Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Background: Endothelial dysfunction is a key factor in the development of vascular complications in insulin resistance and diabetes and recent studies have established that endothelium-derived hyperpolarising factor (EDHF) plays an important physiological role in endothelium-derived relaxation responses, especially in small arteries and arterioles. Objective: This project aimed to identify the role of, and characterise, EDHF in animal models of insulin resistance, including the obese Zucker rat (OZR) as well as the fructose-fed (FF) Sprague-Dawley rat. Methods: Vascular function was studied in third-order mesenteric arteries from male and female Zucker rats using pressure myography, and in lobar arteries from male FF rats using wire myography. Endothelial function was determined by studying responses to the endothelium-dependent dilator acetylcholine (ACh) and the endothelium-independent dilator levcromakalim in the presence of a variety of inhibitors to study the roles of NO, EDHF and gap junctions. The effect of insulin resistance on gap junctions was further assessed by measuring the protein and mRNA expression of vascular connexins. Protein levels were determined by western blotting followed by semi-quantitative analysis of band intensity, whilst mRNA levels were quantified using real-time PCR, in which beta-actin was used as the housekeeping gene. Results: Metabolic parameter comparisons confirmed that male OZRs were type 2 diabetic, whilst female OZRs were insulin resistant. Responses to ACh were reduced in both the male and female OZRs compared with their gender controls, with the male OZR showing a greater degree of endothelial dysfunction. In all Zucker third-order mesenteric arteries, inhibition of NO had no effect; however inhibitors of EDHF abolished relaxation responses to ACh. Inhibitors of gap junctions associated with connexin 40 significantly (p less than 0.05, Student's t-test) attenuated the maximal response to ACh in the LZR, but had no effect in the OZR. Comparison of Western blot band intensity indicated that connexin 40 protein levels in mesenteric vascular homogenates in the OZR were significantly smaller (p less than 0.05, Student's t-test) than in the LZR, with no difference in connexin 43 protein levels. mRNA levels showed a significant (p less than 0.05, Student's t-test) decrease in connexin 40 expression in the OZR compar ed with the LZR, with no change in connexin 43 mRNA expression. Although FF rats did develop insulin resistance, responses to ACh were not altered in the FF rats as compared with their controls, and ACh responses were abolished by NO inhibitors. Conclusion: The findings presented in this thesis demonstrate that endothelial dysfunction is present in third-order mesenteric arteries from insulin-resistant female and type 2 diabetic male OZRs, and is associated with a defect in EDHF. However, endothelial function was not compromised in the insulin-resistant FF rats. Furthermore, the reduction in EDHF-mediated vasodilatation in the mesenteric arteries from female OZRs was associated with the functional absence of connexin 40-related gap junctions as well as a reduction in connexin 40 protein and mRNA levels. This novel finding suggests that gap junctions associated with connexin 40 may be a potential therapeutic target for diabetic vascular disease.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Orientador: Marcos Renato de Assis / Resumo: A expectativa de vida diminui com artrite reumatoide (AR) devido ao aumento da mortalidade por doenças cardiovasculares. Isto se deve, possivelmente, à disfunção endotelial resultante da intensa atividade inflamatória relacionada à AR. Evidências tem apontado para um possível envolvimento do sistema renina-angiotensina (SRA) nos danos cardiovasculares inerentes à AR. Acredita-se que a participação do SRA agrave o comprometimento endotelial decorrente de inflamações sistêmicas através da ativação do receptor de angiotensina II tipo 1 (AT1) pela angiotensina II (Ang II). Por outro lado, a literatura também reporta a influência dos hormônios andrógenos, principalmente da testosterona, nas ações da Ang II sobre os tecidos vasculares. Assim, o objetivo do presente estudo é investigar os efeitos da artrite induzida por adjuvante (AIA) sobre o equilíbrio redox, a função endotelial e as respostas da aorta de ratos à Ang II, bem como, verificar se esses efeitos podem ser influenciados pela redução dos níveis circulantes de testosterona. Para isto, ratos Wistar machos adultos foram submetidos à falsa-castração e falsa-imunização (Controles), castração seguida de falsa-imunização (ORX), falsa-castração seguida de imunização (ORX) e castração seguida de imunização (ORX+AIA). Ao final do experimento, segmentos de aorta torácica foram desafiadas em cubas de órgãos isolados com acetilcolina (ACh), Ang II, KCl e nitroprussiato de sódio e, das curvas concentração resposta obtidas, calculou-se... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Life expectancy of patients with rheumatoid arthritis (RA) is lower due to increased mortality in consequence of cardiovascular diseases. Presumably, this occurs due to endothelial dysfunction resulting from severe inflammatory activity related to RA. Evidence has demonstrated a possible involvement of the renin-angiotensin system (RAS) in the cardiovascular injury inherent to RA. The RAS involvement is considered to worsen the endothelial impairment due to systemic inflammation through angiotensin II type 1 receptor (AT1) activation of Ang II. (Ang II). Additionally, previous studies also observed that androgen hormones, mainly the testosterone, modulate the Ang II actions in cardiovascular tissues. Thus, the aim of the present study is to investigate the effects of adjuvant-induced arthritis (AIA) on systemic redox balance, endothelial function and rat aorta responses to Ang II, as well as, whether these effects may be influenced by the reduction of circulating levels of testosterone. For this, adult male Wistar rats were submitted to false-castration and false-immunization (Controls), castration followed by false-immunization (ORX), false-castration followed by immunization (ORX) and castration followed by immunization (ORX + AIA) . At the end of the experiment, thoracic aorta segments were challenged in isolated organ bath with acetylcholine (ACh), Ang II, KCl and sodium nitroprusside and, from the concentrantion-response curves obtained were calculated pEC50 and maximal ... (Complete abstract click electronic access below) / Doutor

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