Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-02-14T12:00:15Z No. of bitstreams: 1 2016 - Gabriela Mastrangelo Gon?alves.pdf: 1218648 bytes, checksum: d31754b24278f68ec75382b5ff3932c0 (MD5) / Made available in DSpace on 2017-02-14T12:00:15Z (GMT). No. of bitstreams: 1 2016 - Gabriela Mastrangelo Gon?alves.pdf: 1218648 bytes, checksum: d31754b24278f68ec75382b5ff3932c0 (MD5) Previous issue date: 2016-02-23 / Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do RJ - FAPERJ / Several drugs in current use were discovered during experimental tests and by observing animals. When a new compound looks promising, it usually undergoes changes in its chemical structure in order to perfect its selectivity, potency and therapeutic efficacy. The aim of this study was to evaluate the antinociceptive and anti-inflammatory activities of a new synthetic hybrid compound (?)-4-chloro-6-(naphthalen-1-yl)-tetrahydro-2H-pyran-2-yl-methanol (CTHP) prepared from a previous prototype acid, (?) - cis- (6-ethyl-tetrahydropyran-2-yl) Formic. The compound CTHP was evaluated in acute pain induction assays. Oral administration of the compound was able to induce antinociceptive activity in models of writhing induced by acetic acid, formalin (both stages) and tail flick. To elucidate the mechanism of action of the compound, the tail flick model was used. This model was perform by prior administration of naloxone (opioid antagonist non-selective), where we observed the inhibition of the effect produced by the compound. The selective involvement of opioid receptors (?, ? and ?) was then evaluated by prior administration of methylnaltrexone, naltrindol, and nor-binaltorphimine, respectively, where only nor-binaltorphimine was able to reduce the analgesic effect of the compound. To evaluate the possible role of the NO/cGMP/KATP, animals were pretreated with N-nitro-L-arginine methyl ester (L-NAME), 1H- [1,2,4 ] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ) (inhibitor of guanylate cyclase sensitive to nitric oxide), and glibenclamide (blocker of the ATP-regulated potassium channels), where reduction was observed with the administration of analgesic effect prior to all of these. In the tolerance induction test, both morphine and compound developed tolerance, however the compound perform at a slower rate and developed cross-tolerance with morphine. To assess the involvement of serotonin pathway in the activity of the compound, daily administration for 3 days of 4-chloro-DL-phenylalanine (inhibitor of the enzyme tryptophan hydroxylase) was performed. No changes in the analgesic effect of the compound was noted, with regard to the involvement of serotonin pathway. The open field model was used to assess the possibility of interference from motor performance on the analgesic effect, which demonstrated absence of this interference. As for anti-inflammatory activity results in paw edema test indicate anti-oedematogenic effect of compound. There was a decrease in the number of total leukocytes, indicating that the compound was able to reduce existing inflammation in leukocyte migration in the air pouch model. The compound also demonstrated an inhibitory activity on TNF-? production and selective inhibition of COX-2 enzyme. These results indicate significant antinociceptive activity of the compound without evidence of motor impairment. The compound CTHP showed central analgesic effect, which has contribution of opioid systems (selective for the ?-like receptors) and nitrergic in its mechanism of action. It has also showed an anti-inflammatory activity, with inhibition of leukocyte migration, TNF-? production and selective inhibitory activity on COX-2. / Diversos f?rmacos de uso corrente foram descobertos durante ensaios experimentais e mediante a observa??o em animais. Quando um novo composto parece promissor, geralmente este sofre altera??es em sua estrutura qu?mica a fim de aperfei?oar a sua seletividade, pot?ncia e efic?cia terap?utica. O objetivo deste estudo foi avaliar as atividades antinociceptiva e anti-inflamat?ria de um novo composto sint?tico (?)-4-cloro-6-(naftaleno-1-il)-tetrahidro-2h-pirano-2-il-metanol (CTHP) preparado a partir de um prot?tipo anterior, ?cido (?)-cis-(6-etil-tetrahidropirano-2-il) f?rmico. O composto CTHP foi avaliado em ensaios de indu??o de dor aguda. A administra??o oral do composto foi capaz de induzir atividade antinociceptiva nos modelos de contor??es abdominais induzidas por ?cido ac?tico, formalina (em ambas as fases) e retirada da cauda. Para elucida??o do mecanismo de a??o do composto, o modelo de retirada de cauda foi utilizado. Neste modelo foi realizada a administra??o pr?via de naloxona (antagonista opioide n?o-seletivo), em que foi observada a inibi??o do efeito produzido pelo composto. Assim, foi ent?o avaliada a participa??o seletiva de receptores opioides (?, ? e ?), atrav?s de administra??o pr?via de metilnaltrexona, naltrindol e nor-binaltorfimina, respectivamente, onde somente a nor-binaltorfimina foi capaz de reduzir o efeito antinociceptivo do composto. Para avaliar a poss?vel participa??o da via NO/GMPc/KATP, os animais foram pr?-tratados com N-nitro-arginina-L-metil ?ster (L-NAME), 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalina-1-ona (ODQ) (inibidor da guanilato ciclase sens?vel ao ?xido n?trico) e glibenclamida (bloqueador de canais de pot?ssio regulados por ATP), foi observado redu??o do efeito antinociceptivo com a administra??o pr?via de todos estes. No teste de indu??o de toler?ncia, tanto a morfina quanto o composto desenvolveram toler?ncia, por?m o composto de forma mais lenta e houve desenvolvimento de toler?ncia cruzada com a morfina. Para avaliar o envolvimento da via serotonin?rgica na atividade do composto, foi realizada a administra??o di?ria por 3 dias de 4-cloro-DL-fenilalanina (inibidor da enzima triptofano hidroxilase). Nenhuma altera??o no efeito antinociceptivo do composto foi observado, no que diz respeito ao envolvimento da via serotonin?rgica. J? o modelo de campo aberto foi utilizado para avaliar a possibilidade de interfer?ncia da performance motora sobre o efeito antinociceptivo, foi demonstrada aus?ncia desta interfer?ncia. Quanto ? atividade anti-inflamat?ria, o resultado no teste de edema de pata indica efeito antiedematog?nico do composto. Houve uma diminui??o na quantidade de leuc?citos totais, indicando que o composto foi capaz de reduzir a migra??o leucocit?ria na inflama??o existente na bolsa de ar subcut?neo. O composto tamb?m demonstrou atividade inibit?ria sobre a produ??o de TNF-? e inibi??o seletiva da enzima COX-2. Esses resultados indicam atividade antinociceptiva significativa do composto, sem evid?ncias de comprometimento motor. O composto CTHP demonstrou efeito antinociceptivo central, tendo este ?ltimo contribui??o dos sistemas opioide (seletivo para receptores do tipo ?) e nitr?rgico em seu mecanismo de a??o. E ainda, atividade anti-inflamat?ria, com inibi??o da migra??o leucocit?ria, de TNF-? e atividade inibit?ria seletiva sobre COX-2.

Made available in DSpace on 2014-12-17T14:16:34Z (GMT). No. of bitstreams: 1 HeglaynePVS_DISSERT.pdf: 2645894 bytes, checksum: da2c7ec39b2e87b75a199511857a4e83 (MD5) Previous issue date: 2013-03-27 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Of all of the genes associated with the development of Diabetes mellitus type 1 (T1D), the largest contribution comes from the genes in the Human Leukocyte Antigen (HLA) region, mostly the class II DR e DQ genes. Specific combinations of alleles DRB1, DQA1 and DQB1 constituting haplotypes, and further, a combination of more than one haplotype, providing multilocus genotypes are associated with susceptibility, protection and neutrality to DM1. Thus, the aim of present study was to verified the association of polymorphisms of HLA genes class II with susceptibility to type 1 diabetes mellitus (T1D). Ninety-two patients with T1D and 100 individuals normoglycemics (NG) aged between 6 and 20 years were studied. Genomic DNA was obtained from peripheral whole blood, collected in EDTA tube, using the extraction kit Illustra Triple Prep?, GE Healthcare. For HLA typing was used DNA LABType system by One Lambda kit applying Luminex? technology to the method of PCRSSO typing reverse. The alleles DRB1*03:01, *04:05, *04:01, *04:02, DQA1*03:01g, *05:01g, DQB1*02:01g, *03:02, the haplotypes DRB1*03:01-DQA1*05:01-DQB1*02:01, DRB1*04:05-DQA1*03:01g-DQB1*03:02, DRB1*04:02-DQA1*03:01g-DQB1*03:02, DRB1*04:01-DQA1*03:01g-DQB1*03:02 and DR3-DQ2/DR4-DQ8 genotype were significantly associated with the chance of developing T1D. The alleles DRB1*11:01, *15:03, *15:01, *13:01, DQA1*01:02, *04:01g, *01:03, DQB1*06:02, *03:01g, *06:03, *04:02, the haplotypes DRB1*11:01-DQA1*05:01-DQB1*03:01, DRB1*13:01-DQA1*01:03-DQB1*06:03 and DRX-DQX/DRX-DQX genotype, formed by other than the DR3-DQ2 or DR4-DQ8 haplotypes, were significantly associated with T1D protection Despite the major racial Brazilian, even at the regional level, these results are similar to the majority of alleles, genotypes and haplotypes of HLA class II-related susceptibility or resistance to T1D, extensively described in the literature for Caucasian population. Children with age at diagnosis less than 5 years of age had significantly higher frequency of the heterozygous genotype DR3-DQ2/DR4-DQ8 compared to children with age at diagnosis than 5 years old. These results also demonstrate strong association of the genetic profile of the class II HLA for this age group, possibly associated with the severity and rapid progression to the onset of T1D. The knowledge of HLA class II genes may be useful in genetic screens that allow the prediction of T1D / De todos os genes j? relacionados com o desenvolvimento do Diabetes mellitus tipo 1 (DM1), a maior contribui??o vem da regi?o do genoma onde est?o localizados os genes do Ant?geno Leucocit?rio Humano (HLA), sobretudo os genes da classe II do HLA: DR e DQ. Espec?ficas combina??es de alelos DRB1, DQA1 e DQB1 formando hapl?tipos, e ainda, a combina??o de mais de um hapl?tipo, formando gen?tipos multilocus s?o associados com a susceptibilidade, neutralidade e prote??o ao DM1. Dessa forma, o objetivo do estudo foi verificar a associa??o dos polimorfismos dos genes do complexo HLA classe II com a susceptibilidade ao DM1, em pacientes do Rio Grande do Norte. Foram estudados 92 indiv?duos com DM1 e 100 indiv?duos normoglic?micos (NG), com idade entre 6 e 20 anos. O DNA gen?mico foi obtido a partir do sangue total perif?rico, coletado em tubo com EDTA, utilizando o kit de extra??o Illustra Triple Prep?, GE Healthcare. Para a tipagem do HLA foi utilizado o sistema DNA LABType atrav?s de kits One Lambda, que aplica a tecnologia Luminex? ao m?todo de tipagem por PCR-SSO reverso. Os alelos DRB1*03:01, *04:05, *04:01, *04:02; DQA1*03:01g, 05:01g; DQB1*02:01g, *03:02; os hapl?tipos DRB1*03:01-DQA1*05:01-DQB1*02:01, DRB1*04:05-DQA1*03:01g- DQB1*03:02, DRB1*04:02-DQA1*03:01g-DQB1*03:02, DRB1*04:01-DQA1*03:01g- DQB1*03:02 e o gen?tipo heterozigoto, DR3-DQ2/DR4-DQ8 foram significativamente associados com a chance de desenvolvimento do DM1. J? os alelos DRB1*11:01, *15:03, *15:01, *13:01; DQA1*01:02, *04:01g, *01:03; DQB1*06:02, *03:01g, *06:03, *04:02; os hapl?tipos DRB1*11:01-DQA1*05:01-DQB1*03:01, DRB1*13:01-DQA1*01:03-DQB1*06:03 e o gen?tipo DRX-DQX/DRX-DQX, formado por outros hapl?tipos que n?o DR3-DQ2 ou DR4-DQ8, foram significativamente associados a prote??o ao DM1. Apesar da grande miscigena??o racial brasileira, at? em n?vel regional, estes resultados s?o semelhantes a maioria dos alelos, hapl?tipos e gen?tipos de HLA classe II relacionados ? susceptibilidade ou prote??o ao DM1, extensivamente descritos na literatura para a popula??o caucasiana. Crian?as com idade ao diagn?stico inferior a 5 anos de idade apresentaram significativamente maior frequ?ncia do gen?tipo heterozigoto DR3-DQ2/DR4-DQ8, quando comparada ?s crian?as com idade ao diagn?stico superior a 5 anos de idade. Esses resultados demonstram tamb?m forte envolvimento do perfil gen?tico da classe II do HLA para esta faixa et?ria, que estaria relacionada possivelmente com a gravidade e a r?pida progress?o para o in?cio do DM1. O conhecimento dos genes HLA de classe II pode ser ?til em triagens gen?ticas que possibilitem a predi??o do DM1