Non-alcoholic fatty liver disease (NAFLD) is a common consequence of metabolic syndrome (MetS) with the least severe form of NAFLD being hepatic steatosis, which is the accumulation of intrahepatic fat. Omega-3 polyunsaturated fatty acids (n3 PUFAs) are fatty acids in our diets commonly found in marine animals (eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) and certain plants (α-linoleic acid [ALA]). Although past studies have examined the consumption of marine sources or plant sources on hepatic steatosis and MetS parameters, individual n3 PUFA have yet to be compared to each other. Thus fa/fa Zucker rats were provided n3 PUFA diets containing ALA, EPA or DHA for 8 weeks relative to a linoleic acid (LA)-rich n6 PUFA diet provided to fa/fa and lean Zucker rats. Comparisons were to baseline fa/fa Zucker rats. It was shown that DHA prevented the progression of hepatic steatosis and was associated with improvements in insulin resistance.

No description available.

Bone morphogenetic proteins-2/4/7 (BMP-2/4/7) are important cytokines in systemic tissue morphogenesis. It has been demonstrated BMPs may have positive effects on liver repair and regeneration after hepatic injury. However, their function in the liver still remains unclear. D-galactosamine (D-gal) is a hepatotoxin used to induce hepatic failure. We employed D-gal and rat hepatoma cell line (1548) to investigate BMP-2/4/7 expression in hepatic injury induced by D-gal and probe their relations with liver repair and regeneration in hepatic injury. LDH release, mRNA and protein expression were detected. Results indicated that BMP-2/4/7 expression was activated by injury of rat hepatoma cells. It is indicative that repair and regeneration of the liver after hepatic injury and morphogenesis in early embryos seem to proceed through the same process. BMPs may be not only associated with hepatic injury after repair and regeneration, but also involved in chronic liver.

Orientador: Celio Raimundo Machado / Banca: Urara Kawazoe / Banca: Carlos Wilson Gomes Lopes / Banca: Luis Francisco Prata / Banca: Antonio Carlos Paulillo / Resumo: A infecção experimental por Eimeria stiedae em coelhos foi realizada com o objetivo de avaliar os sinais clínicos, alterações hematológicas, metabólicas e anatomopatológicas. Foram utilizados 50 coelhos, raça Nova Zelândia, brancos, com idade entre 40 - 60 dias e de pesos semelhantes. Os animais foram randomizados com relação ao peso e distribuídos em 2 grupos experimentais: grupo infectado, inoculado com 1ml de solução contendo 1x104 oocistos esporulados de E. stiedae; grupo controle, inoculado 1 ml de água destilada. Os animais foram avaliados semanalmente, durante 28 dias, a partir da data de inoculação. Os dados foram avaliados utilizando-se método estatístico não paramétrico pelo teste de Wilcoxon ao nível de 5% de significância. Coelhos infectados com oocistos esporulados de Eimeria stiedae tiveram cirrose hepática que afetou o funcionamento normal do referido órgão repercutindo em produção de proteínas de fase aguda e ocasionando consideráveis alterações metabólicas. / Abstract: The experimental infection by Eimeria stiedae in rabbits was performed to evaluate the clinical signs, hematological, metabolic and pathological changes. Fifty rabbits were used, New Zealand race, white, aged 40 to 60 days and of similar weight. The animals were randomized to the weight and distributed into 2 experimental groups: infected group, inoculated with 1 ml of solution containing 1x104 sporulated oocysts of Eimeria stiedae; control group, inoculated 1 ml of distilled water. The animals were evaluated weekly, for 28 days from the date of inoculation. A statistical was used non-parametric Wilcoxon test method at 5% level of significance. Rabbits infected with sporulated oocysts of Eimeria stiedae had liver cirrhosis that affected the normal functioning of the body resulting in production of acute phase proteins and cause considerable metabolic changes. / Doutor

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-02Bitstream added on 2014-06-13T21:06:05Z : No. of bitstreams: 1 freitas_flc_dr_jabo.pdf: 1139081 bytes, checksum: 856bf3c96a6f7eed40da1a35b470f7ae (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A infecção experimental por Eimeria stiedae em coelhos foi realizada com o objetivo de avaliar os sinais clínicos, alterações hematológicas, metabólicas e anatomopatológicas. Foram utilizados 50 coelhos, raça Nova Zelândia, brancos, com idade entre 40 - 60 dias e de pesos semelhantes. Os animais foram randomizados com relação ao peso e distribuídos em 2 grupos experimentais: grupo infectado, inoculado com 1ml de solução contendo 1x104 oocistos esporulados de E. stiedae; grupo controle, inoculado 1 ml de água destilada. Os animais foram avaliados semanalmente, durante 28 dias, a partir da data de inoculação. Os dados foram avaliados utilizando-se método estatístico não paramétrico pelo teste de Wilcoxon ao nível de 5% de significância. Coelhos infectados com oocistos esporulados de Eimeria stiedae tiveram cirrose hepática que afetou o funcionamento normal do referido órgão repercutindo em produção de proteínas de fase aguda e ocasionando consideráveis alterações metabólicas. / The experimental infection by Eimeria stiedae in rabbits was performed to evaluate the clinical signs, hematological, metabolic and pathological changes. Fifty rabbits were used, New Zealand race, white, aged 40 to 60 days and of similar weight. The animals were randomized to the weight and distributed into 2 experimental groups: infected group, inoculated with 1 ml of solution containing 1x104 sporulated oocysts of Eimeria stiedae; control group, inoculated 1 ml of distilled water. The animals were evaluated weekly, for 28 days from the date of inoculation. A statistical was used non-parametric Wilcoxon test method at 5% level of significance. Rabbits infected with sporulated oocysts of Eimeria stiedae had liver cirrhosis that affected the normal functioning of the body resulting in production of acute phase proteins and cause considerable metabolic changes.

No description available.

Indiana University-Purdue University Indianapolis (IUPUI) / Developmental changes in the liver significantly impact drug disposition. Due to the emergence of microRNAs as important regulators of drug disposition, we hypothesize that age-dependent change in microRNA expression and genetic variants in microRNA target sites contribute to variability in drug disposition. In human liver tissues, expression of 533 microRNAs and over 14,000 genes were measured. In all, 114 microRNAs were upregulated and 72 downregulated from fetal to pediatric, and 2 and 3, respectively, from pediatric to adult. Among these microRNAs, 99 microRNA-mRNA interactions were predicted or have previously been validated to target drug disposition genes and over 1,000 significant negative correlations were observed between miRNA-mRNA pairs. We validated these interactions using various cell culture models. Genetic variants in the promoter and coding regions of drug disposition genes have also been shown to alter enzyme expression and/or activity. However, these variants do not account for all variability in enzyme activity. Emerging evidence has shown that variants in the 3’UTR may explain variable drug response by altering microRNA regulation. Five 3’UTR variants were associated with significantly altered CYP2B6 activity in healthy human volunteers. The rs70950385 (AG>CA) variant was associated with decreased CYP2B6 activity among normal metabolizers. In vitro luciferase assays confirmed that the CA allele altered miR 1275 targeting of CYP2B6 mRNA. Due to the large number of 3’UTR variants predicted to alter microRNA regulation, a high-throughput method, PASSPORT-seq, was developed to test over 100 3’UTR variants simultaneously in different cell lines. Thirty-eight variants resulted in FDR-significant altered expression between wild-type and variant sequences. Our data suggest a mechanism for the marked changes in hepatic gene expression between the fetal and pediatric developmental periods, support a role for these age dependent microRNAs in regulating drug disposition, and provide strong evidence that 3’UTR variants are also an important source of variability in drug disposition.

TGFβ ligands, activin and myostatin have been shown to stimulate insulin production and secretion. Antagonists, Follistatin (FST) and Follistatin like 3 (FSTL3) were partially and fully ablated, respectively, creating hyperinsulinemic mice with fatty liver. Much research has surfaced on the connection between hepatic steatosis and hepatic insulin resistance. We present two different models, each with a different mechanism behind the development of fatty liver. FST288-only mice have increased synthesis of mRNA and proteins responsible for hepatic triglyceride (TG) uptake, while our double mutants have increased synthesis of mRNA and proteins responsible for TG synthesis. This alteration was likely independent of hepatic insulin resistance as livers from both mouse lines were insulin sensitive. Experiments conducted in this study to realize the causal factor of hepatic steatosis can be performed on adipose and muscle tissues in the future to better characterize the phenotype.

Studies have shown that overexpression of C1q TNF-related protein-3 (CTRP3) in mice fed a high alcohol diet can protect the animal from developing alcoholic fatty liver disease (ALD). Preliminary data from a recent study, however, indicated that overexpression of CTRP3 coupled with a high fat diet may cause kidney damage, including the development of (apparent) hyperlipidemic renal tubular vacuolization. If these findings are correct, using CTRP3 as a treatment for ALD would require careful monitoring of the patient diet to insure they limited the amount of fat in their diet. This recent study, however, utilized paraffin embedded kidney tissue. The processing required for paraffin embedding removes lipids/fat from the tissue. Thus, it is possible that what were assumed to be fat vacuoles in the kidney tubules may have been vacuoles unrelated to fat deposition, which represent a different aspect of tissue damage. Adipophilin and/or perilipin are proteins found in the membranes of lipocytes and membrane bound vesicles of fat. Immunohistochemical staining for adipophilin and/or perilipin 1 was used to determine if the damage seen in kidneys from the previous study was related to fat deposition or some other damaging factor. This information can inform further studies to determine how to proceed to safely administer CTRP3 to patients with ALD without damaging their kidneys.

The production of very low density lipoprotein and high density lipoprotein particles by the liver is a tightly regulated process, which begins with synthesis and assembly of core protein components in the rough endoplasmic reticulum. Factors influencing the production and metabolism of these particles are of immediate medical relevance, as their malfunction or hyperactivity can lead to an assortment of disease states. Hepatic lipase is a secreted liver enzyme, with many previously described roles in the metabolism and clearance of both high and low density lipoproteins. Increased production and assembly of this enzyme is an indicator of metabolic dysfunction, while its absence or insufficiency leads to pre-mature atherosclerosis and death. The present study shows that this enzyme’s role in lipoprotein metabolism is not confined to the degradation and clearance of these particles after they have been secreted. Experiments using co-immunoprecipitation targeted at hepatic lipase demonstrate that this protein interacts with ApoA1 and ApoB100, the core protein components of HDL and VLDL respectively, at the ER level in hepatocytes, as part of an enormous multi-subunit protein complex. This interaction with ApoA1 leads to decreased competence of hepatocytes to secrete HDL, which confers a pro-atherogenic phenotype. Analysis of ER to Golgi VLDL transport vesicles, produced with a cell-free in vitro budding assay, has revealed that hepatic lipase is co-secreted between these compartments with immature VLDL particles. Further analysis of cytosol isolated from hepatocytes demonstrates an interaction between hepatic lipase and the LDL-receptor related protein in a post-Golgi vesicle; the significance of which will be investigated in future studies.