mRNA expressions of transcription factors related with hepatic lipid metabolsim were assessed.

<p>Changes of PPARγ2, SREBP1c, ChREBP and PPARα expressions from mice livers were analyzed by RT-PCR (A), and AM significantly inhibited HFD-induced PPARγ2 expression (B).</p

The changes in PPARγ2 protein expression were assessed in mice livers.

<p>Western blot analysis showed increase of PPARγ2 protein expression in HFD group and suppression of the protein in HFD+AM group (A) although the suppression was not statistically significant (B).</p

<i>Aronia melanocarpa</i> Extract Ameliorates Hepatic Lipid Metabolism through PPARγ2 Downregulation

<div><p>Nonalcoholic fatty liver disease (NAFLD) is a hepatic manifestation of metabolic syndrome. Studies have demonstrated that anthocyanin-rich foods may improve hyperlipidemia and ameliorate hepatic steatosis. Here, effects of <i>Aronia melanocarpa</i> (AM), known to be rich of anthocyanins, on hepatic lipid metabolism and adipogenic genes were determined. AM was treated to C57BL/6N mice fed with high fat diet (HFD) or to FL83B cells treated with free fatty acid (FFA). Changes in levels of lipids, enzymes and hormones were observed, and expressions of adipogenic genes involved in hepatic lipid metabolism were detected by PCR, Western blotting and luciferase assay. In mice, AM significantly reduced the body and liver weight, lipid accumulation in the liver, and levels of biochemical markers such as fatty acid synthase, hepatic triglyceride and leptin. Serum transaminases, indicators for hepatocyte injury, were also suppressed, while superoxide dismutase activity and liver antioxidant capacity were significantly increased. In FL83B cells, AM significantly reduced FFA-induced lipid droplet accumulation. Protein synthesis of an adipogenic transcription factor, peroxisome proliferator-activated receptor γ2 (PPARγ2) was inhibited <i>in vivo</i>. Furthermore, transcriptional activity of PPARγ2 was down-regulated <i>in vitro</i>, and mRNA expression of PPARγ2 and its downstream target genes, adipocyte protein 2 and lipoprotein lipase were down-regulated by AM both <i>in vitro</i> and <i>in vivo</i>. These results show beneficial effects of AM against hepatic lipid accumulation through the inhibition of PPARγ2 expression along with improvements in body weight, liver functions, lipid profiles and antioxidant capacity suggesting the potential therapeutic efficacy of AM on NAFLD.</p></div

AM improves HFD-induced redox imbalance.

<p>Hepatic SOD activity (A) and TEAC (B) were decreased in HFD group, but these were significantly increased in HFD+AM group.</p

AM affects HFD-induced lipogenesis, hepatocellular injury and leptin level.

<p>While HFD induced significant elevation in intrahepatic TG (A), FAS (B), serum ALT (C), AST (D) and leptin (E), these were significantly inhibited in HFD+AM group.</p

AM prevents HFD-induced intrahepatic lipid accumulation and weight gain.

<p>Hepatic steatosis was reduced in HFD+AM group both grossly (A) and histologically (B; H&E stain, magnification 100×). HFD-induced increase of body and liver weight were also significantly deterred by AM (C).</p

MicroRNA-27a Modulates HCV Infection in Differentiated Hepatocyte-Like Cells from Adipose Tissue-Derived Mesenchymal Stem Cells

<div><p>Background and Aims</p><p>Despite the discovery of hepatitis C virus (HCV) entry factor, the mechanism by which it is regulated by miRNAs remains unclear. Adipose tissue-derived human mesenchymal stem cells (AT-hMSCs) have been widely used for differentiated hepatocyte-like cells (DHCs). Here, we established an <i>in vitro</i> HCV infection model using DHCs from AT-hMSCs and identified miRNAs that modulate HCV infectivity.</p><p>Methods</p><p>AT-hMSCs were differentiated into DHCs using the conditional media, and evaluated for hepatocyte characteristics using RT-PCR, immunocytochemistry, periodic acid-Schiff staining, and a urea synthesis assay. The expression of HCV candidate receptors was also verified using immunocytochemistry. The levels of candidate miRNAs targeting HCV receptors were then determined by relative quantitative RT-PCR (rqRT-PCR). Finally, DHCs were infected using HCVcc and serum from HCV-infected patients, and infectivity of the virus was measured by rqRT-PCR and transmission electron microscopy (TEM).</p><p>Results</p><p>The expected changes in morphology, function and hepatic gene expression were observed during hepatic differentiation. Moreover, the expression of candidate HCV entry factors and miR-27a were altered during hepatic differentiation. The infection and replication of HCV occurred efficiently in DHCs treated with HCVcc or infected with serum from HCV-infected patients. In addition, HCV infectivity was suppressed in miR-27a-transfected DHCs, due to the inhibition of LDLR expression by miR-27a.</p><p>Conclusions</p><p>Our results demonstrate that AT-hMSCs are a good source of DHCs, which are suitable for in vitro cultivation of HCV. Furthermore, these results suggest that miR-27a modulates HCV infectivity by regulating LDLR expression.</p></div

Nile-red stain was performed to show lipid droplets in FFA treated cells.

<p>FFA-induced lipid droplets were reduced in AM treated cells (bright red spots, A), and fluorometry revealed that AM reduced lipid droplets dose-dependently while FFA treatment increased lipid droplets over twofold of the control.</p

Primer sequences used for RT-PCR.

<p>Primer sequences used for RT-PCR.</p

Expression of candidate HCV entry receptors in DHCs.

<p>To evaluate the possible routes of HCV infection, the mRNA (A) and protein (B) expression of various HCV candidate receptors was assessed in DHCs. (A) RT-PCR analysis of the level of LDLR, CD81, SR-B1, and EGFR in Huh7.5 cells, AT-hMSCs, and DHCs. β-Actin was used as the loading control; Huh7.5 served as the positive control. (B) To detect the protein expression of candidate HCV entry receptors, the cells were stained with Dil-LDL (red), anti-CD81 antibodies (red), anti-SR-B1 antibodies (red), or anti-EGFR antibodies (red), and DAPI (blue). Merging of the images revealed strong expression of candidate HCV entry receptors in the DHCs. Scale bars are each 100 µm (LDLR), 50 µm (CD81 and SR-B1), and 200 µm (EGFR).</p