Monosodium Urate Monohydrate Crystals by Different Herbal Extracts. Am. J. Infect. Dis. 2009, 5(3), 225-230.
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Fiechtner, J. J.; Simkin, P. A. Urate Spherulites in Gout Synovia. JAMA, 1981, 245(15), 1533-1536.
Wilcox, W. R.; Khalaf, A. A. Nucleation of Monosodium Urate Crystals. Ann. Rheum. Dis. 1975, 34(4), 332-339.
Katz, W. A. Deposition of Urate Crystals in Gout. Arthritis Rheum. 1975, 18(S1), 751-756.
Katz, W. A.; Schubert, M. The Interaction of Monosodium Urate with Connective Tissue Components. J. Clin. Invest. 1970, 49(10), 1783-1789.
Seegmiller, J. E.; Rodney Howell, R. The Old and New Concepts of Acute Gouty Arthritis. Arthritis Rheum. 1962, 5(6), 616-623.
Martillo, M. A.; Nazzal, L.; Crittenden, D. B. The Crystallization of Monosodium Urate. Curr. Rheumatol. Rep. 2014, 16(2), 400.
Wilcox, W. R.; Khalaf, A. Solubility of Uric Acid and Monosodium Urate. Med. Biol. Eng. 1972, 10(4), 522-531.
Schubert, G.; Reck, G.; Jancke, H.; Kraus, W.; Patzelt, C. Uric Acid Monohydratea New Urinary Calculus Phase. Urol. Res. 2005, 33(3), 231-238.
Narayana Kalkura, S.; Vaidyan, V. K.; Kanakavel, M.; Ramasamy, P. Crystallization of Uric Acid. J. Cryst. Growth 1993, 132(3-4), 617-620.
Zellelow, A. Z.; Kim, K. H.; Sours, R. E.; Swift, J. A. Solid-State Dehydrate of Uric Acid Dihydrate. Cryst. Growth Des. 2010,10(1), 418-425.
Presores, J. B.; Swift, J. A. Solution-mediated Phase Transformation of Uric Acid Dihydrate. CrystEngComm 2014, 16(31), 7278-7284.
Mandel, N. S.; Mandel, G. S. Monosodium Urate Monohydrate, the Gout Culprit. J. Am. Chem. Soc. 1976, 98(8), 2319-2323.
Wang, Z.; Konigsberger, E. Solubility Equilibria in the Uric Acid-Sodium Urate-Water System. Thermochim. Acta 1998, 310(1-2), 237-242.
Grases, F.; Villacampa, A. I.; Costa-Bauzá, A.; Sӧhnel, O. Uric Acid Calculi Scanning Mircosc. 1999, 13(2-3), 223-234.
Treuhaft, P. S.; McCarty, D. J. Synovial Fluid pH, Lactate, Oxygen and Carbon Dioxide Partial Pressure in Various Joint Disease. Arthritis Rheum. 1971, 14(4), 475-484.
Loeb, J. N. The Influence of Temperature on the Solubility of Monosodium Urate. Arthritis Rheum. 1972, 15(2), 189-191.
Oliviero, F.; Scanu, A.; Punzi, L. Metabolism of Crystal within the Joint. Reumatismo. 2011, 63(4), 221-229.
Calvert, P. D.; Fiddis, R. W.; Vlachos, N. Crystal Growth of Monosodium Urate Monohydrate. Colloids Surf. 1985, 14(1), 97-107.
Khalaf, A. A.; Wilcox, W. R. Solubility and Nucleation of Monosodium Urate in Relation to Gouty Arthritis. J. Cryst. Growth 1973, 20(3), 227-232.
Sheehan, J. K.; Arundel, C.; Phelps, C. F. Effect of the Cations Sodium, Potassium and Calcium on the Interaction of Hyaluronate Chains: a Light Scattering and Viscometric Study. Int. J. Biol. Macromol. 1983, 5(4), 222-228.
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Imberty, A.; Lortat-Jacob, H.; Pérez, S. Structural View of Glycosaminoglycan–Protein Interactions. Carbohydr. Res. 2007, 342(3-4), 430-439.
Presores, J. B.; Cromer, K. E.; Capacci-Daniel, C; Swift, J. A. Calcium Urate Hexahydrate. Cryst. Growth Des. 2013, 13(12), 5162-5164.
Strachan, R. K.; Smith, P.; Gardner, D. L. Hyaluronate in Rheumatology and Orthopaedics: Is There a Role? Ann. Rheum. Dis. 1990, 49(11), 949-952.
Pascual E.; Andres M.; Vela P. Gout Treatment: Should We Aim for Rapid Crystal Dissolution. Ann. Rheum. Dis. 2013, 72(5), 635-637.

Chapter 4
Ascual E.; Andres M.; Vela P. Gout Treatment: Should We Aim for Rapid Crystal Dissolution. Ann. Rheum. Dis. 2013, 72(5), 635-637.
Tamasi, G.; Cini, R.; Gregorkiewitz, M.; Lorenzini, S.; Marcolongo, R.; Cavallo, G. The Dissolution of Monosodium Urate Monohydrate Crystals Formulation of a Biocompatible Buffer Solution with Potential use in the Treatment of Gouty Arthropathies. Rheumatol. Rep. 2013, 5(1), e4.

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