請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32175
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳金洌(Jen-Leih Wu) | |
dc.contributor.author | Chi An Wang | en |
dc.contributor.author | 王啟安 | zh_TW |
dc.date.accessioned | 2021-06-13T03:35:11Z | - |
dc.date.available | 2009-07-31 | |
dc.date.copyright | 2006-07-31 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-07-27 | |
dc.identifier.citation | Ambrose A.M., DeEds F. and Rather L.J., Toxicity of thioacetamide in rats.( 1949) J. Indian Hygiene Toxicol. 31: 158–161
Aseervatham Anusha Amali (2006) Thioacetamide induced liver damage in zebrafish embryo as a disease model for steatohepatitis Journal of Biomedical Science 13: 225–232 Cruz A., Padillo F.J., Torres E., Navarrete C.M., Munoz-Castaneda J.R., Caballero F.J., Briceno J., Marchal T., Tunez I., Montilla P., Pera C. and Muntane J., (2005) Melatonin prevents experimental liver cirrhosis induced by thioacetamide in rats. J. Pineal Res. 39: 143–150 Day C.P.,(2002) Pathogensis of steatohepatitis. Best. Pract. Res. Clin. Gastroenterol. 16, 663–678. Day C.P. & James O.F. (1998) Steatohepatitis: a tale of two ‘hits’? Gastroenterology 114, 842–845. Desvergne B, Wahli W. (1999) Peroxisome proliferator-activated receptors: nuclear control of metabolism. Endocr. Rev ; 20: 649-688. Di-Poi, N., Tan, N. S., Michalik, L., Wahli, W. & Desvergne, B. (2002) Antiapoptotic role of PPARβ in keratinocytes via transcriptional control of the Akt1 signaling pathway. Mol. Cell 10, 721–733. Hevener, A.L., He, W., Barak, Y., Le, J., Bandyopadhyay, G., Olson, P., Wilkes, J., Evans, R.M., and Olefsky, J. (2003). Muscle-specific Pparg deletion causes insulin resistance. Nat. Med. 9, 1491–1497. He, W., Barak, Y., Hevener, A., Olson, P., Liao, D., Le, J., Nelson, M., Ong, E., Olefsky, J.M., and Evans, R.M. (2003). Adipose-specific peroxisome proliferator-activated receptor gamma knockout causes insulin resistance in fat and liver but not in muscle. Proc. Natl. Acad. Sci. USA 100, 15712–15717. Hu K.Q., Kyulo N.L., Esrailian E. et al. (2004) Overweight and obesity, hepatic steatosis, and progression of chronic hepatitis C: a retrospective study on a large cohort of patients in the United States. J. Hepatol. 40, 147–154. Hirshman, M.F., Rosen, E.D., Goodyear, L.J., Gonzalez, F.J., et al.(2003). Muscle-specific PPARgamma-deficient mice develop increased adiposity and insulin resistance but respond to thiazolidinediones. J. Clin. Invest. 112, 608–618. Kimihiko Matsusue, (2003)Liver-specific disruption of PPARγ in leptin-deficient mice improves fatty liver but aggravates diabetic phenotypes J. Clin. Invest. 111:737–747 doi:10.1172/JCI200317223. Kersten, S. & Wahli, W. (2000). Peroxisome proliferator activated receptor agonists. EXS 89, 141–151 Ludwig J., Viggiano T.R., Mcgill D.B., Oh B.J. (1980) Nonalcoholic steatohepatitis: Mayo Clinic experiences with a hitherto unnamed disease. Mayo Clin. Proc. 55, 434–438. Lee SST, Pineau T, Drago J, Lee EJ, Owens JW, Kroetz DL, et al. (1995) Targeted disruption of the α isoform of the peroxisome proliferator-activated receptor gene in mice results in abolishment of the pleiotropic effects of peroxisome proliferators. Mol Cell Biol;15:3012–3022. Memon R.A., Feingold K.R., Moser A.H., Fuller J., Grunfeld C. (1998) Regulation of fatty acid transport protein and fatty acid translocase mRNA levels by endotoxin and cytokines. Am. J. Physiol. 274, E210–E217. Matsusue, K., Haluzik, M., Lambert, G., Yim, S.H., Gavrilova, O., Ward, J.M., Brewer, B., Jr., Reitman, M.L., and Gonzalez, F.J. (2003). Liverspecific disruption of PPARgamma in leptin-deficient mice improves fatty liver but aggravates diabetic phenotypes. J. Clin. Invest. 111, 737–747. Neal R.A. and Halpert J., (1982) Toxicology of thionosulfer compounds. Annu. Rev. Pharmacol. Toxicol. 22: 321–339 Norris, A.W., Chen, L., Fisher, S.J., Szanto, I., Ristow, M., Jozsi, A.C., Powell E.E., Cooksley W.G., Hanson R., Searle J., Halliday J.W., Powell L.W. (1990) The natural history of nonalcoholic steatohepatitis: a follow-up study of forty-two patients for up to 21 years. Hepatology 11, 74–80. Peters JM, Cattley RC, Gonzalez FJ. 1997 Role of PPARα in the mechanism ofaction of the nongenotoxic carcinogen and peroxisome proliferator WY- 14,643. Carcinogenesis; 18: 2029 –2033. Ratziu V., Giral P., Charlotte F. et al. (2000) Liver fibrosis in overweight patients. Gastroenterology 118, 1117–1123. Reddy JK. (2004) Peroxisome proliferators and peroxisome proliferator-activated receptor _. Biotic and xenobiotic sensing. Am J Pathol;164:2305–2321. Sanyal A.J. (2002) AGA technical review on nonalcoholic fatty liver disease. Gastroenterology 123, 1705–1725. Sanyal A.J., Campbell-Sargent C., Mirshahi F. et al. (2001) Nonalcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities. Gastroenterology 120, 1183–1192. Shoelson S.E., Lee J., Yuan M. (2003) Inflammation and the IKK beta/I kappa B/NF-kappa B axis in obesity- and diet-induced insulin resistance. Int. J. Obes. Relat Metab Disord. 27 (Suppl.3), S49–S52. Tan, N. S. et al. (2002) Selective cooperation between fatty acid binding proteins and peroxisome proliferator-activated receptors in regulating transcription. Mol. Cell. Biol. 22, 5114–5127. Unger R.H. and Orci L., (2002) Lipoapoptosis: its mechanism and its diseases. Biochim. Biophy. Acta. 1585: 202–212 Unger RH. (2003)Lipid overload and overflow: metabolic trauma and the metabolic syndrome. Trends Endocrinol Metab; 14:398–403. Yoshimatsu M., Terasaki Y., Sakashita N. et al. (2004) Induction of macrophage scavenger receptor MARCO in nonalcoholic steatohepatitis indicates possible involvement of endotoxin in its pathogenic process. Int. J. Exp. Pathol. 85,335–343. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32175 | - |
dc.description.abstract | 中文摘要:
過氧化酶體增生活化受體γ(PPARγ)被認為是脂肪細胞的一個主要分化調節因子。 在最近的報告中發現,在瘦體素缺乏所導致脂肪肝老鼠模式中(Matsusue et al.,2003),肝臟專一性剔除過氧化酶體增生活化因子γ將改善老鼠脂肪肝的病灶。PPARγ2大量表現在成熟的脂肪細胞並且在脂肪肝動物模式中也有升高的現象。PPARγ調節脂肪生成和堆積在肝細胞 。本篇研究的目的在於肝臟專一性表現PPARγ是否有能力使肝臟脂肪化,以及在脂肪肝形成中扮演什麼樣的角色。為了了解PPARγ在正常及脂肪肝所扮演的角色,首先,選殖出斑馬魚的兩種長度PPARγcDNA分別為1533及1581鹼基對,接下來利用顯微注射的方式,使用肝臟專一性啟動子表現PPARγ在斑馬魚卵。反轉錄聚合鏈反應結果顯示轉殖PPARγ大量表現在肝臟,提高脂肪細胞相關基因,例如:脂肪細胞脂肪酸結合蛋白(aP2)、脂締素(adiponectin)以及脂肪生成相關基因,例如:固醇調控序列結合蛋白一型(SREBP-1c)。第二,利用硫乙胺誘發顯微注射PPARγ5天後之魚苗脂肪肝化。由我們實驗室發表的報告中,指出硫乙胺處理斑馬魚稚魚七天,將造成脂肪肝炎。反轉錄聚合鏈反應結果顯示轉殖PPARγ經TAA處理十天後,在第三有提高的現象,但是從硫乙胺處理控制組的結果顯示第9天時,脂肪生成相關基因才會上升,故PPARγ的表現加強了硫乙胺誘導脂肪肝的形成過程。 | zh_TW |
dc.description.abstract | Abstract:
Peroxisome proliferator-activated receptor gamma (PPARγ) is considered to be one of the master regulators of adipocyte differentiation. In recent report, liver-specific disruption of PPARgamma improves fatty liver in leptin-dificient mice(Matsusue et al.,2003). PPARγ2 is abundantly expressed in mature adipocytes and is elevated in the liver of animals that develop fatty liver. PPARγ regulates lipogenesis and lipid accumulation in steatotic hepatocyte. The aim of this study was to study the role of PPARγin pathogenesis of fatty liver. And to examine over-expression of PPARγ in liver by liver-specific L-FABP promoter and ubiquitous expression by EF-1αpromoter could induce lipogenesis and result in steatosis ? To understand roles of PPARγ in normal and fatty liver in zebrafish, I first cloned two cDNAs with length of 1533 bp and 1581 bp encoding PPARγ1 and PPARγ2, respectively. Liver-specific over-expression of PPARγ in zebrafish larva resulted in up-regulation of adipogenetic genes such as, adipocyte Fatty Acid Binding Protein (AFABP/AP2), adiponectin, and lipogenetic gene such as, SREBP-1. Secondly, I tried to promote hepatosteatosis in PPARγ-injected 5 day post fertilization (dpf) zebrafish larva by hepatotoxin thioacetamid (TAA) treatment. We have previously showed that TAA treatment for 7 days leads to liver steatohepatitis of zebrafish larva at 10 dpf. After injected with pLF2.8-zfPPARγ, zebrafish larva at 5 dpf were treated with TAA for 10 days. The results of RT-PCR showed that zebrafish treated with TAA had adipogenetic and lipogenetic genes up-regulated on the third and 5th day, After injected pLF2.8-zfPPARγ 5-day postfertilization, treaded TAA 10 days. The results of RT-PCR show that zebrafish treaded TAA 10th day made adipogenetic and lipogenetic up-regulated on the 3th and 5th day, because on TAA treated control Data show that lipogenetic gene would up-regulate at 9th day, so overexpression of PPARγ enhance TAA induced fatty liver. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T03:35:11Z (GMT). No. of bitstreams: 1 ntu-95-R93b45006-1.pdf: 4757451 bytes, checksum: 83c90d695dfdcbe25b04ab3395d7a41f (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | 目錄
中文摘要-----------------------------------------1 英文摘要-----------------------------------------3 一、 前言 1. 脂肪肝疾病簡介------------------------------------8 2. 非酒精性脂肪肝斑馬魚模式--------------------------9 3. 過氧化酶體增生活化受體之介紹----------------------10 二、 研究架構------------------------------------11 三、 材料 1. 生物材料------------------------------------------13 2. 引子---------------------------------------14 3. 儀器和器材---------------------------------19 4. 酵素---------------------------------------22 5. 反應試劑-----------------------------------20 6. 生物反應試劑組-----------------------------23 7. 反應溶液及緩衝溶液-------------------------23 8. 化學藥品-----------------------------------24 四、 實驗方法 (一) 膠體萃取--------------------------------26 (二) 銜接反應---------------------------------26 (三) 轉型作用---------------------------------27 (四) 斑馬魚核醣核酸 (RNA) 之抽取--------------27 (五) 斑馬魚的飼養與產卵------------------29 (六) 顯微注射----------------------------30 (七) 螢光顯微鏡的觀察與拍照--------------31 (八) 仔魚冷凍切片------------------------31 (九) 反轉錄聚合酶連鎖反應----------------32 五、實驗結果-------------------------------------35 六、結果討論-------------------------------------41 七、文獻參考-------------------------------------47 八、圖-----------------------------------------53 圖一、Racer 的引子設計 圖二、PPARγ的轉譯序列與其他物種的比較。 圖三、斑馬魚過氧化酶體增生活化受體γ(PPARγ)之DNA及胺基 酸序列 圖四 、在注射不同濃度下,對魚卵的畸形率、死亡率以及紅螢 光表現率的影響 圖五、 pLF2.8-PPARγ-V5/CMV-RFP 和 pT2KXIG-EF-1α-DrPPARγ 圖六、再受精後第五天觀察顯微注射LF2.8-PPARγ-250ng 之魚苗肝 臟發育情形 圖七、觀察顯微注射T2KXIG-PPARγ在受精後第五天之魚苗肝臟發育 情形 圖八、利用共軛焦顯微鏡觀察受精後第九天,肝臟內部細胞型態及綠 螢光表現情形 圖九、利用共軛交顯微鏡觀察並利用軟體統計出兩種細胞大小差異以 及綠螢光表現量差異 圖十、在受精後第九天,利用共軛交顯微鏡觀察並用軟體計算硫乙 胺處理以及合併PPARγ表現之綠螢光的差別 圖十一、利用冷凍切片觀察硫乙胺處理組與硫乙胺合併肝臟專一性表 現PPARγ在處理七天、九天,其肝臟內部型態的差異。 圖十二、利用冷凍切片觀察硫乙胺處理組與硫乙胺合併肝臟專一性表 現PPARγ在處理三天、五天,其肝臟內部型態的差異。 圖十三、利用H&E 染色觀察處理硫乙胺組和PPARγ合併硫乙胺處理 組其肝細胞的型態 圖十四、全身性與肝臟專一性表現在斑馬魚,利用RT-PCR分析脂肪 生成相關基因的變化 圖十五、全身性與肝臟專一性表現在斑馬魚,利用RT-PCR分析肝臟 富含轉錄因子的變化 圖十六、全身性與肝臟專一性表現在斑馬魚,利用RT-PCR分析細胞 凋亡相關基因的變化 圖十七、利用real-time RT-PCR 定出LF2.8-PPARγ表現造成下游基 因表現量的差別 圖十八、肝臟專一性表現合併硫乙胺之處理,利用RT-PCR分析其脂 肪生成相關基因在處理後3、5、7、9天的變化 圖十九、肝臟專一性表現PPARγ合併處理硫乙胺,利用RT-PCR觀察 肝臟富含轉錄因子在處理後3、5、7、9天的基因表現變化 圖二十、肝臟專一性表現PPARγ合併處理硫乙胺,利用RT-PCR分 析分析細胞凋亡相關基因在處理後3、5、7、9天的表現變 化 | |
dc.language.iso | zh-TW | |
dc.title | 以斑馬魚為動物模式探討過氧化酶體活化受體γ因硫乙胺誘發脂肪肝所扮演的角色 | zh_TW |
dc.title | The role of peroxisome proliferater activated receptor γ (PPARγ) in thioacetamide-induced hepato-steatosis of zebrafish(Danio rerio) | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 吳造中(Chau-Chung Wu),李士傑(Shyh-Jye Lee) | |
dc.subject.keyword | 斑馬魚,肝臟, | zh_TW |
dc.subject.keyword | zebrafish,liver, | en |
dc.relation.page | 75 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-07-27 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
顯示於系所單位: | 漁業科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-95-1.pdf 目前未授權公開取用 | 4.65 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。