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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
11

CapAT "An adipose-enriched isoform"

He, Yue 21 June 2021 (has links)
No description available.
12

The independent effects of chronic high-fat feeding and long-term denervation in relation to development of diabetes

Callahan, Zachary J. 02 December 2014 (has links)
No description available.
13

Role of the Sh3 and Cysteine-Rich Domain 3 (STAC3) Gene in Proliferation and Differentiation of Bovine Satellite Cells

Zhang, Yafei 25 September 2013 (has links)
The STAC3 gene is a functionally undefined gene predicted to encode a protein containing two SH3 domains and one cysteine-rich domain. In this study, we determined the potential role of the STAC3 gene in proliferation and differentiation of bovine satellite cells. We isolated satellite cells from skeletal muscle of adult cattle and transfected them with STAC3 small interfering RNA (siRNA) or scrambled siRNA. Cell proliferation assays revealed that STAC3 knockdown had no effect on the proliferation rate of bovine satellite cells. We assessed the differentiation status of bovine satellite cells by quantifying the expression levels of myogenin and myosin heavy chain genes, and by quantifying fusion index. STAC3 knockdown stimulated mRNA and protein expression of myogenin, and myosin heavy chain 3 and 7, and increased fusion index of bovine satellite cells. These data together suggest that STAC3 inhibits differentiation of bovine satellite cells into myotubes. To determine the underlying mechanism, we identified and validated AP1?1 as a STAC3-interacting protein by yeast two-hybrid screening and co-immunoprecipitation. In C2C12 cells, STAC3 knockdown decreased the expression level of AP1?1 protein. In bovine satellite cells, STAC3 knockdown increased the membrane localization of glucose transporter 4 (GLUT4) and glucose uptake. These data together suggest the following mechanism by which STAC3 inhibits differentiation of bovine satellite cells: STAC3 increases AP1?1 stability in cells; a high level of AP1?1 keeps GLUT4 from translocating to the plasma membrane; reduced membrane localization of GLUT4 impedes glucose uptake; and restricted glucose uptake inhibits differentiation of satellite cells into myotubes. / Master of Science
14

Insulina e captação de glicose no corpo lúteo canino / Insulin and glucose uptake in canine corpus luteum

Silva, Renata dos Santos 29 June 2012 (has links)
O diestro é a fase luteínica na cadela caracterizada pelo aumento de progesterona (P4) sérica na primeira metade e por flutuações de 17&#946;-estradiol (E2) na segunda metade. O corpo lúteo (CL) é uma glândula endócrina temporária, que passa por um processo de desenvolvimento, manutenção e regressão, atingindo atividade secretória plena quando sua formação está completa. A insulina é o hormônio anabólico essencial para a manutenção da homeostase de glicose e do crescimento e diferenciação celular, sendo secretado pelas células &#946; pancreáticas. A sinalização intracelular da insulina começa com a sua ligação a um receptor de membrana específico, o que desencadeia uma série de ações metabólicas. Sabe-se que uma das consequências desta ligação é a translocação de transportadores de glicose 4 (GLUT4; gene SLC2A4) para que ocorra a captação de glicose. Nosso grupo demonstrou a expressão de GLUT4 no corpo lúteo de cadelas, expressão esta regulada diferencialmente ao longo do diestro. A presença deste transportador levou-nos a hipotetizar que a insulina seja importante para regulação da função luteínica. Para testar tal hipótese, utilizamos imuno-histoquímica para localizar o receptor de insulina (RI) e outros fatores regulatórios (NFKB e IL6) de GLUT4 no CL canino durante o diestro (dias 10 a >70 pós-ovulação, po) e western blotting para quantificar estas proteínas; investigamos a expressão gênica dos fatores acima mencionados por PCR em tempo real; e por fim, analisamos os efeitos da insulina sobre a expressão gênica de RI e SLC2A4 em células luteínicas nos dias 20 e 40 po e também sobre a captação de glicose destas células. No presente estudo, observou-se que o corpo lúteo canino expressa as proteínas do RI, NFKB e IL6 de maneira distinta ao longo do diestro. A expressão do RNAm do RI apresentou maior expressão nos dias 20 e 70, e diminuição no dia 40. O NFKB apresentou maior expressão no dia 40, enquanto o IL6 apresentou maior expressão do dia 10 ao 40. Observou-se correlação negativa entre o gene RI e os níveis de insulina (r = -0,69; P = 0,006) e positiva com SLC2A4 (r = 0,89; P = 0,01) em todo o diestro, enquanto o IL6 correlacionou-se de maneira positiva com o RI apenas na primeira metade (r = 0,96; P <0,0001) e o NFKB, negativamente com o RI nos dias 30, 40 e 50 (r = -0,57 P <0,05). Após a adição de insulina no meio de cultivo, observou-se que as expressões gênicas de RI e SLC2A4 se comportaram de maneira oposta de acordo com a fase do diestro estudada: células do dia 20 po apresentaram um aumento desta expressão e do dia 40 um declínio. Por fim, através da 12 captação de glicose, observou-se que as células luteínicas caninas são capazes de responder à insulina, aumentando a captação na ordem de 4 vezes (basal: 2,05 ± 0,8; insulina: 5,73 ± 0,5; valor de P <0,01 em cpm/ug proteína; basal: 79,6 ± 35,3; insulina: 212,5 ± 34,5, valor de P <0,05 em cpm/106 células). Esses resultados apontam a insulina, bem como o IL6 e o NFKB como fatores importantes que desempenham um papel na função do CL canino e trazem o CL para o grupo de tecidos que respondem ao estímulo insulínico aumentando a expressão de GLUT4 e consequentemente a captação de glicose. Além disso, estes eventos parecem sofrer controle adicional pelos hormônios esteróides. / Diestrus is the luteal phase in dogs characterized by an increase in progesterone (P4) levels in the first half and fluctuations of 17&#946;-estradiol (E2) in the second half. The corpus luteum (CL) is a temporary endocrine gland, which undergoes a process of development, maintenance and regression, reaching full secretory activity when its formation is complete. Insulin is an anabolic hormone essential for the maintenance of glucose homeostasis, cell growth and differentiation, and is secreted by pancreatic beta cells. Intracellular signaling of insulin begins with its binding to a specific membrane receptor, which triggers a series of metabolic actions. It is known that one consequence of this connection is the translocation of glucose transporters 4 (GLUT4; gene SLC2A4) for glucose uptake. Our group demonstrated the expression of GLUT4 in the canine corpus luteum in a time-related manner throughout diestrus. The presence of this transporter led us to hypothesize that insulin is important for the regulation of luteal function. To test this hypothesis, we used immunohistochemistry to detect the insulin receptor (IR) and possible regulatory factors (IL6 and NFKB) of GLUT4 in canine CL during diestrus (days 10 to > 70 after ovulation, po) and western blotting to quantify these proteins. In addition, we investigated the gene expression of the above mentioned factors by real-time PCR and analyzed the effects of insulin on IR and SLC2A4 gene expression in canine luteal cells at days 20 and 40 po and also on glucose uptake by these cells. Canine CL differentialy expressed RI, NFKB and IL6 during diestrus. The IR expression was higher on days 20 and 70, with and decreased at day 40. NFKB expression was higher on day 40 and IL6 increased on days 10 to 40 po. It was observed a negative correlation between IR expression and insulin levels (r = -0.69 P = 0.006) and positive with SLC2A4 (r = 0.89, P = 0.01) throughout diestrus. IR was positively correlated with IL6 only in the first half of diestrus (r = 0.96, P <0.0001), while it was negatively correlated with NFKB on days 30, 40 and 50 (r = -0.57 P <0.05). After insulin treatment, RI and SLC2A4 expressions behave differently according to the diestrus phase: on day 20, they increased and on day 40 they declined. Finally, we could observe through glucose uptake method that canine luteal cells are able to respond to insulin stimuli, increasing glucose uptake by approximately four folds (basal: 2.05 ± 0.8; insulin: 5.73 ± 0.5 cpm / ug protein, P < 0.01; basal: 79.6 ± 35.3; insulin: 212.5 ± 34.5 cpm/106 cells, P < 0.05). These results point towards a regulatory function 14 exerted by insulin, IL6 and NFKB in the canine CL and place this organ among the insulin sensitive ones, which respond to insulin stimuli increasing GLUT4 expression and glucose uptake. Moreover, these events seems to undergo a further control by steroid hormones.
15

Compostos bioativos com potencial ação no controle da homeostase glicêmica / Bioactive compounds with potential action in the control of glycemic homeostasis.

Souza, Ana Marla Duarte de 18 April 2017 (has links)
Diversos estudos buscam identificar novas moléculas com ações regulatórias sobre a via de sinalização da insulina e consequentemente na homeostase da glicose. Assim, este trabalho visa avaliar o potencial de extratos de frutos no controle da homeostase glicêmica. Os frutos avaliados foram o morango (cv. Toianoca, Camarosa, Oso Grande e Camino Real), a amora-preta e a framboesa vermelha, em dois tempos de amostragem, sendo considerado como tempo A1 e tempo A2. As amostras foram caracterizadas quanto ao seu conteúdo de fenólicos totais, conteúdo de antocianinas monoméricas, capacidade antioxidante, avaliada pelos métodos DPPH e ORAC, ácido elágico total, capacidade de inibição da alfa-glicosidase e captação de glicose e lipólise em tecido adiposo de camundongos (ensaio explante). Dentre os frutos, no primeiro tempo de amostragem, a amora-preta e o morango, cv Oso Grande, foram os que apresentaram maior conteúdo de fenólicos totais (62,36 e 34,89 mg AG/g, respectivamente) no entanto não foram mantidos esses valores no segundo tempo de amostragem, com concentração 30% e 60% inferior, respectivamente; e maior concentração de antocianinas monoméricas (45,33 mg/g e 3,09 mg/g, respectivamente). Em relação a inibição da enzima alfa-glicosidase, avaliado em extrato metanólico, o fruto framboesa vermelha e o morango cv. Camino Real foram as que apresentaram alto potencial inibitório nos dois tempos de amostragem (IC50 0,47 mg FT e IC50 0,57 mg FT para framboesa vermelha e IC50 0,50 mg FT e IC50 0,46 mg FT para Camino Real). Quando avaliado os extratos enriquecidos em fenólicos, o valor de IC50 com maior potencial dentre os frutos avaliados foi da amora-preta, nos dois tempos A1 e A2 (0,0023 mg FT e 0,0021 mg FT, respectivamente). Para captação de glicose em tecido adiposo explate, ao utilizar a insulina para estimular a captação de glicose juntamente com o tratamento (extrato), esse estimulo foi efetivo no aumento da captação de glicose somente com as amostras cv. Camino Real e cv. Oso Grande. Isso pode ser explicado pela alta correlação encontrada de antocianinas identificadas no fruto, como pelargonidina-3-O-glicosídeo. Por outro lado, somente a amora-preta A1 aumentou a lipólise em condição basal, mas nenhuma fruta foi eficiente para reduzir a lipólise em condição estimulada pelo isoproterenol. Sendo assim, frutas vermelhas podem ser boas fontes de compostos bioativos, principalmente antocianinas, as quais podem ter corroborado positivamente com os resultados. / Several studies seek to identify new molecules with regulatory actions on the insulin signaling pathway and consequently on glucose homeostasis. Thus, this work aims to evaluate the potential of fruit extracts in the control of glycemic homeostasis. The fruits evaluated were strawberry (cv. Toianoca, Camarosa, Oso Grande and Camino Real), blackberry and red raspberry, in two sampling times, being considered as time A1 and time A2. Fruits were evaluated for total phenolic, monomeric anthocyanins and contents, antioxidant capacity, evaluated by DPPH and ORAC methods, alpha-glycosidase inhibition capacity and glucose uptake and lipolysis in adipose tissue of mice (explant assay). Among the fruits, in the first sampling period, blackberry and strawberry, cv. Oso Grande, showed the highest total phenolic content (62.36 and 34.89 mg AG / g, respectively), with a decrease of the 30% and 60%, respectively, in the second sampling time; and higher monomeric anthocyanins concentration (45.33 mg/g and 3.09 mg/g, respectively). The methanolic extracts of raspberry and the strawberry cv Camino Real (A1 and A2) presented the highest alpha-glucosidase inhibitory potential. Otherwise, the enriched-polyphenol extract of blackberry (A1 and A2) presented the highest potential among the evaluated fruits. Adipose tissue treated with strawberry cv. Camino Real and cv. Oso Grande was effective in increasing glucose uptake stimulated by insulin. This can be explained by the high correlation with the anthocyanins pelargonidin-3-O-glycoside identified in this fruit. In addition, blackberry A1 was the only sample to increase the lipolysis in basal condition, but all other fruits were not effective to decrease lipolysis in stimulated condition. Thus, berries could be a good sources of bioactive compounds to maintain the glucose homeostasis.
16

The influence of antioxidants on thrombotic risk factors in healthy population

Singh, Indu, indu.singh@rmit.edu.au January 2008 (has links)
Oxidative damage has been suggested to play a key role in the pathogenesis of atherosclerosis and other cardiovascular disease. Increased free radical production induced by oxidative stress can oxidise low density lipoproteins, activates platelets, induces endothelial dysfunction and disturbs glucose transport by consuming endogenous antioxidants. Using a combination, of in vitro and in vivo experimental models, the primary aims of the studies undertaken for this thesis were to examine whether different antioxidants could negate risk factors leading to thrombosis, atherosclerosis and other cardiovascular diseases. The studies utilised the mechanisms involved in platelet activity and glucose uptake by skeletal muscle myotubes. The first study determined if olive leaf extract would attenuate platelet activity in healthy human subjects. Blood samples (n=11) were treated with five different concentrations of extract of Olea europaea L. leaves ranging from 5.4£gg/mL to 54£gg/mL. A significant reduction in platelet activity (pless than0.001) and ATP release from platelets (p=0.02) was observed with 54£gg/mL olive leaf extract. The next crossover study compared the effect of exercise and antioxidant supplementation on platelet function between trained and sedentary individuals. An acute bout of 1 hour exercise (sub maximal cycling at 70% of VO2max) was used to induce oxidative stress in 8 trained and 8 sedentary male subjects, before and after one week supplementation with 236 mg/day of cocoa polyphenols. Baseline platelet count and ATP release increased significantly (pless than0.05) after exercise in all subjects. Baseline platelet numbers in the trained were higher than in the sedentary (235¡Ó37 vs. 208¡Ó34 x109/L, p less than 0.05), whereas platelet activation in trained subjects was lower than sedentary individuals (51¡Ó6 vs. 59¡Ó5%, p less than0.05). Seven days of cocoa polyphenol supplementation did not change platelet activity compared to the placebo group. The third study determined the effect of 5 weeks of either 100mg/day £^-Tocopherol (n=14), 200mg/d £^-Tocopherol (n=13) or placebo (n=12) on platelet function, lipid profile and the inflammatory marker C-reactive protein. Blood £^-tocopherol concentrations increased significantly (pless than0.05) relative to dose. Both doses attenuated platelet activation (pless than0.05). LDL cholesterol, platelet aggregation and mean platelet volume were decreased by 100mg/d £^-tocopherol (all pless than0.05). The final study determined the effect of glucose oxidase induced oxidative stress and £^-tocopherol treatment on glucose transport and insulin signalling in cultured rat L6 muscle cells. One hour treatment with 100mU/mL glucose oxidase significantly decreased glucose uptake both with and without 100nM insulin stimulation (pless than0.05). Pre-treatment with 100ƒÝM and 200ƒÝM £^-tocopherol partially protected cells from the effect of glucose oxidase, whereas 200ƒÝM £^-tocopherol restored both basal and insulin stimulated glucose transport to control levels. Glucose oxidase-induced oxidative stress did not impair basal or insulin stimulated phosphorylation of Akt or AS160, but 200ƒÝM £^-tocopherol improved insulin-stimulated phosphorylation of these proteins. In summary, the results from the studies undertaken for this thesis provide evidence that antioxidant supplementation maintains normal platelet function, exerts a positive effect on blood lipid profile and improves glucose uptake in normal healthy asymptomatic population as well as under conditions of induced oxidative stress. Antioxidants including foods rich in cocoa, olive and gamma tocopherol have the potential to combat oxidative stress induced risk factors leading to cardiovascular diseases.
17

Investigation of phosphatidylinositol 5-phosphate's role in insulin-stimulated glucose uptake in a skeletal muscle cell line

Grainger, Deborah January 2011 (has links)
Phosphatidylinositol 5-phosphate (PtdIns5P) is the least well-characterised member of the phosphoinositide family of essential regulatory phospholipids. PtdIns5P levels are altered within cells in response to a number of stimuli and evidence is accumulating to suggest that it possesses important functions in cellular signalling. However, the physiological role of this lipid remains imperfectly understood. Previous studies have shown that PtdIns5P is elevated in adipocytes in response to insulin, and microinjection of PtdIns5P into these cells promotes plasma membrane insertion of the insulin-regulated glucose transporter GLUT4 (Sbrissa et al., 2004). This finding suggests a potential role of PtdIns5P as a mediator in insulin-stimulated glucose uptake, a process essential for efficient glucose homeostasis. As approximately 75% of postprandial glucose disposal is carried out by skeletal muscle, it is important to investigate the role of PtdIns5P in the response of this tissue to insulin. Therefore, this work has used differentiated myotubes of the rat muscle cell line, L6, to explore the effects of altered PtdIns5P levels on insulin-stimulated glucose uptake. This cell model had not been previously used in the laboratory so it first required characterisation. Here insulin is shown to stimulate a transient increase of PtdIns5P in L6 myotubes, indicative of a signalling role in response to insulin. This project developed several tools to further investigate this potential role for PtdIns5P in the insulin response of myotubes. One such development was the successful overexpression of the PtdIns5P 4-kinase PIP4KIIalpha in these cells, which was able to abolish the insulin-stimulated PtdIns5P rise. This correlated with a loss of insulin-stimulated glucose uptake (upon PIP4KIIalpha expression). Interestingly, artificial elevation of PtdIns5P in L6 myotubes increases glucose uptake in the absence of stimulation. This phenomenon appears to result from the activation of PI3-kinase signalling, as it is abolished by the PI3-kinase inhibitor wortmannin, and involves activation of the PI3-kinase effector Akt. These results are consistent with the idea that insulin-stimulated PtdIns5P production contributes to the robust PI3-kinase/Akt activation necessary for insulin-stimulated glucose uptake in muscle.
18

Insulina e captação de glicose no corpo lúteo canino / Insulin and glucose uptake in canine corpus luteum

Renata dos Santos Silva 29 June 2012 (has links)
O diestro é a fase luteínica na cadela caracterizada pelo aumento de progesterona (P4) sérica na primeira metade e por flutuações de 17&#946;-estradiol (E2) na segunda metade. O corpo lúteo (CL) é uma glândula endócrina temporária, que passa por um processo de desenvolvimento, manutenção e regressão, atingindo atividade secretória plena quando sua formação está completa. A insulina é o hormônio anabólico essencial para a manutenção da homeostase de glicose e do crescimento e diferenciação celular, sendo secretado pelas células &#946; pancreáticas. A sinalização intracelular da insulina começa com a sua ligação a um receptor de membrana específico, o que desencadeia uma série de ações metabólicas. Sabe-se que uma das consequências desta ligação é a translocação de transportadores de glicose 4 (GLUT4; gene SLC2A4) para que ocorra a captação de glicose. Nosso grupo demonstrou a expressão de GLUT4 no corpo lúteo de cadelas, expressão esta regulada diferencialmente ao longo do diestro. A presença deste transportador levou-nos a hipotetizar que a insulina seja importante para regulação da função luteínica. Para testar tal hipótese, utilizamos imuno-histoquímica para localizar o receptor de insulina (RI) e outros fatores regulatórios (NFKB e IL6) de GLUT4 no CL canino durante o diestro (dias 10 a >70 pós-ovulação, po) e western blotting para quantificar estas proteínas; investigamos a expressão gênica dos fatores acima mencionados por PCR em tempo real; e por fim, analisamos os efeitos da insulina sobre a expressão gênica de RI e SLC2A4 em células luteínicas nos dias 20 e 40 po e também sobre a captação de glicose destas células. No presente estudo, observou-se que o corpo lúteo canino expressa as proteínas do RI, NFKB e IL6 de maneira distinta ao longo do diestro. A expressão do RNAm do RI apresentou maior expressão nos dias 20 e 70, e diminuição no dia 40. O NFKB apresentou maior expressão no dia 40, enquanto o IL6 apresentou maior expressão do dia 10 ao 40. Observou-se correlação negativa entre o gene RI e os níveis de insulina (r = -0,69; P = 0,006) e positiva com SLC2A4 (r = 0,89; P = 0,01) em todo o diestro, enquanto o IL6 correlacionou-se de maneira positiva com o RI apenas na primeira metade (r = 0,96; P <0,0001) e o NFKB, negativamente com o RI nos dias 30, 40 e 50 (r = -0,57 P <0,05). Após a adição de insulina no meio de cultivo, observou-se que as expressões gênicas de RI e SLC2A4 se comportaram de maneira oposta de acordo com a fase do diestro estudada: células do dia 20 po apresentaram um aumento desta expressão e do dia 40 um declínio. Por fim, através da 12 captação de glicose, observou-se que as células luteínicas caninas são capazes de responder à insulina, aumentando a captação na ordem de 4 vezes (basal: 2,05 ± 0,8; insulina: 5,73 ± 0,5; valor de P <0,01 em cpm/ug proteína; basal: 79,6 ± 35,3; insulina: 212,5 ± 34,5, valor de P <0,05 em cpm/106 células). Esses resultados apontam a insulina, bem como o IL6 e o NFKB como fatores importantes que desempenham um papel na função do CL canino e trazem o CL para o grupo de tecidos que respondem ao estímulo insulínico aumentando a expressão de GLUT4 e consequentemente a captação de glicose. Além disso, estes eventos parecem sofrer controle adicional pelos hormônios esteróides. / Diestrus is the luteal phase in dogs characterized by an increase in progesterone (P4) levels in the first half and fluctuations of 17&#946;-estradiol (E2) in the second half. The corpus luteum (CL) is a temporary endocrine gland, which undergoes a process of development, maintenance and regression, reaching full secretory activity when its formation is complete. Insulin is an anabolic hormone essential for the maintenance of glucose homeostasis, cell growth and differentiation, and is secreted by pancreatic beta cells. Intracellular signaling of insulin begins with its binding to a specific membrane receptor, which triggers a series of metabolic actions. It is known that one consequence of this connection is the translocation of glucose transporters 4 (GLUT4; gene SLC2A4) for glucose uptake. Our group demonstrated the expression of GLUT4 in the canine corpus luteum in a time-related manner throughout diestrus. The presence of this transporter led us to hypothesize that insulin is important for the regulation of luteal function. To test this hypothesis, we used immunohistochemistry to detect the insulin receptor (IR) and possible regulatory factors (IL6 and NFKB) of GLUT4 in canine CL during diestrus (days 10 to > 70 after ovulation, po) and western blotting to quantify these proteins. In addition, we investigated the gene expression of the above mentioned factors by real-time PCR and analyzed the effects of insulin on IR and SLC2A4 gene expression in canine luteal cells at days 20 and 40 po and also on glucose uptake by these cells. Canine CL differentialy expressed RI, NFKB and IL6 during diestrus. The IR expression was higher on days 20 and 70, with and decreased at day 40. NFKB expression was higher on day 40 and IL6 increased on days 10 to 40 po. It was observed a negative correlation between IR expression and insulin levels (r = -0.69 P = 0.006) and positive with SLC2A4 (r = 0.89, P = 0.01) throughout diestrus. IR was positively correlated with IL6 only in the first half of diestrus (r = 0.96, P <0.0001), while it was negatively correlated with NFKB on days 30, 40 and 50 (r = -0.57 P <0.05). After insulin treatment, RI and SLC2A4 expressions behave differently according to the diestrus phase: on day 20, they increased and on day 40 they declined. Finally, we could observe through glucose uptake method that canine luteal cells are able to respond to insulin stimuli, increasing glucose uptake by approximately four folds (basal: 2.05 ± 0.8; insulin: 5.73 ± 0.5 cpm / ug protein, P < 0.01; basal: 79.6 ± 35.3; insulin: 212.5 ± 34.5 cpm/106 cells, P < 0.05). These results point towards a regulatory function 14 exerted by insulin, IL6 and NFKB in the canine CL and place this organ among the insulin sensitive ones, which respond to insulin stimuli increasing GLUT4 expression and glucose uptake. Moreover, these events seems to undergo a further control by steroid hormones.
19

Extra-Thyroidal Action of TSH on Adipocyte Insulin Signaling

Felske, David January 2015 (has links)
In subclinical hypothyroidism (SH), high levels of circulating thyroid stimulating hormone (TSH) maintain normal thyroid hormone levels, despite mild thyroid failure. SH is associated with cardiovascular disease and insulin resistance, although the underlying pathophysiology is not fully understood. We hypothesized that TSH may inhibit insulin action in adipocytes. To investigate this relationship, we studied primary human differentiated adipocytes. Abdominal subcutaneous adipose tissue samples were obtained (approved by OHSN-REB) from 16 weight-stable patients undergoing elective abdominal surgery. We stimulated adipocytes differentiated from stromal preadipocytes with 5 mU/ml TSH and/or 100 nM insulin, and assessed acute insulin signaling, lipogenesis and glucose uptake. Immunoblot analysis revealed that TSH suppressed insulin-stimulated Akt phosphorylation by 45% (n=5; p = 0.01). When adipocytes were pre-incubated with conventional protein kinase C (cPKC) inhibitor Gö6976, TSH inhibition was blocked. Our data indicate that TSH inhibits insulin-stimulated lipogenesis (up to 37%), but depends on BMI. Insulin-stimulated glucose uptake was enhanced by 36% and also correlated with BMI. This data suggests that TSH can modulate adipocyte insulin signaling.
20

Compostos bioativos com potencial ação no controle da homeostase glicêmica / Bioactive compounds with potential action in the control of glycemic homeostasis.

Ana Marla Duarte de Souza 18 April 2017 (has links)
Diversos estudos buscam identificar novas moléculas com ações regulatórias sobre a via de sinalização da insulina e consequentemente na homeostase da glicose. Assim, este trabalho visa avaliar o potencial de extratos de frutos no controle da homeostase glicêmica. Os frutos avaliados foram o morango (cv. Toianoca, Camarosa, Oso Grande e Camino Real), a amora-preta e a framboesa vermelha, em dois tempos de amostragem, sendo considerado como tempo A1 e tempo A2. As amostras foram caracterizadas quanto ao seu conteúdo de fenólicos totais, conteúdo de antocianinas monoméricas, capacidade antioxidante, avaliada pelos métodos DPPH e ORAC, ácido elágico total, capacidade de inibição da alfa-glicosidase e captação de glicose e lipólise em tecido adiposo de camundongos (ensaio explante). Dentre os frutos, no primeiro tempo de amostragem, a amora-preta e o morango, cv Oso Grande, foram os que apresentaram maior conteúdo de fenólicos totais (62,36 e 34,89 mg AG/g, respectivamente) no entanto não foram mantidos esses valores no segundo tempo de amostragem, com concentração 30% e 60% inferior, respectivamente; e maior concentração de antocianinas monoméricas (45,33 mg/g e 3,09 mg/g, respectivamente). Em relação a inibição da enzima alfa-glicosidase, avaliado em extrato metanólico, o fruto framboesa vermelha e o morango cv. Camino Real foram as que apresentaram alto potencial inibitório nos dois tempos de amostragem (IC50 0,47 mg FT e IC50 0,57 mg FT para framboesa vermelha e IC50 0,50 mg FT e IC50 0,46 mg FT para Camino Real). Quando avaliado os extratos enriquecidos em fenólicos, o valor de IC50 com maior potencial dentre os frutos avaliados foi da amora-preta, nos dois tempos A1 e A2 (0,0023 mg FT e 0,0021 mg FT, respectivamente). Para captação de glicose em tecido adiposo explate, ao utilizar a insulina para estimular a captação de glicose juntamente com o tratamento (extrato), esse estimulo foi efetivo no aumento da captação de glicose somente com as amostras cv. Camino Real e cv. Oso Grande. Isso pode ser explicado pela alta correlação encontrada de antocianinas identificadas no fruto, como pelargonidina-3-O-glicosídeo. Por outro lado, somente a amora-preta A1 aumentou a lipólise em condição basal, mas nenhuma fruta foi eficiente para reduzir a lipólise em condição estimulada pelo isoproterenol. Sendo assim, frutas vermelhas podem ser boas fontes de compostos bioativos, principalmente antocianinas, as quais podem ter corroborado positivamente com os resultados. / Several studies seek to identify new molecules with regulatory actions on the insulin signaling pathway and consequently on glucose homeostasis. Thus, this work aims to evaluate the potential of fruit extracts in the control of glycemic homeostasis. The fruits evaluated were strawberry (cv. Toianoca, Camarosa, Oso Grande and Camino Real), blackberry and red raspberry, in two sampling times, being considered as time A1 and time A2. Fruits were evaluated for total phenolic, monomeric anthocyanins and contents, antioxidant capacity, evaluated by DPPH and ORAC methods, alpha-glycosidase inhibition capacity and glucose uptake and lipolysis in adipose tissue of mice (explant assay). Among the fruits, in the first sampling period, blackberry and strawberry, cv. Oso Grande, showed the highest total phenolic content (62.36 and 34.89 mg AG / g, respectively), with a decrease of the 30% and 60%, respectively, in the second sampling time; and higher monomeric anthocyanins concentration (45.33 mg/g and 3.09 mg/g, respectively). The methanolic extracts of raspberry and the strawberry cv Camino Real (A1 and A2) presented the highest alpha-glucosidase inhibitory potential. Otherwise, the enriched-polyphenol extract of blackberry (A1 and A2) presented the highest potential among the evaluated fruits. Adipose tissue treated with strawberry cv. Camino Real and cv. Oso Grande was effective in increasing glucose uptake stimulated by insulin. This can be explained by the high correlation with the anthocyanins pelargonidin-3-O-glycoside identified in this fruit. In addition, blackberry A1 was the only sample to increase the lipolysis in basal condition, but all other fruits were not effective to decrease lipolysis in stimulated condition. Thus, berries could be a good sources of bioactive compounds to maintain the glucose homeostasis.

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