Project description:To investigate the altered gene expression levels in mouse fibrotic liver tissues, C57BL6/J mice were intraperitoneally injected with CCl4 or vehicle twice every week. After 8 weeks, livers were harvested and RNA was extracted by Trizol. The gene expression levels were analyzed and compared between CCl4 treated group and vehicle treated (control) group.
Project description:To investigate the differences in microRNA expression profiles between fibrotic and normal livers, we performed microRNA microarrays for total RNA extracts isolated from mouse livers treated with carbontetrachloride (CCl4) or corn-oil for 10 weeks (n=3/group). MicroRNAs were considered to have significant differences in expression level when the expression difference showed more than two-fold change between the experimental and control groups at p<0.05. We found that 12 miRNAs were differentially expressed in CCl4-induced fibrotic liver. To induce chronic liver fibrosis, seven-week-old mice received 0.6 ml/kg body weight of carbon-tetrachloride (CCl4) dissolved in corn-oil by intraperitoneal (i.p.) injection, twice a week for 10 weeks (n=3). As a control, same number of mice was injected with equal volume of corn-oil for 10 weeks.
Project description:Purpose: To identify the alterations of genes transcriptional profile of livers obtained from Tgfbr2f/f mice and Tgfbr2HKO mice treated with CCl4. Methods: we generated hepatocyte-specific Tgfbr2 knockout (Tgfbr2HKO) mice by AAV8-TBG-Cre injection via the tail vein of Tgfbr2f/f. Tgfbr2f/f mice and Tgfbr2HKO were intraperitoneally injected with 0.25 ml/kg CCl4 (diluted 1:3 (v/v) in olive oil) twice a week for 4 weeks to induce liver fibrosis.
Project description:Liver firbrosis model of hepatocyte-specific FOXA2 knockout mice. Adeno-associated virus AAV8-TBG-Control or AAV8-TBG-Cre was injected via the tail vein of FOXA2flox/flox (FOXA2f/f) mice 2 weeks prior to CCl4 administration. Hepatic fibrosis was induced by injection of CCl4 twice per week for 4 weeks.
Project description:Background and aims: We aimed to study the pathogenesis of AH in an animal model of acute-on-chronic alcoholic liver disease which combines chronic hepatic fibrosis with intragastric alcohol administration. Methods: Adult male C57BL6/J mice were treated with CCl4 (0.2 ml/kg, 2×weekly by intraperitoneal injections for 6 weeks) to induce chronic liver fibrosis. Then, ethyl alcohol (EtOH) (up to 25 g/kg/day, for 3 weeks) was administered continuously to mice via a gastric feeding tube, with or without one-half dose of CCl4. Liver and serum markers were evaluated to characterize acute-on-chronic-alcoholic liver disease in our model. Results: CCl4 or EtOH treatment alone induced liver fibrosis or steatohepatitis, respectively, findings that were consistent with expected pathology. Combined treatment with CCl4 and EtOH resulted in a marked exacerbation of liver injury, as evident by the development of hepatic inflammation, marked steatosis, and pericellular fibrosis, and by increased serum transaminase levels, compared to mice treated with either treatment alone. Liver transcriptomic changes specific to combined treatment group demonstrated close concordance with pathways perturbed in human severe cases of AH. In addition to gene expression changes, E. coli and Candida species were also significantly more abundant in livers of mice co-treated with CCl4 and EtOH. Conclusions: Mice treated with CCl4 and EtOH displayed several key characteristics of human AH, including pericellular fibrosis, increased hepatic bacterial load, and dysregulation of the same molecular pathways. This model may be useful for developing therapeutics for AH.
Project description:To confirm the mechanism of miR-29a in liver fibrosis healing, we have employed whole genome microarray expression profiling as a discovery platform to identify genes. CCl4 and TAA liver fibrosis model mouse were used for this experiment. After five weeks liver fibrosis induction period, mouse have been observed for one week (1w) or two weeks (2w) and negative control nucleotide (N.C) or miR-29a were injected every 3 days on this period. We used CCl4 1w N.C (n = 1), 1w miR-29a (n = 1), 2w N.C (n = 1), 2w miR-29a (n = 1), and also used TAA model mouse (total n = 8) liver samples for microarray analysis. We can get only one gene (PDGF-c) as a target of miR-29a which relate to liver fibrosis and down-regulated more than 1.5 times in common miR-29a injected group than N.C group. CCl4 and TAA liver fibrosis model mouse were used for this experiment. After five weeks liver fibrosis induction period, mouse have been obserbed for one week (1w) or two weeks (2w) and negative control nucleotide (N.C) or miR-29a were injected every 3 days on this period. We used CCl4 1w N.C (n = 1), 1w miR-29a (n = 1), 2w N.C (n = 1), 2w miR-29a (n = 1), and also used TAA model mouse (total n = 8) liver samples for microarray analysis.
Project description:Livers from wild-type (WT) or Ppara knock-out (Ppara KO) C57Bl6 mice were used to prepare RNA which was then processed for analysis using MoGene-2_0-st Affymetrix microarrays according to standard procedures.
Project description:To investigate the role of hepatic Pck1 in the diethylnitrosamine (DEN)-induced HCC, a Pck1 knockout mouse in the liver was developed with Alb-Cre.HCC was induced in mice by combined treatment with DEN (75 mg/kg) and CCl4 (2 mL/ kg, twice per week for 12 weeks).At 32 weeks, the mice were sacrificed after fasting for 12 hours, and liver tissues with tumors were collected for examination. RNA was extracted from the livers of mice from all treatment groups and used for RNA seqencing. RNAseq data demonstrated that a lack of Pck1 can play an important role in the development of HCC.
Project description:To investigate the differences in microRNA expression profiles between fibrotic and normal livers, we performed microRNA microarrays for total RNA extracts isolated from mouse livers treated with carbontetrachloride (CCl4) or corn-oil for 10 weeks (n=3/group). MicroRNAs were considered to have significant differences in expression level when the expression difference showed more than two-fold change between the experimental and control groups at p<0.05. We found that 12 miRNAs were differentially expressed in CCl4-induced fibrotic liver.
Project description:A source of functioning hepatocytes for liver cell transplantation and liver support is in need. hESCs , when transplanted, generally form teratomas. We studied capacity of hESCs to differentiate to hepatocyte like cells under the effect of in vivo liver regeneration. In SCID-Beige mice hepatocyte replication peaked 48 hours after CCl4 injection; 24 hours earlier 106 hESCs or EBs at different stages of differentiation were transplanted into the spleen. Comparisons were made to teratomas formed in the hind limb of untreated animals. RT-PCR and gene microarray were used for liver and human specific markers. Immunohistochemistry to AFP,AAT, ALB, HEP-PAR I AND CK-18,19 were performed. EBs formed a single large teratoma in the spleen and small teratomas in the liver. Expression of PCR- identified liver specific markers was greater in the spleen than in the liver. Adult hepatocyte specific markers were expressed in the hind limb teratoma excised after 7 weeks. When late EBs were transplanted before CCl4 exposure, no teratomas formed. Rather, an abundance of probably undifferentiated ectodermal origin cells presented. In this descriptive study, transplanted early human EBs formed teratomas that differed in size and molecular markers. Within teratomas, the degree of maturation into hepatocytes correlated better with the time duration in vivo than with growth stimulation. Late EBs formed non differentiated ectodermal cells only in a regenerative microenvironment. 4 samples were analyzed. Clean mouse liver used as neg. control. Mouse liver injected with CCl4 and transplanted with late Ebs, tumor was not observed. Two mouse livers injected with CCl4 and transplanted with late Ebs, tumor was observed.