Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T13:10:33Z No. of bitstreams: 2 dirceu_de_souza_melo.pdf: 1243588 bytes, checksum: e609b2371fe339ca91cc379149b3beca (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T13:13:48Z (GMT) No. of bitstreams: 2 dirceu_de_souza_melo.pdf: 1243588 bytes, checksum: e609b2371fe339ca91cc379149b3beca (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T13:13:44Z (GMT) No. of bitstreams: 2 dirceu_de_souza_melo.pdf: 1243588 bytes, checksum: e609b2371fe339ca91cc379149b3beca (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T13:14:26Z (GMT) No. of bitstreams: 2 dirceu_de_souza_melo.pdf: 1243588 bytes, checksum: e609b2371fe339ca91cc379149b3beca (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Made available in DSpace on 2014-12-17T13:14:27Z (GMT). No. of bitstreams: 2 dirceu_de_souza_melo.pdf: 1243588 bytes, checksum: e609b2371fe339ca91cc379149b3beca (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Previous issue date: 2013 / Evid?ncias recentes sugerem que a restri??o cal?rica intensa (RCI) (>40%) exerce efeitos ben?ficos sobre o cora??o de ratos. No entanto, a maioria destes estudos avaliaram os efeitos da RCI em cora??es de ratos que iniciaram esta restri??o j? na idade adulta. Neste trabalho, investigamos as conseq??ncias de uma restri??o cal?rica de 50% desde o nascimento sobre a fun??o e morfologia card?aca de ratos adultos e avaliamos os poss?veis mecanismos envolvidos nestas adapta??es. Desde o nascimento at? a idade de 90 dias ratos RC50 tiveram sua alimenta??o restrita a 50% do consumo do grupo ad libitum (AL). Durante o per?odo de 90 dias os animais tiveram sua ingest?o alimentar, peso corporal e press?o arterial monitorados. Ap?s este per?odo, foi realizado um teste de capacidade aer?bica m?xima para avaliar indiretamente a fun??o cardiovascular global. Quarenta e oito horas ap?s este teste os animais foram eutanasiados, o sangue foi coletado para an?lise do hemat?crito, bioqu?mica s?rica e estresse oxidativo e o m?sculo s?leo retirado para an?lise do estresse oxidativo. A t?bia foi retirada para aferi??o do comprimento e f?gado, ba?o, supra-renais, test?culos e gordura viceral removidos para aferi??o do peso. O cora??o foi retirado e o ?ndice de desenvolvimento de tens?o m?xima (+dT/dt) e m?nima (-dT/dt) do mioc?rdio foram analisados pela prepara??o de cora??o isolado. Tamb?m foi realizada an?lise do estresse oxidativo card?aco e o di?metro, n?mero e densidade de cardiomi?citos, assim como os n?veis de fibrose card?aca foram obtidos atrav?s da an?lise histol?gica. Mi?citos ventriculares foram isolados para avalia??o do transiente de Ca2+ em microscopia confocal e os n?veis de fosforila??o da Akt e express?o da SERCA2 foram avaliados pela t?cnica de Western blot. Comparado ao grupo AL, os animais RC50 apresentaram menor peso corporal e de ?rg?os, menor comprimento da t?bia, menor glicemia, maior colesterol HDL, menor press?o arterial, menor estresse oxidativo card?aco, maior desempenho aer?bio e da fun??o card?aca, como mostrado pelo aumento ?dT/dt. Apesar do menor di?metro dos cardiomi?citos, ratos RC50 apresentaram aumento na rela??o cora??o/peso corporal, aumento do n?mero e densidade dos cardiomi?citos, e n?veis semelhantes de fibrose card?aca em compara??o aos animais AL. Os n?veis de fosforila??o da Akt foram superiores nos cardiomi?citos dos animais RC50 e n?o houve diferen?as significativas no transiente de Ca2+ e na express?o de SERCA2 entre os cardiomi?citos dos animais RC50 e AL. Em conjunto, estas observa??es revelaram efeitos positivos de uma RCI de 50% desde o nascimento sobre a fun??o, estrutura card?aca e vias de sinaliza??o da sobreviv?ncia de cardiomi?citos em ratos adultos. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2013. / ABSTRACT There has been growing evidence suggesting that a severe caloric restriction (SCR) (above 40%) leads to beneficial effects on heart rats. However, most of the reports are focused on the effects of SCR on hearts of rats which started this restriction at adulthood. In this work, we investigated the consequences of a 50% caloric restriction since birth on cardiac morphology and function in adult rats and evaluated the possible mechanisms involved in these adaptations. From birth up to the age of 90 days CR50 rats were fed restricted at 50% of the Ad Libitum group (AL). During the period of 90 days the animals had their food intake, body weight and blood pressure monitored. After this period, a maximal aerobic test was performed in order to indirectly evaluate the global cardiovascular function. Forty-eight hours after this test the animals were euthanized the blood collected for analysis of hematocrit, serum biochemistry and oxidative stress and soleus muscle collected for oxidative stress analysis. The tibia was removed for measurement of length and liver, spleen, adrenals, testes and visceral fat were removed for measurement of weight. The heart was removed and the index of velocity of myocardial contraction (+dT/dt) and relaxation (-dT/dt) was analyzed by isolated heart preparation. We also carried out analysis of cardiac oxidative stress and cardiomyocyte diameter, number, density and myocardium collagen content were obtained through histological analysis. Ventricular myocytes were isolated by standard methods in order to evaluate phosphorylated AKT levels and the expression of SERCA2a was assessed by Western blot. Compared to AL, CR50 animals had lower body and organs weight, lower tibia length, lower blood glucose, higher HDL cholesterol, lower blood pressure, lower oxidative stress in the heart and blood, increased aerobic performance and cardiac function, as shown by increased ? dT/dt. Despite the smaller cardiomyocyte diameter, CR50 rats presented increased heart/body weight ratio, increased cardiomyocyte density and number, and similar levels of myocardium collagen content compared to AL rats. AKT was hyperphosphorylated in cardiomyocytes from CR50 rats and no significant differences in Ca2+ transient and SERCA2 levels were found between cardiomyocytes of CR50 and AL rats. Collectively, these observations revealed beneficial effects of a 50% caloric restriction from birth on cadiac function, structure and signaling pathways for survival of adult rat cardimyocytes.

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-03-26T12:48:59Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) manuela_ortega_marques_rodrigues.pdf: 2356674 bytes, checksum: e576d7cb84dd67e4068acfffffe22e9f (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-03-29T12:57:38Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) manuela_ortega_marques_rodrigues.pdf: 2356674 bytes, checksum: e576d7cb84dd67e4068acfffffe22e9f (MD5) / Made available in DSpace on 2018-03-29T12:57:38Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) manuela_ortega_marques_rodrigues.pdf: 2356674 bytes, checksum: e576d7cb84dd67e4068acfffffe22e9f (MD5) Previous issue date: 2017 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Funda??o de Amparo ? Pesquisa do Estado de Minas Gerais (FAPEMIG) / A expans?o do tecido adiposo branco na obesidade leva ? express?o alterada de prote?nas em seus adip?citos, bem como a infiltra??o de c?lulas do sistema imune, especialmente macr?fagos, cujas secre??es levam ao desenvolvimento da inflama??o cr?nica de baixo grau, a qual ? considerada subjacente ao desenvolvimento de in?meras comorbidades. Dentre as formas de tratamento da obesidade, dietas de restri??o cal?rica (RC) nutricionalmente balanceadas induzem a perda de peso e melhorias em marcadores sist?micos da inflama??o, mas os efeitos diretos no tecido adiposo visceral ainda s?o controversos. No entanto, existe uma lacuna sobre qual o impacto dessas dietas na inflama??o local, mesmo em condi??es de sobrecarga lip?dica. Assim, o objetivo deste estudo foi avaliar os efeitos da perda de peso corporal induzida por dieta hipolip?dica ad libitum e pela restri??o cal?rica com dieta hiperlip?dica na inflama??o do tecido adiposo visceral de camundongos obesos. Para tal, inicialmente, camundongos C57BL/6 com 12 semanas de idade, machos, foram divididos em dois grupos: LF ? alimentados com dieta controle hipolip?dica ? do ingl?s low fat (10% das calorias, fonte ?leo de soja, rica em ?cidos graxos poli-insaturados); e HF ? alimentados com dieta controle hiperlip?dica ? do ingl?s high fat (60% calorias, fonte banha de porco, rica em ?cidos graxos saturados) para indu??o da obesidade. Ap?s oito semanas, seis animais de cada grupo foram eutanasiados para verifica??o da adiposidade visceral e estado inflamat?rio (dosagens de prote?na C reativa ? PCR s?rica e hep?tica). Em seguida, os animais HF foram aleatoriamente divididos em tr?s grupos HF ? continuaram recebendo dieta HF; LFAL ? submetidos ao emagrecimento pela substitui??o da dieta HF pela LF e acesso livre (ad libitum) e RHF ? submetidos ao emagrecimento por receberem quantidades restritas em calorias da dieta HF para atingir o mesmo peso corporal dos animais LFAL. A partir deste momento, esses grupos foram alimentados, juntamente com os animais LF, por mais sete semanas. Ao final, foram avaliados o ganho/perda de peso corporal, a adiposidade, as concentra??es s?ricas e hep?ticas de PCR, e as concentra??es de leptina, adiponectina, e das citocinas IL-6, TNF e MCP-1 no tecido adiposo retroperitoneal, al?m da morfologia dos adip?citos e a presen?a de infiltrados inflamat?rios no tecido adiposo retroperitoneal. Ao final da fase de indu??o da obesidade, os animais HF estavam obesos e inflamados. Ao final da fase de indu??o da perda de peso, os grupos LFAL e RHF tiveram pesos corporais semelhantes, menores que o HF e se igualaram ao LF. No entanto, houve maior dificuldade em perder peso pelo grupo RHF em compara??o ao LFAL, dado pelas diferen?as significativas entre os deltas de perda de peso, que foram menores para RHF e pelos coeficientes de efici?ncia energ?tica, que foram maiores para o grupo RHF. Os animais LFAL retornaram a adiposidade e a hipertrofia dos adip?citos viscerais a valores semelhantes ao grupo LF. Isto provavelmente foi o que levou ? menor concentra??o de leptina com concomitante aumento da adiponectina e menor infiltra??o de c?lulas inflamat?rias neste tecido, igualando-se tamb?m ao LF. Em consequ?ncia, houve menor concentra??o tecidual de citocinas pr?-inflamat?rias, al?m de menor concentra??o hep?tica e circulante de PCR. J? para os animais RHF, houve apenas atenua??o da adiposidade e da hipertrofia dos adip?citos retroperitoneais. Isso foi suficiente para restabelecer a concentra??o local de leptina a n?veis semelhantes ao grupo LF, embora n?o tenha elevado a concentra??o de adiponectina. Al?m disso, a infiltra??o de c?lulas inflamat?rias menteve-se tamb?m elevada. N?o houve redu??o da concentra??o de citocinas pr?-inflamat?rias, ? exce??o da IL-6, que reduziu levemente. A concentra??o hep?tica de PCR foi atenuada, o que n?o refletiu na concentra??o s?rica dessa prote?na. Concluiu-se que a restri??o cal?rica com dieta hiperlip?dica foi menos eficiente em promover a perda de peso e de adiposidade e n?o melhorou a inflama??o do tecido adiposo visceral, comparada com a dieta hipolip?dica ad libitum. Inferiuse que a ingest?o de dieta com sobrecarga de lip?deos (60% das calorias) e de ?cidos graxos saturados foi mais determinante da inflama??o local do que a restri??o cal?rica per se. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2017. / The expansion of white adipose tissue in obesity leads to altered protein expression in its adipocytes, as well as the infiltration of immune cells, especially macrophages, whose secretions lead to the development of chronic low-grade inflammation, which underlies the development of several comorbidities. Among treatments, caloric restriction (CR) nutritionally balanced diets induce weight loss and ameliorates inflammation systemic markers, but adipose tissue effects are still controversial. Moreover, there is a gap on the impact of these diets on local inflammation, even under lipid overload. Thus, the aim of this study was to evaluate effects of body weight loss induced by a low fat ad libitum diet and a CR in a high fat diet in the visceral adipose tissue inflammation of obese mice. Firstly, 12 weeks of age male C57BL/6 mice were divided into two groups: LF - fed a control low fat diet (10% calories, source soybean oil, high in polyunsaturated fatty acids); and HF - fed a control high fat diet (60% calories, source lard, high in saturated fatty acids) for obesity induction. After eight weeks, six animals from each group were euthanized to verify visceral adiposity and inflammatory status (serum and hepatic C-reactive protein-CRP). Then, HF animals were randomly divided into three groups: HF ? keept at HF diet; LFAL - a weight loss group that was switched from HF to LF and maintained on it ad libitum; RHF - a weight loss group that received restricted amounts of HF to maintain the same body weight as LFAL. Thereafter, these groups were fed, along with the LF animals, for another seven weeks. At end, body weight gain / loss, adiposity, serum and hepatic CRP concentrations, and adipose retroperitoneal tissue concentrations of leptin, adiponectin, IL-6, TNF and MCP-1 were evaluated, as well as adypocite morphology and the presence of inflammatory infiltrates in the retroperitoneal adipose tissue. Obesity was induced, since HF animals had higher weights, adiposity and were inflamed. At the end of the weight loss period, both LFAL and RHF had similar body weight, lower than HF and equal to LF. However, it was more dificcult to loose wheight by the RHF group compared to LFAL, since weight loss deltas were lower for RHF and energy efficiency ratios were higher for RHF group. LFAL animals returned visceral adiposity and retroperitoneal adipocyte hypertrophy similarly to the LF group. Also, there was a lower leptin level with concomitant increase of adiponectin and less infiltration of inflammatory cells in this tissue, also matching to LF. Still, there was a lower tissue concentration of proinflammatory cytokines, and a lower hepatic and serum CRP. For RHF animals, there was only an attenuation in adiposity and visceral adipocyte hypertrophy, although it was sufficient to restore local leptin concentration similarly to LF. However, this regimen was not able to elevate the adiponectin concentration. In addition, the inflammatory cells infiltration was highly elevated. There was no reduction in proinflammatory cytokines concentration, despite IL-6, which was reduced slightly. Hepatic CRP concentration was attenuated, which did not reflect in its serum concentrations. In mice with diet-induced obesity, the weight loss by means a CR in a high fat diet was less effective in promoting wheight and adiposity losses and it did not improve visceral adipose tissue inflammation. It can be inferred that a lipid overload (60% from calories) as well as a saturated fatty acid surplus from the high fat diet were more determinant of local inflammation than caloric restriction per se.