跳到主要內容

臺灣博碩士論文加值系統

(44.220.251.236) 您好!臺灣時間:2024/10/11 04:33
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:陳喜玲
研究生(外文):Hsi-Lin Chen
論文名稱:引朵美辛與葡萄糖胺調控人類軟骨瘤細胞Aggrecan和MMP-3表現之研究
論文名稱(外文):Indomethacin and Glucosamine Hcl modulated the expression of aggrecan and matrix metalloproteinase-3 by human chondrosarcoma cell
指導教授:陳建和陳建和引用關係梁有志梁有志引用關係
指導教授(外文):Chien-Ho ChenYu-Chih Liang
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:醫學技術學系
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:82
中文關鍵詞:骨關節炎引朵美辛葡萄糖胺間質分解酵素醣蛋白聚合物軟骨瘤細胞介白質-1
外文關鍵詞:Indomethacinmatrix metalloproteinasesGlucosamine HclAggrecanchondrosarcomaIL-1βOsteoarthritis
相關次數:
  • 被引用被引用:0
  • 點閱點閱:430
  • 評分評分:
  • 下載下載:70
  • 收藏至我的研究室書目清單書目收藏:0
退化性骨關節炎Osteoarthritis ( OA ) 在目前人口逐漸高齡化的社會中發生率愈來愈高 , 而引朵美辛 ( Indomethacin ) 是一種非類固醇類的止痛抗發炎藥物 ( Non -steroidal anti-inflammatory drugs , NSAIDs ); 葡萄糖胺( Glucosamine ) 是構成關節軟骨glycosaminoglycans( GAG ) 雙醣類的基本結構。研究發現, 退化性 骨關節炎病人的間質分解酵素 ( MMP-3 ) 表現明顯 , 本篇論文主要探討不同劑量的Indomethacin、Glucosamine Hcl對經IL-1β 誘導的軟骨細胞的MMP-3及Aggrecan 的mRNA及蛋白質量表現。我們的實驗發現, 細胞中的MMP-3表現量隨著Indomethacin 劑量的增加而逐漸被抑制,PGE2的分析結果也是同樣被抑制;另外,在Glucosamine部份,隨著劑量的增加, 其MMP-3 表現量會被抑制, 而時間點的增加,Aggrecan的表現量也有增多的趨勢,依我們結果證明Indomethacin 和 Glucosamine Hcl 確實可有效抑制軟骨細胞所產生的MMP-3,PGE2及刺激Aggrecan的產生。總之,依我們結論提供一個有力的證明,對退化性骨關節炎是有幫助的。
The incidence of Osteoarthritis ( OA ) is more higher in progressive elder population now. Indomethacin ,an non-steroid anti-inflammatory drugs ( NSAIDs ) , Glucosamine Hcl is a basic structure of glycosaminoglycan ( GAG ). OA patients obviously have expressed MMP-3 . In this study,we examined the effects of the Indomethacin and Glucosamine Hcl on the mRNA and protein levels of the proteoglycan aggrecan on the activity of matrix metalloproteinase ( MMP-3 ) in cultured human IL-1β induced chondrocyte. The results indicated MMP-3 activity gradually inhibited as the more dose of indomethacin, the same as PGE2 assay. Besides , MMP-3 mRNA will be inhibited as the dose of Glucosamine Hcl and time accumulate , and aggrecan also have this increased trend. Our datas strongly prove Indomethacin and Glucosamine Hcl can inhibit MMP-3 , PGE2 and stimulate aggrecan. These results provide a cogent mechanism to support beneficial effect with Osteoarthritis .
目錄 3
中文摘要 5
Abstract 6
第一章、緒論 8
一、骨關節炎Osteoarthritis( OA )概論 8
二、引朵美辛( Indomethacin ) 11
三、間質分解酵素(matrix metalloproteinases;MMPs)/ TIMP(Tissue inhibitors of metalloproteinases) 13
四、前列腺素( prostaglandin,PG ): 17
五、醣蛋白聚合物 ( Aggrecan ) 18
六、膠原蛋白( Collagen ) 20
七、葡萄糖胺( Glucosamine sulfate,GS ) 21
八、細胞激素(cytokines)概論 23
第二章、實驗儀器與材料 25
一、實驗細胞株 25
二、藥品試劑 25
三、實驗儀器 29
四、常用溶液 30
第三章、實驗方法 32
一、人類軟骨瘤細胞株(human chondrosarcoma cell- SW-1353)培養 32
二、方法 32
三、細胞RNA萃取 (Preparation of cell RNA) 33
四、RNA定量法(RNA assay) 33
五、反轉錄-聚合酶連鎖反應(Reversetranscriptase-polymerase chain reaction) 34
六、電泳 ( electrophoresis) 36
七、細胞蛋白質製備 (preparation of cell lysate) 36
(一)、Protein lysis buffer 36
(二)、方法 37
八、蛋白質定量法 (protein assay) 38
九、西方墨點法 (western blotting) 38
(一)、SDS-PAGE gel preparation 38
(二)、電泳 ( electrophoresis ) 39
(三)、轉印 (transfer) 40
(四)、Immunoblot 40
十、前列腺素分析( PGE2 assay ) 41
第四章、 實驗結果 42
一、Indomethacin抑制軟骨細胞經由IL-1β所誘發的MMP-3在細胞外蛋白質的表現 42
二、Indomethacin抑制軟骨細胞經由IL-1β所誘發的MMP-3在細胞內蛋白質的表現 42
三、Indomethacin抑制軟骨細胞經由IL-1β所誘發的MMP-3基因的表現 43
四、Indomethacin抑制軟骨細胞經由IL-1β所誘發的前列腺素(prostaglandin)的表現 43
五、Indomethacin抑制軟骨細胞經由IL-1β所誘發的MMP-3在細胞外蛋白質的表現 44
六、Indomethacin抑制軟骨細胞經由IL-1β所誘發的MMP-3在細胞內蛋白質的表現 44
七、Indomethacin抑制軟骨細胞經由IL-1β所誘發的MMP-3基因的表現 45
八、Glucosamine抑制軟骨細胞MMP-3基因的表現 45
九、Glucosamine抑制軟骨細胞經由IL-1β所誘發的MMP-3基 因的表現 46
十、Glucosamine可能促使軟骨細胞Aggrecan基因的表現 46
第五章、討論 48
第六章、參考文獻 54
1. Guo-Hua Yuan, Kayo Masuko-Hongo, Tomohiro Kato, and Kusuki
Nishioka. Immunologic Intervention in the Pathogenesis of
Osteoarthritis. Arthritis Rheum 48(3):602–611;2003.
2. Haq I,Murphy E , Dacre J. Osteoarthritis. Postgrad Med J 79
(933):377-383;2003.
3. Walker JM, Helewa A. Physical therapy in arthritis.
Philadelphia, PA:Saunders,1996.

4. Goldring SR, Gravallese EM. Mechanisms of bone loss in
inflammatory arthritis: diagnosis and therapeutic implications.
Arthritis Res 2(1):33-7;2000.
5.Robbins , J. R., B. Thomas , et al. Immortalized
human adult articular chondrocytes maintain cartilage-specific
phenotype and responses to interleukin-1beta. Arthritis Rheum 43
(10): 2189-201;2000.
6.Kojima T,Mwale F,Yasuda T,et al. Early degradation of type IX and
type II collagen with the onset of experimental inflammatory
arthritis. Arthritis Rheum 44(1):120-7;2001.
7.Sandell,L.J., and Aigner, T. Articular cartilage and
changes in arthritis. An introduction: cell biology of
osteoarthritis. Arthritis Res 3(2):107-113;2001.
8.Hideaki Nagase , J Frederick Woessner Jr . Matrix
Metalloproteinases. J.Biol.Chem 274(31):21491-494;1999
9.Dean DD , Martel-Pelletier J, Pelletier J-P, et al. Evidence for
metalloproteinase and metalloproteinase inhibitor imbalance in
human osteoarthritic cartilage. J Clin Invest 84:678–85;1989.
10.W W Bolten. Problem of the atherothrombotic potential of non-
steroidal anti-inflammatory drugs . Ann Rheum Dis
65:7-13;2006
11.Quilley J,McGiffJC. Eicosanoids. In Swales JD ,editor.
Textbook of hypertension. Oxford. Blackwell Scientific
Publications. 314-27;1994.

12. Kenta Matsuda,S Nakamura. Celecoxib inhibits nitric oxide
production in chondrocytes of ligament-damaged osteoarthritic
rat joints. Rheumatol Int. DOI 10.1007 /s00296-006-0107-6;2006.
13. AM. Panicoa,T , V. Cardileb, F. Garufia, et al. Protective
effect of Capparis spinosa on chondrocytes. Life
Sciences 77 :2479–2488;2005.
14. Chris to pher MB, Philip JF, Jennifer D, Rob ert AS, Rob
ert HM, Geoffrey B. Indomethacin potentiates the in
duc tion of HL60 differentiation to neutrophils, by retinoic
acid and granulocyte colony-stimulating factor, and to
Monocytes, by vitamin D3. Leukemia 8(4): 595-604;1994.

15. Venera Cardile. Effect of propolis on human cartilage and
chondrocytes. Life Sciences 73 :1027–1035;2003.
16. Khasigov, P. Z., O. V. Podobed , et al. Role of
matrix metalloproteinases and their inhibitors
in tumor invasion and metastasis. Biochemistry
(Mosc) 68(7): 711-7;2003.
17. Woessner, J. F. Matrix metalloproteinases and their
inhibitors in connective tissue remodeling.FASEB J. 5(8): 2145-
2154;1991.
18. Tokumaru Y., Fujii M., Otani Y., Kaneyama K.,
Imanishi, Y., Igarashi, N., and Kanzaki, J.
Cancer Lett 150:15-21;2000.
19. Matrisian, L. M. The matrix - degrading
metalloproteinases.BioEssays 14(7): 455-463;1992.
20. Quantin B. , Murphy G. , and Breathnach R.
Biochemistry 28: 5327-5334;1989.
21. Von Bredow D. C. , Nagle R. B. , Bowden G. T., and
Cress, A. E. Degradation of fibronectin fibrils by matrilysin and
characterization of the degradation products. Exp. Cell Res. 221
(1):83-91;1995.

22. Uria J. A., Stahle_Backdahl M., Seiki M., Fueyo A.et al
Regulation of collagenase-3 expression in human breast carcinomas
is mediated by stromal-epithelial cell interactions. Cancer Res 57
(21): 4882-4888;1997.
23. Pei, D., and Weiss, S. J. Transmembrane-deletion mutants
of the membrane - type matrix metalloproteinase- 1
process progelatinase A and express intrinsic
matrix-degrading activity.J. Biol. Chem 271(15):
9135-9140;1996.
24. Goetzl, E.J. Specificity of expression and effects of
eicosanoid mediators in normal physiology and human
diseases. FASEB J.9: 1051–1058;1995.
25. Smith, W.L. Pharmacology of prostaglandin endoperoxide
synthase isozymes 1 and 2. Ann. New York Acad. Sci.
714;1994.
26. Poole AR, Rizkalla G, Ionescu M, Reiner A, Brooks E,
Rorabeck C, et al. Osteoarthritis in the human knee:
a dynamic process of cartilage matrix degradation,
synthesis and reorganization. Agents Actions
39(Suppl):3–13;1993.
27. Paulsson M , Morgelin M , Wiedemann H , et al. Extended
and globular protein domains in cartilage proteoglycans. Biochem
J 245(3):763-72;1987.
28. J. D. Sandy Ph.D. A contentious issue .nds some clarity:
on the independent and complementary roles of
aggrecanase activity and MMP activity in human joint
aggrecanolysis .OsteoArthritis and Cartilage 14(2):
95-100;2006.
29. David Eyre . Collagen of articular cartilage.
Arthritis Res 4:30-35;2002.
30. Darko Bosnakovski , Morimichi Mizuno , Gonhyung Kim ,
Et al. Chondrogenic Differentiation of Bovine Bone
Marrow Mesenchymal Stem Cells (MSCs) in Different
Hydrogels : Influence of Collagen Type II
Extracellular Matrix on MSC Chondrogenesis. Wiley InterScience
9,DOI: 10.1002/bit.20828;2006.
31.Hughes R , Carr A . A randomized , double-blind ,
placebo-controlled trial of glucosamine sulphate as
an analgesic in osteoarthritis of the knee.
Rheumatology 41(3):279–84;2002.
32.Oegema TR Jr, Deloria LB, Sandy JD, Hart DA. Effect of oral
glucosamine on cartilage and meniscus in normal and
chymopapaininjected knees of young rabbits. Arthritis Rheum 46
(9): 2495–503;2002.
33. Hua J, Sakamoto K, Nagaoka I. Inhibitory actions of
glucosamine,a therapeutic agent for osteoarthritis, on the
functions of neutrophils.J Leukoc Biol 71: 632–40;2002.
34. Muller-Fassbender H, Bach G , Haase W, Rovati AL:
Glucosamine sulfate compared to ibuprofen in
osteoarthritis of the knee. Osteoarthritis Cart
2:51–59;1994.
35. Karzel K , Domenjoz R : Effects of hexosamine
derivatives and uronic acid derivatives on glycosaminoglycan
metabolism of fibroblast cultures.
Pharmacology 5:337–45;1971.
36. G. R. Dodge Ph.D. *, S. A. Jimenez M.D. Glucosamine sulfate
modulates the levels of aggrecan and matrix metalloproteinase- 3
synthesized by cultured human osteoarthritis articular
chondrocytes. OsteoArthritis and Cartilage 11(6): 424–432;2003.
37. Zhiyong Fan, Brigitte Bau, Huiqing Yang, Stephan Soeder, and
Thomas Aigner. Freshly Isolated Osteoarthritic Chondrocytes Are
Catabolically More Active Than Normal Chondrocytes, but Less
Responsive to Catabolic Stimulation With Interleukin-1β.
Arthritis Rheum 52(1) : 136–143;2005.
38. Chevalier X . Upregulation of enzymatic activity by
interleukin-1 in osteoarthritis.Biomed Pharmacother 51(2):58–
62;1997.
39. M. J. Lo’pez-Armada Ph.D., B. Carame’s Ph.D., M. Lires-
Dea’n B.S. Cytokines, tumor necrosis factor-a and interleukin-
1b, differentially regulate apoptosis in osteoarthritis cultured
human chondrocytes. OsteoArthritis and Cartilage.
doi:10.1016/j.joca.;2006.
40. Stephen Marshall , Owen Nadeau, Kazumitsu Yamasaki .
Glucosamine - induced Activation of Glycogen
Biosynthesis in Isolated Adipocytes . J.Biol.chem
280(12) : 11018–11024;2005.
41. Dong-Ho Han, M M Chen. Glucosamine and glucose induce
insulin resistance by different mechanisms in rat
skeletal muscle.Am J Physiol Endocrinol Metab 285(6):
E1267–E1272; 2003.
42. T. Sadowski and J. Steinmeyer. Effects of non-steroidal
antiinflammatory drugs and dexamethasone on the activity and
expression of matrix metalloproteinase-1, matrix metalloproteinase- 3 and tissue inhibitor of metalloproteinases-1 by bovine
articular chondrocytes. Osteoarthritis and Cartilage. 9(5): 407–
415;2001.
43. R. Largo ,MA Alvarez-Soria. Glucosamine inhibits IL-1β-induced
NFκB activation in human osteoarthritic chondrocytes.
OsteoArthritis and Cartilage 11(4): 290–298;2003.
44. Theodorep, Ciaraldi, Leslie et al. Glucosamine Regulation of
Glucose Metabolism in Cultured Human Skeletal Muscle Cells:
Divergent Effects on GlucoseTransport/Phosphorylation and
Glycogen Synthase in Non-Diabetic and Type 2 Diabetic
Subjects.Endocrinology 140: 3971–3980; 1999.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top