by Patricia Jane Anderson. / Thesis (M. Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 199-215). / List of Figures --- p.i / List of Tables --- p.v / List of Abbreviations --- p.viii / Abstract --- p.1 / Introduction --- p.3 / Chapter Chapter 1 - --- Literature Reviews / Chapter 1.1 --- Pharmacoanthropology and Pharmacogenetics --- p.5 / Chapter 1.1.1 --- Genetic Polymorphisms --- p.7 / Chapter 1.1.2 --- Pharmacogenetics in Asians and Caucasians --- p.13 / Chapter 1.1.2.1 --- ACE-inhibitors in Asians and Caucasians --- p.18 / Chapter 1.2 --- The Renin Angiotensin System --- p.20 / Chapter 1.2.1 --- Discovery of Inhibitors of Angiotensin Converting Enzyme --- p.24 / Chapter 1.3 --- ACE-Inhibiting Drugs --- p.25 / Chapter 1.3.1 --- Pharmacokinetics and Pharmacodynamics of Perindopril --- p.28 / Chapter 1.3.2 --- The Pharmacokinetics and Pharmacodynamics of Cilazapril --- p.32 / Chapter Chapter 2 - --- General Methodology / Chapter 2.1 --- Introduction --- p.38 / Chapter 2.2 --- Subjects --- p.49 / Chapter 2.3 --- Sample Collection --- p.40 / Chapter 2.3.1 --- Blood Samples --- p.40 / Chapter 2.3.2 --- Urine Samples --- p.40 / Chapter 2.4 --- Blood Pressure and Heart Rate Measurements --- p.41 / Chapter 2.5 --- Measurement of Transthoracic Electrical Bioimpedance --- p.41 / Chapter 2.5.1 --- Background --- p.42 / Chapter 2.5.2 --- Practical Details --- p.45 / Chapter 2.6 --- Data Analysis --- p.48 / Chapter 2.6.1. --- Analysis of Pharmacokinetic Parameters --- p.48 / Chapter 2.6.2 --- Analysis of Pharmacodynamic Parameters --- p.59 / Chapter 2.6.3 --- Analysis of Non-Invasive Haemodynamic Monitoring Data --- p.60 / Chapter 2.7 --- Statistical Analysis --- p.64 / Chapter Chapter 3 - --- The Perindopril Study / Chapter 3.1 --- Introduction --- p.67 / Chapter 3.1.1 --- Aims --- p.67 / Chapter 3.2 --- Methodology --- p.68 / Chapter 3.2.1 --- Inclusion Criteria --- p.68 / Chapter 3.2.2 --- Non-Inclusion Criteria --- p.69 / Chapter 3.2.3 --- Study Design --- p.69 / Chapter 3.2.4 --- Blood Sampling --- p.71 / Chapter 3.2.5 --- Urine Sampling --- p.71 / Chapter 3.2.6 --- Blood Pressure and Heart Rate --- p.72 / Chapter 3.2.7 --- Non-invasive Haemodynamic Monitoring --- p.72 / Chapter 3.2.8 --- Analysis of Plasma Samples --- p.73 / Chapter 3.2.9 --- Hormone and Enzyme Assays --- p.74 / Chapter 3.3 --- Data Analysis and Statistical Methods --- p.75 / Chapter 3.3.1 --- Pharmacokinetic Analysis of Plasma --- p.75 / Chapter 3.3.2 --- Pharmacokinetic Analysis of Urine --- p.75 / Chapter 3.3.3 --- Pharmacodynamic Analysis of Hormone Data --- p.75 / Chapter 3.3.4 --- Analysis of Haemodynamic Monitoring Data --- p.76 / Chapter 3.3.5 --- Statistical Analysis --- p.76 / Chapter 3.4 --- Pharmacokinetic Results --- p.77 / Chapter 3.4.1 --- Pharmacokinetics of Perindopril in Plasma --- p.77 / Chapter 3.4.2 --- Pharmacokinetics of Perindopril in Urine --- p.84 / Chapter 3.4.3. --- Pharmacokinetics of Perindoprilat in Plasma --- p.85 / Chapter 3.4.4 --- Pharmacokinetics of Perindoprilat in Urine --- p.89 / Chapter 3.5 --- Pharmacodynamic Results --- p.89 / Chapter 3.5.1 --- Angiotensin Converting Enzyme Inhibition --- p.89 / Chapter 3.5.2 --- Angiotensin I (AI) --- p.102 / Chapter 3.5.3 --- Aldosterone and Plasma Renin Activity (PRA) --- p.102 / Chapter 3.5.4 --- Plasma Protein Binding --- p.102 / Chapter 3.5.5 --- Blood Pressure and Heart Rate --- p.107 / Chapter 3.5.6. --- Safety and Tolerance --- p.108 / Chapter 3.5.7 --- Non-invasive Haemodynamic Monitoring --- p.108 / Chapter 3.6 --- Discussion --- p.120 / Chapter Chapter 4 - --- The Cilazapril Study / Chapter 4.1 --- Introduction --- p.135 / Chapter 4.1.1 --- Aims --- p.135 / Chapter 4.2 --- Methodology --- p.136 / Chapter 4.2.1 --- Inclusion Criteria --- p.136 / Chapter 4.2.2. --- Exclusion Criteria --- p.136 / Chapter 4.2.3 --- Study Design --- p.137 / Chapter 4.2.4 --- Blood Sampling --- p.139 / Chapter 4.2.5 --- Urine Sampling --- p.140 / Chapter 4.2.6 --- Blood Pressure and Heart Rate --- p.140 / Chapter 4.2.7 --- Non-Invasive Haemodynamic Monitoring --- p.140 / Chapter 4.2.8 --- Analysis of Plasma Cilazaprilat Samples --- p.142 / Chapter 4.2.9 --- Hormone and Enzyme Assays --- p.143 / Chapter 4.3 --- Data Analysis and Statistical Methods --- p.143 / Chapter 4.3.1 --- Pharmacokinetic Analysis --- p.143 / Chapter 4.3.2 --- Pharmacodynamic Analysis of Hormone Data --- p.144 / Chapter 4.3.3 --- Analysis of Non-Invasive Haemodynamic Monitoring Data --- p.144 / Chapter 4.3.4 --- Statistical Analysis --- p.146 / Chapter 4.4 --- Pharmacokinetic Results --- p.146 / Chapter 4.4.1 --- Pharmacokinetics of Cilazaprilat in Plasma --- p.146 / Chapter 4.5 --- Pharmacodynamic Results --- p.150 / Chapter 4.5.1 --- Angiotensin Converting Enzyme Inhibition --- p.150 / Chapter 4.5.2 --- Aldosterone and Plasma Renin Activity (PRA) --- p.155 / Chapter 4.5.3 --- Blood Pressure and Heart Rate --- p.155 / Chapter 4.5.4 --- Safety and Tolerance --- p.159 / Chapter 4.5.5 --- Non-Invasive Haemodynamic Monitoring --- p.160 / Chapter 4.6 --- Discussion --- p.182 / Chapter Chapter 5 - --- General Discussion --- p.188 / Appendix --- p.195 / References --- p.199 / Acknowledgements --- p.216

Wei, Lihua. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 166-196). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Tsang, Siu Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 107-121). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.x / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Renin-angiotensin system (RAS) --- p.1 / Chapter 1.1.1 --- Circulating RAS --- p.2 / Chapter 1.1.2 --- Tissue-specific RAS --- p.5 / Chapter 1.2 --- RAS inhibitors --- p.7 / Chapter 1.2.1 --- Angiotensin converting enzyme inhibitor --- p.8 / Chapter 1.2.2 --- Non-specific angiotensin II receptor blocker --- p.9 / Chapter 1.2.3 --- Specific AT1 receptor antagonist --- p.10 / Chapter 1.2.4 --- Specific AT2 receptor antagonist --- p.11 / Chapter 1.3 --- Pancreas and functions of exocrine pancreas --- p.14 / Chapter 1.3.1 --- Structure of pancreas --- p.14 / Chapter 1.3.2 --- Exocrine secretions and pancreatic enzymes --- p.16 / Chapter 1.3.3 --- Regulation of exocrine secretions --- p.17 / Chapter 1.4 --- Pancreatic RAS --- p.18 / Chapter 1.4.1 --- Expression and localization --- p.18 / Chapter 1.4.2 --- Regulation --- p.19 / Chapter 1.4.3 --- Clinical relevance to the pancreas --- p.20 / Chapter 1.5 --- Acute pancreatitis --- p.21 / Chapter 1.5.1 --- Pathogenesis --- p.21 / Chapter 1.5.2 --- Experimental models of acute pancreatitis --- p.22 / Chapter 1.5.3 --- Criteria of acute pancreatitis --- p.23 / Chapter 1.5.4 --- Oxidative stress in acute pancreatitis --- p.24 / Chapter 1.6 --- RAS and acute pancreatitis in exocrine pancreas --- p.26 / Chapter 1.6.1 --- RAS and acute pancreatitis --- p.26 / Chapter 1.6.2 --- RAS and pancreatic microcirculation --- p.26 / Chapter 1.6.3 --- RAS and tissue injury --- p.27 / Chapter 1.6.4 --- Exocrine pancreatic RAS and acute pancreatitis-induced injury --- p.28 / Chapter 1.7 --- Aims of study --- p.29 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Animal models and RAS inhibitors --- p.30 / Chapter 2.1.1 --- Cerulein-induced acute pancreatitis --- p.30 / Chapter 2.1.2 --- Prophylactic treatment with RAS inhibitors --- p.31 / Chapter 2.1.3 --- Therapeutic treatment with RAS inhibitors --- p.32 / Chapter 2.2 --- Evaluation of pancreatic injury --- p.32 / Chapter 2.2.1 --- Assessment of pancreatic water content --- p.33 / Chapter 2.2.2 --- Measurement of α-amylase activity in plasma --- p.33 / Chapter 2.2.3 --- Measurement of lipase activity in plasma --- p.34 / Chapter 2.3 --- Histopathological examinations --- p.34 / Chapter 2.3.1 --- Preparation of paraffin blocks --- p.35 / Chapter 2.3.2 --- Hematoxylin and eosin staining --- p.35 / Chapter 2.4 --- Biochemical assay of pancreatic oxidative status --- p.37 / Chapter 2.4.1 --- Sample preparation --- p.37 / Chapter 2.4.2 --- Quantification of protein content --- p.37 / Chapter 2.4.3 --- Measurement of glutathione levels --- p.38 / Chapter 2.4.4 --- Assessment of protein oxidation --- p.38 / Chapter 2.4.5 --- Assessment of lipid peroxidation --- p.39 / Chapter 2.4.6 --- Measurement of NADPH oxidase activity --- p.40 / Chapter 2.5 --- Studies of pancreatic digestive enzyme secretions from isolated acini --- p.40 / Chapter 2.5.1 --- Dissociation of acini from pancreatic tissue --- p.40 / Chapter 2.5.2 --- Treatment with peptides and RAS inhibitors --- p.42 / Chapter 2.5.3 --- Quantification of protein and DNA contents --- p.43 / Chapter 2.5.4 --- Measurement of a-amylase and lipase secretions --- p.44 / Chapter 2.5.5 --- RT-PCR analysis of RAS components in acinar cells --- p.44 / Chapter 2.6 --- Studies of RAS inhibitors on acute pancreatitis-induced systemic inflammation --- p.45 / Chapter 2.6.1 --- Systemic inflammation treatment --- p.45 / Chapter 2.6.2 --- Measurement of myeloperoxidase activity in lung and liver --- p.46 / Chapter 2.7 --- Statistical analysis --- p.47 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Time-course experiment of acute pancreatitis model --- p.48 / Chapter 3.1.1 --- Effect of acute pancreatitis on tissue injury --- p.48 / Chapter 3.1.2 --- Effects of acute pancreatitis on oxidative status --- p.48 / Chapter 3.2 --- Evaluation of ramiprilat and saralasin on changes of acute pancreatitis- induced pancreatic injury --- p.50 / Chapter 3.2.1 --- Changes in tissue injury and histopathology --- p.50 / Chapter 3.2.2 --- Changes in oxidative status --- p.57 / Chapter 3.3 --- Evaluation of losartan and PD123319 on changes of acute pancreatitis- induced pancreatic injury --- p.61 / Chapter 3.3.1 --- Changes in tissue injury and histopathology --- p.61 / Chapter 3.3.2 --- Changes in oxidative status --- p.68 / Chapter 3.4 --- Evaluation of acinar secretions of digestive enzymes --- p.71 / Chapter 3.4.1 --- Cholecystokinin octapeptide-induced acinar secretions --- p.71 / Chapter 3.4.2 --- Angiotensin II-induced acinar secretions --- p.71 / Chapter 3.4.3 --- Effects of losartan and PD 123319 on α-amylase secretion --- p.74 / Chapter 3.5 --- Existence and regulation of acinar RAS by acute pancreatitis --- p.75 / Chapter 3.5.1 --- Expression of angiotensinogen and its regulation by acute pancreatitis in acini --- p.76 / Chapter 3.5.2 --- Expression of AT1 receptor and its regulation by acute pancreatitis in acini --- p.76 / Chapter 3.5.3 --- Expression of AT2 receptor and its regulation by acute pancreatitis in acini --- p.76 / Chapter 3.5.4 --- Evaluation of RAS inhibitors in acute pancreatitis-induced acinar cells --- p.80 / Chapter 3.6 --- Preliminary data on acute pancreatitis-induced systemic inflammation --- p.81 / Chapter 3.6.1 --- Time-course experiment on lung injury --- p.81 / Chapter 3.6.2 --- Time-course experiment on liver injury --- p.83 / Chapter 3.6.3 --- Evaluation of losartan on systemic inflammation --- p.85 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- "Actions of RAS inhibitors on the changes of tissue injury, oxidative status and histopathology in acute pancreatitis-induced pancreas" --- p.87 / Chapter 4.1.1 --- Differential effects of ramiprilat and saralasin --- p.88 / Chapter 4.1.2 --- Differential effects of losartan and PD123319 --- p.92 / Chapter 4.2 --- Potential functions of RAS in pancreatic acinar secretions --- p.95 / Chapter 4.2.1 --- Potential role of AT1 receptor --- p.96 / Chapter 4.2.2 --- Potential role of AT2 receptor --- p.98 / Chapter 4.3 --- Regulation of RAS in acute pancreatitis-induced acini --- p.98 / Chapter 4.3.1 --- Regulation of RAS components in acinar cells --- p.99 / Chapter 4.3.2 --- Differential actions of losartan and PD123319 --- p.100 / Chapter 4.4 --- Potential role of RAS in acute pancreatitis --- p.102 / Chapter 4.4.1 --- Regulation of RAS components by acute pancreatitis --- p.102 / Chapter 4.4.2 --- Differential functions of AT1 and AT2 receptors in acute pancreatitis --- p.103 / Chapter 4.5 --- Conclusion --- p.104 / Chapter 4.6 --- Further studies --- p.105 / Chapter Chapter 5 --- Bibliography --- p.107

Banca: José Roberto de Oliveira e Silva / Banca:Wilma Pereira Bastos Ramos / Banca: Gerhard Malnic / Banca: Sonia Malheiros Lopes Sanioto / Banca: Frida Zaladek Gil / Resumo: Estudamos o efeito da Angiotensina II sobre a secreção de potásssio em túbulo distal final (segmento conector e duto coletor cortical) através da técnica de microperfusão estacionária in vivo e mensuração da atividade catiônica por meio de microeletrodos contendo resina de troca iônica sensível a K+. A perfusão luminal com ANG II reduziu o fluxo secretório de potássio (JK+) observado no grupo controle de 0.900.19 nmol/cm2.s, n=12, para 0.51±0.05, n=9, (ANG II 10-12M), 0.700.22, n=27 (ANG II 10-11M) e 0.630.08 nmol/cm2.s, n=12 (ANG II 10-9M); (p<0.05 teste t pareado). Entretanto, na presença de dose elevada de ANG II (10-6M) não observamos efeito significante sobre o JK+. A presença de Losartan (10-6M), um bloqueador não peptídico do receptor AT1 reverteu o efeito inibitório da ANG II. No intuito de se avaliar a possibilidade da via PLA2/ácido araquidônico/PGE2 participar deste processo de regulação celular, uma vez que tais agentes participam da inibição de outros mecanismos de transporte que envolve a ativação da sinalização celular mediada por Angiotensina II, perfundimos luminalmente PGE2 o qual inibiu o fluxo secretório de K+ em ambas doses utilizadas no presente trabalho; Jk+ controle = 0.930.08 nmol/cm2.s, n=12 para 0.550.05 nmol/cm2.s, n=12 (PGE2 10-9M) e 0.470.04 nmol/cm2.s, (PGE2 10-6M), n=12, (p<0.01). A perfusão com Indometacina (10-5M), bloqueador inespecífico da via PLA2/Ácido Araquidônico/PGE2 associado a Angiotensina II (10-9M) aumentou o JK+ (0.95±0.12 nmol.cm-2.s-1, n = 13) quando comparado a perfusão isolada de ANG II (10-9M) (Jk+ = 0.630.05 nmol/cm2.s, n = 10); (p<0.05). Concluímos que a ANG II inibiu luminalmente a secreção distal de K+ provavelmente acoplado ao receptor AT1 e este efeito pode ser mediado pela via PLA2/Acido Araquidônico/ /PGE2 / Abstract: The effect of luminal ANGII on K+ secretion by late distal tubule (connecting segment and initial cortical collecting duct) was studied using "in vivo" stationary microperfusion and K-sensitive microelectrode techniques. Luminal perfusion of ANG II reduced Jk+ from a control value of 0.900.19 (n=12) nmol/cm2.s to 0.51±0.05, n=9, (10-12M), 0.700.22, n=27 (10-11M) and 0.630.08, n=12 (10-9M) nmol/cm2.s (p<0.05 by paired t-test). However, high doses of ANGII (10-6M) had no significant effect on Jk+. Losartan 10-6M, a non-peptide AT1 receptor blocker, reverted the inhibitory effect of ANGII. To test the possibility that the PLA2/arachidonic acid/PGE2 pathway, which had been shown to inhibit other transport mechanisms, is involved in ANGII-mediated cellular signaling cascades, PGE2 was perfused luminally (Jk+ control = 0.930.08, n=12 nmol/cm2.s; 10-9M PGE2, Jk+ = 0.550.05, n-12; 10-6M PGE2, Jk+ = 0.470.04), n=12; both doses reduced K+ secretion significantly (p<0.01). Perfusing with Indomethacin, an unspecific blocker of the PLA2/arachidonic acid/PGE2 path, (10-5M), plus ANG II (10-9M), JK+ increased to 0.95±0.12 (n=13) nmol.cm-2.s-1 compared to ANG II alone (Jk+ = 0.630.05, (n=13) nmol/cm2.s, p<0.05). During luminal perfusion with Indomethacin alone, no significant effect on K+ secretion was seen (Jk+ control = 0.730.05 (n=10) nmol/cm2.s, 10-6M INDO Jk+ = 0.630.07 (n=10), P>0.19. In conclusion, ANG II is able to regulate distal K+ secretion when applied to the tubule lumen, probably via AT1 receptors; it is suggested that the signalling path of the inhibitory effect of ANG II may involve PLA2/arachidonic acid/PGE2

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 109-124).

Aging is associated with increases in regional and systemic vascular resistance and impaired ability to increase blood flow to active muscles during exercise. Aging enhances vasoconstrictor responsiveness in both humans and animals, and an increase in Angiotensin II-induced vasoconstriction is one possible mechanism for old age-associated increase in muscle vascular resistance. The purpose of this study was to determine 1) whether aging alters Ang II-induced vasoconstriction, 2) whether exercise training attenuates the age-associated alteration in Ang II-mediated vasoconstriction, and 3) the mechanism(s) through which aging and exercise training alter Ang II-induced vasoconstriction in rat skeletal muscle arterioles. Male Fischer 344 rats were assigned to 4 groups: Young sedentary (YS; 4 months), old sedentary (OS; 24 months), young trained (YT) and old trained (OT). Exercise-trained groups performed treadmill exercises for 60 min/day at 15 m/min, on a 15º incline for 5 days/week for 10-12 weeks. First-order (1A) arterioles were isolated from soleus and gastrocnemius muscles for in vitro experimentation. Intraluminal diameter changes were determined in response to the cumulative addition of Ang II (3×10-11 - 3×10-5 M). Ang II dose responses were then determined following the removal of endothelium and treatment with NG-nitro-L-arginine methyl ester (L-NAME, 10-5 M), a nitric oxide synthase (NOS) inhibitor. Ang II-induced vasoconstriction was augmented in the aged skeletal muscle arterioles, both in soleus and gastrocnemius muscles, and age-associated increases in Ang II-induced vasoconstriction were abolished with the removal of endothelium and with L-NAME. Exercise training ameliorated the age-induced increase in Ang II-vasoconstriction, and this alteration was eliminated by the removal of endothelium and with NOS inhibition. These findings suggest that aging enhances Ang II-induced vasoconstrictor responses in the arterioles from both soleus, high oxidative, and white portion of gastrocnemius, low oxidative glycolytic muscles, and this age-associated change occurs through an endothelium-dependent NOS signaling pathway. These results also demonstrated that exercise training can ameliorate the age-associated increase in Ang II vasoconstriction in the arterioles from both high oxidative and low oxidative glycolytic muscles through an endothelium-mediated NOS mechanism.

The renin-angiotensin system (RAS) is critical for the control of blood pressure (BP) and salt balance in mammals. Studies reveal that local RAS are present in the rat brain and renin is the first effector of the brain RAS for generating angiotensin II (Ang II) which exerts diverse physiological actions in both peripheral and central nervous system. The existence of renin within the brain has now been demonstrated by numerous studies. Previous studies suggest that renin may go through angiotensin-dependent and independent pathway to influence vascular tone, by Ang II type 1 receptor (AT1R) and renin specific (pro)renin receptor (PRR), respectively. Studies also indicate that AT1R and PRR are highly expressed in the nucleus tractus solitarii (NTS), which is important for central feedback regulation of BP. Further studies have shown that Ang II contributes to the release of NO, which plays an important role in cardiovascular regulation in the NTS. These results indicate that renin plays cardiovascular modulatory role in the NTS. However, the mechanisms how renin modulate cardiovascular functions in the NTS remained unclear. In the present study, I investigated the molecular mechanisms of renin-induced cardiovascular effects in the NTS. Unilateral microinjection of renin into the NTS of WKY rats produced prominent depressor and bradycardic effects. Pretreatment with a non-selective NOS inhibitor L-NAME, eNOS specific inhibitor L-NIO, Akt inhibitor IV, and PI3K inhibitor LY294002 significantly attenuated the cardiovascular response evoked by renin, whereas nNOS specific inhibitor Vinyl-L-NIO and MEK inhibitor PD98059 did not cause significant changes. Western blot studies showed renin increased eNOSS1177 and AktS473 phosphorylation instead of nNOSS1416 and ERK1/2T202/Y204 phosphorylation, and pretreatment with LY294002 blocked renin-induced eNOSS1177 and AktS473 phosphorylation. These results indicated that renin might go through PI3K-Akt-eNOS pathway to increase eNOS activity and ultimately result in NO release. The cardiovascular effects of renin were also attenuated by renin specific inhibitor aliskiren, angiotensin converting enzyme inhibitor lisinopril, AT1R antagonist losartan, and intracellular Ca2+ chelator, BAPTA-AM instead of G protein £]£^ subunit inhibitor gallein, PLC inhibitor U73122, calmodulin inhibitor (W-7) and (pro)renin receptor blocker, handle region peptide. These results indicated that renin mainly through AT1R to regulate BP. Therefore, my results indicated that the modulation of cardiovascular effects of renin in the NTS involves AT1R-PI3K-Akt pathway to activate eNOS activation.

Aging is associated with increases in regional and systemic vascular resistance and impaired ability to increase blood flow to active muscles during exercise. Aging enhances vasoconstrictor responsiveness in both humans and animals, and an increase in Angiotensin II-induced vasoconstriction is one possible mechanism for old age-associated increase in muscle vascular resistance. The purpose of this study was to determine 1) whether aging alters Ang II-induced vasoconstriction, 2) whether exercise training attenuates the age-associated alteration in Ang II-mediated vasoconstriction, and 3) the mechanism(s) through which aging and exercise training alter Ang II-induced vasoconstriction in rat skeletal muscle arterioles. Male Fischer 344 rats were assigned to 4 groups: Young sedentary (YS; 4 months), old sedentary (OS; 24 months), young trained (YT) and old trained (OT). Exercise-trained groups performed treadmill exercises for 60 min/day at 15 m/min, on a 15º incline for 5 days/week for 10-12 weeks. First-order (1A) arterioles were isolated from soleus and gastrocnemius muscles for in vitro experimentation. Intraluminal diameter changes were determined in response to the cumulative addition of Ang II (3×10-11 - 3×10-5 M). Ang II dose responses were then determined following the removal of endothelium and treatment with NG-nitro-L-arginine methyl ester (L-NAME, 10-5 M), a nitric oxide synthase (NOS) inhibitor. Ang II-induced vasoconstriction was augmented in the aged skeletal muscle arterioles, both in soleus and gastrocnemius muscles, and age-associated increases in Ang II-induced vasoconstriction were abolished with the removal of endothelium and with L-NAME. Exercise training ameliorated the age-induced increase in Ang II-vasoconstriction, and this alteration was eliminated by the removal of endothelium and with NOS inhibition. These findings suggest that aging enhances Ang II-induced vasoconstrictor responses in the arterioles from both soleus, high oxidative, and white portion of gastrocnemius, low oxidative glycolytic muscles, and this age-associated change occurs through an endothelium-dependent NOS signaling pathway. These results also demonstrated that exercise training can ameliorate the age-associated increase in Ang II vasoconstriction in the arterioles from both high oxidative and low oxidative glycolytic muscles through an endothelium-mediated NOS mechanism.

In this study, we investigated the role of angiotensinergic neurotransmission at nucleus reticularis ventrolateralis (NRVL), and the subtype of angiotensin receptors involved, during experimental endotoxemia induced by E. coli lipopolysaccharide (LPS). In adult male Sprague-Dawley rats maintained under propofol anesthesia (30 mg/kg/h), paralyzed with pancuronium (2 mg/kg/h) and mechanically ventilated (85-95 stroke/min, 2.5-3 ml/stroke), intravenous administration of LPS (15 or 30 mg/kg) induced an immediate hypotension, followed by a rebound increase and a secondary decrease in systemic arterial pressure (SAP). LPS also reduced the power density of the very low-frequency (0-0.25 Hz) and low-frequency (0.25-0.8 Hz) components of SAP signals (Phase ¢¹), which represented the sympathetic vasomotor tone, followed by an increase (Phase ¢º) and a secondary decrease (Phase ¢»). Pretreatment with microinjection of the selective non-peptide AT1 receptor antagonist, losartan (1.6 nmol), or the selective non-peptide AT2 receptor antagonist, PD-123319 (1.6 nmol), into the bilateral NRVL significantly reduced the survival time after the induction of acute experimental endotoxemia. Both pretreatments shortened the duration of Phase ¢º and Phase ¢» in acute endotoxemia, accelerated the secondary hypotension, and excited the power density of the very low-frequency. We conclude that endogenous angiotensin ¢º at the NRVL may play a crucial role in the maintenance of SAP during acute experimental endotoxemia, possibly via an action on both AT1 and AT2 subtype receptors on the very low-frequency component of SAP spectrum.

The rostral ventrolateral medulla (RVLM) is the origin of a ¡§life-and-death¡¨ signal identifies from systemic arterial blood pressure spectrum that reflects failure of central cardiovascular regulation during brain stem death. It is also a target site where endogenous angiotensin II acts on angiotensin II type 1 receptors (AT1R) to increase blood pressure (BP); or on type 2 receptors (AT2R) to inhibit baroreceptor reflex (BRR) response. This study investigated the roles of AT1R and AT2R and their signaling pathways in RVLM for ¡§life-and-death¡¨ signal response during experimental brain stem death, using organophosphate mevinphos (Mev) as the experimental insult. In Sprague-Dawley rats, Mev (640 £gg/kg, i.v.) elicited an increase (pro-life phase) followed by a decrease (pro-death phase). Real-time PCR analysis revealed that whereas AT1R level underwent a 10% increase at pro-life phase, AT2R exhibited a significance increase of up to 40% at pro-death phase. Western blot analysis revealed that whereas AT1R level underwent a 20% increase at pro-life phase, AT2R exhibited a significant increase of up to 50% at pro-death phase. Pretreatment with microinjection of an AT1R antagonist losartan (2 nmol) into RVLM elicited abrupt death because of drastic hypotension through inhibiting NADPH oxidase and its downstream superoxide anion. Pretreatment with NADPH oxidase inhibitor DPI (1.5 nmol) inhibited NADPH oxidase avtiviting and superoxide anion production and decreased ¡§life-and-death¡¨ signal at pro-life phase; using superoxide anion inhibitor tempol (5 nmol) potentiated blood pressure and ¡§life-and-death¡¨ signal at pro-death phase. However, pretreatment with an AT2R antagonist PD123319 (2 nmol) potentiated the ¡§life-and-death¡¨ signal and antagonized hypotension during pro-death phase through inhibiting protein phosphotase 2A (PP2A) then activating extracellular signal-regulated kinase 1/2 (ERK1/2). Similar to AT2R antagonist PD123319, pretreatment with PP2A inhibitor okadaic acid (0.5 fmol) inhibit PP2A, leading to activation of ERK1/2, potentiate ¡§life-and-death¡¨ signal and antagonized hypotension during pro-death phase. These results suggest that AT1R in RVLM plays a ¡§pro-life¡¨ role through NADPH oxidase/superoxide anion during experimental brain stem death by maintaining BP and ¡§life-and-death¡¨ signal; AT2R plays a ¡§pro-death¡¨ role through PP2A/ERK1/2 by inhibiting BP and ¡§life-and-death¡¨ signal, and superoxide may also plays a ¡§pro-life and pro-death¡¨ role at pro-death phase.