<p>Smoking is a worldwide problem and it is the second major cause of death. People often try to quit, but few succeed mainly because of withdrawal symptoms such as irritability, anxiety, increased appetite, hyperventilation and difficulty concentrating.</p><p>The overall aim of this project was to study neurochemical changes in the brain following sensitization to nicotine which could give more information about what causes an individual to go from using drugs to abusing the drugs. Therefore, we investigated messenger ribonucleic acid (mRNA) expression of several genes known to be involved in the mesolimbic dopamine pathway in the nucleus accumbens, caudate putamen, prefrontal cortex and medial prefrontal cortex using real-time polymerase chain reaction (real-time PCR).</p><p>The results showed that in the nucleus accumbens, mRNA expression of gamma-aminobutyric acid (GABA) Aα1 subunit receptor and GABA transporter 3 (GAT-3) were significantly increased following nicotine administration, while in the caudate putamen no difference in expression was observed. In prefrontal cortex, the expression of adrenergic subunit receptor α2A was significantly increased following hexamethonium administration. In medial prefrontal cortex a significant decrease of expression of GAT-1 was shown following nicotine and hexamethonium administration, while a decrease of CART expression only was shown following nicotine administration.</p><p>Overall, these changes in the GABA system may help to explain the mechanism of nicotine sensitization.</p>

Smoking is a worldwide problem and it is the second major cause of death. People often try to quit, but few succeed mainly because of withdrawal symptoms such as irritability, anxiety, increased appetite, hyperventilation and difficulty concentrating. The overall aim of this project was to study neurochemical changes in the brain following sensitization to nicotine which could give more information about what causes an individual to go from using drugs to abusing the drugs. Therefore, we investigated messenger ribonucleic acid (mRNA) expression of several genes known to be involved in the mesolimbic dopamine pathway in the nucleus accumbens, caudate putamen, prefrontal cortex and medial prefrontal cortex using real-time polymerase chain reaction (real-time PCR). The results showed that in the nucleus accumbens, mRNA expression of gamma-aminobutyric acid (GABA) Aα1 subunit receptor and GABA transporter 3 (GAT-3) were significantly increased following nicotine administration, while in the caudate putamen no difference in expression was observed. In prefrontal cortex, the expression of adrenergic subunit receptor α2A was significantly increased following hexamethonium administration. In medial prefrontal cortex a significant decrease of expression of GAT-1 was shown following nicotine and hexamethonium administration, while a decrease of CART expression only was shown following nicotine administration. Overall, these changes in the GABA system may help to explain the mechanism of nicotine sensitization.

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-21T17:52:14Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Ana Paula Gai Fuão.pdf: 1243240 bytes, checksum: d40fbf45a1852ec6eebfa3e33bb9bcc3 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-21T17:53:51Z (GMT) No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Ana Paula Gai Fuão.pdf: 1243240 bytes, checksum: d40fbf45a1852ec6eebfa3e33bb9bcc3 (MD5) / Made available in DSpace on 2016-09-21T17:53:51Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Ana Paula Gai Fuão.pdf: 1243240 bytes, checksum: d40fbf45a1852ec6eebfa3e33bb9bcc3 (MD5) Previous issue date: 2014-08-21 / A hipertensão arterial sistêmica é fundamentada como uma condição clínica multifatorial caracterizada por níveis elevados e sustentados de pressão arterial. A progressão da doença está intimamente ligada com alterações estruturais e funcionais cardíacas e vasculares além de promover danos em alguns órgãos alvo, como os rins, cérebro, além das alterações metabólicas, que produzem um aumento sensível nos riscos de doenças cardiovasculares. De acordo com a Sociedade Brasileira de Cardiologia a hipertensão arterial sistêmica apresenta uma alta prevalência e baixas taxas de controle, tornando-a um dos mais importantes problemas de saúde pública no Brasil. O uso de plantas medicinais para fins terapêuticos é bastante antigo e muitas vezes seu uso é absolutamente empírico, sem nenhum estudo científico que embase essa prática. A Blepharocalyx salicifolius, conhecida como Murta, apresenta várias utilidades terapêuticas baseadas no conhecimento popular, entre elas o uso como digestivo, antibacteriano, antiespasmódico e anti-hipertensivo, entre outros. No presente trabalho foi avaliado o efeito hipotensor do extrato aquoso da B. salicifolius em ratos normotensos, e analisado, através do uso do L-NAME, da losartan e do hexametônio, qual seria seu possível mecanismo de ação. A avaliação dos efeitos hemodinâmicos foi realizada em ratos wistar normotensos, anestesiados com uretana (1,4 mg/Kg i.p.) e via canulação da artéria carótida (para a verificação da PAS, PAD e FC) e da veia jugular (para administração do extrato e drogas). O primeiro protocolo foi realizado para verificação do efeito do extrato sobre os parâmetros hemodinâmicos, utilizando uma curva dose-crescente do extrato (10, 25, 50, 75 e 100 mg/Kg) via intravenosa individualmente (cada grupo contendo 6 animais). O extrato aquoso foi administrado após 30 minutos de estabilização no equipamento, e permaneceram por 60 min. O segundo protocolo foi para avaliar alguns dos possíveis mecanismos envolvidos na resposta hipotensora. Foram utilizados: um inibidor da óxido nítrico sintase - L-NAME (L-NG-nitroarginina metil ester) (30 mg/kg); um antagonista do receptor AT1 - Losartan, (10 mg/Kg), e um bloqueador ganglionar, Hexamêtonio (20mg/Kg). As drogas foram administradas após a estabilização, sendo feito uma nova estabilização para a administração do extrato (concentração 7,5 mg/Kg; volume de 200 μL) e registros realizados por 60 min. O extrato aquoso da B. salicifolius mostrou-se um potente hipotensor, reduzindo a pressão arterial de forma dose-independente, já que as quedas foram semelhantes indiferentemente da dose usada, sem provocar alterações na frequência cardíaca. Na avaliação dos possíveis mecanismos de ação foi observado que o bloqueio do receptor AT1 e administrado o extrato aquoso não houve modificação na hipotensão provocada pelo Losartan bem como após o bloqueio ganglionar com o hexametônio. Já quando induzida a hipertensão por L-NAME, o extrato aquoso causou queda de pressão arterial, sugerindo que a via do óxido nítrico não estaria envolvida. Estes resultados evidenciam pela primeira vez que o efeito hipotensor da administração aguda do extrato aquoso de Blepharocalyx salicifolius, possa ser via bloqueio dos receptores de angiotensina II tipo I e pela possível diminuição da resposta autonômica simpática. / Hypertension is finds as a multifactorial clinical condition characterized by high and sustained levels of blood pressure. Disease progression are intimately linked with structural and functional cardiac and vascular changes, promoting some damage in target organs such as the kidneys, brain, in addition to metabolic changes that produce a significant increase in the risk of cardiovascular diseases. According to the Brazilian Society of Cardiology, hypertension has a high prevalence and low control rates, making it one of the most important public health problems in Brazil. The use of medicinal plants for therapeutic purposes is quite old and often its use is completely empirical, no scientific study that basement this practice. Blepharocalyx salicifolius, known as Murta, has several therapeutic utilities based on popular knowledge, including use as digestive, antibacterial, antispasmodic and antihypertensive, among others. The present study evaluated the hypotensive effect of aqueous extract of B. salicifolius in normotensive rats and analyzed with L-NAME, the losartan and hexamethonium, which would be its possible mechanism of action. The evaluation of the hemodynamic effects was perform in normotensive Wistar rats, anesthetized with urethane (1.4 mg / kg i.p.) and cannulated in the carotid artery (for verification of SBP, DBP and HR) and the jugular vein (for administration of the extract and drugs). The first protocol was performed to verify the effect of the extract on the hemodynamic parameters using a dose increasing curve of the extract (10, 25, 50, 75 and 100 mg/kg) intravenously individually (each group containing six animals). The administration of the aqueous extract was after 30 minutes of stabilization in the equipment, and they remained for 60 min. The second protocol was to evaluate some of the possible mechanisms involved in the hypotensive response. Were used: an inhibitor of nitric oxide synthase - L-NAME (L-NG-L-nitroarginine methyl ester) (30 mg / kg); AT1 receptor antagonist - losartan (10 mg / kg), and a ganglionic blocker - hexamethonium (20mg/kg). Drug administration was performed after stabilization, performing a new stabilization for the administration of the extract and records kept for 60 min. The aqueous extract of B. salicifolius proved a potent hypotensive effect, reducing blood pressure in a dose-independent way since the declines were similar regardless of the dose used, and without effect on heart rate. In evaluation of the possible mechanism of action it was observed, when the AT1 receptor were blockade and the extract was administrated, that there was no change in blood pressure caused by losartan as after ganglionic blockage. When hypertension was induced by L-NAME, the extract caused arterial pressure low, suggesting that the nitric oxide pathway could not be involved. These results show for the first time that the hypotensive effect of acute administration of Blepharocalyx salicifolius aqueous extract may be by blockade of angiotensin II type 1 receptors and the possible reduction in sympathetic autonomic response.

The duodenal mucosa is regularly exposed to acid, digestive enzymes and ingested noxious agents. It is thus critical to maintain a protective barrier to prevent the development of mucosal injury and inflammation, which are often observed in situations when barrier function is impaired. The rate of mucosal bicarbonate secretion, the regulation of epithelial paracellular permeability and motility are each key components of duodenal barrier function. The hormone melatonin is present in high levels in the gastrointestinal tract and it has been hypothesized that melatonin exerts protective properties. This thesis aims to investigate the impact of exogenous melatonin on the regulation of duodenal barrier function and motility in anesthetized rats in vivo. In addition, duodenal tissue was examined histologically and the expression levels of tight junction proteins and melatonin receptors were assessed with qRT-PCR. It was found that melatonin stimulated mucosal bicarbonate secretion and decreased basal paracellular permeability. Exposing the duodenal mucosa to the well-characterized barrier breaker ethanol increased mucosal bicarbonate secretion, paracellular permeability and motility. Omission of luminal Clˉ abolished, while pretreatment with a nicotinic receptor antagonist reduced, the ethanol-induced bicarbonate secretion suggesting that the secretory response to ethanol is meditated via Clˉ/HCO3ˉexchangers and enteric neural pathways. Melatonin reduced the ethanol-induced increases in paracellular permeability and motility either when injected intravenously or when administered in drinking water for two weeks. The actions of melatonin were abolished by the melatonin receptor antagonist luzindole and by nicotinic acetylcholine receptor inhibition. Two weeks oral administration of melatonin up-regulated the expression levels of melatonin receptors, down-regulated the expression of ZO-3 while the expression of ZO-1, ZO-2, claudin 2-4, occludin and myosin light chain kinase were unaffected. Superficial epithelial changes in a few villi were seen in response to ethanol exposure, an effect that was histologically unchanged by melatonin pretreatment. In conclusion, the results suggest that melatonin plays an important role in the neurohumoral regulation of gastrointestinal mucosal barrier function and motility via receptor- and enteric neural-dependent pathways in vivo in rats. Melatonin might be a candidate for treatment of barrier dysfunction in humans.

Abdominal surgery inhibits gastrointestinal motility, a phenomenon referred to as postoperative ileus. Since the postoperative ileus disturbs duodenal physiology it is important to minimize the side effects of this condition. Recent experiments in our laboratory show that treatment of anaesthetised rats with parecoxib, a selective cyclooxygenase-2 inhibitor, prevents duodenal postoperative ileus, increases duodenal mucosal bicarbonate secretion and improves other functions as well. One aim of the thesis was to investigate whether removal of luminal chloride affect the parecoxib- and the vasoactive intestinal peptide (VIP)-induced stimulation of duodenal mucosal bicarbonate secretion. The proximal duodenum of anaesthetised Dark Agouti rats was perfused with isotonic solutions containing zero or low Cl- and the effect on luminal alkalinisation determined. The basal as well as the parecoxib-induced increase in alkalinisation, but not that stimulated by VIP, were markedly reduced in the absence of luminal Cl-. One important function of the duodenum is to adjust luminal osmolality towards that in the blood. It is believed that the adjustment of osmolality in the duodenum is achieved by osmosis and diffusion of electrolytes along their concentration gradients and that these processes occur predominately paracellularly. Another aim of the thesis was to examine whether prevention of postoperative ileus affects the duodenal response to luminal hypertonicity. The proximal duodenum of anaesthetised Dark Agouti and Sprague-Dawley rats were perfused with hypertonic solutions of different composition and osmolality and the effects on duodenal motility, alkaline secretion, transepithelial fluid flux, mucosal permeability and the adjustment of luminal osmolality were determined in absence and presence of parecoxib. It is concluded that COX-2 inhibition increases duodenal mucosal bicarbonate secretion by stimulating apical Cl-/HCO3- exchange in duodenocytes. Furthermore, pretreatment of anaesthetised rats with parecoxib improves a number of duodenal functions in both rat strains that contribute to improve the ability to adjust luminal osmolality. The choice of rat strain is another important feature to consider when interpreting the results because the DA strain was more responsive to luminal hypertonicity than the SD strain. Finally, several evidences are provided to suggest that the adjustment of luminal osmolality in the rat duodenum is a regulated process.