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Title: Gene expression profiles of murine fatty liver induced by the administration of valproic acid

Abstract

Valproic acid (VPA) has been used as anticonvulsants, however, it induces hepatotoxicity such as microvesicular steatosis and necrosis in the liver. To explore the mechanisms of VPA-induced steatosis, we profiled the gene expression patterns of the mouse liver that were altered by treatment with VPA using microarray analysis. VPA was orally administered as a single dose of 100 mg/kg (low-dose) or 1000 mg/kg (high-dose) to ICR mice and the animals were killed at 6, 24, or 72 h after treatment. Serum alanine aminotransferase and aspartate aminotransferase levels were not significantly altered in the experimental animals. However, symptoms of steatosis were observed at 72 h with low-dose and at 24 h and 72 h with high-dose. After microarray data analysis, 1910 genes were selected by two-way ANOVA (P < 0.05) as VPA-responsive genes. Hierarchical clustering revealed that gene expression changes depended on the time rather than the dose of VPA treatment. Gene profiling data showed striking changes in the expression of genes associated with lipid, fatty acid, and steroid metabolism, oncogenesis, signal transduction, and development. Functional categorization of 1156 characteristically up- and down-regulated genes (cutoff > 1.5-fold) revealed that 60 genes were involved in lipid metabolism that was interconnected with biologicalmore » pathways for biosynthesis of triglyceride and cholesterol, catabolism of fatty acid, and lipid transport. This gene expression profile may be associated with the known steatogenic hepatotoxicity of VPA and it may provide useful information for prediction of hepatotoxicity of unknown chemicals or new drug candidates through pattern recognition.« less

Authors:
 [1];  [2];  [1];  [2];  [3];  [2];  [1];  [2];  [3];  [2];  [4];  [2];  [5];  [2];  [6];  [2];  [7];  [2];  [7];  [2] more »;  [8] « less
  1. College of Pharmacy (Korea, Republic of)
  2. (Korea, Republic of)
  3. Seoul National University Biomedical Informatics College of Medicine (Korea, Republic of)
  4. Department of Veterinary Public Health College of Veterinary Medicine, Seoul National University, Seoul 151-742 (Korea, Republic of)
  5. Department of Biochemistry College of Medicine, Ewha Womans University, Seoul 158-710 (Korea, Republic of)
  6. School of Veterinary Medicine, Kangwon National University, Kangwon 200-701 (Korea, Republic of)
  7. Department of Pathology College of Medicine (Korea, Republic of)
  8. College of Pharmacy (Korea, Republic of) and Bio-MAX Institute (Korea, Republic of) and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 (Korea, Republic of). E-mail: molee@snu.ac.kr
Publication Date:
OSTI Identifier:
20976897
Resource Type:
Journal Article
Resource Relation:
Journal Name: Toxicology and Applied Pharmacology; Journal Volume: 220; Journal Issue: 1; Other Information: DOI: 10.1016/j.taap.2006.12.016; PII: S0041-008X(06)00487-X; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ALANINES; ANTICONVULSANTS; BIOLOGICAL PATHWAYS; BIOSYNTHESIS; CATABOLISM; CHOLESTEROL; DATA ANALYSIS; DOSES; GENES; ION CYCLOTRON-RESONANCE; LIVER; MICE; NECROSIS; PATTERN RECOGNITION; SYMPTOMS; TRIGLYCERIDES

Citation Formats

Lee, Min-Ho, Bio-MAX Institute, Hong, Il, Bio-MAX Institute, Kim, Mingoo, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Lee, Byung Hoon, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kim, Ju-Han, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kang, Kyung-Sun, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kim, Hyung-Lae, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Yoon, Byung-Il, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Chung, Heekyoung, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kong, Gu, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, and Lee, Mi-Ock. Gene expression profiles of murine fatty liver induced by the administration of valproic acid. United States: N. p., 2007. Web. doi:10.1016/j.taap.2006.12.016.
Lee, Min-Ho, Bio-MAX Institute, Hong, Il, Bio-MAX Institute, Kim, Mingoo, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Lee, Byung Hoon, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kim, Ju-Han, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kang, Kyung-Sun, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kim, Hyung-Lae, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Yoon, Byung-Il, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Chung, Heekyoung, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kong, Gu, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, & Lee, Mi-Ock. Gene expression profiles of murine fatty liver induced by the administration of valproic acid. United States. doi:10.1016/j.taap.2006.12.016.
Lee, Min-Ho, Bio-MAX Institute, Hong, Il, Bio-MAX Institute, Kim, Mingoo, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Lee, Byung Hoon, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kim, Ju-Han, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kang, Kyung-Sun, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kim, Hyung-Lae, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Yoon, Byung-Il, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Chung, Heekyoung, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, Kong, Gu, Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791, and Lee, Mi-Ock. Sun . "Gene expression profiles of murine fatty liver induced by the administration of valproic acid". United States. doi:10.1016/j.taap.2006.12.016.
@article{osti_20976897,
title = {Gene expression profiles of murine fatty liver induced by the administration of valproic acid},
author = {Lee, Min-Ho and Bio-MAX Institute and Hong, Il and Bio-MAX Institute and Kim, Mingoo and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Lee, Byung Hoon and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Kim, Ju-Han and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Kang, Kyung-Sun and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Kim, Hyung-Lae and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Yoon, Byung-Il and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Chung, Heekyoung and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Kong, Gu and Toxicogenomics Research Consortium, Hanyang University, Seoul 133-791 and Lee, Mi-Ock},
abstractNote = {Valproic acid (VPA) has been used as anticonvulsants, however, it induces hepatotoxicity such as microvesicular steatosis and necrosis in the liver. To explore the mechanisms of VPA-induced steatosis, we profiled the gene expression patterns of the mouse liver that were altered by treatment with VPA using microarray analysis. VPA was orally administered as a single dose of 100 mg/kg (low-dose) or 1000 mg/kg (high-dose) to ICR mice and the animals were killed at 6, 24, or 72 h after treatment. Serum alanine aminotransferase and aspartate aminotransferase levels were not significantly altered in the experimental animals. However, symptoms of steatosis were observed at 72 h with low-dose and at 24 h and 72 h with high-dose. After microarray data analysis, 1910 genes were selected by two-way ANOVA (P < 0.05) as VPA-responsive genes. Hierarchical clustering revealed that gene expression changes depended on the time rather than the dose of VPA treatment. Gene profiling data showed striking changes in the expression of genes associated with lipid, fatty acid, and steroid metabolism, oncogenesis, signal transduction, and development. Functional categorization of 1156 characteristically up- and down-regulated genes (cutoff > 1.5-fold) revealed that 60 genes were involved in lipid metabolism that was interconnected with biological pathways for biosynthesis of triglyceride and cholesterol, catabolism of fatty acid, and lipid transport. This gene expression profile may be associated with the known steatogenic hepatotoxicity of VPA and it may provide useful information for prediction of hepatotoxicity of unknown chemicals or new drug candidates through pattern recognition.},
doi = {10.1016/j.taap.2006.12.016},
journal = {Toxicology and Applied Pharmacology},
number = 1,
volume = 220,
place = {United States},
year = {Sun Apr 01 00:00:00 EDT 2007},
month = {Sun Apr 01 00:00:00 EDT 2007}
}
  • Valproic acid (VPA) is used clinically to treat epilepsy, however it induces hepatotoxicity such as microvesicular steatosis. Acute hepatotoxicity of VPA has been well documented by biochemical studies and microarray analysis, but little is known about the chronic effects of VPA in the liver. In the present investigation, we profiled gene expression patterns in the mouse liver after subchronic treatment with VPA. VPA was administered orally at a dose of 100 mg/kg/day or 500 mg/kg/day to ICR mice, and the livers were obtained after 1, 2, or 4 weeks. The activities of serum liver enzymes did not change, whereas triglyceridemore » concentration increased significantly. Microarray analysis revealed that 1325 genes of a set of 32,996 individual genes were VPA responsive when examined by two-way ANOVA (P < 0.05) and fold change (> 1.5). Consistent with our previous results obtained using an acute VPA exposure model (Lee et al., Toxicol Appl Pharmacol. 220:45-59, 2007), the most significantly over-represented biological terms for these genes included lipid, fatty acid, and steroid metabolism. Biological pathway analysis suggests that the genes responsible for increased biosynthesis of cholesterol and triglyceride, and for decreased fatty acid {beta}-oxidation contribute to the abnormalities in lipid metabolism induced by subchronic VPA treatment. A comparison of the VPA-responsive genes in the acute and subchronic models extracted 15 commonly altered genes, such as Cyp4a14 and Adpn, which may have predictive power to distinguish the mode of action of hepatotoxicants. Our data provide a better understanding of the molecular mechanisms of VPA-induced hepatotoxicity and useful information to predict steatogenic hepatotoxicity.« less
  • Chronic consumption of ethanol can cause cumulative liver damage that can ultimately lead to cirrhosis. To explore the mechanisms of alcoholic steatosis, we investigated the global intrahepatic gene expression profiles of livers from mice administered alcohol. Ethanol was administered by feeding the standard Lieber-DeCarli diet, of which 36% (high dose) and 3.6% (low dose) of the total calories were supplied from ethanol for 1, 2, or 4 weeks. Histopathological evaluation of the liver samples revealed fatty changes and punctate necrosis in the high-dose group and ballooning degeneration in the low-dose group. In total, 292 genes were identified as ethanol responsive,more » and several of these differed significantly in expression compared to those of control mice (two-way ANOVA; p < 0.05). Specifically, the expression levels of genes involved in hepatic lipid transport and metabolism were examined. An overall net increase in gene expression was observed for genes involved in (i) glucose transport and glycolysis, (ii) fatty acid influx and de novo synthesis, (iii) fatty acid esterification to triglycerides, and (iv) cholesterol transport, de novo cholesterol synthesis, and bile acid synthesis. Collectively, these data provide useful information concerning the global gene expression changes that occur due to alcohol intake and provide important insights into the comprehensive mechanisms of chronic alcoholic steatosis.« less
  • Ethanol induces cumulative liver damage including steatosis, steatohepatitis and cirrhosis. The aim of this study is to investigate the global intrahepatic gene expression profile in the mouse liver treated with ethanol. A single oral dose of 0.5 or 5 g/kg ethanol was administered to male ICR mice, and liver samples were obtained after 6, 24 and 72 h. Histopathological evaluation showed typical fatty livers in the high-dose group at 24 h. Microarray analysis identified 28 genes as being ethanol responsive (two-way ANOVA; p < 0.05), after adjustment by the Benjamini-Hochberg multiple testing correction; these genes displayed {>=} 2-fold induction ormore » repression. The expression of genes that are known to be involved in fatty acid synthesis was examined. The transcript for lipogenic transcription factor, sterol regulatory element (SRE)-binding factor 1 (Srebf1), was upregulated by acute ethanol exposure. Of the genes known to contain SRE or SRE-like sequences and to be regulated by SRE-binding protein 1 (SREBP1), those encoding malic enzyme (Mod1), ATP-citrate lyase (Acly), fatty acid synthase (Fasn) and stearyl-CoA desaturase (Scd1) were induced by ethanol. Quantitative real-time PCR confirmed the changes in the expression levels of the selected genes. The change in the Srebf1 mRNA level correlates well with that of the SREBP1 protein expression as well as its binding to the promoters of the target genes. The present study identifies differentially expressed genes that can be applied to the biomarkers for alcohol-binge-induced fatty liver. These results support the hypothesis by which ethanol-induced steatosis in mice is mediated by the fatty acid synthetic pathway regulated by SREBP1.« less
  • Benzo[a]pyrene (BaP) is one of many polycyclic aromatic hydrocarbons that have been identified as major risk factors for developing various cancers. We previously demonstrated that the liver cancer susceptibility gene glycine N-methyltransferase (GNMT) is capable of binding with BaP and protecting cells from BaP-7,8-diol 9,10-epoxide-DNA adduct formation. In this study, we used a cytotoxicity assay to demonstrate that the higher expression level of GNMT, the lower cytotoxicity occurred in the cells treated with BaP. In addition, a cDNA microarray containing 7,597 human genes was used to examine gene expression patterns in BaP-treated HepG2 (a liver cancer cell line that expressesmore » very low levels of GNMT) and SCG2-1-1 (a stable HepG2 clone that expresses high levels of GNMT) cells. The results showed that among 6,018 readable HepG2 genes, 359 (6.0%) were up-regulated more than 1.5-fold and 768 (12.8%) were down-regulated. Overexpression of GNMT in SCG2-1-1 cells resulted in the down-regulation of genes related to the detoxification, kinase/phosphatase pathways, and oncogenes. Furthermore, real-time PCR was used to validate microarray data from 21 genes belonging to the detoxification pathway. Combining both microarray and real-time PCR data, the results showed that among 89 detoxification pathway genes analyzed, 22 (24.7%) were up-regulated and 6 (6.7%) were down-regulated in BaP-treated HepG2 cells, while in the BaP-treated SCG2-1-1 cells, 12 (13.5%) were up-regulated and 26 (29.2%) were down-regulated (P < 0.001). Therefore, GNMT sequesters BaP, diminishes BaP's effects to the liver detoxification pathway and prevents subsequent cytotoxicity.« less
  • Some compounds have structural isomers of which one is apparently carcinogenic, and the other not. Because of the similarity of their chemical structures, comparisons of their effects can allow gene expression elicited in response to the basic skeletons of the isomers to be disregarded. We compared the gene expression profiles of male Fischer 344 rats administered by daily oral gavage up to 28 days using an in-house oligo microarray. 2-Acetylaminofluorene (2-AAF), 2,4-diaminotoluene (2,4-DAT), 2-nitropropane (2-NP), and 2-nitro-p-phenylenediamine (2-NpP) are hepatocarcinogenic. However, their isomers, 4-acetylaminofluorene (4-AAF), 2,6-diaminotoluene (2,6-DAT), 1-nitropropane (1-NP), and 4-nitro-o-phenylenediamine (4-NoP), are non-hepatocarcinogenic. Because of the limited carcinogenicity ofmore » 2-NpP, we attempted to perform two-parametric comparison analyses with (1) a set of 4 isomers: 2-AAF, 2,4-DAT, 2-NP, and 2-NpP as 'carcinogenic', and 4-AAF, 2,6-DAT, 1-NP, and 4-NoP as 'non-carcinogenic'; and (2) a set of 3 isomers: 2-AAF, 2,4-DAT, and 2-NP, as 'carcinogenic', and 4-AAF, 2,6-DAT, and 1-NP as 'non-carcinogenic'. After ratio filtering and Welch's approximate t-test analysis, 54 and 28 genes were selected from comparisons between the sets of 3 and 4 isomers, respectively, for day 28 data. Using hierarchical clustering analysis with the 54 or 28 genes, 2-AAF, 2,4-DAT, and 2-NP clustered into a 'carcinogenic' branch. 2-NpP was in the same cluster as 4-NoP and 4-AAF. This clustering corresponded to the previous finding that 2-NpP is not carcinogenic in male Fischer 344 rats, which indicates that comparing the differences in gene expression elicited by different isomers is an effective method of developing a prediction system for carcinogenicity.« less