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Status |
Public on Oct 04, 2015 |
Title |
Influence of ascorbic acid depletion on gene expression in liver and lipid metabolism |
Organism |
Mus musculus |
Experiment type |
Expression profiling by array
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Summary |
Ascorbic acid (AA) is a powerful antioxidant and play as a cofactor for various enzymes in vivo. In this study, we investigated the effect of AA depletion on gene expression in the liver and lipid metabolism by using SMP30/GNL knockout (KO) mice which are unable to biosynthesis AA. First, we performed microarray analysis. Briefly, SMP30/GNL KO mice were weaned and divided into two groups; AA-depleted and supplemented groups, which mice were free access to water containing 1.5 g/L AA. After 4 weeks, mRNA was isolated and purified from the liver. In this study, Affymetrix® GeneChip® was used for microarray analysis. Actually, AA-depletion altered many gene expressions related to lipid metabolism. Especially, Cytochrome P450 7a1 (Cyp7a1), a late-limiting enzyme of bile acid biosynthesis, gene expression was significantly up-regulated. We also confirmed Cyp7a1 protein levels by Western blotting. Next, we investigated the influence of AA depletion on lipid metabolism. We examined the lipid and bile acid levels in the liver, plasma, and gallbladder from SMP30/GNL KO mice. Amount of total bile acid (TBA), free fatty acid (FA), total cholesterol (TC), triglyceride (TG), and phospholipids (PL) were measured by colorimetric method. AA depletion reduced TBA levels in the liver and gallbladder. However, FA, TC, TG, and PL in the plasma and liver were not changed by AA depletion. Although Cyp7a1 gene expression and protein levels were increased by AA depletion, amount of bile acid were reduced. Conclusively, we have shown that AA depletion reduced bile acid biosynthesis and elevated Cyp7a1 gene expression and protein levels. Thus, AA is an essential for bile acid biosynthesis pathway.
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Overall design |
To investigate SMP30 gene in metabolism of ascorbic acid, liver transcriptome were compared between SMP30/GNL KO mice fed with AA-free diet, AA-supplemented KO mice, and wild type mice.
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Contributor(s) |
Takahashi K, Kishimoto Y, Konishi T, Shimokado K, Maruyama N, Ishigami A, Konishi T |
Citation(s) |
24704458 |
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Submission date |
Oct 05, 2012 |
Last update date |
Mar 04, 2019 |
Contact name |
Tomokazu Konishi |
E-mail(s) |
konishi@akita-pu.ac.jp
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Phone |
+81-18-872-1603
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Organization name |
Akita Prefectural University
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Department |
Bioresource Sciences
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Lab |
Molecular Genetics
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Street address |
Shimoshinjyo Nishi
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City |
Akita |
State/province |
Akita |
ZIP/Postal code |
010-0195 |
Country |
Japan |
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Platforms (1) |
GPL6246 |
[MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version] |
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Samples (6)
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Relations |
BioProject |
PRJNA176691 |
Supplementary file |
Size |
Download |
File type/resource |
GSE41365_RAW.tar |
24.3 Mb |
(http)(custom) |
TAR (of CEL) |
Processed data included within Sample table |
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