Sun, Minghui. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 125-159). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Wu Min. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 87-114). / Abstracts in English and Chinese. / Declaration --- p.i / Acknowledgement --- p.ii / Publication list --- p.iii / Abstract (English) --- p.xi / Abstract (Chinese) --- p.xiv / Abbreviations --- p.xvi / List of figures / tables --- p.xviii / Chapter Chapter 1. --- General Introduction --- p.1 / Chapter 1.1 --- Endothelial function in the regulation of vascular tone --- p.1 / Chapter 1.1.1 --- NO --- p.2 / Chapter 1.1.2 --- PGI2 --- p.5 / Chapter 1.1.3 --- EDHF --- p.6 / Chapter 1.2 --- Alteration of endothelial functions after preservation with cardioplegia /organ preservation solutions in the coronary and pulmonary microcirculations --- p.18 / Chapter 1.2.1 --- Cardioplegia/organ preservation solutions --- p.21 / Chapter 1.2.2 --- Effect of Cardioplegia/organ preservation solutions on endothelial function --- p.22 / Chapter 1.2.2.1 --- Effect of K+ on endothelial function --- p.23 / Chapter 1.2.2.2 --- Effect of other components on endothelial function --- p.24 / Chapter Chapter 2. --- Materials and Methods --- p.26 / Chapter 2.1 --- Isometric force study in coronary/pulmonary resistance arteries --- p.26 / Chapter 2.1.1 --- Preparation of vessels --- p.26 / Chapter 2.1.1.1 --- Preparation of porcine coronary small arteries --- p.26 / Chapter 2.1.1.2 --- Preparation of porcine pulmonary small arteries --- p.26 / Chapter 2.1.2 --- Technique of setting up --- p.29 / Chapter 2.1.2.1 --- Mounting of small vessels --- p.29 / Chapter 2.1.2.2 --- Normalization procedure for small vessels --- p.29 / Chapter 2.1.3 --- EDHF-mediated vasorelaxation --- p.30 / Chapter 2.1.3.1 --- Precontraction and stimuli of EDHF --- p.30 / Chapter 2.1.3.2 --- """True"" response of EDHF" --- p.31 / Chapter 2.1.4 --- Data acquisition and analysis --- p.32 / Chapter 2.2 --- Electrophysiological study --- p.32 / Chapter 2.2.1 --- Preparation of small porcine coronary/pulmonary arteries --- p.32 / Chapter 2.2.2 --- Preparation of microelectrode --- p.32 / Chapter 2.2.3 --- Impaling of microelectrode --- p.33 / Chapter 2.2.4 --- Recording of membrane potential --- p.33 / Chapter 2.3 --- Statistical analysis --- p.34 / Chapter 2.4 --- Chemicals --- p.34 / Chapter Chapter 3. --- Effects of Celsior Solution on Endothelial Function in Resistance Coronary Arteries Compared to St. Thomas' Hospital Solution --- p.37 / Chapter 3.1 --- Abstract --- p.37 / Chapter 3.2 --- Introduction --- p.38 / Chapter 3.3 --- Experimental design and analysis --- p.40 / Chapter 3.3.1 --- Vessel preparation --- p.40 / Chapter 3.3.2 --- Normalization --- p.40 / Chapter 3:3.3 --- "Relaxation study: BK-induced, EDHF-mediated relaxation" --- p.41 / Chapter 3.3.4 --- Cellular electrophysiological study: EDHF-mediated cellular hyperpolarization and associated relaxation --- p.41 / Chapter 3.3.5 --- Data analysis --- p.42 / Chapter 3.4 --- Results --- p.43 / Chapter 3.4.1 --- Relaxation study --- p.43 / Chapter 3.4.1.1 --- Resting force --- p.43 / Chapter 3.4.1.2 --- U46619-induced precontraction --- p.43 / Chapter 3.4.1.3 --- EDHF-mediated relaxation --- p.43 / Chapter 3.4.2 --- Electrophysiological studies --- p.44 / Chapter 3.4.2.1 --- Resting membrane potential --- p.44 / Chapter 3.4.2.2 --- EDHF-mediated cellular hyperpolarization --- p.45 / Chapter 3.4.2.3 --- Cellular hyperpolarization-associated relaxation --- p.45 / Chapter 3.5 --- Discussion --- p.46 / Chapter 3.5.1 --- Effects of Celsior solution on endothelial function --- p.47 / Chapter 3.5.2 --- Effects of ST solution on EDHF-mediated function --- p.48 / Chapter 3.5.3 --- Comparison between Celsior and ST solutions on EDHF-mediated function --- p.48 / Chapter 3.5.4 --- Clinical implications --- p.49 / Chapter Chapter 4. --- Effects of Perfadex and Celsior Solution on Endothelial Function in Resistance Pulmonary Arteries --- p.57 / Chapter 4.1 --- Abstract --- p.57 / Chapter 4.2 --- Introduction --- p.58 / Chapter 4.3 --- Experimental design and analysis --- p.59 / Chapter 4.3.1 --- Vessel Preparation --- p.59 / Chapter 4.3.2 --- Normalization --- p.60 / Chapter 4.3.3 --- Isometric force study --- p.60 / Chapter 4.3.4 --- Electrophysiological studies --- p.61 / Chapter 4.3.5 --- Data analysis --- p.61 / Chapter 4.4 --- Results --- p.62 / Chapter 4.4.1 --- Relaxation study: EDHF-mediated relaxation --- p.62 / Chapter 4.4.1.1 --- Resting force --- p.62 / Chapter 4.4.1.2 --- U46619-induced precontraction --- p.62 / Chapter 4.4.1.3 --- EDHF-mediated relaxation --- p.62 / Chapter 4.4.2 --- Electrophysiological studies --- p.63 / Chapter 4.4.2.1 --- Resting membrane potential --- p.63 / Chapter 4.4.2.2 --- EDHF-mediated cellular hyperpolarization --- p.64 / Chapter 4.4.2.3 --- Cellular hyperpolarization-associated relaxation --- p.64 / Chapter 4.5 --- Discussion --- p.65 / Chapter 4.5.1 --- Effects of Celsior solution on endothelial function during cardiopulmonary surgery --- p.65 / Chapter 4.5.2 --- Effects of Perfadex solution on EDHF-mediated endothelial function --- p.66 / Chapter 4.5.3 --- Comparison between Celsior and Perfadex solutions on EDHF-mediated function --- p.66 / Chapter 4.5.4 --- Clinical implications --- p.67 / Chapter Chapter 5. --- Exploration of the Nature of EDHF - the Effect of H2O2 on the Membrane Potential in the Rat Small Mesenteric Arteries --- p.73 / Chapter Chapter 6. --- General Discussion --- p.75 / Chapter 6.1 --- EDHF-mediated endothelial function in porcine coronary and pulmonary circulation --- p.75 / Chapter 6.1.1 --- Role of EDHF in the regulation of porcine coronary arterial tone --- p.75 / Chapter 6.1.2 --- Role of EDHF in the regulation of porcine pulmonary arterial tone --- p.76 / Chapter 6.2 --- Alteration of EDHF-mediated endothelial functions after exposure to organ preservation solutions --- p.77 / Chapter 6.2.1 --- Effects of hyperkalemic solution on EDHF-mediated endothelial function in coronary and pulmonary circulation --- p.78 / Chapter 6.2.2 --- Effects of low-potassium-based preservation solution on EDHF-mediated endothelial function in pulmonary circulation --- p.79 / Chapter 6.2.3 --- Comparison between hyperkalemic solution and low-potassium-based preservation solution on EDHF-mediated endothelial function --- p.80 / Chapter 6.2.4 --- Effects of other component of organ preservation solutions on EDHF-mediated endothelial function --- p.81 / Chapter 6.3 --- Clinical implications --- p.82 / Chapter 6.4 --- The effect of H202 on the membrane potential in rat small mesenteric arteries --- p.83 / Chapter 6.5 --- Limitation of the study --- p.84 / Chapter 6.6 --- Future investigations --- p.85 / Chapter 6.7 --- Conclusions --- p.85 / References --- p.87

Este estudo avaliou a função vascular em anéis de aorta e no leito vascular mesentérico (LVM) de camundongos com deleção dos receptores <font face=\"symbol\">a2A e <font face=\"symbol\">a2Cadrenérgicos (KO) com 3, 5 e 7 meses, os quais apresentam uma hiperatividade simpática acompanhada de cardiomiopatia. Os KO apresentaram um aumento da freqüência cardíaca em todos os grupos avaliados, e hipertrofia ventricular esquerda aos 5 e 7 meses. Na aorta, o relaxamento dependente (acetilcolina) e independente (nitroprussiato de sódio) do endotélio e da via font face=\"symbol\">a-adrenérgica (isoproterenol), assim como a contração (fenilefrina e serotonina) e a mobilização de Ca2+ não foram alterados nos KO aos 3, 5 e 7 meses. Nos KO aos 3 meses, o relaxamento mediado pelos receptores ?2-adrenérgicos (clonidina) foi reduzido. Tanto a contração (noradrenalina) como o relaxamento (acetilcolina) no LVM dos KO aos 7 meses não foi alterado. Assim, sugere-se que os vasos arteriais parecem ser menos sensíveis do que o coração aos efeitos crônicos da hiperatividade simpática nos camundongos com deleção dos receptores <font face=\"symbol\">a2A e <font face=\"symbol\">a2C adrenérgicos. / This study assed the vascular function in aortic rings and in mesenteric vascular bed (MVB) from mice with disruption of <font face=\"symbol\">a2A and <font face=\"symbol\">a2Cadrenoceptors (KO) with 3, 5 and 7 months of age, that present sympathetic hyperactivity associated with cardiomyopathy. Heart rate was increased in all KO groups, and left ventricular hypertrophy was observed only in 5 and 7 month-old KO. There are no changes in the relaxation induced by acetylcholine (ACh), sodium nitroprusside and isoproterenol in aortic rings from all groups. In addition, the contraction induced by phenylephrine and serotonin, and Ca2+ handling did not change. However, in aorta from 3 month-old KO the relaxation induced by clonidine (<font face=\"symbol\">a2-adrenergic agonist) was reduced. In MVB from 7 month-old KO, neither the contraction (noradrenaline) nor relaxation (ACh) was modified. The results suggest that arterial vessel has been more resistant than heart to chronic effects induced by sympathetic hyperactivity observed in mice with disruption o<font face=\"symbol\">a2A and <font face=\"symbol\">a2C-adrenoceptors.

Dong Yingying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 99-125). / Abstracts in English and Chinese. / Declaration --- p.i / Acknowledgement --- p.ii / Publication list --- p.iii / Abstract (English) --- p.ix / Abstract (Chinese) --- p.xii / Abbreviations --- p.xiv / List of figures / tables --- p.xvi / Chapter Chapter 1. --- General Introduction / Chapter 1.1 --- The role of endothelium in regulating vascular tone --- p.1 / Chapter 1.1.1 --- Nitric oxide (NO) --- p.2 / Chapter 1.1.2 --- Endothelium-derived hyperpolarizing factor (EDHF) --- p.7 / Chapter 1.1.3 --- Prostacyclin (PGI2) --- p.20 / Chapter 1.2 --- EDHF-mediated endothelial function in coronary circulation --- p.22 / Chapter 1.2.1 --- Role of EDHF in coronary microarteries --- p.23 / Chapter 1.2.2 --- Role of EDHF in cardiac veins --- p.24 / Chapter 1.3 --- Effect of ischemia-reperfusion on endothelial function in coronary circulation --- p.25 / Chapter 1.3.1 --- Ischemia-reperfusion injury --- p.26 / Chapter 1.3.2 --- Effect of ischemia-reperfusion on endothelial function in coronary microarteries --- p.28 / Chapter 1.3.3 --- Effect of ischemia-reperfusion on endothelial function in cardiac veins --- p.29 / Chapter 1.4 --- Alteration of endothelial function during cardiac surgery / Chapter 1.4.1 --- Cardioplegia and organ preservation solutions --- p.31 / Chapter 1.4.2 --- Combined effects of hypoxia-reoxygenation and ST solution on endothelial function in coronary microarteries/cardiac veins --- p.34 / Chapter 1.4.3 --- Effect of nicorandil on endothelial function --- p.34 / Chapter Chapter 2. --- Materials and Methods --- p.37 / Chapter 2.1 --- Isometric force study in micro arteries/veins --- p.37 / Chapter 2.1.1 --- Preparation of vessels --- p.37 / Chapter 2.1.1.1 --- Preparation of porcine coronary microarteries --- p.37 / Chapter 2.1.1.2 --- Preparation of porcine cardiac veins --- p.37 / Chapter 2.1.2 --- Technique of setting up --- p.39 / Chapter 2.1.2.1 --- Mounting of microvessels --- p.39 / Chapter 2.1.2.2 --- Normalization procedure for microvessels --- p.39 / Chapter 2.1.3 --- EDHF-mediated vasorelaxation --- p.40 / Chapter 2.1.3.1 --- Precontraction and stimuli of EDHF --- p.40 / Chapter 2.1.3.2. --- “Truéحresponse of EDHF --- p.40 / Chapter 2.1.4 --- Data acquisition and analysis --- p.41 / Chapter 2.2 --- Hypoxia and reoxygenation --- p.41 / Chapter 2.2.1 --- Calibration of 02-special electrode --- p.41 / Chapter 2.2.2 --- Measurement of --- p.02 / Chapter 2.3 --- Statistical analysis --- p.42 / Chapter 2.4 --- Chemicals --- p.43 / Chapter Chapter 3. --- Hypoxia-Reoxygenation in Coronary Microarteries: Combined Effect with St Thomas Cardioplegia and Temperature on the Endothelium- derived Hyperpolarizing Factor and Protective Effect of Nicorandil --- p.44 / Chapter 3.1 --- Abstract --- p.44 / Chapter 3.2 --- Introduction --- p.45 / Chapter 3.3 --- Experimental design and analysis --- p.47 / Chapter 3.3.1 --- Vessel Preparation --- p.47 / Chapter 3.3.2 --- Normalization --- p.48 / Chapter 3.3.3 --- Hypoxia --- p.48 / Chapter 3.3.4 --- Effect of H-R on EDHF-mediated relaxation in coronary microarteries --- p.49 / Chapter 3.3.5 --- Combined effects ofH-R and ST solution on EDHF-mediated relaxation in coronary microarteries --- p.49 / Chapter 3.3.6 --- Effect of addition of nicorandil Krebs or ST solution under H-R on EDHF-mediated relaxation in coronary microarteries --- p.49 / Chapter 3.3.7 --- Data analysis --- p.50 / Chapter 3.4 --- Results --- p.51 / Chapter 3.4.1 --- Resting force --- p.51 / Chapter 3.4.2 --- U46619-induced contraction force --- p.51 / Chapter 3.4.3 --- Partial pressure of oxygen in hypoxia --- p.51 / Chapter 3.4.4 --- EDHF-mediated relaxation in coronary microarteries --- p.51 / Chapter 3.4.4.1 --- Effect of H-R --- p.51 / Chapter 3.4.4.2 --- Combined effects ofH-R and ST solution on EDHF-mediated relaxation --- p.52 / Chapter 3.4.4.3 --- Effects of addition of nicorandil to Krebs or ST solution under H-R on EDHF-mediated relaxation --- p.52 / Chapter 3.5 --- Discussion --- p.53 / Chapter 3.5.1 --- EDHF-mediated relaxation after exposure to H-R --- p.53 / Chapter 3.5.2 --- EDHF-mediated relaxation after H-R in ST solution at different temperature --- p.54 / Chapter 3.5.3 --- Effect of addition of nicorandil to Krebs or ST solution during H-R on EDHF-mediated relaxation --- p.55 / Chapter 3.5.4 --- Clinical implications --- p.56 / Chapter Chapter 4. --- Hypoxia-Reoxygenation in Cardiac Microveins: Combined Effect with Cardioplegia and Temperature on the Endothelial Function --- p.68 / Chapter 4.1 --- Abstract --- p.68 / Chapter 4.2 --- Introduction --- p.69 / Chapter 4.3 --- Experimental design and analysis --- p.73 / Chapter 4.3.1 --- Vessel Preparation --- p.73 / Chapter 4.3.2 --- Normalization --- p.73 / Chapter 4.3.3 --- Hypoxia --- p.73 / Chapter 4.3.4 --- Effect of H-R on EDHF-mediated relaxation in cardiac micro veins --- p.74 / Chapter 4.3.5 --- Combined effects of H-R and ST solution on EDHF-mediated relaxation in cardiac microveins --- p.74 / Chapter 4.3.6 --- Data analysis --- p.75 / Chapter 4.4 --- Results --- p.75 / Chapter 4.4.1 --- Resting force --- p.75 / Chapter 4.4.2 --- U46619-induced contraction force --- p.76 / Chapter 4.4.3 --- Partial pressure of oxygen in hypoxia --- p.76 / Chapter 4.4.4 --- EDHF-mediated relaxation after H-R in Krebs solution at 37°C --- p.76 / Chapter 4.4.5 --- EDHF-mediated relaxation after exposure to H-R in ST solution at different temperatures --- p.77 / Chapter 4.5 --- Discussion --- p.78 / Chapter 4.5.1 --- Effect of H-R on EDHF-mediated relaxation --- p.78 / Chapter 4.5.2 --- Combined effects of H-R with ST solution on EDHF-mediated relaxation --- p.80 / Chapter 4.5.3 --- Clinical implications / Chapter Chapter 5. --- General Discussion --- p.89 / Chapter 5.1 --- EDHF-mediated endothelial function in porcine coronary circulation --- p.89 / Chapter 5.1.1 --- EDHF in porcine coronary microarteries --- p.92 / Chapter 5.1.2 --- EDHF in porcine cardiac veins --- p.90 / Chapter 5.2 --- Alteration of EDHF-mediated function after exposure to H-R --- p.91 / Chapter 5.2.1 --- In coronary microarteries --- p.91 / Chapter 5.2.2 --- In cardiac veins --- p.92 / Chapter 5.3 --- Alteration of EDHF-mediated function after exposure to ST solution under H-R --- p.92 / Chapter 5.3.1 --- In coronary microarteries --- p.93 / Chapter 5.3.2 --- In cardiac veins --- p.93 / Chapter 5.4 --- EDHF-mediated function in nicorandil-supplemented ST solution under H-R in coronary microarteries --- p.93 / Chapter 5.5 --- Clinical implications / Chapter 5.5.1 --- H-R injury --- p.94 / Chapter 5.5.2 --- H-R injury and cardioplegic solution --- p.95 / Chapter 5.5.2 --- Nicorandil-supplementation in cardioplegic solution --- p.95 / Chapter 5.6 --- Limitation of the study --- p.96 / Chapter 5.7 --- Future investigations --- p.96 / Chapter 5.8 --- Conclusions --- p.97 / References --- p.99

Chronic treatment with enalapril and valsartan significantly improved endothelium-dependent relaxations of aortas from ovariectomized rats. The present results clearly point to that chronic treatment with enalapril or valsartan reduced expression and function of RAS and associated oxidative stress, thereby augmented NO bioavailability and improved endothelium-dependent relaxations. These results provided novel evidence supporting a potential application of ACEI and ARB in the treatment of endothelial dysfunction-associated vascular complications in postmenopausal women. / Functional studies showed that acetylcholine-induced relaxations in isolated aortas were impaired in a time-dependent manner, from the 4th-week to the 12th-week after ovariectomy. The impaired relaxations were partially restored by acute treatment with losartan [angiotensin II type 1 receptor (AT1R) blocker] and apocynin [NAD(P)H oxidase inhibitor]. The present results demonstrate that estrogen deficiency blunted endothelium-dependent relaxations due to impaired the NO bioavailability, which is closely associated with the reduced eNOS activity and elevated RAS expression and associated NAD(P)H oxidase-mediated oxidative stress in the vascular wall. / The present study shows that chronic consumption of cranberry juice restored the endothelium-dependent relaxations in aortas from ovariectomized rats. In ovariectomized rats, the phenylephrine-induced a higher active vascular tension; which was prevented by chronic consumption of cranberry juice. The present data also shows that cranberry juice administration significantly reduces the elevated serum levels of total cholesterol, triglyceride, high density lipoprotein (HDL) cholesterol, non-HDL (nHDL) cholesterol, and nHDL/HDL. The active ingredients in the cranberry juice organic extract accounting for the vascular benefit remain to be further examined even though the extract causes endothelial NO-dependent relaxations in normal rat aortas and contains several bioactive compounds, some of which may protect the vascular function. This study provides the first line of evidence concerning a significant vascular benefit of chronic consumption of cranberry juice during estrogen deficiency. (Abstract shortened by UMI.) / The present study used ovariectomized female rats that mimic the "equivalent" state of menopause in human and investigated whether dysregulation of RAS components contribute to endothelial dysfunction and whether chronic treatment with ACEI (enalapril) or ARB (valsartan) could restore endothelial function in ovariectomized rats. / The second objective of the present study was to investigate whether or not consumption of cranberry juice, a popular drink in Western countries, could restore endothelial function during estrogen deficiency and to elucidate the cellular mechanisms underlying the improved endothelial function. / Yung, Lai Ming. / Adviser: Huana Yu. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3252. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 148-168). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Zhang Rongzhen. / "July 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 130-176). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

O choque séptico é uma síndrome inflamatória sistêmica secundária a um processo infeccioso, no qual as disfunções das células endoteliais e do músculo liso vascular contribuem para suprimento sanguíneo insuficiente a órgãos vitais, com consequente hipotensão sistêmica, insuficiência múltipla de órgãos e morte. Em geral, nos pacientes com choque séptico, existe um desequilíbrio dos fatores hemodinâmicos, levando ao baixo débito cardíaco e vasodilatação, além da redução da resposta contrátil aos diferentes agonistas, nos quais a participação do óxido nítrico (NO), de NO-sintases (NOS) e do estresse oxidativo são evidentes. A contribuição do sistema de peptídeos natriuréticos é evidenciada nos pacientes e em animais submetidos à sepse severa ou ao choque séptico. O peptídeo natriurético tipo C (CNP) é um agente vasodilatador que leva ao relaxamento vascular pela produção de GMPc intracelular, NO, hiperpolarização de membrana das células do músculo liso vascular e redução da concentração citosólica de cálcio. O tratamento com antagonista de receptores NPR-A/B tornou os animais menos susceptíveis à sepse, com menor resposta hipotensora, melhora da contração a agonistas e redução do processo inflamatório e NO plasmático. Portanto, a hipótese do presente trabalho é que no modelo de choque séptico induzido por ligação e perfuração cecal (CLP) em ratos, ocorreria menor resposta vasoconstritora ao agonista seletivo 1-adrenérgico Fenilefrina (PE) e este efeito seria atenuado pela inibição da sinalização desencadeada pelo CNP e modulado pelo estresse oxidativo. A sobrevivência dos ratos CLP foi menor do que a observada em ratos controle-operados (Sham). Os parâmetros cardiovasculares foram mais prejudicados nos ratos CLP do que em ratos Sham. A PE apresentou efeito pressórico menor em ratos CLP comparados aos ratos Sham. O CNP induziu efeito hipotensor em ambos os grupos, porém com maior aumento de frequência cardíaca nos ratos CLP. A contração induzida pela PE foi menor em aorta e artéria mesentérica de resistência, isoladas de ratos CLP, cujo efeito foi modulado pelo endotélio vascular. A vasodilatação induzida pelo CNP foi menor em aorta de ratos CLP. Além disso, o CNP apresentou efeito modulador negativo sobre a contração da PE em aortas com ou sem endotélio, mas não em artérias mesentéricas de resistência isoladas de ratos Sham e CLP. A menor resposta contrátil induzida pela PE em aortas de ratos CLP foi aumentada pelo antagonista de receptor NPR-B, pela inibição das enzimas NOS, xantina oxidase e metaloproteinases da matriz extracelular (MMPs) em aortas com endotélio. O efeito modulador negativo do CNP sobre a contração da PE foi revertido pelo antagonista de receptor NPR-B, inibição de NOS, redução da disponibilidade de O2- e pela degradação de H2O2, em ambos os grupos. Além disso, o estresse oxidativo e a concentração citosólica de H2O2 foram maiores no músculo liso vascular de ratos CLP. A expressão protéica de CNP endógeno foi menor no endotélio e maior no músculo liso da aorta de ratos CLP do que de Sham. Porém, a expressão protéica dos receptores 1-adrenérgicos e NPR-B não foram diferentes entre os grupos, mas a expressão protéica do receptor NPR-C foi menor na aorta de ratos CLP. A mobilização de cálcio intracelular foi menor no músculo liso da aorta de ratos CLP e a alta concentração de potássio extracelular não foi suficiente para despolarizar a membrana das células musculares da aorta de ratos CLP. A expressão protéica dos receptores NPR-B e NPR-C não foram diferentes no coração de ratos CLP. Entretanto, o potencial de membrana do ventrículo esquerdo (VE) dos ratos CLP foi menor que aquele de ratos Sham. Além disso, a concentração citosólica de cálcio no VE de ratos CLP foi menor que em ratos Sham, mas a redução de cálcio citoplasmático induzida pelo CNP foi maior no VE de ratos CLP comparada aquela em ratos Sham. Ocorreu disfunção cardíaca nos ratos CLP. No modelo de choque séptico induzido por cirurgia CLP em ratos, a participação do sistema de peptídeos natriuréticos, sobretudo do CNP, contribui para a menor resposta contrátil à fenilefrina e representa potencial via de intervenção na sepse severa e no choque séptico. / Septic shock is a systemic inflammatory syndrome secondary to an infection which the vascular dysfunction leads to an insufficient blood flow to vital organs with systemic hypotension, multiple organ injury and death. Usually, in septic shock patients there is an imbalance on hemodynamic factors, leading to low cardiac output and vasodilation, with decrease on contractile responses to several agonists, which the contribution of nitric oxide (NO), NO-synthases (NOS) and oxidative stress are evident. The contribution of natriuretic peptide system is observed on patients and animals submitted to severe sepsis or septic shock. C-type natriuretic peptide (CNP) is a vasodilator that leads to vascular relaxation through cGMP, NO, vascular smooth muscle hyperpolarization and reduction in cytosolic calcium. Treatment of animals with NPR-A/B antagonist, as well as knockout mice to NPR-A receptor are less susceptible to sepsis, with less hypotension, enhanced contraction to different agonists and reduction in inflammation and plasma NO. Therefore, the hypothesis of the present work is in septic shock induced by cecal ligation and puncture (CLP) model in rats there is low vasoconstriction to the selective 1- adrenoceptor agonist Phenylephrine (PE) and this effect could be attenuated by the inhibition of CNP signaling, strongly modulated by the oxidative stress. CLP survival was lower than the control rats (Sham) and cardiovascular parameters were impaired in CLP compared to the Sham rats. PE had positive pressure effect lower in CLP than in Sham rats. CNP induced hypotension in both groups with greater increases of cardiac rate in CLP. PE-induced contraction was decreased in aorta and resistance mesenteric artery isolated from CLP rats and this effect was modulated by the vascular endothelium. CNP-induced vasodilation was lower in rat aorta of CLP. Moreover, CNP had a negative modulator effect over the PE contraction on aortas with or without endothelium, but not on resistance mesenteric artery isolated from Sham and CLP rats. The low contractile response induced by PE on CLP aortas was enhanced by NPR-B antagonist, and by NOS, xanthine oxidase or extracellular matrix metalloproteinases (MMPs) inhibition on aortas with endothelium. The negative modulation induced by CNP over the PE contraction was reversed by the presence of NPR-B antagonist, NOS inhibition, and by the decrease on O2- availability and H2O2 degradation in both groups. Furthermore, oxidative stress and H2O2 intracellular content were greater on vascular smooth muscle of CLP rats. Protein expression of endogenous CNP was lower on endothelium and greater on smooth muscle of aortas isolated from CLP compared to Sham rats. However, the protein expression of 1-adrenoceptor and NPR-B were not different between the groups, but the protein expression of NPR-C was lower on smooth muscle of CLP aortas. Intracellular calcium mobilization was decreased on vascular smooth muscle of CLP aortas and extracellular high potassium solution was not able to depolarize smooth muscle layer of CLP aortas. Protein expression of NPR-B and NPR-C were not different on CLP hearts. However, membrane potential of left ventricle (LV) of CLP was lower than in Sham rats. Besides, intracellular calcium content of LV of CLP was lower than Sham rats but the decrease on cytosolic calcium induced by CNP was greater on LV of CLP compared to the Sham rats. There was cardiac dysfunction on CLP rats. In rat CLP septic shock model, the role of natriuretic peptide, mainly CNP, is of great importance in the decreased 1-adrenoceptor contraction, representing a potential via of intervention on severe sepsis and septic shock.

Yung, Lai Hang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 93-115). / Abstract also in Chinese. / Declaration --- p.i / Acknowledgements --- p.ii / Abbreviations --- p.iii / Abstract in English --- p.v / Abstract in Chinese --- p.ix / Contents --- p.xi / Chapter Chapter I - --- Introduction / Chapter 1.1) --- Endothelial cells function --- p.1 / Chapter 1.2) --- Oxidative stress in the vascular wall --- p.2 / Chapter 1.2.1) --- Sources of ROS --- p.3 / Chapter 1.2.2) --- Actions of ROS --- p.3 / Chapter 1.2.2.1) --- Impaired endothelium-dependent vasodilatation --- p.3 / Chapter 1.2.2.2) --- VSMC migration --- p.4 / Chapter 1.2.2.3) --- Programmed cell death (cell apoptosis) --- p.4 / Chapter 1.3) --- Endothelial cell apoptosis --- p.7 / Chapter 1.3.1) --- Apoptosis and cardiovascular diseases --- p.7 / Chapter 1.3.2) --- Mechanisms of endothelial cells apoptosis --- p.7 / Chapter 1.3.2.1) --- What are caspases? --- p.8 / Chapter 1.3.2.2) --- Death receptor-mediated apoptosis --- p.9 / Chapter 1.3.2.3) --- Mitochondria-dependent pathway --- p.9 / Chapter 1.3.3) --- Regulations of endothelial cells apoptosis --- p.10 / Chapter 1.3.3.1) --- Oxidative stress --- p.10 / Chapter 1.3.3.2) --- Shear Stress --- p.11 / Chapter 1.3.3.3) --- Growth factors --- p.12 / Chapter 1.3.3.4) --- NO --- p.12 / Chapter 1.3.3.5) --- Inflammatory mediators --- p.13 / Chapter 1.4) --- Mitogen activated kinases signaling in apoptosis --- p.15 / Chapter 1.5) --- Bone morphogenic proteins (BMPs) --- p.17 / Chapter 1.5.1) --- BMPs functions and cardiovascular system --- p.17 / Chapter 1.5.2) --- BMPs signaling pathways --- p.18 / Chapter 1.5.2.1) --- Smad-dependent pathway --- p.18 / Chapter 1.5.2.2) --- MAPKs and SAPKs pathways --- p.19 / Chapter 1.5.2.3) --- Antagonists of BMPs signaling --- p.20 / Chapter 1.5.3) --- BMP4 and cardiovascular diseases --- p.20 / Chapter 1.6) --- "Justification, long-term significance and objectives of the present project" --- p.23 / Chapter Chapter II - --- Methods and Materials / Chapter 2.1) --- Animal handling --- p.24 / Chapter 2.2) --- Endothelial cell isolation and culture --- p.24 / Chapter 2.2.1) --- Primary culture of rat endothelial cells --- p.24 / Chapter 2.2.2) --- Culture of human umbilical cord vein endothelial cells… --- p.25 / Chapter 2.3) --- Apoptosis assessment --- p.25 / Chapter 2.3.1) --- Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay --- p.25 / Chapter 2.3.2) --- Cell death detection ELISA kit --- p.26 / Chapter 2.3.3) --- Flow cytometry --- p.27 / Chapter 2.4) --- Western blot analysis --- p.28 / Chapter 2.4.1) --- Sample preparation --- p.28 / Chapter 2.4.2) --- SDS-PAGE and transfer --- p.28 / Chapter 2.5) --- DHE fluorescence --- p.29 / Chapter 2.6) --- "Drugs, chemicals and other reagents" --- p.30 / Chapter 2.6.1) --- Drugs and chemicals used in the present experiments --- p.30 / Chapter 2.6.2) --- Reagents for Western blot analysis --- p.30 / Chapter 2.6.3) --- Primary antibodies --- p.33 / Chapter 2.7) --- Small interfering RNA experiment --- p.34 / Chapter 2.8) --- Statistical analysis --- p.34 / Chapter Chapter III - --- BMP4 induces endothelial cell apoptosis in ROS related p38 MAPK and JNK mediated caspase-3 dependent pathway / Chapter 3.1) --- Introduction --- p.35 / Chapter 3.2) --- Methods and materials --- p.39 / Chapter 3.2.1) --- Isolation and culture of endothelial cells --- p.39 / Chapter 3.2.2) --- Drugs treatment --- p.39 / Chapter 3.2.3) --- Assay for cell apoptosis --- p.40 / Chapter 3.2.3.1) --- Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay --- p.40 / Chapter 3.2.3.2) --- Cell death detection ELISA kit --- p.41 / Chapter 3.2.3.3) --- Flow cytometric analysis --- p.41 / Chapter 3.2.4) --- Western blot analysis --- p.41 / Chapter 3.2.5) --- Dihydroethidium (DHE) staining --- p.42 / Chapter 3.2.6) --- Statistical analysis --- p.42 / Chapter 3.3) --- Results --- p.43 / Chapter 3.3.1) --- Dose- and time-dependent effect of BMP4 --- p.43 / Chapter 3.3.2) --- Role of caspases in apoptosis of RAECs and HUVECs --- p.43 / Chapter 3.3.3) --- Roles of BMP4 and ROS in endothelial cell apoptosis --- p.44 / Chapter 3.3.3.1) --- Noggin antagonism of BMP4-induced effect --- p.44 / Chapter 3.3.3.2) --- NAD(P)H oxidase-mediated ROS production --- p.44 / Chapter 3.3.3.3) --- Inhibition of endothelial cell apoptosis by ROS scavengers --- p.45 / Chapter 3.3.4) --- Roles of MAPKs/SAPKs in BMP4-induced endothelial cell apoptosis --- p.45 / Chapter 3.3.5) --- Relationship between ROS and MAPKs/SAPKs --- p.46 / Chapter 3.3.6) --- Relationship between p38 MAPK and JNK --- p.46 / Chapter 3.4) --- Discussion --- p.82 / Chapter 3.4.1) --- Caspase-dependent pathways --- p.82 / Chapter 3.4.2) --- Oxidative stress --- p.85 / Chapter 3.4.3) --- Role of MAPKs activation in BMP4-induced endothelial cell apoptosis --- p.87 / Chapter 3.4.4) --- ROS mediates BMP4-induced activation of MAPKs --- p.88 / Chapter 3.4.5) --- Role of p38 MAPK in the activation of JNK 1 --- p.89 / Chapter 3.5) --- Concluding remarks --- p.91 / References --- p.93 / Publications and Awards --- p.116

Atherosclerosis is an inflammatory disease which develops focally in regions of the vasculature where there is dysfunction of endothelial cells modulated in part by shear stress from flowing blood. To address the clinical crisis of atherosclerosis, tissue engineering has focused on development of a living blood vessel substitute for use as a vascular graft in bypass surgery. Despite substantial progress in understanding the biological basis and developing clinical treatments for cardiovascular disease, critical challenges remain. As a novel strategy to improve understanding of basic human vascular biology and develop superior tissue engineered grafts, this dissertation combines the scientific and clinical approaches by using a tissue engineered blood vessel as a more physiologic in vitro model to study endothelial cell biology. Through the use of transcriptional profiling, results demonstrate significant changes in endothelial cell gene expression using the tissue engineered blood vessel model. Furthermore, the presence of a more physiologic substrate alters the cellular response to shear stress which is a critical mediator of vascular pathology. A case study of endothelial cell function in this system focuses on cell-cell communication through gap junctions. Endothelial cell connexins which form gap junctions are shown to be differentially regulated by substrate and shear stress. Moreover, gap junction communication between endothelial cells is modulated by the mechanical environment. Studies using RNA interference to knockdown expression of individual connexin isotypes demonstrate integrated regulation of connexins yet unique roles in endothelial cell function. Collectively, results exemplify the sensitivity of endothelial cell phenotype to substrate and shear stress and underline the importance of using more physiologic models in the study of basic cell biology.

Thesis (Ph. D.)--University of Missouri--Columbia, 2000. / Typescript. Vita. Includes bibliographical references (leaves 186-210). Also available on the Internet.