綠茶表沒食子酸酯型唲茶素酸酯對胰島素刺激前脂肪細胞增生的抑制

以作者查詢圖書館館藏

、以作者查詢臺灣博碩士

、以作者查詢全國書目

、勘誤回報

、線上人數：41

、訪客IP：3.144.29.103

姓名

劉咸君(Hsien-Chun Liu) 查詢紙本館藏

畢業系所

生命科學系

論文名稱

綠茶表沒食子酸酯型唲茶素酸酯對胰島素刺激前脂肪細胞增生的抑制
(Inhibitory effect of green tea (-)-epigallocatechin gallate on insulin-stimulated mitogenesis of 3T3-L1 preadipocytes)

相關論文

★ 中華鱉腦垂體甘丙氨激素之研究：cDNA選殖、表現及調控	★ 辛基苯酚對3T3-L1脂肪細胞中resistin的調節作用
★ FoxO1 調節抗胰島素激素基因的表現	★ 綠茶表沒食子唲茶素沒食子酸酯受器對於人類乳癌細胞株MCF7生長的影響
★ 綠茶表沒食子酸酯型唲茶素酸酯抑制第一型内皮素作用於脂肪細胞上攝入葡萄糖的訊息機制	★ 綠茶表兒茶素藉由microRNA-494路徑改善橫向主動脈繃紮術誘導型小鼠的心臟疾病
★ 內皮素誘導前脂肪細胞生長的訊息路徑	★ 綠茶對前脂肪細胞生長的影響
★ 綠茶唲茶素對由第一型類胰島素所調節前脂肪細胞生長的影響	★ 綠茶唲茶素對於前脂肪細胞分化的影響
★ Cdk2在綠茶唲茶素調節3T3-L1前脂肪細胞的生長和細胞凋亡扮演著必要性的角色	★ 綠茶唲茶素透過MAPK相關途徑抑制3T3-L1前脂肪細胞的生長
★ 第一型類胰島素生長因子、綠茶唲茶素及雌性素對3T3-L1脂肪細胞中resistin的基因表達有不同的調節效果	★ 綠茶唲茶素對前脂肪細胞內活性氧及榖胱甘肽的影響
★ 胰島素接受器受質在綠茶唲茶素對胰島素刺激前脂肪細胞生長作用中扮演的角色	★ 賜諾殺穩定性及東方果實蠅費洛蒙之研究

檔案

[Endnote RIS 格式]

[Bibtex 格式]

[相關文章]

[文章引用]

[完整記錄]

[館藏目錄]

至系統瀏覽論文 ( 永不開放)

摘要(中)

肥胖是一種常見的疾病，與癌症，糖尿病，高血壓以及心血管疾病都有相關。造成肥胖的原因是脂肪細胞數目的增加，或脂肪細胞內三酸甘油酯的堆積(泛稱脂肪細胞分化)使然。根據研究指出，內分泌與營養都會影響到肥胖的調節作用，其中胰島素在細胞分化時期會刺激細胞進行mitotic clonal expansion及adipogenesis，並促使細胞進行脂肪生成和抑制脂肪分解作用。然而綠茶唲茶素(亦稱維生素P)，尤其是綠茶表沒食子酸酯型唲茶素酸酯(英文名稱和簡稱分別為 (-)-epigallocatechin gallate和EGCG)，已被證實可以抑制脂肪細胞的增生與分化，並可促使前脂肪細胞和脂肪細胞的apoptosis，以及促進脂肪氧化作用，因此可視為預防肥胖的保健物質。有鑑於此，EGCG是否可調節胰島素刺激前脂肪細胞的增生仍然不清楚。在本論文中，使用3T3-L1前脂肪細胞株做為研究素材，我發現胰島素濃度在1-1000 nM，細胞數目和BrdU攝入皆有增加的情形，可以得知胰島素會刺激前脂肪細胞的增生。而胰島素的有效濃度EC50-100 在2- 4天的處理為100 nM。此外，由於胰島素可分別增加胰島素接受器、IRS-1和 Erk三種蛋白的磷酸化，可見胰島素分別增加胰島素接受器、IRS-1和MEK-1的活性。然而前處理EGCG (10-50 μM)於前脂肪細胞可以明顯抑制胰島素所促進的細胞數目、BrdU攝入、胰島素接受器的活性、IRS-1的活性、 MEK-1的活性及胰島素接受器與IRS-1兩者之間的結合作用。以上種種的跡象顯示，EGCG可透過阻礙胰島素的訊息傳導路徑而抑制前脂肪細胞的增生。

摘要(英)

Obesity, a common disease resulting from mitogenesis and lipid accumulation (so-called adipogenesis) of fat cells, is associated with the risks of cancers, diabetes, hypertension, and cardiovascular disease. Development of obesity can be regulated by endocrine and nutritional cues based on a variety of laboratory studies. Insulin (INS) was reported to stimulate mitotic clonal expansion and adipogenesis during fat cell differentiation in additional to stimulating lipogenesis and inhibiting lipolysis. In contrast, green tea polyphenols (the so-called vitamin P), especially the catechin, (-)-epigallocatechin gallate (EGCG), was found to inhibit proliferation and differentiation of fat cells, to induce the apoptosis of preadipocytes and adipocytes, to stimulate fat oxidation, and to be proposed as an obesity chemopreventative. But, determining whether EGCG acts to regulate the stimulatory effect of INS on the mitogenesis of fat cells is still not clear. In this thesis using 3T3-L1 preadipocytes as the research cell model, I found that INS at 1-1000 nM could stimulate the growth of 3T3-L1 preadipocytes as indicated by increased cell number and BrdU incorporation. The EC50-100 of INS was about 100 nM after 2~4 days of treatment. Also, INS stimulated activities of INS receptor (INSR), insulin receptor substrate 1 (IRS-1), and MEK1, respectively, as indicated by increased phosphorylation of INSR, IRS-1, and Erk proteins. However, pretreatment of preadipocytes with EGCG (10-50 μM) resulted in significant decreases in the INS-induced increases in their cell number, BrdU incorporation, activities of INSR, IRS-1, and MEK-1, and the association of INSR with IRS-1. These data suggest that EGCG can inhibit preadipocyte mitogenesis by reduction of INS-mediated signaling in these cells.

關鍵字(中)

★ 綠茶表沒食子酸酯型唲茶素酸酯
★ 前脂肪細胞
★ 胰島素

關鍵字(英)

★ (-)-epigallocatechin gallate
★ insulin
★ 3T3-L1 preadipocytes

論文目次

中文摘要 I
Abstract II
縮寫與全名對照 III
目錄 IV
圖目錄 VI
第一章：緒論 1
一、胰島素(Insulin) 1
二、胰島素接受器(insulin receptor,簡寫為INSR) 1
三、胰島素訊息傳導途徑 2
四、胰島素與前脂肪細胞 3
五、綠茶唲茶素與前脂肪細胞 4
六、動機與目的 5
第二章：材料與方法 6
一.實驗材料 6
二.實驗方法 7
1. 細胞培養 7
2. 細胞處理 7
3. 細胞計數方法 7
4. DNA合成5-bromo-2’-deoxyuridine (BrdU)試驗 8
5. 蛋白質濃度定量 8
6. 抑制劑(PD98059, LY294002, U0126, SB203580）的處理 9
7. 西方點墨法 (Western blotting) 9
8. 免疫沉析法 (Immunoprecipitation) 10
9. 人類胰島素的配製 10
10.生統分析 11
第三章：結果 12
一、胰島素刺激前脂肪細胞的增生 12
二、胰島素刺激前脂肪細胞的增生與MEK-1活性有關 12
三、胰島素刺激前脂肪細胞的增生受到綠茶唲茶素EGCG抑制 13
四、綠茶唲茶素EGCG對胰島素刺激MEK-1活性的影響 13
五、不同種類綠茶唲茶素影響胰島素刺激前脂肪細胞的生長和MEK-1活性 14
六、胰島素刺激前脂肪細胞的胰島素接受器和下游IRS-1蛋白的活性 15
七、綠茶唲茶素EGCG抑制胰島素刺激前脂肪細胞的胰島素接受器和下游IRS-1蛋白的活性 15
八、綠茶唲茶素EGCG抑制胰島素刺激前脂肪細胞之胰島素接受器和下游IRS-1之間的結合能力 16
第四章：討論 17
第五章：結論與展望 20
第六章：參考文獻 21
第七章：附錄 38

參考文獻

1. Banting FG and Best GH. The internal secretion of the pancreas. J Lab Clin Med 7: 241-66, 1992.
2. Proud CG and Denton RM. Molecular mechanisms for the control of translation by insulin. Biochem J 328: 329-42, 1997.
3. Holman GD and Kasuga M. From receptor to transporter: Insulin signaling to glucose transport. Diabetologia 40: 991-1003, 1997.
4. White MF. The IRS-signalling system: a network of docking proteins that mediate insulin action. Mol Cell Biochem 182: 3–11, 1998.
5. O’Neill T, Craparo A and Gustafson T. Characterization of an interaction between insulin receptor substrate 1 and the insulin receptor by using the two-hybrid system. Mol Cell Biol 14: 6433–6442, 1994.
6. He W, Craparo A, Zhu Y, O’Neill T, Wang LM, Pierce J and Gustafson T. Interaction of insulin receptor substrate-2 (IRS-2) with the insulin and insulin-like growth factor I receptors. Evidence for two distinct phosphotyrosine-dependent interaction domains within IRS-2. J Biol Chem 271: 11641–11645, 1996.
7. Burks DJ, Wang J, Towery H, Ishibashi O, Lowe D, Riedel H and White MF. IRS pleckstrin homology domains bind to acidic motifs in proteins. J Biol Chem 273: 31061–31067, 1998.
8. Stark F, Golla R and Nachmias VT. Formation and contraction of a microfilamentous shell in saponin-permeabilized platelets. J Cell Biol 112: 903–913, 1991.
9. Franke WW, Hergt M and Grund C. Rearrangement of the vimentin cytoskeleton during adipose conversion: formation of an intermediate filament cage around lipid globules. Cell 49: 131–141, 1987.
10. Kops GJ and Burgering BM. Forkhead transcription factors are targets of signalling by the proto-oncogene PKB (C-AKT). J Anat 197: 571-574, 2000.
11. Zhang JW, Tang QQ, Vinson Charles and Lane MD. Dominant-negative C/EBP disrupts mitotic clonal expansion and differentiation of 3T3-L1 preadipocytes. PNAS 101: 43–47, 2004.
12. Craig W J. Health-promoting properties of common herbs. Am J Clin Nutr 70: 491S-499S, 1999.
13. Hollman PC. Bioavailability of flavonoids. Eur J Clin Nutr 51: S66-S69, 1997.
14. Guo Q, Zhao B, Shen S, Hou J, Hu J, and Xin W. ESR study on the structure-antioxidant activity relationship of tea catechins and their epimers. Biochim Biophys Acta 1427: 13-23, 1999.
15. Katiyar SK and Mukhtar H. Tea antioxidants in cancer chemoprevention. J Cell Biochem Suppl 27: 59-67, 1997.
16. Yang CS and Wang ZY. Tea and cancer. J Natl Cancer Inst 85: 1038-1049, 1993.
17. Cao Y and Cao R. Angiogenesis inhibited by drinking tea. Nature (Lond.) 398: 381, 1999
18. Nakayama M, Suzuki K, Toda M, Okubo S and Hara Y. Inhibition of the infectivity of influenza virus by tea polyphenols. Antivir Res 21: 289-299, 1993.
19. Kao YH, Hiipakka RA and Liao S. Modulation of Endocrine Systems and Food Intake by Green Tea Epigallocatechin Gallate. Endocrinology 141: 980-987, 2000.
20. Wu BT, Hung PF, Chen HC, Huang RN, Chang HH, Kao YH. The apoptotic effect of green tea (-)-epigallocatechin gallate on 3T3-L1 preadipocytes depends on the Cdk2 pathway. J Agric Food Chem 53: 5695-701, 2005.
21. Hung PF, Wu BT, Chen HC, Chen YH, Chen CL, Wu MH, Liu HC, Lee MJ, Kao YH. Antimitogenic effect of green tea (-)-epigallocatechin gallate on 3T3-L1 preadipocytes depends on the ERK and Cdk2 pathways. Am J Physiol Cell Physiol 288: C1094-108, 2005
22. Lin J, Della-Fera MA, Baile CA. Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res 13: 982-90, 2005
23. Shimizu M, Deguchi A, Joe A, McKoy J, Moriwaki H and Weinstein IB. EGCG inhibits activation of HER3 and expression of cyclooxygenase-2 in human colon cancer cells. J Exper Ther Oncol 5: 69–78, 2005.
24. Masuda M, Suzui M and Weinstein IB. Effects of epigallocatechin-3-gallate on growth, epidermal growth factor receptor signaling pathways, gene expression, and chemosensitivity in human head and neck squamous cell carcinoma cell lines. Clin Cancer Res 7: 4220–4229, 2001.
25. Masuda M, Suzui M, Lim JT and Weinstein IB. Epigallocatechin-3-gallate inhibits activation of HER-2/neu and downstream signaling pathways in human head and neck and breast carcinoma cells. Clin Cancer Res 9: 3486–3491, 2003
26. Shimizu M, Deguchi A, Lim JT, Moriwaki H, Kopelovich L and Weinstein IB. EGCG and polyphenon E inhibit growth and activation of the EGFR and HER2 signaling pathways in human colon cancer cells. Clinical Cancer Res 9: 3486–3491, 2003.
27. Shimizu M and Weinstein IB. Modulation of signal transduction by tea catechins and related phytochemicals. Mutat Res (in press).
28. Liang YC, Lin-shiau SY, Chen CF and Lin JK. Suppression of extracellular signals and cell proliferation through EGF receptor binding by (−)-epigallocatechin gallate in human A431 epidermoid carcinoma cells. J Cell Biochem 67: 55–65, 1997.
29. Sachinidis A, Seul C and Seewald S. Green tea compounds inhibit tyrosine phosphorylation of PDGF beta-receptor and transformation of A172 human glioblastoma. FEBS Lett 471: 41–55, 2000.
30. Wilson JD and Foster DW. Williams Textbook of Endocrinology 8th edition, 1096-1106, 1992,
31. Fujimura Y, Tachibana H and Yamada K. Lipid raft-associated catechin suppresses the FcepsilonR1 expression by inhibiting phosphorylation of the extracellular signal-regulated kinase 1/2. FEBS Lett 556: 204–210, 2004.
32. Amelia EP and Kahn CR. Differential Roles of the Insulin and Insulin-like Growth Factor-I (IGF-I) Receptors in Response to Insulin and IGF-I. JBC 279: 38016-38024, 2004.
33. Tang HR, Covington AD and Hancock RA. Structure-activity relationships in the hydrophobic interactions of polyphenols with cellulose and collagen. Biopolymers 70: 403-13, 2003.
34. Tachibana H, Koga K, Fujimura Y and Yamada K. A receptor for green tea polyphenol EGCG. Nat Struct Mol Biol 11: 380-381, 2004.
35. Liu HS, Chen YH, Hung PF and Kao YH. Inhibitory effect of green tea (-)-epigallocatechin gallate on resistin gene expression in 3T3-L1 adipocytes depends on the ERK pathway. Am J Physiol Endocrinol Metab 290: E273-81

指導教授

高永旭(Yung-Hsi Kao)

審核日期

2006-1-24

推文