臺灣博碩士論文加值系統

肺動脈高壓(pulmonary arterial hypertension ,PAH)是一種肺小動脈疾病，其病程會持續的進展且預後差，疾病的特徵為平均肺動脈壓以及肺動脈血管阻力不斷的上升，若不經過治療，會導致右心室肥大甚至心臟衰竭而死亡，是屬於一種高死亡率的疾病。研究指出，肺動脈高壓為不明原因的肺血管不正常增生以及重塑引起血管阻力上升而造成，因此降低肺小動脈過度增生重塑是一個有利方向。7,8-Dihydroxyflavone(7,8-DHF)是黃酮類化合物的親屬，在水果以及蔬菜中具有相當豐富的成份，過去文獻證實7,8-DHF在多種神經疾病具保護療效，也具抗氧化壓力、發炎作用，同時文獻指出7,8-DHF在血管舒張、減輕高血壓亦具療效。因此本研究探討在肺動脈高壓的動物模式中評估給予7,8-DHF治療是否有保護效果並探討相關機制。本實驗設計為在左肺葉切除手術後的第7天皮下注射野百合花鹼(60 mg/kg)，之後於第14天給予7,8-DHF (20,40 mg/kg)腹腔注射，每隔兩天投與一次，直至第35天犧牲。實驗結果顯示，7,8-DHF可以降低右心室壓力以及減輕右心室肥大，也減輕肺小動脈管壁增厚之情形，同時發現降低肺組織活性氧化物(super oxide)產量、iNOS、ET-1、ETAR表現量、β-catenin活化以及GSK3β、ERK1/2之磷酸化，並增加eNOS、HO-1、Nrf2。因此，推論7,8-DHF減輕肺動脈高壓動物模式，可能是藉由抗氧化作用減少自由基生成以及降低促細胞增生蛋白表現，同時7,8-DHF顯著增加HO-1表現和Nrf2活性，而達到改善改善肺動脈高壓。

Pulmonary arterial hypertension (PAH) is characterized by abnormal increase in pulmonary arteries (PAs) resistance and right ventricular (RV) hypertrophy. 7, 8-Dihydroxyflavone (7, 8-DHF) is a neuroprotective effect, and that has anti-inflammatory and antioxidant activities. However, whether 7,8-DHF improves the PAH remains unknown. Therefore, the aim of this study was to investigate whether 7, 8-DHF has beneficial effect on pneumonectomy and monocrotaline (P/M)-induced PAH in rats. The PAH model was established in rats injected with monocrotaline (60 mg/kg) at Day 7 after pneumonectomy. The PAH rats were treated intraperitoneally with vehicle or 7, 8- DHF (20 and 40 mg/kg/2 day, respectively) from Day 14 to Day 35 after pneumonectomy. We found that therapeutic administration with 7,8-DHF attenuated the development of PAH, as shown by lower values for RV systolic pressure and hypertrophy, and histopathological change of vascular remodeling of PAs including vascular wall thickening. The decreased endothelial nitric oxide synthase (eNOS), increased inducible NOS, endothelin-1(ET-1) level and ETA receptor (ETAR), and overproduction of superoxide observed in the lung of PAH rats, were marklly decreased by 7,8-DHF. PAH also induced glycogen synthase kinase-3β(GSK3β) phosphorylation in pulmonary artery smooth muscle cell and β-catenin, accompanied by ERK 1/2 phosphorylation of the lung, which was inhibited by 7,8-DHF. In addition 7,8-DHF increased H0-1(heme oxygenase-1,HO-1) and Nrf2 (nuclear factor erythroid-2-related factor, Nrf2)expression. In conclusion, we demonstrate for the first time that treatment with 7,8DHF exerts a beneficial effect in P/M induced PAH through inhibition GSK3β/β-catenin but enhancement of Nrf2/HO-1induction attenuated PAH

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目錄--------------------------------------------------------------------------------Ⅰ
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中文摘要--------------------------------------------------------------------------------V
英文摘要----------------------------------------------------------------------VII
英文縮寫對照表---------------------------------------------------------------VIII
第一章緒論----------------------------------------------------------------------1
第二章研究目的-----------------------------------------------------------------12
第三章材料與方法---------------------------------------------------------------13
第四章實驗結果-----------------------------------------------------------------27
第五章討論---------------------------------------------------------------------35
第六章結論---------------------------------------------------------------------42
圖次-------------------------------------------------------------------------------44
參考文獻-----------------------------------------------------------------------58

1.Developing new guidelines for the diagnosis and treatment of pulmonary hypertension.Eur Heart J, 2009. 30(20): p. 2416-8.
2.Schermuly, R.T., et al., Mechanisms of disease: pulmonary arterial hypertension.Nat Rev Cardiol, 2011. 8(8): p. 443-55.
3.Farber, H.W. and J. Loscalzo, Pulmonary arterial hypertension.N Engl J Med, 2004. 351(16): p. 1655-65.
4.Graham, B.B. and R.M. Tuder, [Pathogenesis of pulmonary hypertension].Zhonghua Yi Xue Za Zhi, 2009. 89(30): p.2091-3.
5.Budhiraja, R., R.M. Tuder, and P.M. Hassoun, Endothelial dysfunction in pulmonary hypertension.Circulation, 2004. 109(2): p. 159-65.
6.Giaid, A. and D. Saleh, Reduced expression of endothelial nitric oxide synthase in the lungs of patients with pulmonary hypertension.N Engl J Med, 1995. 333(4): p. 214-21.
7.Stasch, J.P., P. Pacher, and O.V. Evgenov, Soluble guanylate cyclase as an emerging therapeutic target in cardiopulmonary disease.Circulation, 2011. 123(20): p. 2263-73.
8.Barst, R.J., A review of pulmonary arterial hypertension: role of ambrisentan.Vasc Health Risk Manag, 2007. 3(1): p. 11-22.
9.Peng, X., et al., Inducible nitric oxide synthase contributes to ventilator-induced lung injury.Am J Respir Crit Care Med, 2005. 172(4): p. 470-9.
10.Howard, P.G., C. Plumpton, and A.P. Davenport, Anatomical localization and pharmacological activity of mature endothelins and their precursors in human vascular tissue.J Hypertens, 1992. 10(11): p. 1379-86.
11.Masuda, Y., et al., Two differentforms of endothelin receptors in rat lung.FEBS Lett, 1989. 257(2): p. 208-10.
12.Levin, E.R., Endothelins.N Engl J Med, 1995. 333(6): p. 356-63.
13.Konior, A., et al., NADPH oxidases in vascular pathology.Antioxid Redox Signal, 2014. 20(17): p. 2794-814.
14.Kamezaki, F., et al., Gene transfer of extracellular superoxide dismutase ameliorates pulmonary hypertension in rats.Am J Respir Crit Care Med, 2008. 177(2): p. 219-26.
15.Sanders, K.A. and J.R. Hoidal, The NOX on pulmonary hypertension.Circ Res, 2007. 101(3): p. 224-6.
16.Sklepkiewicz, P., et al., Glycogen synthase kinase 3beta contributes to 57

proliferation of arterial smooth muscle cells in pulmonary hypertension.PLoS One, 2011. 6(4): p. e18883.
17.Mao, S.Z., et al., Intermedin modulates hypoxic pulmonary vascular remodeling by inhibiting pulmonary artery smooth muscle cell proliferation.Pulm Pharmacol Ther, 2014. 27(1): p. 1-9.
18..
19.Constantin, M., et al., Therapeutic potential of heme oxygenase-1/carbon monoxide in lung disease.Int J Hypertens, 2012. 2012: p. 859235.
20.Morita, T., et al., Carbon monoxide controls the proliferation of hypoxic vascular smooth muscle cells.J Biol Chem, 1997. 272(52): p. 32804-9.
21.Pae, H.O. and H.T. Chung, Heme oxygenase-1: its therapeutic roles in inflammatory diseases.Immune Netw, 2009. 9(1): p. 12-9.
22.Dong, Q., et al., The Nrf2-ARE pathway is associated with Schisandrin b attenuating benzo(a)pyrene-Induced HTR cells damages in vitro.Environ Toxicol, 2015. 6(10): p. 22149.
23..
24.Visner, G.A., Rapamycin Induces Heme Oxygenase-1 in Human Pulmonary Vascular Cells: Implications in the Antiproliferative Response to Rapamycin.Circulation, 2003. 107(6): p. 911-916.
25.Hsu, H.H., et al., Simvastatin ameliorates established pulmonary hypertension through a heme oxygenase-1 dependent pathway in rats.Respir Res, 2009. 10: p. 32.
26.Okada, K., et al., Pulmonary hemodynamics modify the rat pulmonary artery response to injury. A neointimal model of pulmonary hypertension.Am J Pathol, 1997. 151(4): p. 1019-25.
27.Dorfmuller, P., et al., Increased oxidative stress and severe arterial remodeling induced by permanent high-flow challenge in experimental pulmonary hypertension.Respir Res, 2011. 12: p. 119.
28.Park, H.Y., et al., 7,8-Dihydroxyflavone exhibits anti-inflammatory properties by downregulating the NF-kappaB and MAPK signaling pathways in lipopolysaccharide-treated RAW264.7 cells.Int J Mol Med, 2012. 29(6): p. 1146-52.
29.Ryu, M.J., et al., Effect of 7, 8-dihydroxyflavone on the up-regulation of Nrf2-mediated heme oxygenase-1 expression in hamster lung fibroblasts.In Vitro Cell Dev Biol Anim, 2014. 50(6): p. 549-54.
30.Huai, R., et al., Vasorelaxing and antihypertensive effects of 7,8-dihydroxyflavone.Am J Hypertens, 2014. 27(5): p. 750-60.58

31.Galie, N., A. Manes, and A. Branzi, The endothelin system in pulmonary arterial hypertension.Cardiovasc Res, 2004. 61(2): p. 227-37.
32.Deng, H., et al., Pulmonary artery smooth muscle hypertrophy: roles of glycogen synthase kinase-3beta and p70 ribosomal S6 kinase.Am J Physiol Lung Cell Mol Physiol, 2010. 298(6): p. L793-803.
33.Atkins, R.J., et al., Regulation of glycogen synthase kinase-3 beta (GSK-3beta) by the Akt pathway in gliomas.J Clin Neurosci, 2012. 19(11): p. 1558-63.
34.Crosswhite, P. and Z. Sun, Nitric oxide, oxidative stress and inflammation in pulmonary arterial hypertension.J Hypertens, 2010. 28(2): p. 201-12.
35.Wedgwood, S. and S.M. Black, Endothelin-1 decreases endothelial NOS expression and activity through ETA receptor-mediated generation of hydrogen peroxide.Am J Physiol Lung Cell Mol Physiol, 2005. 288(3): p. L480-7.
36.Hampl, V., et al., Pulmonary vascular iNOS induction participates in the onset of chronic hypoxic pulmonary hypertension.Am J Physiol Lung Cell Mol Physiol, 2006. 290(1): p. L11-20.
37.Ryter, S.W., J. Alam, and A.M. Choi, Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications.Physiol Rev, 2006. 86(2): p. 583-650.
38.Yet, S.F., et al., Hypoxia induces severe right ventricular dilatation and infarction in heme oxygenase-1 null mice.J Clin Invest, 1999. 103(8): p. R23-9.
39.Minamino,T., et al., Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia.Proc Natl Acad Sci U S A, 2001. 98(15): p. 8798-803.
40.Goto, J., et al., Heme oxygenase-1 reduces murine monocrotaline-induced pulmonary inflammatory responses and resultant right ventricular overload.Antioxid Redox Signal, 2002. 4(4): p. 563-8.