Fang Yujing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 148-182). / Abstracts in English and Chinese. / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Overview of the study --- p.1 / Chapter 1.2 --- Overview of Hypertension --- p.4 / Chapter 1.3 --- Overview of Obesity-Related Hypertension --- p.9 / Chapter 1.3.1 --- Body fat distribution --- p.11 / Chapter 1.3.2 --- Insulin resistance and Hyperinsulinaemia --- p.12 / Chapter 1.3.3 --- Sympathetic nervous system activity --- p.13 / Chapter 1.3.4 --- Genetics of Obesity --- p.15 / Chapter 1.3.4.1 --- Brown adipose tissue (BAT) --- p.15 / Chapter 1.3.4.2 --- Uncoupling protein --- p.16 / Chapter 1.3.4.3 --- Uncoupling Protein 1 Gene --- p.17 / Chapter 1.3.4.4 --- Association of the UCP1 Polymorphism and Weight Gain in Obesity --- p.18 / Chapter 1.4 --- Overview of Genetics of Hypertension --- p.19 / Chapter 1.4.1 --- The Renin-Angiotensin System --- p.19 / Chapter 1.4.1.1 --- Functions of Renin-Angiotensin System --- p.20 / Chapter 1.4.1.2 --- The Renin-Angiotensin System and Hypertension --- p.21 / Chapter 1.4.2 --- Renin --- p.22 / Chapter 1.4.3 --- Angiotensinogen --- p.25 / Chapter 1.4.4 --- Angiotensin-Converting Enzyme (ACE) --- p.29 / Chapter 1.4.4.1 --- Angiotensin-Converting Enzyme Gene --- p.29 / Chapter 1.4.4.2 --- Association of the ACE I/D Polymorphism with Hypertension --- p.30 / Chapter 1.4.4.3 --- Association of the ACE I/D Polymorphism with Other disease --- p.32 / Chapter 1.4.5 --- The Angiotensin II Receptor --- p.35 / Chapter 1.4.5.1 --- Type 1 Angiotensin II Receptor --- p.35 / Chapter 1.4.5.2 --- The Type 1 Angiotensin Receptor Gene --- p.36 / Chapter 1.4.6 --- Dopamine --- p.39 / Chapter 1.4.6.1 --- Dopamine Receptors --- p.42 / Chapter 1.4.6.2 --- The Dopamine D2 Receptor Gene --- p.45 / Chapter 2 --- Aims --- p.47 / Chapter 3 --- Materials and methodology --- p.48 / Chapter 3.1 --- Patient recruitment protocol --- p.48 / Chapter 3.2 --- Subjects --- p.49 / Chapter 3.2.1 --- Classification of Hypertension --- p.50 / Chapter 3.2.2 --- Definition of Dyslipidaemia --- p.51 / Chapter 3.2.3 --- Classification of Diabetes Mellitus --- p.52 / Chapter 3.2.4 --- Definition of Obesity --- p.53 / Chapter 3.2.5 --- Exclusion Criteria --- p.54 / Chapter 3.3 --- Routine Assessment --- p.54 / Chapter 3.3.1 --- Blood Pressure --- p.54 / Chapter 3.3.2 --- Measurements of obesity --- p.55 / Chapter 3.3.2.1 --- Body mass index --- p.55 / Chapter 3.3.2.2 --- Waist to hip ratio --- p.55 / Chapter 3.3.2.3 --- Skin-Fold Thickness --- p.55 / Chapter 3.3.2.4 --- Skinfold Percentage Fat --- p.56 / Chapter 3.3.3 --- Biochemical measurements --- p.56 / Chapter 3.3.3.1 --- Assays measuring biochemical factors from plasma --- p.57 / Chapter 3.3.3.1.1 --- Plasma electrolytes --- p.57 / Chapter 3.3.3.1.2 --- Plasma urate --- p.57 / Chapter 3.3.3.1.3 --- Plasma creatinine --- p.57 / Chapter 3.3.3.1.4 --- Fasting plasma glucose --- p.57 / Chapter 3.3.3.1.5 --- Fasting plasma cholesterol --- p.57 / Chapter 3.3.3.1.6 --- Fasting plasma triglyceride --- p.58 / Chapter 3.3.3.2 --- Assays measuring biochemical factors from urine --- p.58 / Chapter 3.3.3.2.1 --- Urinary electrolytes --- p.58 / Chapter 3.3.3.2.2 --- Urinary creatinine --- p.58 / Chapter 3.3.3.2.3 --- Urinary albumin concentration --- p.58 / Chapter 3.4 --- Extraction of DNA from blood specimen --- p.59 / Chapter 3.5 --- Polymerase Chain Amplification protocols --- p.60 / Chapter 3.5.1 --- Uncoupling protein 1 gene polymorphism --- p.60 / Chapter 3.5.2 --- Angiotensin-Converting Enzyme insertion-deletion polymorphism --- p.62 / Chapter 3.5.3 --- Angiotensin type 1 receptor gene A1166C polymorphism --- p.64 / Chapter 3.5.4 --- Dopamine D2 receptor TaqI polymorphism --- p.66 / Chapter 3.5.5 --- Dopamine D2 receptor TaqI polymorphism --- p.66 / Chapter 3.6 --- Statistical analysis --- p.68 / Chapter 3.6.1 --- Paired sample T test --- p.68 / Chapter 3.6.2 --- Conditional Logistic Regression --- p.68 / Chapter 3.6.3 --- Linkage analysis --- p.69 / Chapter 3.6.3.1 --- Allelic frequency and genotypic distribution --- p.69 / Chapter 3.6.3.2 --- Hardy- Weinberg equilibrium --- p.69 / Chapter 3.6.3.3 --- Parametric analysis --- p.71 / Chapter 3.6.3.4 --- Nonparametric analysis --- p.71 / Chapter 3.6.3.4.1 --- The affected sib pair (ASP) method --- p.74 / Chapter 3.6.3.4.2 --- The affected pedigree member (APM) method of linkage analysis --- p.76 / Chapter 3.6.3.4.3 --- Quantitative traits linkage analysis --- p.79 / Chapter 4 --- Results --- p.81 / Chapter 4.1 --- Description of the characteristics of in siblings --- p.81 / Chapter 4.1.1 --- Siblings and sib-pairs --- p.81 / Chapter 4.1.2 --- Demographic characteristics --- p.81 / Chapter 4.1.3 --- Relationship to age and gender --- p.83 / Chapter 4.1.3.1 --- Hypertension versus age and gender --- p.83 / Chapter 4.1.3.2 --- Central obesity versus age and gender --- p.83 / Chapter 4.1.3.3 --- General obesity versus age and gender --- p.84 / Chapter 4.1.3.4 --- Hypertension-central obesity versus age and gender --- p.84 / Chapter 4.1.3.5 --- Hypertension- general obesity versus age and gender --- p.85 / Chapter 4.1.4 --- Relationship to anthropometric indices --- p.85 / Chapter 4.1.4.1 --- Large proportion of obesity --- p.85 / Chapter 4.1.4.2 --- Hypertension versus anthropometric indices --- p.86 / Chapter 4.1.5 --- Relationship to biochemistry indices --- p.87 / Chapter 4.1.5.1 --- Large proportion of dyslipidaemia --- p.87 / Chapter 4.2 --- Association between disease traits and covariates in discordant sib pairs --- p.87 / Chapter 4.2.1 --- Association between blood pressure and covariates in discordant sib-pairs --- p.87 / Chapter 4.2.2 --- Association between general obesity and covariates in discordant sib-pairs --- p.89 / Chapter 4.2.3 --- Association between obesity related hypertension and covariates in combined discordant sib-pairs --- p.91 / Chapter 4.3 --- Description of the analysis of the polymorphisms of 4 genes which might be related to hypertension and obesity --- p.93 / Chapter 4.3.1 --- The uncoupling protein 1 gene --- p.93 / Chapter 4.3.1.1 --- Comparison of A-G polymorphism in terms of hypertension or obesity --- p.94 / Chapter 4.3.1.2 --- Comparison of A-G polymorphism in terms of HT and obesity --- p.97 / Chapter 4.3.1.3 --- Comparison of characteristics among different genotypes --- p.99 / Chapter 4.3.2 --- The angiotensin-converting enzyme gene --- p.99 / Chapter 4.3.2.1 --- Comparison of the ACE I/D polymorphism in terms of HT --- p.100 / Chapter 4.3.2.2 --- Comparison of characteristics among different genotypes --- p.101 / Chapter 4.3.3 --- The angiotensin type 1 receptor gene --- p.104 / Chapter 4.3.3.1 --- Comparison of the A T1R A1166C polymorphism in terms of HT --- p.104 / Chapter 4.3.3.2 --- Comparison of characteristics among different genotypes --- p.105 / Chapter 4.3.4 --- The Dopamine D2 receptor gene --- p.105 / Chapter 4.3.4.1 --- Comparison of the DRD2 gene TaqI polymorphism in terms of HT --- p.106 / Chapter 4.3.4.2 --- Comparison of the DRD2 gene TaqI polymorphism in terms of general obesity or central obesity --- p.108 / Chapter 4.3.4.3 --- Comparison of the DRD2 gene TaqI polymorphism in terms of general obesity/central obesity and HT --- p.110 / Chapter 4.4 --- Sib pair linkage analysis --- p.113 / Chapter 4.4.1 --- Linkage between each gene and hypertension in our data --- p.114 / Chapter 4.4.1.1 --- Genetic linkage of the marker near the UCP1 gene locus to hypertension --- p.114 / Chapter 4.4.1.2 --- Genetic linkage of the angiotens in-converting enzyme gene locus to hypertension --- p.116 / Chapter 4.4.1.3 --- Genetic linkage of the angiotensin type 1 (AT1) receptor gene locus to hypertension --- p.117 / Chapter 4.4.1.4 --- Genetic linkage of the dopamine D2 receptor gene locus to hypertension --- p.118 / Chapter 4.4.2 --- Linkage between each gene locus and obesity in Hong Kong hypertensive Chinese families --- p.120 / Chapter 4.4.2.1 --- Genetic linkage of the uncoupling protein 1 gene locus to obesity with hypertensive family history --- p.120 / Chapter 4.4.2.2 --- Genetic linkage of the angiotens in-converting enzyme gene locus to obesity with hypertensive family history --- p.123 / Chapter 4.4.2.3 --- Genetic linkage of the angiotensin type 1 receptor gene locus to obesity with hypertensive family history --- p.124 / Chapter 4.4.2.4 --- Genetic linkage of the dopamine D2 gene locus to obesity --- p.127 / Chapter 5 --- Discussion --- p.129 / Chapter 5.1 --- Age-related anomalies --- p.129 / Chapter 5.2 --- Gender-related anomalies --- p.129 / Chapter 5.3 --- Obesity- related hypertension --- p.130 / Chapter 5.4 --- Abnormal biochemical parameters in hypertension --- p.131 / Chapter 5.5 --- Genetic parameters involved in the pathogenesis of hypertension and obesity. --- p.132 / Chapter 5.6 --- The uncoupling protein gene --- p.132 / Chapter 5.6.1 --- Higher frequency in central obese males --- p.132 / Chapter 5.6.2 --- Linkage of systolic blood pressure with G allele --- p.134 / Chapter 5.6.3 --- Metabolic link --- p.135 / Chapter 5.7 --- The angiotensin-converting enzyme gene --- p.137 / Chapter 5.7.1 --- The ACE D allele and hypertension in previous studies --- p.137 / Chapter 5.7.2 --- Positive role of the ACE DD genotype found in our data --- p.139 / Chapter 5.8 --- Angiotensin II type 1 receptor gene --- p.141 / Chapter 5.8.1 --- No linkage with HT --- p.141 / Chapter 5.9 --- Dopamine D2 gene --- p.143 / Chapter 5.9.1 --- Dopamine and obesity --- p.143 / Chapter 5.9.2 --- Linkage with obese and HT --- p.144 / Chapter 5.10 --- Summary of the study --- p.146 / Chapter 5.11 --- Possible further developments in this study --- p.146 / Chapter 6 --- References --- p.148

Although essential hypertension has long been recognized to involve a strong genetic predisposition, the genes that increase susceptibility remain virtually unknown. With recent advances in molecular biology and statistical methods, it has become feasible to study candidate genes which may contribute to the pathogenesis of essential hypertension in humans. We identified the polymorphisms of five genes by applying a micoarray genotyping system for multiplex analysis of a panel of single nucleotide polymorphisms (SNPs) in genes involved in the regulation of blood pressure, then determined whether specific SNPs in genes were related to blood pressure in Hong Kong Chinese. / Based on the study results, we conclude that blood pressure levels are determined by complex interactions between genetic and environmental factors. The AGT gene 235T and the DD1R gene -48G alleles strongly predicted the development of hypertension in Hong Kong Chinese. The variants of the DD2R gene and the UCP1 gene A-3826G variant only weakly impacted upon blood regulation. However, the three variants of the INSR gene and the L10F variant of the AGT gene were not detected in Hong Kong Chinese. Although success in identifying single genes contributing to hypertension has been limited, the use of intermediate phenotypes and dense mapping of candidate genes shows the influence of gene-gene interaction on hypertension or obesity-related hypertension in our hypertensive families of Hong Kong Chinese. / Families were recruited if the proband was found to have hypertension and had siblings resident in Hong Kong. We identified 126 families with at least one hypertensive sibling. A total of 434 siblings were studied. All subjects underwent clinical and biochemical investigation to exclude those with either secondary hypertension, impaired glucose tolerance or type 2 diabetes mellitus. / Twelve SNPs in five candidate genes, which included the M235T, T174M, G-217A and L10F polymorphisms of the angiotensinogen gene (AGT); A-48G polymorphism of the dopamine D1 receptor gene (DD1R); the TaqI A, -141C Ins/Del and A-241G polymorphisms of the dopamine D2 receptor gene (DD2R); Phe382Val, Lys460Glu, and Gly1008Val polymorphisms of the insulin receptor gene (INSR); and the A-3826G polymorphism of the uncoupling protein 1 gene (UCP1). / by Fang Yujing. / "January 2006." / Adviser: Brian Tomlinson. / Source: Dissertation Abstracts International, Volume: 67-11, Section: B, page: 6298. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 148-181). / 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.

研究背景和目的 / 高血壓和糖尿病是人群中常見的疾病，兩者常共同存在，其共存的病理生理機制非常複雜，其中腎素血管景張素系統功能紊亂起重要作用。多個研究表明血管緊張素轉化晦抑制劑和血管緊張素II 1 型受體阻滯劑通過調節不同基因的表達，發揮其保護心血管和腎臟功能的效用。然而，目前仍缺乏遠兩類藥物影響全基因表達譜的全面調查。因此，本研究應用全基因表達譜晶片技術，檢測分析了高血壓和糖尿病並發的病人在服用安慰劑、雷米普利(ramipril)和替米沙坦(telmisartan)後的全基因表達譜的變化，從而全面評估了血管緊張素轉化臨抑制劑和血管繁張素II 1 型受體阻滯劑對相關基因的轉錄調控作用。 / 方法 / 11 名患有高血壓和糖尿病的病人(男性5 名)在服用安慰劑最少2 星期后，以隨機吹序接受為期各6 星期的雷米普利和替米沙坦治療，並分別在安慰劑期和2 個藥物治療期結束后提取心A 進行全基因表達譜分析。 / 結果 / 與服用安慰劑時的全基因表達譜相比，雷米普利治療后有267 個基因的表達降低， 99 個基因的表達增強。表達差異幅度為-2.0 至1.3 (P < 0.05) 。表達下降的基因主要與血管平滑肌收縮、炎症反應和氧化壓力相關。表達增強的基因主要與心血管炎症反應負調節相關。基因共表達網絡分析表明， 2 個共表達基因組與雷米普利的降血壓作用相闕， 3 個共表達基因組與肥胖相關。 / 與服用安慰劑時的全基因表達譜相比， 替米拉)、坦治療后有55 個基因表達降低， 158 個基因的表達增強。表達差異幅度為-1. 9 至1.3 (P < 0.05) 。表達增強的基因主要與脂質代謝、糖代謝和抗炎症因子作用相關。基因共表達網絡分析表明， 2 個共表達基因組與替米沙坦對24 小時舒張壓負荷量的作用相關， 2 個共表達基因組則與總膽固醇， 低密度脂蛋白膽固醇和C 反應蛋白相關。 / 結論 / 本論文描述了高血壓和2 型糖尿病病患全基因組表達的總體模式及經藥物治療後表達譜的相應改變， 為今後進一步研究腎素血管緊張素系統抑制劑和高血壓、糖尿病發展進程的相互作用提供了方向。 / Background and aim: Pathophysiological mechanisms underpinning the coexistence of hypertension and type 2 diabetes are complex systemic responses involving dysregulation of the renin-angiotensin system (RAS). We conducted this study to investigate the genome wide gene expression changes in patients with both hypertension and diabetes at three treatment stages, including placebo, ramipril and telmisartan. This study aimed to obtain a panoramic view of interactions between gene transcription and antihypertensive therapy by RAS inhibition. / Methods: 11 diabetic patients (S men) with hypertension were treated with placebo for at least 2 weeks followed by 6 weeks randomised crossover treatment with ramipril Smg daily and telmisartan 40mg daily, respectively. Total RNA were extracted from leukocytes at the end of placebo and each treatment period, and were hybridized to the whole transcript microarray. The limma package for R was used to identify differentially expressed genes between placebo and the 2 active treatments. The weighted gene coexpression network analysis (WGCNA) was applied to identify groups of genes (modules) highly correlated to a common biological function in pathogenesis and progression of hypertension and diabetes. / Results: There were 267 genes down-regulated and 99 genes up-regulated with ramipril. Fold changes of gene expression were ranged from -2.0 to 1.3 (P < 0.05). The down-regulated genes were involved in vascular signalling pathways responsible for vascular smooth muscle contraction, inflammation and oxidative stress. The up-regulated genes were associated with negative regulation of cardiovascular inflammation. The WGCNA identified 17 coexpression gene modules related to ramipril. The midnight blue (57 genes, r < -0.44, P < 0.05) and magenta (190 genes, r < -0.44, P < 0.05) modules were significantly correlated to blood pressure differences between placebo and ramipril. / There were 55 genes down-regulated and 158 genes up-regulated with telmisartan. Fold changes of gene expression were ranged from -1.9 to 1.3 (P < 0.05). The down-regulated genes were mainly associated with cardiovascular inflammation and oxidative stress. The up-regulated genes were associated with lipid and glucose metabolism and anti-inflammatory actions. The WGCNA identified 8 coexpression gene modules related to telmisartan. The black (56 genes, r = 0.46, P = 0.03) and turquoise (1340 genes, r = -0.48, P = 0.02) modules were correlated with diastolic blood pressure load. The blue (1027 genes) module was enriched with genes correlated with total cholesterol (r = - 0.52, P = 0.01), LDL-C (r = - 0.58, P = 0.004), and hsCRP (r = -0.57, P = 0.006). The green module (272 genes) was significantly correlated with LDL-C (r = - 0.44, P = 0.04) and hsCRP (r = - 0.59, P = 0.004). / Conclusion: Genome wide gene expression profiling in this study describes the general pattern and treatment responses in patients with hypertension and type 2 diabetes, which suggests future directions for further investigations on the interaction between actions of the RAS blockers and disease progression. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Deng, Hanbing. / "December 2011." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 198-256). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Declaration --- p.i / Publications --- p.ii / Abstract --- p.iv / 論文摘要 --- p.vi / Acknowledgements --- p.viii / Table of Contents --- p.x / List of tables --- p.xiv / List of figures --- p.xv / List of appendices --- p.xvii / List of abbreviations --- p.xviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Overview --- p.1 / Chapter 1.2 --- Epidemiology --- p.6 / Chapter 1.2.1 --- Epidemiology of hypertension --- p.9 / Chapter 1.2.2 --- Epidemiology of type 2 diabetes --- p.10 / Chapter 1.3 --- Aetiology --- p.13 / Chapter 1.3.1 --- Ageing --- p.13 / Chapter 1.3.1.1 --- Age-induced artery stiffness --- p.14 / Chapter 1.3.1.2 --- Age-related endothelial dysfunction --- p.14 / Chapter 1.3.2 --- The renin-angiotensin system (RAS) --- p.16 / Chapter 1.3.2.1 --- The local RAS --- p.20 / Chapter 1.3.2.2 --- The RAS and insulin resistance --- p.22 / Chapter 1.3.2.3 --- The RAS and inflammation --- p.26 / Chapter 1.3.2.4 --- The RAS and oxidative stress --- p.28 / Chapter 1.3.3 --- Obesity --- p.31 / Chapter 1.3.3.1 --- Obesity and renin-angiotensin system (RAS) --- p.33 / Chapter 1.3.3.2 --- Obesity and insulin resistance --- p.36 / Chapter 1.3.3.3 --- Obesity and oxidative stress --- p.38 / Chapter 1.3.3.4 --- Obesity and sympathetic nervous system (SNS) --- p.38 / Chapter 1.4 --- Pharmacogenomics of antihypertensive therapy --- p.39 / Chapter 1.4.1 --- Angiotensin-converting enzyme inhibitors (ACEIs) --- p.41 / Chapter 1.4.2 --- Angiotensin II type 1 receptor blockers (ARBs) --- p.43 / Chapter Chapter 2 --- Aim --- p.59 / Chapter Chapter 3 --- Methods --- p.60 / Chapter 3.1 --- Subjects --- p.60 / Chapter 3.1.1 --- Subject recruitment protocol --- p.60 / Chapter 3.1.2 --- Definition of type 2 diabetes --- p.62 / Chapter 3.1.3 --- Definition of obesity --- p.62 / Chapter 3.1.4 --- Definition of dyslipidaemia --- p.63 / Chapter 3.2 --- Study design and procedure --- p.64 / Chapter 3.2.1 --- Blood pressure assessments --- p.65 / Chapter 3.2.2 --- Anthropometric measurements --- p.68 / Chapter 3.2.3 --- Medical history, life style and side effect evaluation --- p.68 / Chapter 3.2.4 --- RNA isolation --- p.68 / Chapter 3.2.5 --- RNA quality assessment --- p.70 / Chapter 3.2.6 --- Oligonucleotide microarrays --- p.71 / Chapter 3.2.7 --- DNA extraction --- p.75 / Chapter 3.2.8 --- Biomedical measurements --- p.76 / Chapter 3.2.8.1 --- Glycosylated haemoglobin Alc (HbA₁c) --- p.77 / Chapter 3.2.8.2 --- Fasting plasma glucose (FP G) --- p.77 / Chapter 3.2.8.3 --- Fasting insulin --- p.77 / Chapter 3.2.8.4 --- Plasma urate --- p.77 / Chapter 3.2.8.5 --- High sensitive C-reactive protein (hsCRP) --- p.78 / Chapter 3.2.8.6 --- Fasting plasma triglycerides (TG) --- p.78 / Chapter 3.2.8.7 --- Fasting plasma cholesterols --- p.78 / Chapter 3.2.8.8 --- Renal and liver functions --- p.78 / Chapter 3.2.8.9 --- Urinary parameters --- p.79 / Chapter 3.3 --- Statistical Analysis --- p.79 / Chapter 3.3.1 --- Statistical analysis of clinical and biomedical data --- p.79 / Chapter 3.3.2 --- Analysis of microarray data --- p.80 / Chapter 3.3.2.1 --- Raw data assessment --- p.80 / Chapter 3.3.2.2 --- Data normalisation --- p.92 / Chapter 3.3.2.3 --- Data filtering --- p.96 / Chapter 3.3.2.4 --- Linear models for assessment of differential expression --- p.96 / Chapter 3.3.2.5 --- Weighted gene coexpression network analysis --- p.101 / Chapter 3.3.2.6 --- Network visualisation and gene ontology analysis --- p.102 / Chapter 3.3.3 --- Sample size calculation --- p.103 / Chapter Chapter 4 --- Results --- p.104 / Chapter 4.1 --- Demographic and biomedical characteristics at baseline --- p.104 / Chapter 4.1.1 --- Hypertension and diabetes status at baseline --- p.108 / Chapter 4.1.2 --- Prevalence of dyslipidaemia --- p.108 / Chapter 4.1.3 --- Prevalence of obesity --- p.109 / Chapter 4.1.4 --- Prevalence of metabolic syndrome --- p.109 / Chapter 4.1.5 --- Inflammation markers --- p.110 / Chapter 4.2 --- Blood pressure response to the RAS blockers --- p.110 / Chapter 4.2.1 --- Clinic blood pressure --- p.110 / Chapter 4.2.2 --- 24-hour ambulatory blood pressure --- p.112 / Chapter 4.3 --- Biomedical characteristics --- p.118 / Chapter 4.4 --- Compliance, side effects and adverse events --- p.120 / Chapter 4.5 --- Gene expression differences between treatments --- p.121 / Chapter 4.5.1 --- Gene expression differences between placebo and ramipril --- p.121 / Chapter 4.5.1.1 --- Expression changes in genes related to regulation of transcription with ramipril --- p.122 / Chapter 4.5.1.2 --- Expression changes with ramipril in genes related to molecular mechanism of cardiovascular changes in hypertension --- p.125 / Chapter 4.5.1.3 --- Expression changes in genes related to blood pressure with ramipril --- p.128 / Chapter 4.5.1.4 --- Expression changes in genes related to fatty acid metabolism with ramipril --- p.130 / Chapter 4.5.1.5 --- Expression changes in genes related to inflammation with ramipril --- p.130 / Chapter 4.5.1.6 --- Expression changes in genes related to oxidative stress with ramipril --- p.133 / Chapter 4.5.1.7 --- Power estimation --- p.133 / Chapter 4.5.2 --- Gene expression differences between placebo and telmisartan --- p.135 / Chapter 4.5.2.1 --- Changes in regulation oftranscription with telmisartan --- p.137 / Chapter 4.5.2.2 --- Expression changes in genes related to glucose metabolism with telmisartan --- p.141 / Chapter 4.5.2.3 --- Expression changes in genes related to lipid metabolism with telmisartan --- p.143 / Chapter 4.5.2.4 --- Expression changes in genes related to inflammation with telmisartan --- p.143 / Chapter 4.5.2.5 --- Power estimation --- p.145 / Chapter 4.5.3 --- WGCNA for comparison between placebo and ramipriI --- p.147 / Chapter 4.5.3.1 --- Midnight blue module and clinical responses to ramipril --- p.152 / Chapter 4.5.3.2 --- Magenta module and blood pressure responses to ramipril --- p.154 / Chapter 4.5.3.3 --- Yellow module and clinical responses to ramipril --- p.158 / Chapter 4.5.3.4 --- Red module and clinical responses to ramipril --- p.161 / Chapter 4.5.3.5 --- Salmon module and clinical responses to ramipril --- p.163 / Chapter 4.5.4 --- WGCNA for comparison between placebo and telmisaItan --- p.168 / Chapter 4.5.4.1 --- Diastolic blood pressure load and gene coexpression modules --- p.168 / Chapter 4.5.4.2 --- Lipids, hsCRP and gene coexpression modules --- p.172 / Chapter Chapter 5 --- Discussion --- p.176 / Chapter 5.1 --- Gene expression changes related to ramipril --- p.177 / Chapter 5.1.1 --- Gene expression changes and blood pressure reduction by ramipri1 --- p.177 / Chapter 5.1.2 --- Gene expression changes and vascular protection by ramipri1 --- p.181 / Chapter 5.1.3 --- Obesity and gene expression changes by ramipril --- p.183 / Chapter 5.2 --- Gene expression changes related to telmisartan --- p.185 / Chapter 5.2.1 --- Blood pressure and coexpressed gene modules with telmisartan --- p.185 / Chapter 5.2.2 --- Lipid metabolism and gene expression changes by telmisartan --- p.187 / Chapter 5.2.3 --- Glucose metabolism and gene expression changes by telmisartan --- p.189 / Chapter 5.2.4 --- hsCRP and gene expression changes by telmisartan --- p.190 / Chapter 5.3 --- Limitations of this study and future directions of research --- p.191 / Chapter Chapter 6 --- Conclusion --- p.194 / References --- p.198 / Appendices --- p.257

Para avaliar as repercussões da insuflação prolongada da cavidade peritoneal com gás carbônico sobre a hipertensão arterial essencial, utilizou-se ratos machos espontaneamente hipertensos (SHR) e como normotensos ratos machos Wistar-Kioto (WKY). No total foram utilizados 34 animais, sendo 22 SHRs e 12 WKYs, onde os ratos SHR foram distribuídos aleatoriamente aos grupos G1 e G3. O primeiro grupo (G1) com 12 animais SHRs e o segundo (G2) com 12 animais WKYs foram expostos a pneumoperitônio com dióxido de carbono por 120 minutos, enquanto que o terceiro grupo (G3) com 10 animais SHRs, passou por insuflação da cavidade peritoneal, seguida de punção com trocarte e esvaziamento do pneumoperitônio. Os animais deste grupo permaneceram anestesiados e com o abdome puncionado por 2 horas. Previamente a confecção do pneumoperitônio, a artéria e veia femorais direita foram dissecadas e canuladas. A artéria foi conectada ao transdutor de pressão para o registro contínuo da pressão arterial (PA), após a coleta inicial de 0,2 ml para dosagem da gasometria basal e 0,8 ml para as dosagens de uréia (U) e creatinina (Cr) basais. A veia femoral foi uttilizada para a expansão volêmica lenta com 10 ml de solução fisiológica após a coleta inicial de 1,0 ml de sangue arterial. Feito isto, procedeu-se a insuflação e punção abdominal mantendo ou não o pnemoperitônio, conforme o grupo. Foram feitas medidas da pressão arterial a cada 15 minutos e 5 minutos após o esvaziamento do abdome. Após a última aferição, foi colhido aproximandamente 3 ml de sangue arterial e 1 ml para a gasometria mais dosagem da U e Cr. A análise multivariada para medidas repetidas ao longo do tempo permitiu concluir que: nos cinco minutos após a desinsuflação, houve diferença estatística significante (p<0,0001) nas pressões arteriais sistólica, diastólica e média no G1 com uma curva ascendente em relação ao G2 e G3; O pH diminuiu (p<0,0001) de maneira similar nos três grupos de intervenção, enquanto a pCO2 aumentou (p<0,0001) de maneira similar nos três grupos de intervenção; não houve mudanças significativas na creatinina (p=0,3232); a uréia apresentou um efeito de momento com significância estatística (p<0,0001) e a atividade da renina plasmática foi significativamente maior no G2 em relação aos outros dois grupos / To assess the effects of prolonged peritoneal cavity insufflation with carbon dioxide on the essential hypertension, a experimental study was designed using male spontaneously hypertensive rats (SHR) and male normotensive Wistar-Kyoto (WKY). Thirty-four animals were used, 22 SHRs and 12 WKYs, where SHR rats were randomly assigned to groups G1 and G3. The first group (G1) with 12 animals SHRs and second group (G2) with 12 animals WKYs were exposed to pneumoperitoneum with carbon dioxide for 120 minutes, while the third group (G3) with 10 animals SHRs, had the peritoneal cavity insufflated, followed by puncture with trocarte and released the pneumoperitoneum. The animals of this group remained anesthetized and the abdomen punctured by 2 hours. Before making the pneumoperitoneum, right femoral artery and vein were dissected and cannulated. The artery was connected to the transducer pressure for the continuous recording of blood pressure (BP), after the initials blood samples: 0.2 ml for blood gases measurement and 0.8 ml for urea (U) and creatinine (Cr ). The femoral vein was used to volume expansion with 10 ml of saline solution after the initial sample of 1.0 ml arterial blood. Afterwards, a pnemoperitoneum insufflation and maintaining is done or not, depending on group. Blood pressure was recorded every 15 minutes and 5 minutes after pnemoperitoneum released. After last blood pressure record, a 3.0 ml blood sample was collected to measure plasma renin activity (PRA), and 1.0 ml for blood gases measurement, urea (U) and creatinine (Cr). The multivariate analysis for repeated measurements over time has concluded that: five minutes after pnemoperitoneum released, systolic, diastolic and mean blood pressure has significant statistic differences (p <0.0001) in G1 with an upward curve in relation to G2 and G3; The pH decreased (p <0.0001) in a similar way in the three groups of intervention, while pCO2 increased (p <0.0001) in a similar way in the three groups of intervention, with no significant changes in creatinine (p = 0.3232), but the urea had a moment effect with statistical significance (p <0.0001) and the plasma renin activity (PRA) was significantly higher in G2 compared with the other two groups

O emprego de \"Total Genome Scan\" em modelos genéticos de doenças complexas tem sido fundamental para seleção de regiões cromossômicas envolvidas com traços complexos. Em nosso laboratório, identificamos cinco regiões cromossômicas associadas ao traço quantitativo pressão arterial (BP-QTL) que explicam 43% da variação da pressão arterial numa progênie obtida a partir de animais espontaneamente hipertensos (SHR) e \"Brown Norway\" (BN). Os BP-QTLs foram, então, validados por desenvolvimento de linhagens congênicas, incluindo uma para o cromossomo 4 (SHR.BN4) cuja substituição das sequências SHR pelo do animal BN levou a redução da pressão arterial sistólica basal (~14 mm Hg). O objetivo deste trabalho foi identificar as variantes genéticas candidatas neste intervalo cromossômico com base em diferenças no padrão de expressão gênica e na presença de alterações genéticas não sinônimas \"missense\" ou em regiões regulatórias conservadas que possam estar envolvidas na gênese da hipertensão. Identificamos 533 genes com expressão renal, dentre os 682 do intervalo, sendo que 28 apresentaram padrão de expressão diferente entre amostras de animais adultos (congênico vs. SHR) e seis apresentaram alterações não sinônimas \"missense\". É importante salientar que dos genes diferentemente expressos, encontramos alterações estruturais em regiões conservadas com potencial de participar na regulação em 11. Em conjunto, utilizamos uma plataforma integrada para selecionar 34 genes candidatos no cromossomo 4, dos quais 17 genes serão priorizados, para ser investigados quanto sua contribuição na hipertensão arterial do SHR e na hipertensão primária humana / Total genome scan in genetic models of complex diseases have been instrumental to select candidate genes underlying complex traits. We previously mapped 5 blood pressure related quantitative trait loci (BP-QTLs) that explain about 43% of the BP variance in a progeny derived from Spontaneous Hypertensive Rat (SHR) and Brown Norway (BN) rats. The BP-QTLs were then validated by derivation of congenic strains, including one for chromosome 4 (SHR.BN4) in which a segment from BN replaced the SHR sequences reducing basal systolic BP (~14 mm Hg). The aim of this project is to identify the candidate genetic variants within the chromosome interval based on differences in renal gene expression patterns and structural changes in both non-synonymous missense or within adjacent regulatory sequences that may contribute to hypertension. We identified 533 genes with renal expression, out of 682 in the interval, in which 28 presented differences in expression pattern in adult samples (congenic vs. SHR) and six presented non-synonymous missense alterations. In addition, 11 out of 28 differentially expressed genes showed structural alterations in adjacent conserved regions that potentially contribute to gene regulation. Taken together, using the proposed combination of strategies, we selected 34 hypertensive candidate genes in chromosome 4, in which 17 will be prioritized, to be further explored to assess their contribution to hypertension in the SHR and to essential hypertension in humans

O emprego de \"Total Genome Scan\" em modelos genéticos de doenças complexas tem sido fundamental para seleção de regiões cromossômicas envolvidas com traços complexos. Em nosso laboratório, identificamos cinco regiões cromossômicas associadas ao traço quantitativo pressão arterial (BP-QTL) que explicam 43% da variação da pressão arterial numa progênie obtida a partir de animais espontaneamente hipertensos (SHR) e \"Brown Norway\" (BN). Os BP-QTLs foram, então, validados por desenvolvimento de linhagens congênicas, incluindo uma para o cromossomo 4 (SHR.BN4) cuja substituição das sequências SHR pelo do animal BN levou a redução da pressão arterial sistólica basal (~14 mm Hg). O objetivo deste trabalho foi identificar as variantes genéticas candidatas neste intervalo cromossômico com base em diferenças no padrão de expressão gênica e na presença de alterações genéticas não sinônimas \"missense\" ou em regiões regulatórias conservadas que possam estar envolvidas na gênese da hipertensão. Identificamos 533 genes com expressão renal, dentre os 682 do intervalo, sendo que 28 apresentaram padrão de expressão diferente entre amostras de animais adultos (congênico vs. SHR) e seis apresentaram alterações não sinônimas \"missense\". É importante salientar que dos genes diferentemente expressos, encontramos alterações estruturais em regiões conservadas com potencial de participar na regulação em 11. Em conjunto, utilizamos uma plataforma integrada para selecionar 34 genes candidatos no cromossomo 4, dos quais 17 genes serão priorizados, para ser investigados quanto sua contribuição na hipertensão arterial do SHR e na hipertensão primária humana / Total genome scan in genetic models of complex diseases have been instrumental to select candidate genes underlying complex traits. We previously mapped 5 blood pressure related quantitative trait loci (BP-QTLs) that explain about 43% of the BP variance in a progeny derived from Spontaneous Hypertensive Rat (SHR) and Brown Norway (BN) rats. The BP-QTLs were then validated by derivation of congenic strains, including one for chromosome 4 (SHR.BN4) in which a segment from BN replaced the SHR sequences reducing basal systolic BP (~14 mm Hg). The aim of this project is to identify the candidate genetic variants within the chromosome interval based on differences in renal gene expression patterns and structural changes in both non-synonymous missense or within adjacent regulatory sequences that may contribute to hypertension. We identified 533 genes with renal expression, out of 682 in the interval, in which 28 presented differences in expression pattern in adult samples (congenic vs. SHR) and six presented non-synonymous missense alterations. In addition, 11 out of 28 differentially expressed genes showed structural alterations in adjacent conserved regions that potentially contribute to gene regulation. Taken together, using the proposed combination of strategies, we selected 34 hypertensive candidate genes in chromosome 4, in which 17 will be prioritized, to be further explored to assess their contribution to hypertension in the SHR and to essential hypertension in humans

Para avaliar as repercussões da insuflação prolongada da cavidade peritoneal com gás carbônico sobre a hipertensão arterial essencial, utilizou-se ratos machos espontaneamente hipertensos (SHR) e como normotensos ratos machos Wistar-Kioto (WKY). No total foram utilizados 34 animais, sendo 22 SHRs e 12 WKYs, onde os ratos SHR foram distribuídos aleatoriamente aos grupos G1 e G3. O primeiro grupo (G1) com 12 animais SHRs e o segundo (G2) com 12 animais WKYs foram expostos a pneumoperitônio com dióxido de carbono por 120 minutos, enquanto que o terceiro grupo (G3) com 10 animais SHRs, passou por insuflação da cavidade peritoneal, seguida de punção com trocarte e esvaziamento do pneumoperitônio. Os animais deste grupo permaneceram anestesiados e com o abdome puncionado por 2 horas. Previamente a confecção do pneumoperitônio, a artéria e veia femorais direita foram dissecadas e canuladas. A artéria foi conectada ao transdutor de pressão para o registro contínuo da pressão arterial (PA), após a coleta inicial de 0,2 ml para dosagem da gasometria basal e 0,8 ml para as dosagens de uréia (U) e creatinina (Cr) basais. A veia femoral foi uttilizada para a expansão volêmica lenta com 10 ml de solução fisiológica após a coleta inicial de 1,0 ml de sangue arterial. Feito isto, procedeu-se a insuflação e punção abdominal mantendo ou não o pnemoperitônio, conforme o grupo. Foram feitas medidas da pressão arterial a cada 15 minutos e 5 minutos após o esvaziamento do abdome. Após a última aferição, foi colhido aproximandamente 3 ml de sangue arterial e 1 ml para a gasometria mais dosagem da U e Cr. A análise multivariada para medidas repetidas ao longo do tempo permitiu concluir que: nos cinco minutos após a desinsuflação, houve diferença estatística significante (p<0,0001) nas pressões arteriais sistólica, diastólica e média no G1 com uma curva ascendente em relação ao G2 e G3; O pH diminuiu (p<0,0001) de maneira similar nos três grupos de intervenção, enquanto a pCO2 aumentou (p<0,0001) de maneira similar nos três grupos de intervenção; não houve mudanças significativas na creatinina (p=0,3232); a uréia apresentou um efeito de momento com significância estatística (p<0,0001) e a atividade da renina plasmática foi significativamente maior no G2 em relação aos outros dois grupos / To assess the effects of prolonged peritoneal cavity insufflation with carbon dioxide on the essential hypertension, a experimental study was designed using male spontaneously hypertensive rats (SHR) and male normotensive Wistar-Kyoto (WKY). Thirty-four animals were used, 22 SHRs and 12 WKYs, where SHR rats were randomly assigned to groups G1 and G3. The first group (G1) with 12 animals SHRs and second group (G2) with 12 animals WKYs were exposed to pneumoperitoneum with carbon dioxide for 120 minutes, while the third group (G3) with 10 animals SHRs, had the peritoneal cavity insufflated, followed by puncture with trocarte and released the pneumoperitoneum. The animals of this group remained anesthetized and the abdomen punctured by 2 hours. Before making the pneumoperitoneum, right femoral artery and vein were dissected and cannulated. The artery was connected to the transducer pressure for the continuous recording of blood pressure (BP), after the initials blood samples: 0.2 ml for blood gases measurement and 0.8 ml for urea (U) and creatinine (Cr ). The femoral vein was used to volume expansion with 10 ml of saline solution after the initial sample of 1.0 ml arterial blood. Afterwards, a pnemoperitoneum insufflation and maintaining is done or not, depending on group. Blood pressure was recorded every 15 minutes and 5 minutes after pnemoperitoneum released. After last blood pressure record, a 3.0 ml blood sample was collected to measure plasma renin activity (PRA), and 1.0 ml for blood gases measurement, urea (U) and creatinine (Cr). The multivariate analysis for repeated measurements over time has concluded that: five minutes after pnemoperitoneum released, systolic, diastolic and mean blood pressure has significant statistic differences (p <0.0001) in G1 with an upward curve in relation to G2 and G3; The pH decreased (p <0.0001) in a similar way in the three groups of intervention, while pCO2 increased (p <0.0001) in a similar way in the three groups of intervention, with no significant changes in creatinine (p = 0.3232), but the urea had a moment effect with statistical significance (p <0.0001) and the plasma renin activity (PRA) was significantly higher in G2 compared with the other two groups