Objetivo: Foi investigada a hipótese da hiperprolactinemia modular a resposta inflamatória alérgica pulmonar em ratos machos e em fêmas lactantes sem tratamento de domperidona. Métodos: Em ratos machos, a hiperprolactinemia foi de curta duração (5 dias) induzida pela domperidona (5,1 mg.kg-1 por dia, i.p). A resposta alérgica foi gerada por sensibilização e desafios inalatórios com ovoalbumina. Foi feita contagem de leucócitos totais e diferenciados do lavado bronco alveolar (BAL), lavado medular femoral (BFL) e sangue; a percentagem de produção de muco e colageno no pulmão, níveis de corticosterona e prolactina e citocinas TNF-α, IL-4, IL-6, IL-10, em explantes de pulmão e IFNg no BAL, foram medidos. Pela citometria foram avaliadaos os receptores de prolactina; Resultados: Hiperprolactinemia de curta duração feita antes do desafio inalatório disminuiu a resposta alérgica pulmonar na contagem de leucócitos no lavado broncoalveolar. Esse tratamento reduziu a celularidade no BFL e a percentagem de muco e aumentou a expressão de citocinas IL-4, IL-6, IL-10, TNFα e da expressão do IFNg. Níveis altos de prolactina diminuiram o número de eosinófilos ao pulmão no BAL. Pela citometria revelou-se que além de ter menor número de granulócitos migrados ao pulmão, estes apresentaram maior expressão do número de receptores por granulócito para prolactina no grupo tratado com domperidona. Alterações similares foram reveladas em fêmeas lactantes como foi a diminuição nos leucócitos do BAL, e no número de células do BFL. O tratamento profilático diminuiu a resposta alérgica tanto no grupo hiperprolactinêmico como no grupo veículo. O tratamento feito após o desafio inalatório não evidenciou alterações relevantes nas variáveis medidas. Conclusões: A hiperprolactinemia de curta duração, feita após a sensibilização e antes da inalação diminui a resposta inflamatória no pulmão em ratos. Os resultados deste estudo demonstram que a hiperprolactinemia induzida antes do desafio antigênico diminue a inflamação alérgica pulmonar. Assim, é provável que a prolactina endógena tenha um papel relevante como um imunomodulador da asma. Este estudo aponta a possibilidade futura do uso da domperidona para pacientes asmáticos. Durante a primavera muitas espécies de mamíferos têm seus filhotes e ocorre abundância de fatores alergenos no ar. Logo, um fator endógeno que favoreça a proteção de fêmeas durante a lactação, tal como a hiperprolactinemia, tem elevado valor adaptativo / Objective: It was investigated if hyperprolactinemia has modulatory actions on lung allergic inflammatory response in male rats. Lactating female rats were tested for pulmonary allergy as well. Methods: In male rats, short-term (5 days) hyperprolactinemia was induced by domperidone (5.1 mg.kg-1 per day, ip). Allergic response was generated by sensitization and inhalation challenge with ovalbumin. Total and differential leukocytes bronchoalveolar lavage (BAL), femoral medullary lavage (BFL) and blood; the percentage of collagen and mucus production in the lungs, plasma levels of corticosterone and prolactin cytokines and TNF-α, IL-4, IL-6, IL-10, IFNg explants lung and BAL, were measured. Flow cytometry was used to evaluate prolactin receptor; Results: Short-term hyperprolactinemia made before the inhaled challenge reduced the pulmonary allergic response in white blood cell counts in BAL. This treatment reduced the cellularity in BFL and the percentage of mucus and increased expression of cytokines IL-4, IL-6, IL-10, TNFa and IFNg expression. High prolactin levels decreased the number of eosinophils to the lung in BAL. There were fewer granulocytes migrated to the lung. These granulocytes showed higher expression prolactin receptors in hyperprolactinemia animals. Similar changes were revealed in lactating females. In these animals, there was a reduction in BAL leukocyte, and the number of cells BFL. Prophylactic treatment decreased the allergic response in both hyperprolactinemic and vehicle groups. The treatment made after inhalational challenge did not induce significant changes in the variables measured in this study. Conclusions: Short-term hyperprolactinemia, made after sensitization and before inhalation, decreases the inflammatory response in the lung of rats. The results of this study demonstrate that hyperprolactinemia, induced before antigen challenge, decreases pulmonary allergic inflammation. Thus, it is probable that the endogenous prolactin has an important role as an immunomodulator of asthma. This study points out the prospect of a future use of domperidone for asthmatic patients. For various mammalian species, parturition occurs during springtime. Pollen in the air might be an abundant allergic factor during springtime. Thus, protecting lactating females against this type allergy might have high adaptive value

Objetivo: Foi investigada a hipótese da hiperprolactinemia modular a resposta inflamatória alérgica pulmonar em ratos machos e em fêmas lactantes sem tratamento de domperidona. Métodos: Em ratos machos, a hiperprolactinemia foi de curta duração (5 dias) induzida pela domperidona (5,1 mg.kg-1 por dia, i.p). A resposta alérgica foi gerada por sensibilização e desafios inalatórios com ovoalbumina. Foi feita contagem de leucócitos totais e diferenciados do lavado bronco alveolar (BAL), lavado medular femoral (BFL) e sangue; a percentagem de produção de muco e colageno no pulmão, níveis de corticosterona e prolactina e citocinas TNF-α, IL-4, IL-6, IL-10, em explantes de pulmão e IFNg no BAL, foram medidos. Pela citometria foram avaliadaos os receptores de prolactina; Resultados: Hiperprolactinemia de curta duração feita antes do desafio inalatório disminuiu a resposta alérgica pulmonar na contagem de leucócitos no lavado broncoalveolar. Esse tratamento reduziu a celularidade no BFL e a percentagem de muco e aumentou a expressão de citocinas IL-4, IL-6, IL-10, TNFα e da expressão do IFNg. Níveis altos de prolactina diminuiram o número de eosinófilos ao pulmão no BAL. Pela citometria revelou-se que além de ter menor número de granulócitos migrados ao pulmão, estes apresentaram maior expressão do número de receptores por granulócito para prolactina no grupo tratado com domperidona. Alterações similares foram reveladas em fêmeas lactantes como foi a diminuição nos leucócitos do BAL, e no número de células do BFL. O tratamento profilático diminuiu a resposta alérgica tanto no grupo hiperprolactinêmico como no grupo veículo. O tratamento feito após o desafio inalatório não evidenciou alterações relevantes nas variáveis medidas. Conclusões: A hiperprolactinemia de curta duração, feita após a sensibilização e antes da inalação diminui a resposta inflamatória no pulmão em ratos. Os resultados deste estudo demonstram que a hiperprolactinemia induzida antes do desafio antigênico diminue a inflamação alérgica pulmonar. Assim, é provável que a prolactina endógena tenha um papel relevante como um imunomodulador da asma. Este estudo aponta a possibilidade futura do uso da domperidona para pacientes asmáticos. Durante a primavera muitas espécies de mamíferos têm seus filhotes e ocorre abundância de fatores alergenos no ar. Logo, um fator endógeno que favoreça a proteção de fêmeas durante a lactação, tal como a hiperprolactinemia, tem elevado valor adaptativo / Objective: It was investigated if hyperprolactinemia has modulatory actions on lung allergic inflammatory response in male rats. Lactating female rats were tested for pulmonary allergy as well. Methods: In male rats, short-term (5 days) hyperprolactinemia was induced by domperidone (5.1 mg.kg-1 per day, ip). Allergic response was generated by sensitization and inhalation challenge with ovalbumin. Total and differential leukocytes bronchoalveolar lavage (BAL), femoral medullary lavage (BFL) and blood; the percentage of collagen and mucus production in the lungs, plasma levels of corticosterone and prolactin cytokines and TNF-α, IL-4, IL-6, IL-10, IFNg explants lung and BAL, were measured. Flow cytometry was used to evaluate prolactin receptor; Results: Short-term hyperprolactinemia made before the inhaled challenge reduced the pulmonary allergic response in white blood cell counts in BAL. This treatment reduced the cellularity in BFL and the percentage of mucus and increased expression of cytokines IL-4, IL-6, IL-10, TNFa and IFNg expression. High prolactin levels decreased the number of eosinophils to the lung in BAL. There were fewer granulocytes migrated to the lung. These granulocytes showed higher expression prolactin receptors in hyperprolactinemia animals. Similar changes were revealed in lactating females. In these animals, there was a reduction in BAL leukocyte, and the number of cells BFL. Prophylactic treatment decreased the allergic response in both hyperprolactinemic and vehicle groups. The treatment made after inhalational challenge did not induce significant changes in the variables measured in this study. Conclusions: Short-term hyperprolactinemia, made after sensitization and before inhalation, decreases the inflammatory response in the lung of rats. The results of this study demonstrate that hyperprolactinemia, induced before antigen challenge, decreases pulmonary allergic inflammation. Thus, it is probable that the endogenous prolactin has an important role as an immunomodulator of asthma. This study points out the prospect of a future use of domperidone for asthmatic patients. For various mammalian species, parturition occurs during springtime. Pollen in the air might be an abundant allergic factor during springtime. Thus, protecting lactating females against this type allergy might have high adaptive value

Gastroparesis is a disorder characterized by delayed gastric emptying due to chronic abnormal gastric motility. Prokinetic agents such as domperidone and metoclopramide are the cornerstone in treatment of gastroparesis. Although these medications have been used for decades, essential information about their metabolism is not available. Lack of knowledge about the metabolites formed in the body upon administration of the aforementioned medications as well as the enzymes involved in their metabolism limits key information needed to make sound medical decisions. Accurate and comprehensive identification of the metabolites along with reaction phenotyping of prokinetic agents will ensure safe and effective use of these drugs and hence enhance the clinical outcome. The thesis starts with an introductory chapter which comprises a comprehensive literature review on gastroparesis and the available pharmacological treatment options. The chapter also emphasizes the importance of metabolic profiling of prokinetic agents (domperidone and metoclopramide) and its impact on enhancing the safety and efficacy of these medications. Chapter 2 of this project was aimed to determine phase oxidative and conjugative metabolites of domperidone in the plasma and urine of gastroparesis patients using tandem mass spectrometry. First, the metabolites were identified in in-vitro human subcellular fractions. The knowledge gained in this experiment helped identifying the metabolites in the biological fluids of patients. In total, 12 metabolites including 7 new metabolites were identified, 5 of which were not reported previously. Chapter 3 aimed to identify the cytochrome P450 (CYP) enzymes responsible for the metabolism of metoclopramide. The parent depletion approach was used and a novel LC-MS/MS method was developed and validated to enable metoclopramide quantification. CYP2D6 was showed to the predominant isoform in metoclopramide metabolism; other isoforms also contribute to a minor extent. Chapter 4 discusses the possibility of potential drug-drug interaction (DDI) in the current management practice of gastroparesis. We identified and investigated some frequently used drug combinations that are known to share common metabolic pathways. Domperidone in combination with pioglitazone and ondansetron was evaluated. Results showed that pioglitazone inhibited domperidone metabolism in-vitro. Our experiments did not predict a DDI for the domperidone - ondansetron combination. In summary, the ultimate goal of this thesis was to improve the management of gastroparesis by increasing information about the metabolic disposition of prokinetic agents and to investigate the magnitude of putative drug combinations. The knowledge provided by this work will help in making more effective and less hazardous clinical decisions which will ultimately lead to more successful gastroparesis management. / Pharmaceutical Sciences

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal