title: Mirtoselect, an anthocyanin-rich bilberry extract, attenuates non-alcoholic steatohepatitis and associated fibrosis in ApoE(∗)3Leiden mice.
abstract: OBJECTIVE
  Anthocyanins may have beneficial effects on lipid metabolism and inflammation and are demonstrated to have hepatoprotective properties in models of restraint-stress- and chemically-induced liver damage. However, their potential to protect against non-alcoholic steatohepatitis (NASH) under conditions relevant for human pathogenesis remains unclear. Therefore, we studied the effects of the standardised anthocyanin-rich extract Mirtoselect on diet-induced NASH in a translational model of disease.
  
  
  METHODS
  ApoE(∗)3Leiden mice were fed a Western-type cholesterol-containing diet without (HC) or with 0.1% (w/w) Mirtoselect (HCM) for 20weeks to study the effects on diet-induced NASH.
  
  
  RESULTS
  Mirtoselect attenuated HC-induced hepatic steatosis, as observed by decreased macro- and microvesicular hepatocellular lipid accumulation and reduced hepatic cholesteryl ester content. This anti-steatotic effect was accompanied by local anti-inflammatory effects in liver, as demonstrated by reduced inflammatory cell clusters and reduced neutrophil infiltration in HCM. On a molecular level, HC diet significantly induced hepatic expression of pro-inflammatory genes Tnf, Emr1, Ccl2, Mpo, Cxcl1, and Cxcl2 while this induction was less pronounced or significantly decreased in HCM. A similar quenching effect was observed for HC-induced pro-fibrotic genes, Acta2 and Col1a1 and this anti-fibrotic effect of Mirtoselect was confirmed histologically. Many of the pro-inflammatory and pro-fibrotic parameters positively correlated with intrahepatic free cholesterol levels. Mirtoselect significantly reduced accumulation and crystallisation of intrahepatic free cholesterol, providing a possible mechanism for the observed hepatoprotective effects.
  
  
  CONCLUSIONS
  Mirtoselect attenuates development of NASH, reducing hepatic lipid accumulation, inflammation and fibrosis, possibly mediated by local anti-inflammatory effects associated with reduced accumulation and crystallisation of intrahepatic free cholesterol.
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands; Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Hanzeplein 1 (EA11), 9713 GZ Groningen, The Netherlands; Top Institute Food and Nutrition, Nieuwe Kanaal 9A, 6709 PA Wageningen, The Netherlands. Electronic address: martine.morrison@tno.nl.
authors.forename: Martine C
authors.initials: MC
authors.lastname: Morrison
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Wen
authors.initials: W
authors.lastname: Liang
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Petra
authors.initials: P
authors.lastname: Mulder
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Lars
authors.initials: L
authors.lastname: Verschuren
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Elsbet
authors.initials: E
authors.lastname: Pieterman
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Karin
authors.initials: K
authors.lastname: Toet
authors.affiliation: Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Hanzeplein 1 (EA11), 9713 GZ Groningen, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Heeringa
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Peter Y
authors.initials: PY
authors.lastname: Wielinga
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Teake
authors.initials: T
authors.lastname: Kooistra
authors.affiliation: Department of Metabolic Health Research, Netherlands Organisation for Applied Scientific Research (TNO), Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Robert
authors.initials: R
authors.lastname: Kleemann
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D000872:Anthocyanins / Q000494:pharmacology*; D000890:Anti-Infective Agents / Q000494:pharmacology; D002135:Calcium-Binding Proteins; D054360:Chemokine CXCL1 / Q000378:metabolism; D002788:Cholesterol Esters / Q000378:metabolism; D002791:Cholesterol, Dietary / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D019610:Cytoprotection; D066273:Diet, Western; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008103:Liver Cirrhosis / Q000209:etiology / Q000503:physiopathology / Q000517:prevention & control*; D051379:Mice; D065626:Non-alcoholic Fatty Liver Disease / Q000150:complications / Q000188:drug therapy / Q000503:physiopathology; D010936:Plant Extracts; D011956:Receptors, Cell Surface / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D016896:Treatment Outcome; D029800:Vaccinium myrtillus / Q000737:chemistry*
chemical_list: C558396:Acta2 protein, mouse; D000199:Actins; C000634132:Adgre1 protein, mouse; D000872:Anthocyanins; D000890:Anti-Infective Agents; D002135:Calcium-Binding Proteins; D054360:Chemokine CXCL1; D002788:Cholesterol Esters; D002791:Cholesterol, Dietary; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C475895:Cxcl1 protein, mouse; D010936:Plant Extracts; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C073305:Vaccinium myrtillus extract
keywords: Anthocyanins; Bilberry; Cholesterol crystals; Fibrosis; Free cholesterol; Non-alcoholic steatohepatitis; Polyphenols; Steatosis