Please use this identifier to cite or link to this item:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64034
Full metadata record
???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
---|---|---|
dc.contributor.advisor | 丁詩同 | |
dc.contributor.author | Jui-Ting Yang | en |
dc.contributor.author | 楊瑞婷 | zh_TW |
dc.date.accessioned | 2021-06-16T17:27:20Z | - |
dc.date.available | 2014-12-31 | |
dc.date.copyright | 2012-08-28 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-15 | |
dc.identifier.citation | Chapter 6 References
Anderson, D. B. and R. G. Kauffman. 1972. Cellular and enzymatic changes in porcine adipose tissue during growth. J. Lipid Res. 14: 160-168. Arellano-Garcia, M. E., R. Li, X. Liu, Y. Xie, X. Yan, J. A. Loo and S. Hu. 2010. Identification of tetranectin as a potential biomarker for metastatic oral cancer. Int. J. Mol. Sci. 11: 3106-3121. Arvanitis, D. L., E. F. Kamper, L. Kopeikina, A. Stavridou, M. N. Sgantzos, V. Kallioras, E. Athanasiou and P. Kanavaros. 2002. Tetranectin expression in gastric adenocarcinomas. Histol. Histopathol. 17: 471-475. Banga, A., R. Unal, P. Tripathi, I. Pokrovskaya, R. J. Owens, P. A. Kern and G. Ranganathan. 2009. Adiponectin translation is increased by the PPARgamma agonists pioglitazone and omega-3 fatty acids. Am. J. Physiol. Endocrinol. Metab. 296: 480-489. Begg, D. P. and S. C. Woods. 2012. The central insulin system and energy balance. Handb. Exp. Pharmacol. 209: 111-29. Begum, F. D., E. Høgdall, S. K. Kjaer, J. Blaakaer, I. J. Christensen, L. Christensen and C. Høgdall. 2009. Preoperative serum tetranectin, CA125 and menopausal status used as single markers in screening and in a risk assessment inDEX (RAI) in discriminating between benign and malignant ovarian tumors. Gynecol. Oncol. 113: 221-227. Berglund, L. and T. E. Petersen. 1992. The gone structure of tetranectin, a plasminogen binding protein. FASEB 309: 15-19. Bevan, D., E. Gherardi, T. P. Fan, D. Edwards and R. Warn. 2004. Diverse and potent activities of HGF/SF in skin wound repair. J. Pathol. 203: 831-838. Brohi, K., M. J. Cohen and R. A.Davenport. 2007. Acute coagulopathy of trauma: mechanism, identification and effect. Curr. Opin. Crit. Care 13: 680-685. Brunner, A., C. Ensinger, M. Christiansen, S. Heiss, I. Verdorfer, G. Mikuz and A. Tzankov. 2007. Expression and prognostic significance of Tetranectin in invasive and non-invasive bladder cancer. Virchows. Arch. 450: 659-664. Carey, N. L. and A. R. Saltiel. 2011. Inflammatory links between obesity and metabolic disease. Clin. Invest. 121: 2111-2117. Carlos, P., J. T. Huff and K. R. Yamamoto. 2008. Glucocorticoid signaling defines a novel commitment state during adipogenesis in vitro. Mol. Biol. Cell. 19: 4032-4041. Chaves, V. E., D. Frasson and N. H. Kawashita. 2011. Several agents and pathways regulate lipolysis in adipocytes. Biochimie 93: 1631-1640. Chen, Y. J., C. C. Chen, T. K. Li, P. H. Wang, L. R. Liu, F. Y. Chang, Y. C. Wang, Y. H. Yu, S. P. Lin, H. J. Mersmann and S. T. Ding. 2012. Docosahexaenoic acid suppresses the expression of FoxO and its target genes. J. Nutr. Biochem. [Epub ahead of print] Clemmensen, I., L. C. Petersen and C. Kluft. 1986. Purification and characterization of a novel, oligomeric, plasminogen kringle 4 binding protein from human plasma: tetranectin. Eur. J. Biochem. 156: 327-333. Cristancho, A. G. and M. A. Lazar. 2011. Forming functional fat: a growing understanding of adipocyte differentiation. Nat. Rev. Mol. Cell Biol. 12: 722-734. De Taeye, B., L. H. Smith and D. E. Vaughan. 2005. Plasminogen activator inhibitor-1: a common denominator in obesity, diabetes and cardiovascular disease. Curr. Opin. Pharmacol. 5: 149-154. Dominiczak, M. H. and M. J.Caslake. 2011. Apolipoproteins: metabolic role and clinical biochemistry applications. Ann. Clin. Biochem. 48: 498-515. Fernández-Alvarez, A., M. S. Alvarez, R. Gonzalez, C. Cucarella, J. Muntané and M. Casado. 2011. Human SREBP1c expression in liver is directly regulated by peroxisome proliferator-activated receptor alpha (PPARalpha). J. Biol. Chem. 286: 21466-21477. Fernández-Puente, P., J. Mateos, C. Fernández-Costa, N. Oreiro, C. Fernández-López, C. Ruiz-Romero and F. J. Blanco. 2011. Identification of a panel of novel serum osteoarthritis biomarkers. J. Proteome Res. 10: 5095-5101. Gaillard, D., R. Negrel, M. Lagarde and G. Ailhaud. 1989. Requirement and role of arachidonic acid in the differentiation of pre-adipose cells. Biochem. J. 257: 389-397. Gani, O. A. 2008. Are fish oil omega-3 long-chain fatty acids and their derivatives peroxisome proliferator-activated receptor agonists? Cardiovasc. Diabetol. 7: 6. Gathercole, L. L., S. A. Morgan, I. J. Bujalska, D. Hauton, P. M. Stewart and J. W. Tomlinson. 2011. Regulation of lipogenesis by glucocorticoids and insulin in human adipose tissue. Plos One 6: e26223. Gebbink, M. F. 2011. Tissue-type plasminogen activator-mediated plasminogen activation and contact activation, implications in and beyond haemostasis. J. Thromb. Haemost. 1: 174-181. Ghim, C. M., S. K. Lee, S. Takayama and R. J. Mitchell. 2010. The art of reporter proteins in science: past, present and future applications. BMB Rep. 43: 451-460. Hansen, C., A. Fu, C. Li, W. T. Dixon, R. Christopherson and S. S. Moore. 2004. Global gene expression patterns spanning 3T3-L1 preadipocyte differentiation. Can. J. Anim. Sci. 84: 367-376. Hirsch, J. and J. L. Knittle. 1970. Cellularity of obese and nonobese human adipose tissue. Federation Proc. 29: 1516-1521. Høgdall, C. K., I. J. Christensen, R. W. Stephens, S. Sørensen, B. Nørgaard-Pedersen and H. J. Nielsen. 2002. Serum tetranectin is an independent prognostic marker in colorectal cancer and weakly correlated with plasma suPAR, plasma PAI-1 and serum CEA. APMIS 110: 630-638. Holtet, T. L., J. H. Graversen, I. Clemmensen, H. C. Thøgersen and M. Etzerodt. 1997. Tetranectin, a trimeric plasminogen-binding C-type lectin. Protein Sci. 6: 1511-1515. Hotamisligil, G. S. 2006. Inflammation and metabolic disorders. Nature 444: 860-867. Howell, G. 3rd., X. Deng, C. Yellaturu, E. A. Park, H. G. Wilcox, R. Raghow and M. B. Elam. 2009. N-3 polyunsaturated fatty acids suppress insulin-induced SREBP-1c transcription via reduced trans-activating capacity of LXRalpha. Biochim. Biophys. Acta. 1791: 1190-1196. Iba, K., H. Chiba, T. Yamashita, S. Ishii and N. Sawada. 2001a. Phase-independent inhibition by retinoic acid of mineralization correlated with loss of tetranectin expression in a human osteoblastic cell line. Cell Struct. Funct. 26: 227-233. Iba, K., M. E. Durkin, L. Johnsen, E. Hunziker, K. Damgaard-Pedersen, H. Zhang, E. Engvall, R. Albrechtsen, U. M. Wewer. 2001b. Mice with a targeted deletion of the tetranectin gene exhibit a spinal deformity. Mol. Cell Biol. 21: 7817-7825. Iba, K., N. Hatakeyama, T. Kojima, M. Murata, T. Matsumura, U. M. Wewer, T. Wada, N. Sawada and T. Yamashita. 2009. Impaired cutaneous wound healing in mice lacking tetranectin. Wound Repair Regen. 17: 108-112. Iba, K., N. Sawada, H. Chiba, U. M. Wewer, S. Ishii and M. Mori. 1995. Transforming growth factor-beta 1 downregulates Dexamethasone-induced tetranectin gene expression during the in vitro mineralization of the human osteoblastic cell line SV-HFO. FEBS Lett. 373: 1-4. Jensen, B. A. and I. Clemmensen. 1988. Plasma tetranectin is reduced in cancer and related to metastasia. Cancer. 62: 869-872. Jiang, D., H. W. Jarrett and W. E. Haskins. 2009. Methods for proteomic analysis of transcription factors. J. Chromatogr. A. 1216: 6881-6889. Kaplan, Z. S. and S. P. Jackson. 2011. The role of platelets in atherothrombosis. Hematol. Am. Soc. Hematol. Educ. Program. 2011: 51-61. Kluft, C., A. F. Jie, P. Los, E. de Wit, and L. Havekes. 1989a. Functional analogy between lipoprotein(a) and plasminogen in the binding to the kringle 4 binding protein, tetranectin. Biochem. Biophys. Res. Commun. 161: 427-433. Kluft, C., P. Los and I. Clemmensen. 1989b. Calcium-dependent binding of tetranectin to fibrin. Thromb Res. 55: 233–238. Kim, H. J., M. Takahashi and O. Ezaki. 1999. Fish oil feeding decreases mature sterol regulatory element-binding protein 1 (SREBP-1) by downregulation of SREBP-1c mRNA in mouse liver. A possible mechanism for downregulation of lipogenic enzyme mRNAs. J. Biol. Chem. 274: 25892-25898. Kim, P. G. Suh, S. H. Ryu and T. G. Lee. 2010. Comparative analysis of the secretory proteome of human adipose stromal vascular fraction cells during adipogenesis. Proteomics. 10: 394-405. Kunau, W. H., V. Dommes and H. Schulz. 1995. Beta-oxidation of fatty acids in mitochondria, peroxisomes, and bacteria: a century of continued progress. Prog. Lipid Res. 34: 267-342. Lay, S. L., I. Lefrere, C. Trautwein, I. Dugail and S. Krief. 2002. Insulin and sterol-regulatory element-binding protein-1c (SREBP-1C) regulation of gene expression in 3T3-L1 adipocytes. Identification of CCAAT/enhancer-binding protein beta as an SREBP-1c target. J. Biol. Chem. 277: 35625-35634. Liu, B. H., P. H. Wang, Y. C. Wang, W. M. Cheng, H. J. Mersmann and S. T. Ding. 2008. Fasting regulates the expression of adiponectin receptors in young growing pigs. J. Anim. Sci. 86: 3377-3384. Madsen, L., L. M. Pedersen, B. Liaset, T. Ma, R. K. Petersen, S. van den Berg, J. Pan, K. Müller-Decker, E. D. Dülsner, R. Kleemann, T. Kooistra, S. O. Døskeland and K. Kristiansen. 2008. cAMP-dependent signaling regulates the adipogenic effect of n-6 polyunsaturated fatty acids. J. Biol. Chem. 283: 7196-7205. McGarry, J. D. 2002. Banting lecture 2001: dysregulation of fatty acid metabolism in the etiology of type 2 diabetes. Diabetes 51: 7-18. Miller, C. W., D. A. Casimir and J. M. Ntambi. 1996. The mechanism of inhibition of 3T3-L1 preadipocyte differentiation by prostaglandin F2alpha. Endocrinology 137: 5641-5650. Mogues, T., M. Etzerodt, C. Hall, G. Engelich, J. H. Graversen and K. L. Hartshorn. 2004. Tetranectin binds to the kringle 1-4 form of angiostatin and modifies its functional activity. J. Biomed. Biotechnol. 2004: 73-78. Morrison, R. F. and S. R. Farmer. 2000. Hormonal signaling and transcriptional control of adipocyte differentiation. J. Nutr. 130: 3116-3121. Neame, P. J., C. N. Young and J. T. Treep. 1992. Primary structure of a protein isolated from reef shark (Carcharhinus springeri) cartilage that is similar to the mammalian C-type lectin homolog, tetranectin. Protein Sci. 1: 161-168. Nielbo, S., J. K. Thomsen, J. H. Graversen, P. H. Jensen, M. Etzerodt, F. M. Poulsen and H. C.Thøgersen. 2004. Structure of the plasminogen kringle 4 binding calcium-free form of the C-type lectin-like domain of tetranectin. Biochemistry. 43: 8636-8643. Nielsen, B. B., J. S. Kastrup, H. Rasmussen, T. L. Holtet, J. H. Graversen, M. Etzerodt, H. C. Thøgersen and I. K. Larsen. 1997. Crystal structure of tetranectin, a trimeric plasminogen-binding protein with an alpha-helical coiled coil. FEBS Lett. 412: 388-396. Nielsen, H. 2011. The transcriptional landscape. Methods Mol. Biol. 703: 3-14. Obrist, P., G. Spizzo, C. Ensinger, D. Fong, T. Brunhuber, G. Schäfer, M. Varga, R. Margreiter, A. Amberger, G. Gastl and M. Christiansen. 2004. Aberrant tetranectin expression in human breast carcinomas as a predictor of survival. J. Clin. Pathol. 57: 417-421. Petersen, K. F., D. Laurent, D. L. Rothman, G. W. Cline and G. I. Shulman. 1998. Mechanism by which glucose and insulin inhibit net hepatic glycogenolysis in humans. J. Clin. Invest. 101: 1203-1209. Pfaffl, M. W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29: e45. Prasad, H., D. A. Ryan, M. F. Celzo and D. Stapleton. 2012. Metabolic syndrome: definition and therapeutic implications. Postgrad. Med. 124: 21-30. Rossetti, L. and A. Giaccari. 1990. Relative contribution of glycogen synthesis and glycolysis to insulin-mediated glucose uptake. A dose-response euglycemic clamp study in normal and diabetic rats. J. Clin. Invest. 85: 1785-1792. Samad, F., L. Badeanlou, C. Shah and G. Yang. 2011. Adipose tissue and ceramide biosynthesis in the pathogenesis of obesity. Adv. Exp. Med. Biol. 721: 67-86. Shimano, H. 2001. Sterol regulatory element-binding proteins (SREBPs): transcriptional regulators of lipid synthetic genes. Prog. Lipid Res. 40: 439-452. Smas, C. M., L. Chen, L. Zhao, M. J. Latasa and H. S. Sul. 1999. Transcriptional repression of pref-1 by glucocorticoids promotes 3T3-L1 adipocyte differentiation. J. Biol. Chem. Apr. 274: 12632-12641. Sørensen, B., T. Mariann, L. Ole H, P. N. Laursen and C. Fenger-Eriksen and C. J. Rea. 2011. The role of fibrinogen: a new paradigm in the treatment of coagulopathic bleeding. Thromb. Res. 1: 13-16. Tai, C. C., C. Y. Chen, H. S. Lee, Y. C. Wang, T. K. Li, H. J. Mersamm, S. T. Ding and P. H. Wang. 2009. Docosahexaenoic acid enhances hepatic serum amyloid A expression via protein kinase A-dependent mechanism. J. Biol. Chem. 284: 32239-32247. Tang, Q. Q. and M. D. Lane. 1999. Activation and centromeric localization of CCAAT/enhancer-binding proteins during the mitotic clonal expansion of adipocyte differentiation. Genes Dev. 13: 2231-2241. Valdes, A. M., D. J. Hart, K. A. Jones, G. Surdulescu, P. Swarbrick, D. V. Doyle, A. J. Schafer and T. D. Spector. 2004. Association study of candidate genes for the prevalence and progression of knee osteoarthritis. Arthritis. Rheum. 50: 2497-2507. Vannini, A. and P. Cramer. 2012. Conservation between the RNA polymerase I, II, and III transcription initiation machineries. Mol. Cell. 45: 439-46. Vedel, V. and I. Scotti. Promoting the promoter. 2011. Plant. Sci. 180: 182-189. Westergaard, U. B., M. H. Andersen, C. W. Heegaard, S. N. Fedosov and T. E. Petersen. 2003. Tetranectin binds hepatocyte growth factor and tissue-type plasminogen activator. Eur. J. Biochem. 270: 1850-1854. Wewer, U. M., K. Iba, M. E. Durkin, F. C. Nielsen, F. Loechel, B. J. Gilpin, W. Kuang, E. Engvall and R. Albrechtsen. 1998. Tetranectin is a novel marker for myogenesis during embryonic development, muscle regeneration, and muscle cell differentiation in vitro. Dev. Biol. 200: 247-259. Wewer, U. M., K. Iba, P. Schj¢rring, M. E. Durkin, M. E. Young and R. Albrechtsen. 1994. A potential role for tetranectin in mineralization during osteogenesis. J. Cell Biol. 127: 1767-1775. Xu, X., B. Gilpin, K. Iba, A. Maier, E. Engvall, R. Albrechtsen and U. M. Wewer. 2001. Tetranectin in slow intra- and extrafusal chicken muscle fibers. J. Muscle Res. Cell Motil. 22: 121-132. Yang, X., P. A. Jansson, I. Nagaev, M. M. Jack, E. Carvalho, K. S. Sunnerhagen, M. C. Cam, S. W. Cushman and U. Smith. 2004. Evidence of impaired adipogenesis in insulin resistance. Biochem. Biophys. Res. Commun. 317: 1045-1051. Yki-Jarvinen, H., E. Helve, T. Sane, N. Nurjhan and M. R. Taskinen. 1989. Insulin inhibition of overnight glucose production and gluconeogenesis from lactate in NIDDM. Am. J. Physiol. 256: 732-739. Yoshikawa, T., H. Shimano, N. Yahagi, T. Ide, M. Amemiya-Kudo, T. Matsuzaka, M. Nakakuki, S. Tomita, H. Okazaki, Y. Tamura, Y. Iizuka, K. Ohashi, A. Takahashi, H. Sone, Ji. J. Osuga, T. Gotoda, S. Ishibashi and N. Yamada. 2002. Polyunsaturated fatty acids suppress sterol regulatory element-binding protein 1c promoter activity by inhibition of liver X receptor (LXR) binding to LXR response elements. J. Biol. Chem. 277: 1705-1711. Zinder, O. and B. Shapiro. 1971. Effect of cell size on epinephrine- and ACTH-induced fatty acid release from isolated fat cells. J. Lipid Res. 12: 91-95. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64034 | - |
dc.description.abstract | 結合四素 (tetranectin) 會參與溶血作用 (fibrinolysis),並與骨母細胞分化(osteoblast differentiation) 有關。其可藉由與組織型纖維蛋白溶酶原激活物 (tissue-type plasminogen activator) 結合,將無活性的纖溶酶原(plasminogen) 轉變成具活性的纖溶酶 (plasmin) 以裂解纖維蛋白(fibrin) (Clemmensen et al., 1986)。然而,近來本研究室與他人分別做出新奇發現而指出結合四素竟為人類初代脂肪細胞所分泌之脂泌素 (adipokine),並與脂肪細胞生成 (adipogenesis) 有關 (丁博士研究室未發表資料)。
為瞭解結合四素在脂肪代謝上所扮演的角色及其基因調控機制,本試驗成功選殖豬隻結合四素的部分互補DNA序列 (463 bp),以其設計引子並執行即時量化聚合脢連鎖反應(Real-time quantitative PCR) 以偵測先前收集之三十頭去勢保育豬組織樣本。三十頭保育豬隨機分配至三種飼糧處理組,每組十頭豬分別餵飼30天之2%大豆油 (soybean oil)、二十二碳六烯酸油 (DHA oil) 與牛油 (beef tallow) 之結合四素基因表現量。結果發現二十二碳六烯酸油飼糧處理降低結合四素在豬隻脂肪組織中之基因表現量 (P < 0.05),在肝臟與肺臟中雖不具統計顯著性,但亦具相似之趨勢 (P = 0.08)。 再者,為確認二十二碳六烯酸油可降低結合四素的表現量,本試驗進一步分離豬隻初代脂肪細胞並分析100 μΜ二十二碳六烯酸及棕櫚油酸 (palmitic acid) 對結合四素表現量之影響。同樣地,結果顯示二十二碳六烯酸確實可降低其基因表現量,這些研究結果指出二十二碳六烯酸似可藉由轉錄調控的方式影響結合四素之基因表現量。 最後,為瞭解二十二碳六烯酸對結合四素轉錄調控的機制,本試驗成功選殖長度為1956 bp之豬隻結合四素假定啟動子 (putative promoter),並藉由網路線上軟體TFsitescan預測此啟動子之轉錄因子調結結合位 (regulatory element) 並設計逐列截除啟動子片段所需之引子。並將此啟動子片段轉殖 (subclone) 至冷光酶報導載體 (luciferase reporter vector) 並轉染 (transfect) 至 HEK293T細胞以分析二十二碳六烯酸對結合四素啟動子轉錄活性之影響。結果顯示結合四素啟動子上有兩個區段會受二十二碳六烯酸所抑制,分別是 -1956 至 -1576 與 -559 至 -310處。同時網路線上軟體也預測另一膽固醇調節因子結合位 (Sterol regulatory element, SRE) 落於 -825 至 -559區間。若進一步將此膽固醇調節因子結合位突變則可削弱二十二碳六烯酸之抑制作用,此結果表示二十二碳六烯酸可藉由此調節因子抑制結合四素之基因表現。 綜合上述,本試驗研究結果顯示結合四素在脂肪細胞分化中扮演相關角色;而具抗肥胖與代謝症候群之二十二碳六烯酸則可藉由結合四素啟動子上之膽固醇調節因子結合位,抑制其基因表現量,但連結二十二碳六烯酸至膽固醇調節因子間之詳細訊息傳導機制則有待未來進一步之探討。 | zh_TW |
dc.description.abstract | While tetranectin (TN) is a plasminogen-binding protein originally found to be involved in fibrinolysis and bone formation, it was recently identified by our group and others as an adipokine in human adipocyte culture media and implicated in adipogenesis and lipid metabolism.
To elucidate its role and regulation in lipid metabolism, we firstly cloned the partial cDNA of porcine TN and used the sequence information to analyze the previously collected tissue samples of weaned piglets fed with 2% soybean oil, DHA oil or beef tallow (as-fed basis) by real-time qPCR and Western blotting analysis. Our results showed that the expression of TN was reduced in the adipose (P < 0.05), liver (P = 0.08) and lung (P = 0.08), from DHA-fed group (with lowered serum levels of triacylglycerol and cholesterol) compared to the beef tallow group, which was confirmed in primary porcine adipocyte culture treated with DHA. These results indicated that TN is involved in lipid metabolism and negatively regulated by DHA. To further investigate the transcriptional mechanism underlying DHA-suppressed TN expression, we cloned a 1956-bp putative porcine TN promoter by using the NCBI genomic sequences and “TFsitescan” software, transfected 293T cells with the luciferase gene constructs of serially-deleted TN promoter fragments and tested its transactivating activities by luciferase assay. Interestingly, two DHA-sensitive fragments from nucleotide position -1956 to -1576 and -559 to -310 were identified. In addition, a sterol regulatory element (SRE) was predicted within -825 to -559 by an on-line website program. Mutation on this SRE reduced its transactivating activity and rendered it resistant to DHA’s inhibitory effect, indicating its potential role in mediating DHA’s action. In conclusion, our current study suggests that TN plays a role in adipogenesis and lipid metabolism, and, for the first time, identified an SRE in TN’s promoter region, which is responsible for the negative effects of DHA on TN expression. The upstream events relaying DHA to SRE-mediated TN gene expression await future investigation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T17:27:20Z (GMT). No. of bitstreams: 1 ntu-101-R99626022-1.pdf: 2042999 bytes, checksum: e11210965de7cc77a1440b17f3369d7f (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | Contents
Page Acknowledgements…………………………………………………..…...I 摘要……………………………………...………………………......II Abstract..………………………………………………………………....IV Contents………………………………………………………………….VI List of Figures………………………………………………………..VIII List of Tables…………………………………………………………….X Chapter 1 Literature review……………………………………………….1 1.1 Introduction……………………………………………………….1 1.2 Lipid metabolism and metabolic syndrome………………….…...1 1.3 Adipocyte differentiation and its regulation: 3T3-L1 as a study model for adipogenesis………………...……………..…………..6 1.4 The role of docosahexaenoic acid (DHA) in lipolysis…………9 1.5 Tetranectin: identification and biological functions………...…..13 1.6 Transcriptional regulation of gene expression: transcription initiation and promoters……………………………………..…..21 1.7 Techniques for the study of transcriptional regulation of gene expression……………………………….……………………....23 Chapter 2 The aim of this study………………………………….………26 Chapter 3 Materials and methods...…………………………...…………27 3.1 Animal tussues………………….…………………..…..……….27 3.2 Isolation of porcine stromal vascular cells…………..…....……27 3.3 Cell culture and differentiation of porcine adipocytes...……...…28 3.4 Cloning of porcine TN cDNA………………………………...…29 3.5 Real-time qPCR analysis……………………………………..…30 3.6 Western blotting………..……………………………………..…31 3.7 Lung cancer cells and culture media…..…………………..….…33 3.8 Hepatoma cell, embryonic kidney cell and culture media.……...33 3.9 Mouse preadipocyte and adipocyte culture media…...……….…34 3.10 DHA, insulin, dexamethasone and isobutylmethylxanthine treatments………………………………………………………..35 3.11 Cloning of putative porcine TN promoters to pGL3-basic vector…………………………………………………………..36 3.12 Subcloning of porcine TN promoter to phrGFP vector…..…37 3.13 Transient transfection and dual luciferase assay……..….....37 3.14 Statistical analysis………………..………………………..…38 Chapter 4 Results………………………………………………….….44 Chapter 5 Discussions……………………………………………...…….64 Chapter 6 References..…………………..……………………………….70 List of Figures Page Figure 1. Propsed scheme of how increased adipose tissue mass and enlarge adipocytes contribute to metabolic and cardiovascular risk in obesity……..…………………………………………….5 Figure 2. Two main type of promoters in human genome………...……..22 Figure 3. The map of pJET1.2 vector..………..…………………………42 Figure 4. The map of cloning serially deleted TN promoter into pGL3 vector…………...……..……………...………………………..42 Figure 5. Diagram of normal and mutated SRE site-containing porcine TN promoter luciferase vectors………………………...………….43 Figure 6. The map of phrGFP vector…………………………..………...43 Figure 7. Sequence alignment of TN genes……………………………..44 Figure 8. Effects of fat supplementation on the expression of TN mRNA in various tissues of weaned piglets…………………………...…..46 Figure 9. Effects of fat supplementation on the protein levels of TN in adipose tissues of weaned piglets…………………………..…..47 Figure 10. Effects of free fatty acids on TN expression in porcine adipocytes……………………………………..………………..48 Figure 11. Screening of cell lines for the study of DHA-regulated TN gene expression………………………………………..……………..51 Figure 12. TN gene expression during adipogenesis in 3T3-L1….....…...53 Figure 13. Effects of insulin on TN gene expression………………...…..54 Figure 14. Effects of DEX (dexamethasone) and MIX (isobutylmethylxanthine) on TN gene expression in HEK293T cell………………………………………………………………55 Figure 15. Sequence alignment of the putative porcineTN promoter by clustalW program………………...…………………………..57 Figure 16. The transactivating activity of porcine TN promoter drived GFP expression in HEK293T cells……………………..…………61 Figure 17. Identification of DHA-responsive elements on TN promoter in HEK293T cells…………………...…………………………..62 Figure 18. Effects of sterol-regulatory element (SRE) mutation on DHA responsiveness of TN promoter…………………..……….…63 Figure 19. Propsed scheme of the DHA inhibitory effect on TN gene expression.………………...…………………………………68 List of Tables Page Table 1. Primers used for porcine TN cDNA cloning………...……….....39 Table 2. Gene-specific primers used for real-time qPCR…..…………..39 Table 3. Primers used for TN promoter cloning...…..…………………...40 Table 4. Primers used for cloning of TN promoter into phrGFP vector..41 | |
dc.language.iso | zh-TW | |
dc.title | 二十二碳六烯酸透過轉錄調控抑制結合四素的基因表達 | zh_TW |
dc.title | DHA suppresses the expression of tetranectin through transcriptional regulation | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 王佩華 | |
dc.contributor.oralexamcommittee | 陳洵一,陳珠亮 | |
dc.subject.keyword | 結合四素,二十二碳六烯酸,膽固醇調節因子結合位,膽固醇調節因子結合蛋白,脂肪分化, | zh_TW |
dc.subject.keyword | Tetranectin,DHA,Sterol regulatory element,Sterol regulatory element- binding protein,Adipogenesis, | en |
dc.relation.page | 77 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-08-16 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
Appears in Collections: | 動物科學技術學系 |
Files in This Item:
File | Size | Format | |
---|---|---|---|
ntu-101-1.pdf Restricted Access | 2 MB | Adobe PDF |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.