請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65358
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張震東 | |
dc.contributor.author | Chih-I Wu | en |
dc.contributor.author | 吳致誼 | zh_TW |
dc.date.accessioned | 2021-06-16T23:38:25Z | - |
dc.date.available | 2013-08-01 | |
dc.date.copyright | 2012-08-01 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-07-25 | |
dc.identifier.citation | 參考文獻
1. Buchakjian MR, Kornbluth S: The engine driving the ship: metabolic steering of cell proliferation and death. Nature reviews Molecular cell biology 2010, 11(10):715-727. 2. Sutton-McDowall ML, Gilchrist RB, Thompson JG: The pivotal role of glucose metabolism in determining oocyte developmental competence. Reproduction 2010, 139(4):685-695. 3. Chung SSM: Contribution of Polyol Pathway to Diabetes-Induced Oxidative Stress. Journal of the American Society of Nephrology 2003, 14(90003):233S-236. 4. Porat S, Weinberg-Corem N, Tornovsky-Babaey S, Schyr-Ben-Haroush R, Hija A, Stolovich-Rain M, Dadon D, Granot Z, Ben-Hur V, White P et al: Control of pancreatic beta cell regeneration by glucose metabolism. Cell metabolism 2011, 13(4):440-449. 5. Groppi S, Belotti F, Brandao RL, Martegani E, Tisi R: Glucose-induced calcium influx in budding yeast involves a novel calcium transport system and can activate calcineurin. Cell calcium 2011, 49(6):376-386. 6. Wang Y, Pierce M, Schneper L, Guldal CG, Zhang X, Tavazoie S, Broach JR: Ras and Gpa2 mediate one branch of a redundant glucose signaling pathway in yeast. PLoS biology 2004, 2(5):E128. 7. Zaman S, Lippman SI, Schneper L, Slonim N, Broach JR: Glucose regulates transcription in yeast through a network of signaling pathways. Molecular systems biology 2009, 5:245. 8. Haneda M, Kikkawa R, Sugimoto T, Koya D, Araki S, Togawa M, Shigeta Y: Abnormalities in protein kinase C and MAP kinase cascade in mesangial cells cultured under high glucose conditions. Journal of diabetes and its complications 1995, 9(4):246-248. 9. Huang J, Siragy HM: Regulation of (pro)renin receptor expression by glucose-induced mitogen-activated protein kinase, nuclear factor-kappaB, and activator protein-1 signaling pathways. Endocrinology 2010, 151(7):3317-3325. 10. Zhang Y, Peng F, Gao B, Ingram AJ, Krepinsky JC: High glucose-induced RhoA activation requires caveolae and PKCbeta1-mediated ROS generation. American journal of physiology Renal physiology 2012, 302(1):F159-172. 11. Natarajan R, Gonzales N, Xu L, Nadler JL: Vascular smooth muscle cells exhibit increased growth in response to elevated glucose. Biochemical and biophysical research communications 1992, 187(1):552-560. 12. Ho FM, Lin WW, Chen BC, Chao CM, Yang CR, Lin LY, Lai CC, Liu SH, Liau CS: High glucose-induced apoptosis in human vascular endothelial cells is mediated through NF-kappaB and c-Jun NH2-terminal kinase pathway and prevented by PI3K/Akt/eNOS pathway. Cellular signalling 2006, 18(3):391-399. 13. Price SA, Hounsom L, Purves-Tyson TD, Fernyhough P, Tomlinson DR: Activation of JNK in sensory neurons protects against sensory neuron cell death in diabetes and on exposure to glucose/oxidative stress in vitro. Annals of the New York Academy of Sciences 2003, 1010:95-99. 14. Sakuma H, Yamamoto M, Okumura M, Kojima T, Maruyama T, Yasuda K: High glucose inhibits apoptosis in human coronary artery smooth muscle cells by increasing bcl-xL and bfl-1/A1. American journal of physiology Cell physiology 2002, 283(2):C422-428. 15. Marani M, Hancock D, Lopes R, Tenev T, Downward J, Lemoine NR: Role of Bim in the survival pathway induced by Raf in epithelial cells. Oncogene 2004, 23(14):2431-2441. 16. Roudier E, Perrin A: Considering the role of pyruvate in tumor cells during hypoxia. Biochimica et biophysica acta 2009, 1796(2):55-62. 17. Nordberg J, Arner ES: Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free radical biology & medicine 2001, 31(11):1287-1312. 18. Chatham JC, Gilbert HF, Radda GK: The metabolic consequences of hydroperoxide perfusion on the isolated rat heart. European journal of biochemistry / FEBS 1989, 184(3):657-662. 19. Janero DR, Hreniuk D, Sharif HM: Hydroperoxide-induced oxidative stress impairs heart muscle cell carbohydrate metabolism. The American journal of physiology 1994, 266(1 Pt 1):C179-188. 20. Vaage J, Antonelli M, Bufi M, Irtun O, DeBlasi RA, Corbucci GG, Gasparetto A, Semb AG: Exogenous reactive oxygen species deplete the isolated rat heart of antioxidants. Free radical biology & medicine 1997, 22(1-2):85-92. 21. Constantopoulos G, Barranger JA: Nonenzymatic decarboxylation of pyruvate. Anal Biochem 1984, 139(2):353-358. 22. Comte B, Vincent G, Bouchard B, Jette M, Cordeau S, Rosiers CD: A 13C mass isotopomer study of anaplerotic pyruvate carboxylation in perfused rat hearts. The Journal of biological chemistry 1997, 272(42):26125-26131. 23. Mallet RT, Sun J: Antioxidant properties of myocardial fuels. Molecular and cellular biochemistry 2003, 253(1-2):103-111. 24. Schafer FQ, Buettner GR: Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free radical biology & medicine 2001, 30(11):1191-1212. 25. Bindoli A, Barzon E, Rigobello MP: Inhibitory effect of pyruvate on release of glutathione and swelling of rat heart mitochondria. Cardiovascular research 1995, 30(5):821-824. 26. Lee YJ, Kang IJ, Bunger R, Kang YH: Enhanced survival effect of pyruvate correlates MAPK and NF-kappaB activation in hydrogen peroxide-treated human endothelial cells. J Appl Physiol 2004, 96(2):793-801; discussion 792. 27. Mallet RT, Sun J: Mitochondrial metabolism of pyruvate is required for its enhancement of cardiac function and energetics. Cardiovascular research 1999, 42(1):149-161. 28. Ochiai K, Zhang J, Gong G, Zhang Y, Liu J, Ye Y, Wu X, Liu H, Murakami Y, Bache RJ et al: Effects of augmented delivery of pyruvate on myocardial high-energy phosphate metabolism at high workstate. American journal of physiology Heart and circulatory physiology 2001, 281(4):H1823-1832. 29. Mongan PD, Capacchione J, Fontana JL, West S, Bunger R: Pyruvate improves cerebral metabolism during hemorrhagic shock. American journal of physiology Heart and circulatory physiology 2001, 281(2):H854-864. 30. Martin BJ, Valdivia HH, Bunger R, Lasley RD, Mentzer RM, Jr.: Pyruvate augments calcium transients and cell shortening in rat ventricular myocytes. The American journal of physiology 1998, 274(1 Pt 2):H8-17. 31. Montgomery CM, Fairhurst AS, Webb JL: Metabolic studies on heart mitochondria. III. The action of parapyruvate on alpha-ketoglutaric oxidase. The Journal of biological chemistry 1956, 221(1):369-376. 32. Montgomery CM, Webb JL: Metabolic studies on heart mitochondria. II. The inhibitory action of parapyruvate on the tricarboxylic acid cycle. The Journal of biological chemistry 1956, 221(1):359-368. 33. Majerus PW, Connolly TM, Deckmyn H, Ross TS, Bross TE, Ishii H, Bansal VS, Wilson DB: The metabolism of phosphoinositide-derived messenger molecules. Science 1986, 234(4783):1519-1526. 34. Singer WD, Brown HA, Sternweis PC: Regulation of eukaryotic phosphatidylinositol-specific phospholipase C and phospholipase D. Annual review of biochemistry 1997, 66:475-509. 35. Rhee SG: Regulation of phosphoinositide-specific phospholipase C. Annual review of biochemistry 2001, 70:281-312. 36. Hofmann SL, Majerus PW: Identification and properties of two distinct phosphatidylinositol-specific phospholipase C enzymes from sheep seminal vesicular glands. The Journal of biological chemistry 1982, 257(11):6461-6469. 37. Ryu SH, Cho KS, Lee KY, Suh PG, Rhee SG: Two forms of phosphatidylinositol-specific phospholipase C from bovine brain. Biochemical and biophysical research communications 1986, 141(1):137-144. 38. Ryu SH, Cho KS, Lee KY, Suh PG, Rhee SG: Purification and characterization of two immunologically distinct phosphoinositide-specific phospholipases C from bovine brain. The Journal of biological chemistry 1987, 262(26):12511-12518. 39. Ryu SH, Suh PG, Cho KS, Lee KY, Rhee SG: Bovine brain cytosol contains three immunologically distinct forms of inositolphospholipid-specific phospholipase C. Proceedings of the National Academy of Sciences of the United States of America 1987, 84(19):6649-6653. 40. Suh PG, Ryu SH, Moon KH, Suh HW, Rhee SG: Cloning and sequence of multiple forms of phospholipase C. Cell 1988, 54(2):161-169. 41. Harden TK, Sondek J: Regulation of phospholipase C isozymes by ras superfamily GTPases. Annual review of pharmacology and toxicology 2006, 46:355-379. 42. Gresset A, Sondek J, Harden TK: The phospholipase C isozymes and their regulation. Sub-cellular biochemistry 2012, 58:61-94. 43. Essen LO, Perisic O, Cheung R, Katan M, Williams RL: Crystal structure of a mammalian phosphoinositide-specific phospholipase C delta. Nature 1996, 380(6575):595-602. 44. Paterson HF, Savopoulos JW, Perisic O, Cheung R, Ellis MV, Williams RL, Katan M: Phospholipase C delta 1 requires a pleckstrin homology domain for interaction with the plasma membrane. The Biochemical journal 1995, 312 ( Pt 3):661-666. 45. Wang T, Dowal L, El-Maghrabi MR, Rebecchi M, Scarlata S: The pleckstrin homology domain of phospholipase C-beta(2) links the binding of gbetagamma to activation of the catalytic core. The Journal of biological chemistry 2000, 275(11):7466-7469. 46. Otterhag L, Sommarin M, Pical C: N-terminal EF-hand-like domain is required for phosphoinositide-specific phospholipase C activity in Arabidopsis thaliana. FEBS letters 2001, 497(2-3):165-170. 47. Bahk YY, Song H, Baek SH, Park BY, Kim H, Ryu SH, Suh PG: Localization of two forms of phospholipase C-beta1, a and b, in C6Bu-1 cells. Biochimica et biophysica acta 1998, 1389(1):76-80. 48. Park D, Jhon DY, Lee CW, Ryu SH, Rhee SG: Removal of the carboxyl-terminal region of phospholipase C-beta 1 by calpain abolishes activation by G alpha q. The Journal of biological chemistry 1993, 268(5):3710-3714. 49. Smrcka AV, Sternweis PC: Regulation of purified subtypes of phosphatidylinositol-specific phospholipase C beta by G protein alpha and beta gamma subunits. The Journal of biological chemistry 1993, 268(13):9667-9674. 50. Lee CW, Lee KH, Lee SB, Park D, Rhee SG: Regulation of phospholipase C-beta 4 by ribonucleotides and the alpha subunit of Gq. The Journal of biological chemistry 1994, 269(41):25335-25338. 51. Camps M, Carozzi A, Schnabel P, Scheer A, Parker PJ, Gierschik P: Isozyme-selective stimulation of phospholipase C-beta 2 by G protein beta gamma-subunits. Nature 1992, 360(6405):684-686. 52. Runnels LW, Scarlata SF: Determination of the affinities between heterotrimeric G protein subunits and their phospholipase C-beta effectors. Biochemistry 1999, 38(5):1488-1496. 53. Paulssen RH, Woodson J, Liu Z, Ross EM: Carboxyl-terminal fragments of phospholipase C-beta1 with intrinsic Gq GTPase-activating protein (GAP) activity. The Journal of biological chemistry 1996, 271(43):26622-26629. 54. Martelli AM, Gilmour RS, Bertagnolo V, Neri LM, Manzoli L, Cocco L: Nuclear localization and signalling activity of phosphoinositidase C beta in Swiss 3T3 cells. Nature 1992, 358(6383):242-245. 55. Kim CG, Park D, Rhee SG: The role of carboxyl-terminal basic amino acids in Gqalpha-dependent activation, particulate association, and nuclear localization of phospholipase C-beta1. The Journal of biological chemistry 1996, 271(35):21187-21192. 56. Tabellini G, Bortul R, Santi S, Riccio M, Baldini G, Cappellini A, Billi AM, Berezney R, Ruggeri A, Cocco L et al: Diacylglycerol kinase-θ is localized in the speckle domains of the nucleus. Experimental Cell Research 2003, 287(1):143-154. 57. Xu A, Suh PG, Marmy-Conus N, Pearson RB, Seok OY, Cocco L, Gilmour RS: Phosphorylation of nuclear phospholipase C beta1 by extracellular signal-regulated kinase mediates the mitogenic action of insulin-like growth factor I. Molecular and cellular biology 2001, 21(9):2981-2990. 58. Neri LM, Borgatti P, Capitani S, Martelli AM: Nuclear diacylglycerol produced by phosphoinositide-specific phospholipase C is responsible for nuclear translocation of protein kinase C-alpha. The Journal of biological chemistry 1998, 273(45):29738-29744. 59. Avazeri N, Courtot AM, Lefevre B: Regulation of spontaneous meiosis resumption in mouse oocytes by various conventional PKC isozymes depends on cellular compartmentalization. Journal of cell science 2004, 117(Pt 21):4969-4978. 60. Xu A, Wang Y, Xu LY, Gilmour RS: Protein kinase C alpha -mediated negative feedback regulation is responsible for the termination of insulin-like growth factor I-induced activation of nuclear phospholipase C beta1 in Swiss 3T3 cells. The Journal of biological chemistry 2001, 276(18):14980-14986. 61. Lukinovic-Skudar V, Matkovic K, Banfic H, Visnjic D: Two waves of the nuclear phospholipase C activity in serum-stimulated HL-60 cells during G(1) phase of the cell cycle. Biochimica et biophysica acta 2007, 1771(4):514-521. 62. Visnjic D, Banfic H: Nuclear phospholipid signaling: phosphatidylinositol-specific phospholipase C and phosphoinositide 3-kinase. Pflugers Archiv : European journal of physiology 2007, 455(1):19-30. 63. Hocevar BA, Fields AP: Selective translocation of beta II-protein kinase C to the nucleus of human promyelocytic (HL60) leukemia cells. The Journal of biological chemistry 1991, 266(1):28-33. 64. Hocevar BA, Burns DJ, Fields AP: Identification of protein kinase C (PKC) phosphorylation sites on human lamin B. Potential role of PKC in nuclear lamina structural dynamics. The Journal of biological chemistry 1993, 268(10):7545-7552. 65. Goss VL, Hocevar BA, Thompson LJ, Stratton CA, Burns DJ, Fields AP: Identification of nuclear beta II protein kinase C as a mitotic lamin kinase. The Journal of biological chemistry 1994, 269(29):19074-19080. 66. Walker SD, Murray NR, Burns DJ, Fields AP: Protein kinase C chimeras: catalytic domains of alpha and beta II protein kinase C contain determinants for isotype-specific function. Proceedings of the National Academy of Sciences of the United States of America 1995, 92(20):9156-9160. 67. Sun B, Murray NR, Fields AP: A role for nuclear phosphatidylinositol-specific phospholipase C in the G2/M phase transition. The Journal of biological chemistry 1997, 272(42):26313-26317. 68. Gokmen-Polar Y, Fields AP: Mapping of a molecular determinant for protein kinase C betaII isozyme function. The Journal of biological chemistry 1998, 273(32):20261-20266. 69. Faenza I, Matteucci A, Manzoli L, Billi AM, Aluigi M, Peruzzi D, Vitale M, Castorina S, Suh PG, Cocco L: A role for nuclear phospholipase Cbeta 1 in cell cycle control. The Journal of biological chemistry 2000, 275(39):30520-30524. 70. Divecha N, Letcher AJ, Banfic HH, Rhee SG, Irvine RF: Changes in the components of a nuclear inositide cycle during differentiation in murine erythroleukaemia cells. The Biochemical journal 1995, 312 ( Pt 1):63-67. 71. Hinchliffe KA, Irvine RF, Divecha N: Regulation of PtdIns4P 5-kinase C by thrombin-stimulated changes in its phosphorylation state in human platelets. The Biochemical journal 1998, 329 ( Pt 1):115-119. 72. Faenza I, Bavelloni A, Fiume R, Lattanzi G, Maraldi NM, Gilmour RS, Martelli AM, Suh PG, Billi AM, Cocco L: Up-regulation of nuclear PLCbeta1 in myogenic differentiation. Journal of cellular physiology 2003, 195(3):446-452. 73. Scott BL, Deeg HJ: Myelodysplastic syndromes. Annual review of medicine 2010, 61:345-358. 74. Follo MY, Bosi C, Finelli C, Fiume R, Faenza I, Ramazzotti G, Gaboardi GC, Manzoli L, Cocco L: Real-time PCR as a tool for quantitative analysis of PI-PLCbeta1 gene expression in myelodysplastic syndrome. International journal of molecular medicine 2006, 18(2):267-271. 75. Martelli AM, Faenza I, Billi AM, Fala F, Cocco L, Manzoli L: Nuclear protein kinase C isoforms: key players in multiple cell functions? Histology and histopathology 2003, 18(4):1301-1312. 76. Manzoli L, Martelli AM, Billi AM, Faenza I, Fiume R, Cocco L: Nuclear phospholipase C: involvement in signal transduction. Progress in lipid research 2005, 44(4):185-206. 77. Martelli AM, Evangelisti C, Nyakern M, Manzoli FA: Nuclear protein kinase C. Biochimica et biophysica acta 2006, 1761(5-6):542-551. 78. Allen DA, Yaqoob MM, Harwood SM: Mechanisms of high glucose-induced apoptosis and its relationship to diabetic complications. The Journal of nutritional biochemistry 2005, 16(12):705-713. 79. Ferguson KM, Berger MB, Mendrola JM, Cho HS, Leahy DJ, Lemmon MA: EGF activates its receptor by removing interactions that autoinhibit ectodomain dimerization. Molecular cell 2003, 11(2):507-517. 80. Herbst RS: Review of epidermal growth factor receptor biology. International journal of radiation oncology, biology, physics 2004, 59(2 Suppl):21-26. 81. Uniewicz KA, Ori A, Xu R, Ahmed Y, Wilkinson MC, Fernig DG, Yates EA: Differential scanning fluorimetry measurement of protein stability changes upon binding to glycosaminoglycans: a screening test for binding specificity. Analytical chemistry 2010, 82(9):3796-3802. 82. Divecha N, Banfic H, Irvine RF: The polyphosphoinositide cycle exists in the nuclei of Swiss 3T3 cells under the control of a receptor (for IGF-I) in the plasma membrane, and stimulation of the cycle increases nuclear diacylglycerol and apparently induces translocation of protein kinase C to the nucleus. The EMBO journal 1991, 10(11):3207-3214. 83. Avruch J, Khokhlatchev A, Kyriakis JM, Luo Z, Tzivion G, Vavvas D, Zhang XF: Ras activation of the Raf kinase: tyrosine kinase recruitment of the MAP kinase cascade. Recent progress in hormone research 2001, 56:127-155. 84. Satoh T, Nakafuku M, Kaziro Y: Function of Ras as a molecular switch in signal transduction. The Journal of biological chemistry 1992, 267(34):24149-24152. 85. Turner JL, Bierman EL: Effects of glucose and sorbitol on proliferation of cultured human skin fibroblasts and arterial smooth-muscle cells. Diabetes 1978, 27(5):583-588. 86. Dhanasekaran M, Indumathi S, Rajkumar JS, Sudarsanam D: Effect of high glucose on extensive culturing of mesenchymal stem cells derived from subcutaneous fat, omentum fat and bone marrow. Cell biochemistry and function 2012. 87. Gadau S: Nitrosative stress induces proliferation and viability changes in high glucose-exposed rat Schwannoma cells. Neuro Endocrinol Lett 2012, 33(3):279-283. 88. Hardie DG: Minireview: the AMP-activated protein kinase cascade: the key sensor of cellular energy status. Endocrinology 2003, 144(12):5179-5183. 89. Motoshima H, Goldstein BJ, Igata M, Araki E: AMPK and cell proliferation--AMPK as a therapeutic target for atherosclerosis and cancer. The Journal of physiology 2006, 574(Pt 1):63-71. 90. Du J, Guan T, Zhang H, Xia Y, Liu F, Zhang Y: Inhibitory crosstalk between ERK and AMPK in the growth and proliferation of cardiac fibroblasts. Biochemical and biophysical research communications 2008, 368(2):402-407. 91. Brownlee M: Biochemistry and molecular cell biology of diabetic complications. Nature 2001, 414(6865):813-820. 92. Sattler UG, Mueller-Klieser W: The anti-oxidant capacity of tumour glycolysis. International journal of radiation biology 2009, 85(11):963-971. 93. Mallet RT, Sun J, Knott EM, Sharma AB, Olivencia-Yurvati AH: Metabolic cardioprotection by pyruvate: recent progress. Exp Biol Med (Maywood) 2005, 230(7):435-443. 94. Suh PG, Park JI, Manzoli L, Cocco L, Peak JC, Katan M, Fukami K, Kataoka T, Yun S, Ryu SH: Multiple roles of phosphoinositide-specific phospholipase C isozymes. BMB reports 2008, 41(6):415-434. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65358 | - |
dc.description.abstract | 丙酮酸(Pyruvate)為糖解(Glycolysis)作用的終產物,並可以再繼續進行發酵反應產生乳酸或乙醇,或進入粒線體進行檸檬酸循環產生更大的能量。如同其他的α酮酸(α-keto acids),丙酮酸具有抗氧化的性質,例如以非酵素作用方式與過氧化氫作用產生乙酸、二氧化碳及水。過去認為丙酮酸是醣類代謝的重要調控分子,藉由異位調節(Allosteric regulation)來調控糖解作用及檸檬酸循環的速率。隨著近年的代謝體學發展,許多代謝小分子被發現除了在本身代謝途徑中的調控,同時也具有其他生理上的意義。我們藉由串連疏水性層析及質譜儀分析,發現丙酮酸和PLC-β1可能具有極佳之結合力,當HEK(Human embryonic kidney)細胞經過glucose starvation並以丙酮酸處理後,會造成PLC-β1活化,並由細胞質轉移至細胞核中,在核內活化下游的PKC pathway。此一現象與HEK細胞經不同濃度之葡萄糖處理後之表現相符,也佐證了葡萄糖代謝會引發特定訊息傳導的想法,而且丙酮酸可能是媒介為醣類代謝訊息傳導的關鍵物。同時外加入的丙酮酸,也會活化ERK-MAPK pathway,並可能在核內活化PLC-β1,利用抑制劑,我們確認丙酮酸是同時活化PLC-β1及ERK-MAPK pathway,而非單一的藉由ERK 1/2來活化核內的PLC-β1。當外加丙酮酸時,不論是在正常細胞或過度表現PLC-β1的細胞,都擁有較佳的存活率,而過度表現PLC-β1的細胞又稍高於正常細胞,因此由丙酮酸所誘發的PLC-β1活化路徑,可能對於細胞生存上有著正面意義。 | zh_TW |
dc.description.abstract | Pyruvate is the end product of glycolysis. It may undergo the fermentation process leading to lactate or ethanol production, or enter the TCA cycle in the mitochondria to produce more energy. Like the other α-keto acids, pyruvate has the antioxidative property in which pyruvate reacts with hydrogen peroxide in a non-enzymatic way yielding acetate, carbon dioxide and water. Pyruvate has been considered as an important regulatory molecule in the carbohydrate metabolism affecting the velocity of glycolysis and TCA cycle by allosteric regulation. Recently many metabolites are considered to exert other physiological functions besides serving as metabolic intermediates. Using Affinity Elution from Tandem Hydrophobic Interaction Chromatography(AETHIC) and MS analysis, we discovered that PLC-β1 may be a pyruvate-binding protein. In addition, pyruvate induces PLC-β1 activation and translocation from cytosol to nucleus in glucose-starved HEK cells, leading to PKC activation. Similar result have been found in starved cells treated with glucose. These data confirmed that glucose-induced signal transduction may be mediated by pyruvate and PLC-β1. Treatment of pyruvate also activates the ERK-MAPK pathway. However we have found that activation of PLC-β1 and ERK are two independent pathways. In the presence of exogenous pyruvate, both normal cells and cells over-expressing PLC-β1 survive better than cells without pyruvate. Therefore, pyruvate serves as a signal molecules in cells with a high flux of glycolysis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T23:38:25Z (GMT). No. of bitstreams: 1 ntu-101-R99B46024-1.pdf: 2646895 bytes, checksum: 35e784bac231f41b666e7ccca4f1efea (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 目錄
誌謝 Ⅰ 中文摘要 Ⅱ 英文摘要 Ⅲ 縮寫表 Ⅳ 目錄 Ⅵ 第一章 前言 1.1 葡萄糖代謝誘發訊息傳導 1.1.1 葡萄糖代謝所引發之生理意義 1 1.1.2 葡萄糖在細胞內引起訊息傳導 1 1.1.3 MAPK 在葡萄糖誘發之細胞凋亡扮演的角色 2 1.2 丙酮酸在葡萄糖代謝扮演關鍵角色 1.2.1 丙酮酸連結葡萄糖代謝相關路徑 3 1.2.2 丙酮酸的抗氧化性質 3 1.2.3 丙酮酸有效地改善器官在缺血環境下所造成的後遺症 4 1.3 磷脂酶在細胞內引發多樣性的訊息 1.3.1 Phosphoinositide-specific phospholipase C 5 1.3.2 PLC-β1在細胞膜受到G protein刺激而活化 5 1.3.3 PLC-β1在細胞核內的活化及調控 6 1.3.4 PLC-β1對於細胞生長分化及生理的意義 6 1.4 研究動機 7 第二章 材料與實驗方法 2.1 質譜儀樣本製備 2.1.1 Affinity Elution from Tandam Hydrophobic Interaction Chromatography 9 2.1.2 In-Gel Digestion 9 2.2 細胞實驗方法 2.2.1 細胞培養 10 2.2.2 繼代培養 10 2.2.3 細胞冷凍及解凍 10 2.2.4 細胞均質液 (whole cell lysate)萃取 11 2.2.5 核膜質萃取(Differential Detergent Fractionation,DDF) 11 2.2.6 核質萃取 11 2.2.7 MTT assay 11 2.3重組人類 PLC-β1 蛋白 2.3.1 引子設計 (Primer Designing) 12 2.3.2 聚合酶連鎖反應 (Polymerase Chain Reaction, PCR) 12 2.3.3 DNA純化 (purify) 12 2.3.4 核酸限制酶切割 (Digestion of Restriction Enzyme) 12 2.3.5 pCMV14 vector與DNA的接合反應 (Ligation) 13 2.3.6瓊脂膠電泳分析 (Agarose Gel Electrophoresis) 13 2.3.7細胞轉染 (Transfection) 13 2.4 蛋白質實驗方法 2.4.1 BCA 蛋白質定量 13 2.4.2 SDS聚丙烯胺凝膠電泳(Sodium Dodecyl Sulfate- PolyAcrylamide Gel Electrophoresis,SDS-PAGE) 14 2.4.3 Coomassie Brilliant Blue G-250 染色 15 2.4.4 銀染色法(Silver stainning) 15 2.4.5 電泳轉印 15 2.4.6 西方點墨法 (Western Blotting) 15 2.4.7 免疫螢光染色 (Immunofluorescence Staining) 16 2.5 藥物配置及處理 16 第三章 結果 3.1 辨識細胞內以丙酮酸為受質可能引發之訊息傳遞 17 3.2 丙酮酸誘發 PLC-β1 於細胞內之位移 17 3.3 丙酮酸同時活化 MEK/ERK MAPK pathway 18 3.4 丙酮酸的活化路徑其受器並非透過 EGFR 19 3.5 PLC-β1受丙酮酸活化路徑與MEK/ERK MAPK pathway活化 為兩條路徑 19 3.6 丙酮酸具有增加細胞增殖的功效 20 第四章 討論 4.1 細胞對於環境養份之生存策略 21 4.2 丙酮酸誘發胞內 PLC-β1 位移 21 4.3 丙酮酸活化 ERK-MAPK 路徑 23 4.4 丙酮酸誘導的PLC-β1路徑促進 HEK 細胞增殖 24 第五章 結語 26 第六章 實驗結果圖表 圖1 以銀染色法對親和性沖提串聯疏水性作用層析(AETHIC) 結果進行確認 27 表1 Octyl-sepharose管柱第2管質譜鑑定protein ID結果 30 圖2 選用HEK細胞株做為研究模式系統 31 圖3 PLC-β1 受丙酮酸刺激產生位移 32 圖4 核內 PLC-β1 下游蛋白質PKCα訊息受丙酮酸刺激而活化 33 圖5 免疫螢光染色丙酮酸處理之 HEK 細胞 35 圖6 PLC-β1與PKCα的共同易位受U73122抑制 36 圖7 由丙酮酸所引起的PLC-β1訊息傳導與葡萄糖處理相符 37 圖8 丙酮酸同時誘導 ERK-MAPK 路徑的活化 38 圖9 由丙酮酸誘導的 ERK-MAPK 路徑活化並非透過EGFR做為受器 40 圖10 PLC-β1在核內的活化非單一受pERK活化 41 圖11 丙酮酸所誘發的 PLC-β1 活化路徑幫助細胞維持生存 42 第七章 參考文獻 43 第八章 附錄 附圖一 葡萄糖代謝路徑 51 附圖二 polyol pathway造成氧化壓力上升 52 附圖三 葡萄糖刺激β細胞進行有絲分裂 53 附圖四 葡萄糖引發不同的MAPK pathway影響細胞凋亡 54 附圖五 丙酮酸透過非酵素的方式與過氧化氫結合 55 附圖六 丙酮酸非直接的提升體內還原態榖胱甘肽 56 附圖七 PLC催化DAG及IP3產生 57 附圖八 PLC isoform 58 附圖九 PLCβ 同時受Gα及Gβγ活化 59 附圖十 丙酮酸活化 PLC-β1 訊號路徑示意圖 60 | |
dc.language.iso | zh-TW | |
dc.title | PLC-β1及丙酮酸媒介部分葡萄糖誘發之訊息傳導 | zh_TW |
dc.title | PLC-β1 and pyruvate mediating part of the glucose-induced signaling pathway | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李明亭,陳宏文,張茂山 | |
dc.subject.keyword | 葡萄糖代謝,丙酮酸,PLC-β1,PKC,ERK 1/2, | zh_TW |
dc.subject.keyword | glucose metablosim,pyruvate,PLC-β1,PKC,ERK 1/2, | en |
dc.relation.page | 60 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-07-26 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-101-1.pdf 目前未授權公開取用 | 2.58 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。