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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 林思洸(Sze-Kwan Lin) | |
dc.contributor.author | Ling-Hsiu Chao | en |
dc.contributor.author | 趙怜琇 | zh_TW |
dc.date.accessioned | 2021-06-16T08:41:35Z | - |
dc.date.available | 2013-09-24 | |
dc.date.copyright | 2013-09-24 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-09-06 | |
dc.identifier.citation | 1. Scott DL, Wolfe F, Huizinga TW. Rheumatoid arthritis. Lancet. 2010 ;376(9746): 1094-108.
2. Symmons D, Turner G, Webb R, et al. The prevalence of rheumatoid arthritis in the United Kingdom: new estimates for a new century. Rheumatology (Oxford) 2002; 41: 793-800. 3. van der Woude D, Houwing-Duistermaat JJ, Toes RE, et al. Quantitative heritability of anti-citrullinated protein antibody-positive and anti-citrullinated protein antibody-negative rheumatoid arthritis. Arthritis Rheum 2009; 60: 916-23. 4. Carlens C, Hergens MP, Grunewald J, et al. Smoking, use of moist snuff , and risk of chronic infl ammatory diseases. Am J Respir Crit Care Med 2010; 181: 1217-22. 5. Brigstock, D.R., The CCN family: a new stimulus package. J Endocrinol, 2003. 178(2): p. 169-75. 6. Chen CC, Lau LF. Functions and mechanisms of action of CCN matricellular proteins. Int J Biochem Cell Biol 2009;41:771–83. 7. Yang, G.P. and L.F. Lau, Cyr61, product of a growth factor-inducible immediate early gene, is associated with the extracellular matrix and the cell surface. Cell Growth Differ, 1991. 2(7): p. 351-7. 8. Leask A, Abraham DJ. All in the CCN family: essential matricellular signaling modulators emerge from the bunker. J Cell Sci 2006;119:4803–10. 9. Jun JI, Lau LF. Taking aim at the extracellular matrix: CCN proteins as emerging therapeutic targets. Nat Rev Drug Discov. 2011;10(12):945-63. 10. Chen CC, Mo FE, Lau LF. The angiogenic factor cyr61 activates a genetic program for wound healing in human skin fibroblasts. J Biol Chem 2001;276:47329–37. 11. O’Brien TP, Lau LF. Expression of the growth factor-inducible immediate early gene Cyr61 correlates with chondrogenesis during mouse embryonic development. Cell Growth Differ 1992;3: 645–54. 12. Schober JM, Chen N, Grzeszkiewicz TM, Jovanovic I, Emeson EE, Ugarova TP, et al. Identification of integrin alpha(M)beta(2) as an adhesion receptor on peripheral blood monocytes for Cyr61 (CCN1) and connective tissue growth factor (CCN2): immediate-early gene products expressed in atherosclerotic lesions. Blood. 2002;99(12):4457-65. 13. Grzeszkiewicz TM, Kirschling DJ, Chen N, Lau LF. CYR61 stimulates human skin fibroblast migration through Integrin alpha vbeta 5 and enhances mitogenesis through integrin alpha vbeta 3, independent of its carboxyl-terminal domain. J Biol Chem 2001;276:21943–50. 14. Kireeva ML, Mo FE, Yang GP, Lau LF. Cyr61, a product of a growth factor-inducible immediate-early gene, promotes cell proliferation, migration, and adhesion. Mol Cell Biol 1996;16: 1326–34. 15. Menendez JA, Vellon L, Mehmi I, Teng PK, Griggs DW, Lupu R. A novel CYR61-triggered 'CYR61-alphavbeta3 integrin loop' regulates breast cancer cell survival and chemosensitivity through activation of ERK1/ERK2 MAPK signaling pathway. Oncogene 2005;24:761–79. 16. Haas CS, Creighton CJ, Pi X, Maine I, Koch AE, Haines GK III, et al. Identification of genes modulated in rheumatoid arthritis using complementary DNA microarray analysis of lymphoblastoid B cell lines from disease-discordant monozygotic twins. Arthritis Rheum 2006;54:2047–60. 17. Zhang Q, Wu J, Cao Q, Xiao L, Wang L, He D, et al. A critical role of Cyr61 in interleukin-17–dependent proliferation of fibroblast-like synoviocytes in rheumatoid arthritis. Arthritis Rheum 2009;60:3602–12. 18. Kok SH, Hou KL, Hong CY, Wang JS, Liang PC, Chang CC, Hsiao M, Yang H, Lai EH, Lin SK. Simvastatin inhibits cytokine-stimulated Cyr61 expression in osteoblastic cells: a therapeutic benefit for arthritis. Arthritis Rheum. 2011;63(4):1010-20. 19. Schutze N, Rucker N, Muller J, Adamski J, Jakob F. 5' flanking sequence of the human immediate early responsive gene ccn1 (cyr61) and mapping of polymorphic CA repeat sequence motifs in the human ccn1 (cyr61) locusMol Pathol. 2001;54(3):170-5. 20. Mayr B, Montminy M. Transcriptional regulation by the phosphorylation- dependent factor CREB. Nat Rev Mol Cell Biol. 2001;2(8):599-609. 21. Conkright MD, Guzman E, Flechner L, Su AI, Hogenesch JB, Montminy M. Genome-wide analysis of CREB target genes reveals a core promoter requirement for cAMP responsiveness. Mol Cell. 2003 Apr;11(4):1101-8. 22. Montminy MR, Sevarino KA, Wagner JA, Mandel G, Goodman RH. Identification of a cyclic-AMP-responsive element within the rat somatostatin gene. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6682-6. 23. Han JS, Macarak E, Rosenbloom J, Chung KC, Chaqour B. Regulation of Cyr61/CCN1 gene expression through RhoA GTPase and p38MAPK signaling pathways. Eur J Biochem. 2003 Aug;270(16):3408-21. 24. Yamamoto, K.K., et al., Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature, 1988. 334(6182): p. 494-8. 25. Gonzalez, G.A. and M.R. Montminy, Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell, 1989. 59(4): p. 675-80. 26. Gonzalez, G.A., et al., A cluster of phosphorylation sites on the cyclic AMP-regulated nuclear factor CREB predicted by its sequence. Nature, 1989. 337(6209): p. 749-52. 27. Montminy, M.R. and L.M. Bilezikjian, Binding of a nuclear protein to the cyclic-AMP response element of the somatostatin gene. Nature, 1987. 328(6126): p. 175-8. 28. Assaily W, Benchimol S. Differential utilization of two ATP-generating pathways is regulated by p53. Cancer Cell. 2006 Jul;10(1):4-6. 29. Henderson B, Bitensky L, Chayen J. Glycoltic activity in human synovial lining cells in rheumatoid arthritis. Ann Rheum Dis 1979;38:63–7. 30. Ciurtin C, Cojocaru VM, Miron IM, et al. Correlation between different components of synovial fluid and pathogenesis of rheumatic diseases. Rom J Intern Med. 2006;44:171–81. 31. Muz B, Khan MN, Kiriakidis S, Paleolog EM. The role of hypoxia and HIF-dependent signalling events in rheumatoid arthritis. Arthritis Res Ther. 2009;11(1):201. doi: 10.1186/ar2568. Epub 2009 Jan 20. 32. Naughton D, Whelan M, Smith EC, Williams R, Blake DR, Grootveld M () An investigation of the abnormal metabolic status of synovial fluid from patients with rheumatoid arthritis by high field proton nuclear magnetic resonance spectroscopy. FEBS Lett. 1993;317:135–8. 33. Larsen H, Akhavani MA, Raatz Y, Paleolog EM: Gene expression studies to investigate disease mechanisms in rheumatoid arthritis: does angiogenesis play a role? Curr Rheumatol Rev 2007, 3:243-251. 34. Blake DR, Merry P, Unsworth J, Kidd BL, Outhwaite JM, Ballard R, Morris CJ, Gray L, Lunec J: Hypoxic-reperfusion injury in the inflamed human joint. Lancet 1989, 1:289-293. 35. Hitchon CA, El-Gabalawy HS, Bezabeh T. Characterization of synovial tissue from arthritis patients: a proton magnetic resonance spectroscopic investigation. Rheumatol Int. 2009;29:1205–11. 36. Kawauchi K, Araki K, Tobiume K, Tanaka N. p53 regulates glucose metabolism through an IKK-NF-kappaB pathway and inhibits cell transformation. Nat Cell Biol. 2008;10, 611–8. 37. Kawauchi K, Araki K, Tobiume K, Tanaka N. Loss of p53 enhances catalytic activity of IKK {beta} through O-linked {beta}-N-acetyl glucosamine modification. Proc Natl Acad Sci USA. 2009;106, 3431–6. 38. Bolan˜ os JP, Delgado-Esteban M, Herrero-Mendez A, Fernandez- Fernandez S, Almeida A. Regulation of glycolysis and pentose–phosphate pathway by nitric oxide: impact on neuronal survival. Biochim Biophys Acta. 2008; 1777, 789–93. 39. Chang X, Wei C. Glycolysis and rheumatoid arthritis. Int J Rheum Dis. 2011;14(3):217-22. 40. Frye RA. Phylogenetic classification of prokaryotic and eukaryotic Sir2-like proteins. Biochem Biophys Res Commun. 2000 Jul 5; 273(2):793-8. 41. Imai, S.-i., Armstrong, C. M., Kaeberlein, M. & Guarente, L. Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. Nature 2000;403: 795–800 42. Sauve, A. A., Wolberger, C., Schramm, V. L. & Boeke, J. D. The biochemistry of sirtuins. Annu. Rev. Biochem. 2006;75:435–465 43. Michan, S. & Sinclair, D. Sirtuins in mammals: insights into their biological function. Biochem. J. 2007;404:1–13 44. Liszt G, Ford E, Kurtev M, Guarente L. Mouse Sir2 homolog SIRT6 is a nuclear ADPribosyltransferase. J Biol Chem 2005;280:21313-20. 45. Kawahara TL, Michishita E, Adler AS, Damian M, Berber E, Lin M, et al. SIRT6 links histone H3 lysine 9 deacetylation to NF-κB-dependent gene expression and organismal life span. Cell 2009;136:62-74. 46. Michishita E, McCord RA, Boxer LD, Barber MF, Hong T, Gozani O, et al. Cell cycledependent deacetylation of telomeric histone H3 lysine K56 by human SIRT6. Cell Cycle 2009;8:2664-6. 47. Yang B, Zwaans BM, Eckersdorff M, Lombard DB. The sirtuin SIRT6 deacetylates H3 K56Ac in vivo to promote genomic stability. Cell Cycle 2009;8(16):2662-3. 48. Lombard DB, Schwer B, Alt FW, Mostoslavsky R. SIRT6 in DNA repair, metabolism and ageing. J Intern Med. 2008;263(2):128-41. 49. Zhong L, D'Urso A, Toiber D, Sebastian C, Henry RE, Vadysirisack DD, Guimaraes A, Marinelli B, Wikstrom JD, Nir T, Clish CB, Vaitheesvaran B, Iliopoulos O, Kurland I, Dor Y, Weissleder R, Shirihai OS, Ellisen LW, Espinosa JM, Mostoslavsky R. The histone deacetylase Sirt6 regulates glucose homeostasis via Hif1alpha. Cell. 2010;140(2):280-9. 50. Zhong L, Mostoslavsky R. SIRT6: a master epigenetic gatekeeper of glucose metabolism. Transcription. 2010;1(1):17-21 51. Lee HS, Ka SO, Lee SM, Lee SI, Park JW, Park BH. Overexpression of SIRT6 suppresses inflammatory responses and bone destruction in collagen-induced arthritic mice. Arthritis Rheum. 2013. 52. Xiao C, Wang RH, Lahusen TJ, Park O, Bertola A, Maruyama T, Reynolds D, Chen Q, Xu X, Young HA, Chen WJ, Gao B, Deng CX. Progression of chronic liver inflammation and fibrosis driven by activation of c-JUN signaling in Sirt6 mutant mice. J Biol Chem. 2012;287(50):41903-13. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58966 | - |
dc.description.abstract | 類風濕性關節炎(rheumatoid arthritis, RA)是一種慢性的全身性免疫疾病,持續發炎造成RA關節長期處於一個缺氧(hypoxia)的狀態,過去文獻證實RA synovial tissue確實具有高度的糖解作用活性,過多的糖解作用所導致的代謝失衡,除了會刺激細胞造成不正常增生,糖解作用產物─乳酸的過度堆積也會導致細胞毒性,使得關節發炎的情形更加惡化。所以,如何改善這樣失衡的葡萄糖代謝情形,是一個值得探討的主題。
SIRT6為Sirtuins七個成員之一,目前已知和維持基因組的穩定性、防止老化、代謝以及發炎反應有關,尤其在維持生物體糖類代謝方面具有重要性。目前關於SIRT6能否調控類風溼性關節炎中失衡的糖類代謝,甚至藉此減緩發炎反應,都還是未知,這也是本研究所欲探討的主要目標。 在本次研究中,我們首先收集並分析不同病程進展的RA患者關節組織切片,初步證實SIRT6的表現與發炎程度具有臨床相關性。接著透過西方墨點法(Western blot)及乳酸含量檢測套組(Lactate assay kit)分析,我們發現SIRT6會減緩人類成骨細胞在缺氧環境下失衡的糖解作用,包含抑制乳酸脫氫脢(LDH; Lactate dehydrogenase)及降低乳酸 (Lactate)產量。我們更進一步發現SIRT6可以藉此機制去抑制Cyr61(cysteine rich protein 61) ,一個在RA患者的滑膜組織、FLS和關節液中均高度表達的蛋白質的表現。 為了更深入了解這之間的詳細機轉,根據本實驗室過去的實驗結果─Cyr61表現會受到轉錄因子CREB的調控,透過螢光素酶檢測法(Luciferase assay)及染色質免疫沉澱法(Chromatin Immunoprecipitation) 分析,我們發現在缺氧刺激下SIRT6會透過降低CREB結合到Cyr61啟動子的親和力去抑制Cyr61啟動子的活性。我們更進一步證明,乳酸會促進CREB對Cyr61啟動子的親和力,而SIRT6會透過抑制乳酸去抑制此一作用。 最後,我們透過動物實驗模式印證了在細胞實驗中所得到的結果。在膠原誘導關節炎大鼠模型中,我們發現SIRT6能抑制成骨細胞中 LDH及Cyr61的表現,並且能降低巨噬細胞的浸潤,具有抑制發炎進展、減緩骨吸收的作用。 總結本研究實驗結果,SIRT6可經由調控缺氧刺激下成骨細胞中失衡的葡萄糖代謝來降低Cyr61表現,藉此減緩關節炎的病程進展。期許此機制能夠成為RA治療上一個有潛力的新途徑。 | zh_TW |
dc.description.abstract | Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease, characterized by persistent synovitis. Evidence indicates that RA synovial tissues have increased glycolytic activity, which is related to the nature of hypoxia in RA synovium. Elevated production of lactate caused by unbalanced glycolysis stimulates abnormal cell proliferation and leads to joint destruction. Therefore, how to improve the situation of glucose metabolism is a topic worth exploring.
Within the seven members of the sirtuin family, SIRT6 (sirtuin 6) is particularly involved in maintaining genomic stability, preventing age-related disorders, regulating metabolic processes and attenuating inflammation. It is unclear whether SIRT6 can play a role in regulating unbalanced glucose metabolism in RA. In this study, we demonstrated that SIRT6 suppressed Cyr61 expression via modulating increased glycolysis stimulated by hypoxia in osteoblastic cells. We also found that SIRT6 inhibited hypoxia-induced CREB phosphorylation and CREB-DNA binding. Hypoxia-induced Cyr61 promoter activation was dependent on CRE-CREB interaction and is inhibited by SIRT6. In rat collagen-induced arthritis (CIA), marked expression of Cyr61, LDH, CD68 was noted in the severe joint destruction group. In contrast, obvious SIRT6 was found in the group of mild severity. In conclusion, we have demonstrated that SIRT6 attenuates hypoxia-induced Cyr61 expression via modulating unbalanced glycolysis in osteoblastic cells and suppresses disease progression in rat CIA model. This finding may provide a new perspective on the treatment of RA. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T08:41:35Z (GMT). No. of bitstreams: 1 ntu-102-R99422018-1.pdf: 2207841 bytes, checksum: 4cb3c3a0718e2f80e891085a7e139760 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 口試委員會審定書--------------------------------------------------------------------------------I
誌謝-------------------------------------------------------------------------------------------------II 中文摘要------------------------------------------------------------------------------------------III 英文摘要-------------------------------------------------------------------------------------------V 第一章 導論--------------------------------------------------------------------------------------1 1.1 類風濕性關節炎---------------------------------------------------------------------1 1.2 Cyr61與類風濕性關節炎----------------------------------------------------------2 1.3 Cyr61的轉錄調控(transcriptional regulation) ----------------------------------3 1.3.1 轉錄因子CREB-------------------------------------------------------------3 1.3.2 CREB的活化----------------------------------------------------------------3 1.4 糖解作用與類風濕性關節炎------------------------------------------------------4 1.5 Sirtuin 6與葡萄糖代謝-------------------------------------------------------------5 1.6 Sirtuin 6與類風濕性關節炎-------------------------------------------------------6 第二章 實驗目的-------------------------------------------------------------------------------8 第三章 材料與方法-----------------------------------------------------------------------------9 3.1 實驗細胞株---------------------------------------------------------------------------9 3.2 SIRT6穩定表達細胞株的建立(hSIRT stable lines) ---------------------------------------------------9 3.3 細胞加藥處理------------------------------------------------------------------------9 3.4 細胞內蛋白質的萃取--------------------------------------------------------------10 3.5 西方點墨法(Western blot) --------------------------------------------------------11 3.6 乳酸含量檢測(Lactate assay) ----------------------------------------------------12 3.7 暫時性轉染(transient transfection) ----------------------------------------------12 3.8 螢光素酶檢測法(Luciferase assay) ---------------------------------------------12 3.9 染色質免疫沉澱法(Chromatin Immunoprecipitation) -----------------------13 3.10 膠原誘導關節炎大鼠模型(Rat Collagen-Induced Arthritis model) ------13 3.11 免疫組織化學染色. (Immunohistochemistry, IHC) -------------------------14 第四章 實驗結果------------------------------------------------------------------------------15 4.1 化學缺氧劑氯化鈷誘發HIF-1α蛋白質表現---------------------------------15 4.2 SIRT6抑制缺氧刺激下失衡的糖解作用-------------------------------------15 4.2.1 SIRT6抑制缺氧刺激下的LDH蛋白質表現-------------------------15 4.2.2 SIRT6降低缺氧刺激下的乳酸產量-----------------------------------16 4.3 SIRT6藉由調節糖解作用影響缺氧刺激下Cyr61蛋白質表現------------16 4.3.1 SIRT6抑制缺氧刺激下Cyr61蛋白質表現---------------------------16 4.3.2 糖解作用之產物─乳酸促使Cyr61蛋白質表現增加---------------17 4.3.3 乳酸恢復SIRT6所抑制的Cyr61蛋白質表現------------------------17 4.4 SIRT6可透過調控CREB對Cyr61啟動子的結合親和力去抑制缺氧刺激下Cyr61的表現--------------------------------------------------------------------17 4.4.1 SIRT6抑制缺氧刺激所促使的CREB phosphorylation-------------17 4.4.2 SIRT6透過CREB抑制缺氧刺激下Cyr61蛋白質的表現-------18 4.5 SIRT6可透過抑制乳酸影響CREB活化並藉此調控Cyr61的表現-------18 4.5.1 乳酸促進CREB活化----------------------------------------------------18 4.5.2 乳酸促進CREB對Cyr61啟動子的結合親和力---------------------18 4.6 膠原誘導關節炎大鼠模型中SIRT6 , LDH, Cyr61, CD68的表現-------19 第五章 討論------------------------------------------------------------------------------------20 5.1 缺氧刺激下SIRT6對糖解作用的影響-----------------------------------------20 5.2 缺氧刺激下SIRT6對Cyr61蛋白質表現的影響------------------------------21 5.2.1 缺氧刺激誘導Cyr61蛋白質表現--------------------------------------21 5.2.2 SIRT6抑制缺氧刺激下Cyr61蛋白質表現-------------------------21 第六章 結論------------------------------------------------------------------------------------23 參考文獻------------------------------------------------------------------------------------------24 附錄------------------------------------------------------------------------------------------------29 | |
dc.language.iso | zh-TW | |
dc.title | SIRT6可經由調控人類成骨細胞中失衡的葡萄糖代謝來減緩關節炎的進展 | zh_TW |
dc.title | SIRT6 Attenuates Hypoxia-Induced Cyr61 Expression via Modulating Unbalanced Glycolysis in Human Osteoblastic Cells: A Therapeutic Potential for Arthritis | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 郭生興(Sang-Heng Kok),洪志遠(Chi-Yuan Hong) | |
dc.subject.keyword | 類風濕性關節炎,缺氧,葡萄糖代謝,糖解作用,Cyr61,SIRT6, | zh_TW |
dc.subject.keyword | rheumatoid arthritis,hypoxia,glucose metabolism,glycolysis,Cyr61,SIRT6, | en |
dc.relation.page | 39 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-09-06 | |
dc.contributor.author-college | 牙醫專業學院 | zh_TW |
dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
顯示於系所單位: | 臨床牙醫學研究所 |
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