研究生: |
林家漢 Chia-Han Lin |
---|---|
論文名稱: |
豬關節軟骨中的蛋白質以及醣胺素之生化分析 Biochemical analysis of porcine articular cartilage |
指導教授: |
張兗君
Yen-Chung Chang |
口試委員: | |
學位類別: |
碩士 Master |
系所名稱: |
生命科學暨醫學院 - 分子醫學研究所 Institute of Molecular Medicine |
論文出版年: | 2004 |
畢業學年度: | 92 |
語文別: | 中文 |
論文頁數: | 70 |
中文關鍵詞: | 醣胺素 、軟骨 、蛋白體學 、二維電泳 |
外文關鍵詞: | glycosaminoglycan, cartilage, proteomics, 2D-PAGE |
相關次數: | 點閱:2 下載:0 |
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我們使用豬的關節軟骨做為實驗模型,以不同溫度及時間對軟骨組織中的蛋白質以及醣胺素進行萃取,進而對萃取物中之蛋白質以及醣胺素成分進行定量及定性分析。
目前,我們發現將軟骨碎片冷凍乾燥後以 guanidine HCl 並加溫至60℃以上,可以快速萃取出大量以及完整之蛋白,目前已針對這些萃取蛋白進行蛋白體學之分析。正在進行中的工作包括:探討萃取醣胺素之最佳條件,以及建立分析醣胺素之方法學。預定在上述方法學建立完備之後,進而分析正常與病變之人類軟骨組織,以期對軟骨之各種組成的功能有更深入的了解,並能提供組織工程在人工軟骨研發上有用的資訊。
We use porcine articular cartilage as an experimental model to extract proteins and glycosaminoglycans to optimize the conditions for extraction of proteins and GAGs from cartilage and the procedures for carrying out biochemical analyses of the extractable materials.
Currently, we can extract sufficiently large amount of proteins after treating the lyophilized cartilage slices with guanidine HCl at
60℃, and we have conducted proteomic analysis of these extractable proteins. The work still in progress include: to optimize the extract conditions for glycosaminoglycans and to establish the methodology for glycosaminoglycans analysis. We will analyze normal and pathological human cartilage after establishing above methodologies in order to understanding the function of various cartilage components and to apply useful information for the tissue engineers to develop artificial cartilage.
Adams ME, Hype about glucosamine. The Lancet 354, 353-354 (1999).
Aigner T, McKenna L. Molecular pathology and pathobiology of osteoarthritis cartilage. Cell. Mol.Life.Sci. 59, 5-18 (2002).
Aigner T, Loos S, Muller S, Sandell LJ, Unni KK, Kirchner T. Cell differentiation and matrix gene expression in mesenchymal chondrosarcomas. Am. J. Patho. 156, 1327-1335 (2000).
Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P, Molecular Biology of the Cell, Fourth Edition, 1065 (Garland Science, 2002)
Berkelman T, Stenstedt T, 2-D Electrophoresis using immobilized pH gradients- Principles and Methods, (Amersham Bioscience, 1998)
Carney SL, in Proteoglycans, (ed. Chaplin MF and Kennedy JF.), Carbohydrate analysis : a practical approach, 97-141 (IRL press, 1987)
Chevanne M, Caldini R, Manao G, Ruggiero M. Heparin binding peptides co-purify with glycosaminoglycans from human plasma. FEBS Letters 463, 121-124 (1999).
Fife RS, Beckerman BK, Houser DS, Proctor C, Liepnieks J, Masudall I, McCarty DJ, Ryanll LM. Evidence that a 550,000-Dalton cartilage matrix glycoprotein is a chondrocyte membrane-associated protein closely related to ceruloplasmin. J. Biol. Chem. 268, 4407-4411 (1993).
Fisher LW, Robey PG, Young MF, Termine JD. in Bone glycoproteins, (ed. Cunningham, L. W.), Methods in enzymology ;V.145,269-272 (Academic Press, 1987).
Geng Y, Blanco FJ, Cornlisson M, Loyz M. Regulation of cyclooxygenase-2 expression in normal human articular chondrocyte. J Immunol. 155, 796-801 (1995).
Hascall VC, Calabro A, Midura RJ, Yanagishita M. in Isolation and characterization of proteoglycans, (ed. Lennarz, W. J. & Hart, G. W.), Methods in enzymology V.230, 403-404 (Academic Press, 1994)
Heinegard D, Sommarin Y, Hedbom E, Wieslander J, Larsson B. Assay of proteoglycan population using ararose-polyacryamide gel electrophoresis. Anal Biochem. 151, 41-48 (1985).
Heinegard D, Larsson T, Sommarin Y, Franzen A, Paulsson M, Hedbom E. Two novel matrix proteins isolated from articular cartilage show wide distributions among connective tissues. J Biol Chem. 261, 13866-13872 (1986).
Van den Hoogen BM, van Weeren PR, Lopes-Cardozo M, van Golde LMG, Barneveld A, van de lest CHA. A microtiter plate assay for the determination of uronic acids. Anal Biochem. 257, 107-111 (1998).
Hou WS, Li Z, Gordon RE, Chan K, Klein MJ, Levy R, Keysser M, Keyszer G, Bromme D. Cathepsin K is a critical protease in synovial fibroblast-mediated collagen degradation. Am J Pathology 159, 2167-2177 (2001).
Houpt J, McMillan R, Wein C, Paget-Dellio SD. Effect of glucosamine hydrochloride in the treatment of pain on osteoarthritis of the knee. J Rheumatol. 26, 2423-2430 (1999).
Hyttinen MM, Arokoski JPA, Parkkinen JJ, Lammi MJ, Lapvetelainen T, Mauranen K, Kiraly K, Tammi MI, Helminen HJ. Age matters: collagen birefringence of superficial articular cartilage is increased in young guinea-pigs but decreased in older animals after identical physiological type of joint loading. Osteoarthritis and Cartilage 9, 694-701 (2001).
Kendrew J. The encyclopedia of molecular biology, p366 (Blackwell science, 1994).
van de Lest CHA, Versteeg EMM, Veerkamp JH, van Kuppevelt TH. Quantification and characterization of glycosaminoglycans at the nanogram level by a combined azure A-silver staining in agarose gels. Anal Biochem. 221, 356-361(1994).
Lovekamp J, Vyavahare N. Periodate-mediated glycosaminoglycan stabilization in bioprosthetic heart valves. J Biomed Mater Res., 478-486 (2001).
Moller HJ, Heinegard D, Poulsen JH. Combined alcian blue and silver staining of subnanogram quantities of proteoglycans and glycosaminoglycan in sodium dodecyl sulfate-polyacryamide gels. Anal. Biochem. 209, 169-175, (1993).
Muir, H. The chondrocyte,architect of cartilage. Bio eaasys 17 no.12, 1039-1048 (1995).
Neame PJ, Treep JT, Young CN. An 18 kDa Glycoprotein from Bovine Nasal Cartilage, J. Biol. Chem. 265, 9628-9633 (1990).
Reginster JY, Deroisy R, Rovati LC, Lee RL, Lejeune E, Bruyere O, Giacovelli G, Henrotin Y, Dacre JE, Gossett C. Long-term effects of glucosamine sulphate on osteoarthritis progression: a randomized placebo-controlled clinical trial. The Lancet 357, 251-256 (2001).
Ruoslahti E, Engvall E. Extracellular matrix components, Methods in enzymology v.245, (Academic Press ,1994).
Rohm K. Color Atlas of Biochemistry, 314-315 (Thieme ,1996).
Sugumaran G, Vertel BM. in Biosynthesis of chondrotin sulfate and dermatan sulfate proteoglycans, (ed. Ernst B and Hart GW.) Carbohydrates in chemistry and biology, Vol 4, p377 (Wiley-Vch., 2000).
Tran NT, Carbanes-Macheteau M, Taverna M. in Analysis of Glycoproteins and their glycopeptide and glyco fragments by electrophoresis and capillary electrophoresis. (ed.Rassi ZE.) Carbohydrate analysis by modern chromatography and electrophoresis, 693-703, (Elsevier, 2002)
Vannucchi S, Fibbi G, Pasquali F, Rosso MD, Cappelletti R, Chiarugi V. Adhesion-dependent heparin production by platelets. Nature 296, 352-353, (1982).
Venkatesan N, Ebihara T, Roughley PJ, Ludwig MS. Alterations in large and small proteoglycans in bleomycin-induced pulmonary fibrosis in rats. Am J Respir Crit Care Med. 161, 2066-2073 (2000).
www.ukcte.org/ images/cartilage.jpg
Yu WH, Woessner JF Jr. Heparan sulfate proteoglycans as extracellular docking molecules for Matrilysin. J. Biol. Chem. 275, 4183-4191 (2000).
湯湘芸, 學界科專計畫第一年成果報告, (2003)
王純怡, 膠原蛋白基質結構及多醣類組成對於細胞移入之研究, 國立台灣大學化學工程研究所碩士論文, (1997)
李兆偉, 退化性膝關節炎病人服用葡萄胺酸治療後之生物力學分析,國立成功大學醫學工程研究所碩士論文, ( 2002)
李昭怡, CC 趨化激素 Eotaxin-1 與骨關節炎病程相關性之研討,國立台北醫學大學生物醫學技術研究所碩士論文, (2002)
張雲鵬, 利用電腦模擬及顯微鏡技術研究蛇毒與具胺基葡萄糖之間的作用模式,國立清華大學生命科學研究所碩士論文, (1998)
章世豪, 膠原蛋白改質聚α氫氧酸支架應用於軟骨修復的研究,國立中興大學化學工程研究所碩士論文, (2001)
曾建榮, 經濟部技術處技術尖兵雜誌, 經濟部技術處發行, (2002)