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Accession IconGSE41365

Influence of ascorbic acid depletion on gene expression in liver and lipid metabolism

Organism Icon Mus musculus
Sample Icon 6 Downloadable Samples
Technology Badge Icon Affymetrix Mouse Gene 1.0 ST Array (mogene10st)

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Description
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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