Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Impaired Hepatic Vitamin A Metabolism in NAFLD Mice Leading to Vitamin A Accumulation in Hepatocytes

doi: 10.1016/j.jcmgh.2020.07.006

Figure Lengend Snippet: Hepatic fat accumulation and steatohepatitis in HFC-fed mice . Mice fed chow or HFC diet for 12 or 20 weeks were analyzed for ( A ) body weight, ( B ) liver weight, ( C ) liver total cholesterol levels, ( D ) liver triglyceride (TG) levels, ( E ) liver free cholesterol levels, F ) plasma total cholesterol levels, ( G ) plasma free cholesterol levels, ( H ) plasma insulin levels, and ( I ) hematoxylin and eosin (H&E) staining, Oil Red O staining, and CD68 immunohistochemistry. Quantification of (immuno)histochemistry is described in the Methods and Materials. ∗ P ≤ .05, ∗∗ P ≤ .01, ∗∗ ∗P ≤ .001.

Article Snippet: Immunohistochemistry for CD68 (1:300; rabbit anti-mouse CD68; #137002; Biolegio, Nijmegen, the Netherlands) and LRAT (1:200 rabbit anti-mouse LRAT; #28075; Takara Immuno-Biological Laboratories, Gunma, Japan) was performed on snap-frozen liver sections as previously described.

Techniques: Clinical Proteomics, Staining, Immunohistochemistry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Impaired Hepatic Vitamin A Metabolism in NAFLD Mice Leading to Vitamin A Accumulation in Hepatocytes

doi: 10.1016/j.jcmgh.2020.07.006

Figure Lengend Snippet: Expression of hepatic genes involved in lipid uptake, lipid synthesis, inflammation, and fibrosis in HFC-fed mice. Mice fed chow or HFC diet for 12 or 20 weeks were analyzed by Q-PCR for hepatic expression of genes involved in ( A ) hepatic lipid uptake ( Srb1 , Cd36 ) and synthesis ( Scd1 , Fasn , Acc1 ), ( B ) hepatic inflammation ( Cd68 , Tnf-α , Nos2 , Ccl2 , Il-1β , Il-6 ), and ( C ) liver fibrosis ( Coll1a1 , Acta2 , Tgf-β , and Timp1 ). Hepatic lipid uptake and synthesis, hepatic inflammation, and hepatic fibrosis were strongly increased in HFC-fed mice. ∗ P ≤ .05, ∗∗ P ≤ .01, ∗∗ ∗P ≤ .001.

Article Snippet: Immunohistochemistry for CD68 (1:300; rabbit anti-mouse CD68; #137002; Biolegio, Nijmegen, the Netherlands) and LRAT (1:200 rabbit anti-mouse LRAT; #28075; Takara Immuno-Biological Laboratories, Gunma, Japan) was performed on snap-frozen liver sections as previously described.

Techniques: Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Impaired Hepatic Vitamin A Metabolism in NAFLD Mice Leading to Vitamin A Accumulation in Hepatocytes

doi: 10.1016/j.jcmgh.2020.07.006

Figure Lengend Snippet: Accumulation of vitamin A in lipid-loaded hepatocytes in vivo and in vitro. ( A ) Oil Red O staining (left panels) and vitamin A–specific autofluorescence (middle panels) immunohistochemistry of LRAT (right panels) of liver sections of chow-fed (top panels) and HFC-fed (bottom panels) mice. Vitamin A–specific autofluorescence was strongly increased in livers of HFC-fed mice and was located predominantly in hepatocytes compared with its location in sparsely present HSCs in livers of chow-fed mice. ( B ) Freshly isolated and 4-hour attached primary rat hepatocytes and quiescent primary HSCs were and immediately treated with and without palmitate (PA) for 24 hours. Cells were harvested and analyzed by Q-PCR analysis for Lrat and Pnpla3 expression. The gene expression is presented in 2 -ΔΔCT and normalized to 18S. ( C ) Freshly isolated and 4-hour attached primary rat hepatocytes were treated for 48 hours with and without PA and retinol (ROH), followed by analysis of cellular retinyl PA and ROH levels. ∗ P ≤ .05, ∗∗ P ≤ .01.

Article Snippet: Immunohistochemistry for CD68 (1:300; rabbit anti-mouse CD68; #137002; Biolegio, Nijmegen, the Netherlands) and LRAT (1:200 rabbit anti-mouse LRAT; #28075; Takara Immuno-Biological Laboratories, Gunma, Japan) was performed on snap-frozen liver sections as previously described.

Techniques: In Vivo, In Vitro, Staining, Immunohistochemistry, Isolation, Expressing, Gene Expression

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Impaired Hepatic Vitamin A Metabolism in NAFLD Mice Leading to Vitamin A Accumulation in Hepatocytes

doi: 10.1016/j.jcmgh.2020.07.006

Figure Lengend Snippet: Primers and Probes Used in Study for Analysis of Target Genes

Article Snippet: Immunohistochemistry for CD68 (1:300; rabbit anti-mouse CD68; #137002; Biolegio, Nijmegen, the Netherlands) and LRAT (1:200 rabbit anti-mouse LRAT; #28075; Takara Immuno-Biological Laboratories, Gunma, Japan) was performed on snap-frozen liver sections as previously described.

Techniques: