Project description:In order to study the physiological consequences of a high-copper diet on hepatic gene expression, 6 mM CuCl2 was added to the drinking water for a period of 1 month. After this period, livers of seven control mice and eight copper-treated mice were isolated and were subjected to microarray analysis and copper measurements. The hepatic gene expression profile of copper-treated mice was compared to non-treated mice using a pooled reference.
Project description:Mice developing diethylnitrosamine (DEN)-induced hepatocellular carcinoma were transfected in vivo with Gas1 by hydrodynamic gene delivery. The treatment significantly (p < 0.05) reduced the number of large tumors (more than 2 mm diameter) and histological analyses revealed that the number of carcinoma foci in liver was reduced in favor of less malignant lesions, such as hyperplastic areas, and that the number of lung metastases was also diminished. To study the effects of DEN and the subsequent transfection with Gas1, an expression microarray experiment was designed in which the transcription of 20758 genes in liver was determined. A triangular analysis of microarray data shows that transfection with Gas1 restores the level of transcription level of 182 out of the 698 genes, whose expression was affected by DEN. Therefore, transfection of liver tumor-bearing animals with Gas1 significantly reduces the size and proliferative activity of tumors and restores the transcriptional profile of the liver. Here we studied effects of diethylnitrosamine treatment of mice on the transcription of 20758 liver genes and the consequences of transfecting the livers with Gas1.
Project description:Retinoid-related orphan receptors RORalpha and RORgamma are both expressed in liver; however, their physiological functions have not yet been clearly defined. RORalpha1 and RORgamma1, but not RORalpha4, show an oscillatory pattern of expression during circadian rhythm. To obtain insight into the physiological functions of ROR receptors in liver, we analyzed the gene expression profiles of livers from WT, RORalpha-deficient staggerer mice (RORalphasg/sg), RORgamma-/-, and RORalpha sg/sgRORgamma-/- double knockout (DKO) mice by microarray analysis. DKO mice were generated to study functional redundancy between RORalpha and RORgamma. These analyses demonstrated that RORalpha and RORgamma affect the expression of a number of genes. RORalpha and RORalpha are particularly important in the regulation of genes encoding several Phase I and Phase II metabolic enzymes, including several 3beta-hydroxysteroid dehydrogenases (Hsd3b), the cytochrome P450 (Cyp) enzymes Cyp7b1, Cyp4a14, and Cyp8b1, and the sulfotransferases Sult1e1 and Sult2a1. Our data show that RORalpha and RORgamma can influence gene expression positively as well as negatively. In addition, our results indicate that RORalpha and RORgamma each affect the expression of a specific set of genes but also exhibit functional redundancy. Our study shows that RORalpha and RORgamma receptors influence the regulation of several metabolic pathways, including those involved in the metabolism of steroids, bile acids, and xenobiotics, suggesting that RORs are important in the control of metabolic homeostasis. Experiment Overall Design: We generated RORalpha and RORgamma double knockout (DKO) mice and compared the gene expression profiles of livers from wild type (WT), RORalphasg/sg, RORgamma-/-, and DKO mice by microarray analysis
Project description:In order to explore the potential mechanism of HNF4α in ICC, RNA-seq was performed to obtain differentially expressed genes (DEGs) between HuCCT-1 cells infected with AdGFP or AdHNF4α
Project description:Harsh environmental conditions including microgravity and radiation during prolonged spaceflights are known to alter hepatic metabolism. Our studies have focused on the analysis of possible changes in metabolic pathways in livers of mice which experienced 30 days of spaceflight with and without an additional re-adaption period of 7 days compared to control mice on Earth. Utilizing shotgun mass spectrometry and label-free quantification, we performed proteomic profiling to investigate mice livers from the spaceflight project “Bion-M 1”. No significant alterations in protein levels were observed between control mice liver and spaceflight mice which is possibly caused by insufficient fold change detection combined with high variances within the groups. In contrast, our results show that more than a third of the quantified protein levels are altered comparing the liver proteome of mice with and without re-adaption time after their spaceflight. Proteins related to amino acid metabolism showed higher levels after re-adaption, which may indicate higher rates of gluconeogenesis. Members of the peroxisome proliferator-activated receptor pathway reconstitute their level after 7 days due to a decrease in fold change, which indicates decreased signs of non-alcoholic fatty liver disease. Moreover, bile acid secretion regenerates on Earth due to reconstitution of related transmembrane proteins and elevated levels of the drug-metabolising enzymes belonging to the CYP superfamily decrease 7 days after the spaceflight. Thus, our study demonstrates reconstitution of pharmacological response and early signs of non-alcoholic fatty liver disease recover within 7 days, whereas glucose uptake should be monitored due to alterations in gluconeogenesis.
Project description:Purpose: The goal of this study was to determine biological consequences during liver regeneration following partial hepatectomy in mice by next-generation sequencing. A particular interest was to compare mice with either a floxed b-PDGFR allele to mice that harbored a deletion of b-PDGFR in hepatic stellate cells (HSCs), by crossing b-PDGFR fl/fl mice with transgenic GFAP-Cre mice. Methods: b-PDGFR fl/fl mice or mice with a HSC-specific deletion of b-PDGFR underwent either sham operation or 70% partial hepatectomy. Following 72 hours, livers were collected and total RNA was extracted using tizol, followed by a purification using Quiagen spin columns including an on-column DNAse digestion step. Conclusion: Our study represents a detailed analysis of hepatic transcriptome, with biologic replicates, generated by RNA-seq technology of livers following sham operation or partial hepatectomy in b-PDGFR fl/fl mice or b-PDGFRfl/fl/GRAP-Cre mice. Whole liver mRNA profiles of sham operated livers or livers collected 72hours after partial hepatectomy of beta-PDGFR fl/fl and beta-PDGFR fl/fl/GFAP-Cre (creating a hepatic stellate cell-specific deletion of b-PDGFR) mice were generated by deep sequencing, in duplicate, using Illumina HiSeq2000.
Project description:We used NGS-derived transcriptome profiling (RNA-seq) to compare the transcriptional difference between human aortic smooth muscle cells (HASMCs) infected with 20MOI adenovirus encoding human BAF60a (AdBAF60a) or GFP (AdGFP)
Project description:BACKGOUND: Drinking water can be contaminated with pharmaceuticals. However, it is uncertain whether this contamination can have harmful consequences for the liver, especially in the context of obesity. OBJECTIVES: To determine whether chronic, low dose exposure to pharmaceuticals could have deleterious effects in livers of lean and obese mice. METHODS: Lean and ob/ob male mice (5-week-old) were treated for 4 months with a mixture of 11 drugs (acetaminophen, caffeine, carbamazepine, cotinine, diclofenac, erythromycin, ibuprofen, phenazone, roxithromycin, salicylic acid and sulfamethoxazole) provided in drinking water at a concentration of 1 mg/L (for each drug). At the end of the treatment, investigations were performed in liver and plasma. RESULTS: Some liver and plasma abnormalities were observed in ob/ob mice treated with the cocktail containing 1 mg/L of each drug. For this dosage, a gene expression analysis by microarray showed altered expression of circadian genes (e.g. Bmal1, Dbp, Cry1) in lean and obese mice. RT-qPCR analyses carried out in all groups of animals indicated that expression of 8 different circadian genes was significantly modified in a dose-dependent manner. For some genes, a significant modification was observed for dosages as low as 100-1,000 ng/L. Drug mixture and obesity presented an additive effect on circadian gene expression. These data were confirmed in an independent study performed in female mice. CONCLUSIONS: Chronic, low dose exposure to pharmaceuticals disturbed hepatic expression of circadian genes, especially in obese mice. Because some of the 11 drugs can be found in the drinking water at such concentrations (e.g. acetaminophen, carbamazepine, ibuprofen) our data could be relevant in environmental toxicology, in particular for obese individuals exposed to these contaminants. C57BL/6J lean and ob/ob male mice (5-week-old) were treated for 4 months with a mixture of 11 drugs provided in drinking water at a concentration of 1 mg/L (for each drug). 4 groups were designed: untreated versus treated WT and ob/ob mice (n=6 mice per group).