臺灣博碩士論文加值系統

海洋深層水意指海平面以下200公尺的海水，台灣具有獨特的地理位置來採用海洋深層水，因為在台灣東岸50公里處，海水的深度就已達4公里。許多生技公司宣稱，海洋深層水(deep sea water , DSW)可以保護心臟及促進免疫能力。然而，查詢相關的文獻，發現甚少有文章探討，海洋深層水保護我們的心血管系統的機轉。而本研究的目的即為了進一步評估不同處理的海洋深層水對高膽固醇引起的心肌凋亡的影響及機轉。本實驗採用24隻七週齡ICR雄性小鼠以隨機分成四組，所有組別的小鼠皆餵以1%高膽固醇飲食，再分別餵以蒸餾水(normal distilled water , NDW)、逆滲透處理之海洋深層水(Reverse osmosis deep sea water , RO-DSW)、電透析處理之海洋深層水(Electrodialysis Reversal deep sea water, ER-DSW)、10%稀釋之海洋深層水(10% DSW+90% ddH2O , 10%DSW)，為期共八周。緊接著將左心室游離，以組織染色切片，西方墨點法，及TUNEL分析。結果顯示,各種處理之海洋深層水均具有降低1%高膽固醇引起之心肌肥大，BNP上升及TNF-alpha,同時抑制了death-receptor pathway(Fas Ligan，Fas，FADD，caspase 8，及t-bid)蛋白，並降低bak、bax(促凋亡蛋白)，壓制活化態caspase 9及caspase 3的表現。更進一步促進了survival路徑相關蛋白，如IGF1 receptor，pAkt及Bcl-2(抑凋亡蛋白)的表現。結果顯示，RO-DSW比ER-DSW及10% DSW組的抑制凋亡及促survival效果差。以上實驗提供了飲用海洋深層水的利多，並比較出不同處理方法對心肌保護的差異。所以我們可藉由飲用ER及10%稀釋之海洋深層水，來作為預防或是治療改善高膽固醇飲食造成之心臟異常的發生。

Deep sea water refers to water 200m under the sea surface. Taiwan have a special geographical location for using deep sea water, because it is 50km off Taiwan''s east coast, the seabed drops to a depth of 4km.Many companies in Taiwan said that deep sea water can protect our heart and improve the immune system. However, we have searched the related papers, there is still few study to investigate the mechanisms of the deep sea water protect our cardiovascular system.The purpose of this study was to evaluate the effects of differently pretreated DSW on suppressing the high cholesterol-induced cardiac apoptosis. Twenty-four ICR mice at 7 weeks of age were randomly divided into four groups, all groups of mice were fed 1% high cholesterol, and treated as pure water (control),reverse osmosis DSW (RO-DSW), and electrodialysis reversal DSW(ER-DSW),and 10% diluted DSW groups for 8 weeks. The excised left ventricle from mice were measured by histological analysis, western blotting , and TUNEL assays. High cholesterol-induced cardiac abnormalities including hypertrophy,abnormal myocardial architecture, more cardiac TUNEL-positive apoptotic cells and slightly fibrosis. All treatments decreased high cholesterol-induced BNP hypertrophic marker, tumor necrosis factor death receptors,Fas-dependent apoptotic proteins,caspase 8 , t-bid , Bak , Bax , activated caspase 9 and activated caspase 3, but increased IGF1 receptor, pAkt,and anti-apoptotic protein(Bcl-2). However , RO-DSW group showed less anti-apoptotic effects than ER-DSW and 10% DSW groups.Therefore,ER-DSW and 10% DSW highly activated the cardiac IGF-1 survival pathway and suppressed high cholesterol-induced cardiac death receptor-dependent and mitochondria-dependent apoptotic pathway activities.The findings may provide a possible approach by drinking for potentially treating deep sea water to prevent cardiac abnormalities.

中文摘要----------------II
英文摘要----------------IV
誌謝--------------------VI
第一章 前言---------------1
第一節 研究背景-----------1
第二節 研究目的--------16
第二章 研究方法--------17
第一節 研究材料--------17
第二節 研究設計--------21
第三章 研究結果--------27
第四章 討論------------33
第一節 結果討論--------33
第二節 其他相關性討論--36
第五章 結論------------39
參考文獻---------------41
實驗圖表---------------48
附錄圖表---------------63

1.Katsuda, S., et al., Deep-sea water improves cardiovascular hemodynamics in Kurosawa and Kusanagi-Hypercholesterolemic (KHC) rabbits. Biol Pharm Bull, 2008. 31(1):p.38-44.
2.Ruilope, L.M. and R.E. Schmieder, Left ventricular hypertrophy and clinical outcomes in hypertensive patients. Am J Hypertens, 2008. 21(5): p.500-8.
3.Kubota, T., et al., Overexpression of tumor necrosis factor- alpha activates both anti- and pro-apoptotic pathways in the myocardium. J Mol Cell Cardiol, 2001. 33(7): p. 1331-44.
4.Brummer, E., et al., Production of IL-6, in contrast to other cytokines and chemokines, in macrophage innate immune responses: effect of serum and fungal (Blastomyces) challenge. Cytokine, 2007. 39(3): p.163-70.
5.Unal, S., et al., Interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) levels and IL-6, TNF-polymorphisms in children with thrombosis. J Pediatr Hematol Oncol, 2008. 30(1): p.26-31.
6.Wakefield, T.W., et al., P-selectin and TNF inhibition reduce venous thrombosis inflammation. J Surg Res, 1996. 64(1): p.26-31.
7.Yang, J. and G.R. Stark, Roles of unphosphorylated STATs in signaling. Cell Res, 2008. 18(4): p. 443-51.
8.Lu, Y., et al., JAK/STAT and PI3K/AKT pathways form a mutual transactivation loop and afford resistance to oxidative stress-induced apoptosis in cardiomyocytes. Cell Physiol Biochem, 2008. 21(4): p.305-14.
9.Stephanou, A. and D.S. Latchman, STAT-1: a novel regulator of apoptosis. Int J Exp Pathol, 2003. 84(6): p. 239-44.
10.Fischer, P. and D. Hilfiker-Kleiner, Survival pathways in hypertrophy and heart failure: the gp130-STAT axis. Basic Res Cardiol, 2007. 102(5): p. 393-411.
11.Nobori, K. and Y. Munehisa, [Intracellular signaling pathways for cardiac hypertrophy: ERK, JAK-STAT, S6 kinase]. Nippon Rinsho, 2007. 65 Suppl 4: p. 196-200.
12.Brown, M.D. and D.B. Sacks, Compartmentalised MAPK Pathways, in Handb Exp Pharmacol. 2008. p. 205-235.
13.Dai, X., et al., The NF-{kappa}B, p38 MAPK and STAT1 pathways differentially regulate the dsRNA-mediated innate immune responses of epidermal keratinocytes. Int Immunol, 2008.
14.Deng, J., et al., XGPR3 is a Constitutively Active Cell Surface G Protein-Coupled Receptor that Participates in Maintaining Meiotic Arrest in Xenopus Laevis Oocytes. Mol Endocrinol, 2008.
15.Schenning, M., et al., The anti-apoptotic activity associated with phosphatidylinositol transfer protein alpha activates the MAPK and Akt/PKB pathway. Biochim Biophys Acta, 2008.
16.Regan, C.P., et al., Erk5 null mice display multiple extraembryonic vascular and embryonic cardiovascular defects. Proc Natl Acad Sci U S A, 2002. 99(14): p. 9248-53.
17.Wojcikiewicz, R.J., Regulated ubiquitination of proteins in GPCR-initiated signaling pathways. Trends Pharmacol Sci, 2004. 25(1): p. 35-41.
18.Nicol, R.L., et al., Activated MEK5 induces serial assembly of sarcomeres and eccentric cardiac hypertrophy. Embo J, 2001. 20(11): p. 2757-67.
19.De, S., et al., IL-1 beta and IL-6 induce hyperplasia and hypertrophy of cultured guinea pig airway smooth muscle cells. J Appl Physiol, 1995. 78(4): p. 1555-63.
20.Haugen, E., et al., Parallel gene expressions of IL-6 and BNP during cardiac hypertrophy complicated with diastolic dysfunction in spontaneously hypertensive rats. Int J Cardiol, 2007. 115(1): p. 24-8.
21.Yu, C., et al., Opposing effects of proteasomes and lysosomes on LIFR: modulation by TNF. J Mol Neurosci, 2007. 32(1): p. 80-9.
22.Frantz, S., et al., Role of p38 mitogen-activated protein kinase in cardiac remodelling. Br J Pharmacol, 2007. 150(2): p. 130-5.
23.Kankaanranta, H., et al., SB 203580, an inhibitor of p38 mitogen-activated protein kinase, enhances constitutive apoptosis of cytokine-deprived human eosinophils. J Pharmacol Exp Ther, 1999. 290(2): p. 621-8.
24.Obata, K. and K. Noguchi, [Contribution of the primary sensory neurons and spinal glial cells to the pathomechanisms of neuropathic pain]. Brain Nerve, 2008. 60(5): p.483-92.
25.Dhingra, S., et al., p38 and ERK1/2 MAPKs mediate the interplay of TNF-alpha and IL-10 in regulating oxidative stress and cardiac myocyte apoptosis. Am J Physiol Heart Circ Physiol, 2007. 293(6): p. H3524-31.
26.Wang, Y., et al., Cardiac muscle cell hypertrophy and apoptosis induced by distinct members of the p38 mitogen-activated protein kinase family. J Biol Chem, 1998. 273(4): p. 2161-8.
27.Zechner, D., et al., A role for the p38 mitogen-activated protein kinase pathway in myocardial cell growth, sarcomeric organization, and cardiac-specific gene expression. J Cell Biol, 1997. 139(1): p. 115-27.
28.Ndebele, K., et al., Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induced mitochondrial pathway to apoptosis and caspase activation is potentiated by phospholipid scramblase-3. Apoptosis, 2008.
29.Araki, S., et al., Apoptosis of vascular endothelial cells by fibroblast growth factor deprivation. Biochem Biophys Res Commun, 1990. 168(3): p. 1194-200.
30.Scheid, M.P., K.M. Schubert, and V. Duronio, Regulation of bad phosphorylation and association with Bcl-x(L) by the MAPK/Erk kinase. J Biol Chem, 1999. 274(43): p. 31108-13.
31.Pistilli, E.E., J.R. Jackson, and S.E. Alway, Death receptor-associated pro-apoptotic signaling in aged skeletal muscle. Apoptosis, 2006. 11(12): p. 2115-26.
32.Chen, P.Y., et al., Possible involvement of MAP kinase pathways in acquired metal-tolerance induced by heat in plants. Planta, 2008.
33.Milner, A.E., et al., Induction of apoptosis by chemotherapeutic drugs: the role of FADD in activation of caspase-8 and synergy with death receptor ligands in ovarian carcinoma cells. Cell Death Differ, 2002. 9(3): p. 287-300.
34.Desagher, S. and J.C. Martinou, Mitochondria as the central control point of apoptosis. Trends Cell Biol, 2000. 10(9): p. 369-77.
35.Ploner, C., et al., The BCL2 rheostat in glucocorticoid-induced apoptosis of acute lymphoblastic leukemia. Leukemia, 2008. 22(2): p. 370-7.
36.Nakamura, M. and Y. Tanigawa, Characterization of ubiquitin-like polypeptide acceptor protein, a novel pro-apoptotic member of the Bcl2 family. Eur J Biochem, 2003. 270(20): p.4052-8.
37.Gal, A., et al., Bcl-2 or Bcl-XL gene therapy reduces apoptosis and increases plasticity protein GAP-43 in PC12 cells. Brain Res Bull, 2008. 76(4): p. 349-53.
38.Karki, P., et al., Intracellular K(+) inhibits apoptosis by suppressing the Apaf-1 apoptosome formation and subsequent downstream pathways but not cytochrome c release. Cell Death Differ, 2007. 14(12): p. 2068-75.
39.Levine, B., S. Sinha, and G. Kroemer, Bcl-2 family members: dual regulators of apoptosis and autophagy. Autophagy, 2008. 4(5).
40.Ekert, P.G., et al., Apaf-1 and caspase-9 accelerate apoptosis, but do not determine whether factor-deprived or drug-treated cells die. J Cell Biol, 2004. 165(6): p.835-42.
41.Li, Z., et al., Down-regulation of 14-3-3zeta suppresses anchorage-independent growth of lung cancer cells through anoikis activation. Proc Natl Acad Sci U S A, 2008. 105(1): p.162-7.
42.Racz, B., et al., PKA-Bad-14-3-3 and Akt-Bad-14-3-3 signaling pathways are involved in the protective effects of PACAP against ischemia/reperfusion-induced cardiomyocyte apoptosis. Regul Pept, 2008. 145(1-3): p. 105-15.
43.Adam, A.L., G. Kohut, and L. Hornok, Fphog1, a HOG-type MAP kinase gene, is involved in multistress response in Fusarium proliferatum. J Basic Microbiol, 2008. 48(3): p. 151-9.
44.Won, C.K., et al., Estradiol prevents the injury-induced decrease of Akt activation and Bad phosphorylation. Neurosci Lett, 2005. 387(2): p. 115-9.
45.Chen, D.B., L. Wang, and P.H. Wang, Insulin-like growth factor I retards apoptotic signaling induced by ethanol in cardiomyocytes. Life Sci, 2000. 67(14): p. 1683-93.
46.Velloso, C.P., Regulation of muscle mass by growth hormone and IGF-I. Br J Pharmacol, 2008. 154(3): p. 557-68.
47.Fernandez-Cancio, M., et al., IGF-I and not IGF-II expression is regulated by glucocorticoids in human fetal epiphyseal chondrocytes. Growth Horm IGF Res, 2008.
48.Li, Z., T.X. Gu, and Y.H. Zhang, Hepatocyte growth factor combined with insulin like growth factor-1 improves expression of GATA-4 in mesenchymal stem cells cocultured with cardiomyocytes. Chin Med J (Engl), 2008. 121(4): p. 336-40.
49.Thornburg, K.L., S. Louey, and G.D. Giraud, The role of growth in heart development. Nestle Nutr Workshop Ser Pediatr Program, 2008. 61: p. 39-51.
50.Isgaard, J. and C.H. Bergh, Clinical potential of growth hormone in the treatment of congestive heart failure. BioDrugs, 1999. 12(4): p. 245-50.
51.Yamamura, T., et al., IGF-I differentially regulates Bcl-xL and Bax and confers myocardial protection in the rat heart. Am J Physiol Heart Circ Physiol, 2001. 280(3): p. H1191-200.
52.Samarel, A.M., IGF-1 Overexpression rescues the failing heart. Circ Res, 2002. 90(6): p. 631-3.
53.Li, Q., et al., Insulin-like growth factor I deficiency prolongs survival and antagonizes paraquat-induced cardiomyocyte dysfunction: role of oxidative stress. Rejuvenation Res, 2007. 10(4): p. 501-12.
54.Suleiman, M.S., R.J. Singh, and C.E. Stewart, Apoptosis and the cardiac action of insulin-like growth factor I. Pharmacol Ther, 2007. 114(3): p. 278-94.
55.Aikawa, R., et al., Insulin prevents cardiomyocytes from oxidative stress-induced apoptosis through activation of PI3 kinase/Akt. Circulation, 2000. 102(23): p. 2873-9.
56.Tavridou, A., et al., Antiatherosclerotic Properties of EP2302, a Novel Squalene Synthase Inhibitor, in the Cholesterol-fed Rabbit. J Cardiovasc Pharmacol, 2008.
57.Yang, J., et al., Effect of niacin on adipocyte leptin in hypercholesterolemic rabbits. Cardiovasc Pathol, 2008.
58.Prasad, K., Regression of hypercholesterolemic atherosclerosis in rabbits by secoisolariciresinol diglucoside isolated from flaxseed. Atherosclerosis, 2008. 197(1): p. 34-42.
59.Ehara, S., et al., Small coronary calcium deposits and elevated plasma levels of oxidized low density lipoprotein are characteristic of acute myocardial infarction. J Atheroscler Thromb, 2008. 15(2): p. 75-81.
60.Mohty, D., et al., Association between plasma LDL particle size, valvular accumulation of oxidized LDL, and inflammation in patients with aortic stenosis. Arterioscler Thromb Vasc Biol, 2008. 28(1): p. 187-93.
61.Guan, S. and B. Wang, Effects of fosinopril and valsartan on expressions of ICAM-1 and NO in human umbilical vein endothelial cells. Chin Med J (Engl), 2003. 116(6): p. 923-7.
62.Napoli, C., Low density lipoprotein oxidation and atherogenesis: from experimental models to clinical studies. G Ital Cardiol, 1997. 27(12): p. 1302-14.
63.Zhao, S.P. and D.Y. Xu, Oxidized lipoprotein(a) enhanced the expression of P-selectin in cultured human umbilical vein endothelial cells. Thromb Res, 2000. 100(6): p. 501-10.
64.Wang, J.S., et al., Exercise paradoxically modulates oxidized low density lipoprotein-induced adhesion molecules expression and trans-endothelial migration of monocyte in men. Thromb Haemost, 2005. 94(4): p. 846-52.
65.Clinton, S.K., et al., Macrophage colony-stimulating factor gene expression in vascular cells and in experimental and human atherosclerosis. Am J Pathol, 1992. 140(2): p. 301-16.
66.dos Santos, M.G., et al., Risk factors for the development of atherosclerosis in childhood and adolescence. Arq Bras Cardiol, 2008. 90(4): p. 276-283.
67.Gudmundsdottir, I.J., et al., Role of the endothelium in the vascular effects of the thrombin receptor (protease-activated receptor type 1) in humans. J Am Coll Cardiol, 2008. 51(18): p. 1749-56.
68.Getz, G.S. and C.A. Reardon, Diet and murine atherosclerosis. Arterioscler Thromb Vasc Biol, 2006. 26(2): p. 242-9.
69.Benner, J., et al., Ozonation of reverse osmosis concentrate: Kinetics and efficiency of beta blocker oxidation. Water Res, 2008.
70.Pronk, W., M. Biebow, and M. Boller, Electrodialysis for recovering salts from a urine solution containing micropollutants. Environ Sci Technol, 2006. 40(7): p.2414-20.
71.Pantos, C., et al., Long-term thyroid hormone administration reshapes left ventricular chamber and improves cardiac function after myocardial infarction in rats. Basic Res Cardiol, 2008. 103(4): p. 308-18.
72.Xu, Z., et al., Pravastatin attenuates left ventricular remodeling and diastolic dysfunction in angiotensin II-induced hypertensive mice. J Cardiovasc Pharmacol, 2008. 51(1): p.62-70.
73.Kapoun, A.M., et al., B-type natriuretic peptide exerts broad functional opposition to transforming growth factor-beta in primary human cardiac fibroblasts: fibrosis, myofibroblast conversion, proliferation, and inflammation. Circ Res, 2004. 94(4): p. 453-61.
74.Matsuda, F., et al., cFLIP Regulates Death Receptor-mediated Apoptosis in an Ovarian Granulosa Cell Line by Inhibiting Procaspase-8 Cleavage. J Reprod Dev, 2008.
75.Chen, X.L., et al., Involvement of PI3K/AKT/GSK3beta pathway in tetrandrine-induced G1 arrest and apoptosis. Cancer Biol Ther, 2008. 7(7).
76.Durlach, J., M. Bara, and A. Guiet-Bara, Magnesium level in drinking water and cardiovascular risk factor: a hypothesis. Magnesium, 1985. 4(1): p. 5-15.
77.Hewitt, D. and L.C. Neri, Development of the ''Water Story'': some recent Canadian studies. J Environ Pathol Toxicol, 1980. 4(2-3): p. 51-63.