博碩士論文 102224003 詳細資訊




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:7 、訪客IP:54.198.52.8
姓名 陳毅(Yi Chen)  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兒茶素酸酯在3T3-L1脂肪細胞分化過程中調節FoxO家族基因的表現
(Green tea epigallocatechin gallate regulates the expression of the FoxO family during differentiation of 3T3-L1 fat cells)
檔案 [Endnote RIS 格式]    [Bibtex 格式]    至系統瀏覽論文 (2022-1-24以後開放)
摘要(中) Forkhead box class O (FoxO)家族基因包含有FoxO1、FoxO3、FoxO4和FoxO6四種,過去有許多文獻分別指出各FoxO基因在細胞內調控了包括分化、葡萄醣代謝、DNA修復和細胞凋亡在內的許多功能。在本實驗中我們使用了3T3-L1前脂肪細胞分化為脂肪細胞的系統,透過處理Dexamethasone、Insulin和3-isobutyl-1-methylxanthine等3種藥物所組成之分化劑,促使細胞進行為期14天的分化。在分化的過程中,細胞的數目有明顯的增加,而細胞內會有三酸甘油脂的堆積。在我們的實驗中發現,相較於沒有處理EGCG的組別,在分化期間處理EGCG會使細胞內油滴的數量減少。當沒有處理EGCG時,細胞的FoxO1、FoxO3、FoxO4和FoxO6的mRNA表現量在分化過程的第2~6天之間會有增加的趨勢,而在第6~14天之間的表現量則會有降低的趨勢。在14天的分化期間,EGCG會大幅降低FoxO1、FoxO3、FoxO4和FoxO6的mRNA表現量。此外ATF-3的mRNA表現量則會有增加的趨勢,其表現量並不會受到EGCG的影響。這些實驗結果顯示EGCG會透過調節FoxO的表現量去影響3T3-L1前脂肪細胞的分化。
摘要(英) The forkhead box class O (FoxO) family of mammals contained 4 members: FoxO1, FoxO3, FoxO4 and FoxO6. Previous studies have revealed that FoxO genes regulate many functions in cells, including differentiation, glucose metabolism, DNA repair and apoptosis. In this study, we used the preadipocyte-adipocyte differentiation system of 3T3-L1 cells, induced by dexamethasone, insulin and 3-isobutyl-1-methylxanthine for a period of 14 days. During the differentiation, the cell number increased, and the TG droplet increased in cells. Our study also found that the TG droplet decreased when 3T3-L1 cells were treated EGCG, compared to control. In the absence of EGCG, FoxO1, FoxO3, FoxO4, and FoxO6 tended to increase their mRNA expressions from Day 2 to Day 6 of differentiation and decrease from Day 6 to Day 14 of differentiation. EGCG, at 100 μM, but not at 25 or 50 μM, significantly decreased levels of FoxO1, FoxO3, FoxO4, and FoxO6 mRNAs during a 14-day period of differentiation. Moreover, activating transcription factor (ATF)-3 tended to increase its mRNA expression and was not altered by EGCG treatment. These data suggest that EGCG regulates 3T3-L1 adipocyte differentiation through modulations of FoxO but not ATF-3 levels.
關鍵字(中) ★ 脂肪細胞
★ 兒茶素
★ 細胞分化
關鍵字(英) ★ FoxO
★ EGCG
★ adipocyte
★ differentiation
★ ATF-3
論文目次 電子檔授權書……………………………………………………………………………i
紙本延後上架申請書……………………………………………………………..…….ii
推薦書…………………………………………………………………………….…….iii
審定書…………………………………………………………………………….…….iv
中文摘要………………………….……………………………………………………v
英文摘要……………………….………………...……………………………………..vi
致謝…………………………………………….………………………………………vii
使用藥劑之縮寫與全名對照表………………...…….………………………………viii
目錄…………………………………………………..……………………………….ix
一、 前言……………………………………………….……………………….……..1
I. FoxO轉錄因子…………………………………..…………….…………….1
II. ATF-3轉錄因子……………...……………………………………….…….3
III. 脂肪細胞與細胞分化………………………………………….……………..3
IV. 綠茶兒茶素EGCG……………………...…………………………………..5
V. 研究動機與目的……..…………………………………..…………………7
二、 材料與方法………………………….…………..……….………………………8
I. 實驗材料…………..……………………………….…………………………8
II. 實驗方法…………….………………………………..…………………….8
三、 實驗結果………………………………………….……….……………………12
I. 前脂肪細胞分化方式的建立……………………..………………………12
II. 綠茶兒茶素EGCG對於脂肪細胞分化的影響…..….……..………….....12
III. 綠茶兒茶素EGCG在分化過程中對各FoxO基因表現量的影響………13
IV. 不同濃度的EGCG在分化過程中對ATF-3 mRNA表現量的影響…......…15
四、 討論………………………………………………….………………………..…16
I. 新的分化劑濃度可使細胞進入分化………….……………….………….16
II. 分化過程中EGCG對脂肪細胞的影響…………………………….…….16
III. EGCG在分化過程中對各FoxO mRNA表現量變化的影響…………….17
IV. EGCG在分化過程中對ATF3 mRNA表現量變化的影…….…………...19
V. 未來可能進行的實驗……………………………………….….…………19
五、結論………………………………………………………………...………………21
六、參考文獻……………………………………………………….………………...22
七、附表…………………………………...…………………….……………………29
八、附圖………………………………………...…………….………………………31
九、附錄……………………………………………...…….…………………………40
參考文獻 1. Barthel A, Schmoll D and Unterman TG. (2005) FoxO proteins in insulin action and metabolism. Trends Endocrinol Metab 16: 183-189.
2. Belguise K, Guo S and Sonenshein GE. (2007) Activation of FOXO3a by green tea polyphenol epigallocatechin-3-gallate induces estrogen receptor alpha expression reserving invasive phenotype of breast cancer cells. Cancer Res 67:5793-5670.
3. Biggs WH 3rd, Cavenee WK and Arden KC. (2001) Identification and characterization of members of the FKHR (FOX O) subclass of winged-helix transcription factors in the mouse. Mamm Gename 12:416-425.
4. Birkenkamp KU and Coffer PJ. (2003) FOXO transcription factors as regulators of immune hemeostasis: molecules to die for? J Immunol 171:1623-1629
5. Cannon B, Hedin A and Nedergaard J. (1982) Exclusive occurrence of thermogenin antigen in brown adipose tissue. FEBS Lett 150:129-132.
6. Cannon B and Nedergaard J (2004) Brown adipose tissue: function and physiological significance. Physiol Rev 84:277-359.
7. Chen YJ, Chen CC, Li TK, Wang PH, Liu LR, Chang FY, Wang YC, Yu YH, Lin SP, Mersmann HJ and Ding ST. (2012) Docosahexaenoic acid suppresses the expression of FoxO and its target genes. J Nutr Biochem 23:1609-1616.
8. Dulloo A, G and Duret C. (1999) Effecacy of a green tea extract rich in catechin polyphenols and caffeine in increasing 24-h expenditure and fat oxidation in humans. Am. J Clin Nutr 70:1040-1045.
9. Elberg G, Gimble JM and Tsai SY. (2000) Modulation of the murine peroxisome proliferator-activated receptor γ2 promoter activity by CCAAT/enhancer-binding proteins. J Biol Chem 275:27815-27822.
10. Gregoire FM, Smas CM and Sul HS. (1998) Understanding adipocyte differentiation. Physiol Rev. 78:783-809.
11. Frayn KN. (2002) Adipose tissue as a buffer for daily lipid flux. Diabetologia 45:1201-1210.
12. Furuyashiki T, Nagayasu H, Aoki Y, Bessho H, Hashimoto T, Kanazawa K and Ashida H. (2004) Tea catechin suppresses adipocyte differentiation accompanied by down-regulation of PPARγ2 and C/EBPα in 3T3-L1 cells. Biosci Biotechnol Biochem 68:2353-2359.
13. Greer EL and Brunet A. (2005) FOXO transcription factors at the interface between longevity and tumor suppression. Oncogene 24:7410-7425
14. Gregoire FM, Smas CM and Sul HS. (1998) Understanding adipocyte differentiation. Physiol Rev. 78:783-809.
15. Han J, Hajjar DP, Zhou X, Gotto AM, Jr and Nicholson AC. (2002) Regulation of peroxisome proliferator-activated receptor-γ-mediated gene expression. J Biol Chem 277:23582-23586
16. Higuchi M, Dusting GJ, Peshavariya H, Jiang F, Hsiao ST, Chan EC and Liu GS. (2013) Differentiation of human adipose-derived stem cells into fat involves reactive oxygen species and Forkhead box O1 mediated upregulation of antioxidant enzymes. Stem Cells Dev 22:878-888.
17. Hotamisligil GS, Shargill NS, and Spiegelman BM. (1993) Adipose expression of tumor necrosis factor-α: direct role in obesity-linked insulin resistance. Science 259:87-91.
18. Hung PF, Wu BT, Chen HC, Chen YH, Chen CL, Wu MH, Liu HC, Lee MJ and Kao YH. (2005) Antimitogenic effect of green tea (-)-epigallocatechin gallate on 3T3-L1 preadipocytes depends on the ERK and Cdk2 pathways. Am J Physiol Cell Physiol 288:C1094-1108.
19. Hwang JT, Park IJ, Shin JI, Lee YK, Lee SK, Baik HW, Ha J and Park OJ. (2005) Genistein, EGCG, and capsaicin inhibit adipocyte differentiation process via activating AMP-activated protein kinase. Biochem Biophys Res Commun 338:694-699.
20. Jacobs FM, van der Heide LP, Wijchers PJ, Burbach JP, Hoekman MF and Smidt MP. (2003) FoxO6, a novel number of the FoxO class of transcription factors with distinct shuttling dynamics. J Biol Chem 278:35959-35967.
21. Jang MK, Kim CH, Seong JK, Jung MH. (2012) ATF3 inhibits adipocyte differentiation of 3T3-L1 cells. Biochem Biophys Res Commun 421:38-43.
22. Kao YH, Hiipakka RA and Liao S. (2000a) Modulation of endocrine systems and food intake by green tea epigallocatechin gallate. Endocrinology 141:980-987.
23. Kao YH, Hiipakka RA and Liao S (2000b) Modulation of obesity by a green tea catechin. Am J Clin Nutr 72:1232-1234.
24. Kao YH, Chang HH, Lee MJ and Chen CL. (2006) Tea, obesity, and diabetes. Mol Nutr Food Res 50:188-210.
25. Kim DH, Zhang T, Lee S, Calabuig-Navarro V, Yamauchi J, Piccirillo A, Fan Y, Uppala R, Goetzman E and Dong HH. (2014) FoxO6 intergrates insulin signaling with MTP for regulating VLDL production in the liver. Endocrinology. 155:1255-1267.
26. Kim H, Hiraishi A, Tsuchiya K and Sakamoto K. (2010) (-)Epigallocatechin gallate suppresses the differentiation of 3T3-L1 preadipocytes through transcription factors FoxO1 and SREBP1c. Cytotechnology 62:245-255.
27. Kim S, Kim Y, Lee J and Chung J. (2010) Regulation of FOXO1 by TAK1-Nemo-like kinase pathway. J Biol Chem 285:8122-8129.
28. Ku HC, Chang HH, Liu HC, Hsiao CH, Lee MJ, Hung PF, Liu CW and Kao YH. (2009) Green tea (-)-epigallocatechin gallate inhibits insulin stimulation of 3T3-L1 preadipocyte mitogenesis via the 67-kDa laminin receptor pathway. Am J Physiol Cell Physiol 297:121-132.
29. Lea-Currie YR, Monroe D and McIntosh MK. (1999) Dehydroepiandrosterone and relates steroids alter 3T3-L1 preadipocyte proliferation and differentiation. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol123:17-25.
30. Lehmann OJ, Sowden JC, Carlsson P, Jordan T and Bhattacharya SS. (2003) Fox’s in development and disease. Trends in Genetics 19:339-344.
31. Lin J, Della-Fera MA and Baile CA. (2005) Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res 13:982-990.
32. Liu CW, Yang SY, Lin CK, Liu HS, Ho LT, Wu LY, Lee MJ, Ku HC, Chang HH, Huang RN and Kao YH. (2014) The forkhead transcription factor FOXO1 stimulates the expression of the adipocyte resistin gene. Gen Comp Endocrinol 196:41-51.
33. Matsuzaki H, Daitoku H, Hatta M, Tanaka K and Fukamizu A. (2003) Insulin-induced phosphorylation of FKHR (Foxo1) targets to proteasomal degradation. Proc Natl Acad Sci USA. 100:11285-11290.
34. Mandrup S, Loftus TM, MacDougald OA, Kuhajda FP and Lane MD. (1997) Obese gene expression at in vivo levels by fat pads derived from s.c. implanted 3T3-F442A preadipocytes. Proc Natl Acad Sci USA. 94:4300-4305.
35. Moo HS, Chung CS, Lee HG, Kim TG, Choi YJ and Cho CS. (2007) Inhibitory effect of (-)-epigallocatechin-3-gallate on lipid accumulation of 3T3-L1 cells. Obesity 15:2571-2582.
36. Munekata K and Sakamoto K. (2009) Forkhead transcription factors Foxo1 is essential for adipocyte differentiation. In Vitro Cell Dev Biol Anim 45:642-651.
37. Murase T, Misawa K, Haramizu S and Hase T. (2009) Catechin-induced activation of the LKB1/AMP-activated protein kinase pathway. Biochem Pharmacol 78:78-84.
38. Nakae J, Kitamura T, Kitamura Y, Biggs WH. 3rd, Arden KC and Accili D. (2003) The forkhead transcription factor Foxo1 regulates adipocyte differentiation.
39. Obsil T and Obsilova V. (2008) Structure/ function relationships underlying regulation of FOXO transcription factors. Oncogene. 27:2263-2275.
40. Oellerich MF and Potente M. (2012) FOXOs and sirtuins in vascular growth, maintenance, and aging. Cric Res 110:1238-1251.
41. Reznikoff CA, Brankow DW and Heideberger C. (1973) Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res 33:3231-3238.
42. Ricquier D, Casteilla L and Bouillaud F. (1991) Molecular studies of the uncoupling protein. FASEB J 5:2237-2242.
43. Sakurai N, Mochizuki K, Kameji H, Shimada M and Goda T. (2009) (-)-epigallocatechin gallate enhances the expression ofgenes related to insulin sensitivity and adipocyte differentiation in 3T3-L1 adipocytes at an early stage of differentiation. Nutrition 25:1047-1056.
44. Smith PJ, Wise LS, Berkowitz R, Wan C and Robin CS. (1998) Insulin-like growth factor-1 is a essential regulator of differentiation of 3T3-L1 adipocyte. J Biol Chem 263:9402-9408.
45. Student AK, Hsu RYa and Lane MD. (1980) Induction of fatty acid synthetase synthesisin differentiating 3T3-L1 preadipocytes. J Biol Chem. 255:4745-4750.
46. Sung HY, Hong CG, Suh YS, Cho HC, Park JH, Bae JH, Park WK, Han J and Song DK. (2010) Role of (-)-epigallocatechin-3-gallate in cell viability, lipogenesis, and retinol-binding protein 4 expression in adipocytes. Naunyn-Schmied Arch Pharmacol 382:303-310.
47. Tang QQ, Otto TC and Lane MD. (2004) Commitment of C3H10T1/2 pluripotent stem cells to the adipocyte lineage. Proc Natl Acad Sci USA. 101:9607-9611.
48. Tamura T, Nakatani K and Yau KW. (1991) Calcium feedback an sensitivity regulation in primate rods. J Gen Physiol 98:95-130.
49. Van der Heide LP, Jacobs FM, Burbach JP, Hoekman MF and Smidt MP. (2005) FoxO6 transcriptional activity is regulated by Thr26 and Ser184, independent of nucleo-cytoplasmic shuttling. Biochem J. 391:623-629.
50. Van der Hoest A and Burgering BM. (2007) Stressing the role of FoxO proteins in lifespan and disease. Nat Rev Mol Cell Biol 8:440-450.
51. Watanabe J and Niki R. (1998) Isolation and identification of acetyl-coA carboxylase inhibitors from green tea. Biosci Biophys Biochem 62:532-534.
52. Wei dinger CI, Krause K, Klagge A, Karger S and Fuhrer D. (2008) Forkhead box-O transcriptional factor: critical conductor of cancer’s fate. Endocri Relat Cancer 15:917-929.
53. Wolfram S, Raederstorff D, Wang Y, Teixeira SR, Elste V and Weber P. (2005) TEAVIGO (epigallocatechin gallate) supplementation prevents obesity in rodents by redents by reducing adipocyte tissue mass. Ann Nutr Metab 49:54-63.
54. Yu S, Matsusue K, Kashireddy p, Cao WQ, Yeldandi V, Yeldandi AV, Rao MS, Gonzalez FJ and Reddy jk. (2002) Adipocyte-specific gene expression and adipogenic Steatosis in the mouse liver due to peroxisome proliferator-activated receptor γ1 (PPARγ1) overexpression. J Biol Chem 278:498-505.
55. Zhang F, Chen Y, Heiman M and Dimarchi R. (2005) Leptin: structure, function and biology. Vitam Horm 71:345-372.
56. Zhang T, Yang D, Fan Y, Xie P and Li H. (2009) Epigallocatechin-3-gallate enhances ischemia/reperfusion-induced apoptosis in human umbilical vein endothelial cells via AKT and MAPK pathways. Apoptosis. 14:1245-1254.
57. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L and Friedman JM. (1994) Positional cloning of the mouse obese gene and its human homologue. Nature 372:425-432.
58. Zhu J, Mounzih K, Chehab EF, Mitro N, Saez E and Chehab FF. (2010) Effects of FoxO4 overexpression on cholesterol biosynthesis, triacylglycerol accumulation, and glucose uptake. J Lipid Res 51:1312-1324.
指導教授 高永旭(Yung-Hsi Kao) 審核日期 2017-1-25
推文 facebook   plurk   twitter   funp   google   live   udn   HD   myshare   reddit   netvibes   friend   youpush   delicious   baidu   
網路書籤 Google bookmarks   del.icio.us   hemidemi   myshare   

若有論文相關問題,請聯絡國立中央大學圖書館推廣服務組 TEL:(03)422-7151轉57407,或E-mail聯絡  - 隱私權政策聲明