24 Further clarification of the potential role of iron in disordered lipid metabolism is required. To examine this, we studied the effects of iron status on hepatic cholesterol synthesis in mice

with iron burdens ranging from deficient to overloaded. We show that increasing iron burden in mice results in an increase in the transcripts of approximately half of the enzymes of the cholesterol Selumetinib biosynthetic pathway, resulting in an increase in hepatic total cholesterol. These results provide a new and potentially important additional mechanism by which iron could contribute to the development of NAFLD or lipotoxicity. Abc, adenosine triphosphate-binding cassette; Apo, apolipoprotein; Bhmt2, betaine-homocysteine methyltransferase 2; C/EBPα, CCAAT/enhancer binding protein α; Cyp51, lanosterol-14α demethylase; Cyp27b1, 25-hydroxyvitamin D3-1α-hydroxylase; Cyp7a1, cholesterol 7α-monooxygenase; Ebp, cholestenol-Δ-isomerase; Ggcx, gamma-glutamyl carboxylase; Ggps1, geranylgeranyl diphosphate synthase 1; GSEA, gene set enrichment analysis; Hmgcr, 3-hydroxy-3-methylglutarate-coenzymeA reductase; Hnf4a, hepatocyte nuclear factor 4α;

selleck products Hsd17b7, 3-keto-steroid reductase; Hsd3b7, hydroxy-Δ5-steroid dehydrogenase; Idi1, isopentenyl-diphosphate-Δ-isomerase; mRNA, messenger RNA; Mvk, mevalonate kinase; NAFLD, nonalcoholic fatty liver disease; NASH, nonalcoholic steatohepatitis; Nqo1, NAD(P)H dehydrogenase (quinone) 1; Nr1h3, nuclear receptor 1H3; Nsdhl, sterol-4α-carboxylate 3-dehydrogenase; Pmvk, phosphomevalonate kinase; Psap, prosaponin; RT-PCR, real-time polymerase chain reaction; Sc5d, lathosterol oxidase; Srebf2, sterol-regulatory element binding factor 2; Tm7sf2, Δ14-sterol reductase; Tmem97, transmembrane protein 97; Vkorc1, vitamin

selleck kinase inhibitor K epoxide reductase complex (subunit 1); VLDL, very low density lipoprotein; Vrk3, vaccinia-related kinase 3. Male AKR mice (Animal Resources Centre, Murdoch, Australia) were fed a diet of normal mouse chow containing 0.01% iron (normal iron diet; Specialty Feeds, Glen Forrest, Australia) ad libitum. A second group of mice were fed a diet supplemented with 2% carbonyl iron (Sigma, Sydney, Australia; iron-loaded) for 3 weeks, and a third group were fed a diet containing no added iron (0.001% iron; iron-deficient) for 7 weeks from 3 weeks of age. Mice were sacrificed at 10 weeks of age following an overnight fast. Organs were perfused with isotonic saline in situ; livers were harvested and snap-frozen in liquid nitrogen. All procedures were approved by the Animal Ethics Committee of the University of Western Australia. Total RNA was extracted from the livers of 12 mice (four from each group) using Tri-Reagent (Invitrogen, Sydney, Australia) and treated with deoxyribonuclease I (Ambion, Austin, TX). RNA used for microarray analysis was further purified using an RNeasy kit (Qiagen, Sydney, Australia).