title: Identification of novel regulatory cholesterol metabolite, 5-cholesten, 3β,25-diol, disulfate.
abstract: Oxysterol sulfation plays an important role in regulation of lipid metabolism and inflammatory responses. In the present study, we report the discovery of a novel regulatory sulfated oxysterol in nuclei of primary rat hepatocytes after overexpression of the gene encoding mitochondrial cholesterol delivery protein (StarD1). Forty-eight hours after infection of the hepatocytes with recombinant StarD1 adenovirus, a water-soluble oxysterol product was isolated and purified by chemical extraction and reverse-phase HPLC. Tandem mass spectrometry analysis identified the oxysterol as 5-cholesten-3β, 25-diol, disulfate (25HCDS), and confirmed the structure by comparing with a chemically synthesized compound. Administration of 25HCDS to human THP-1-derived macrophages or HepG2 cells significantly inhibited cholesterol synthesis and markedly decreased lipid levels in vivo in NAFLD mouse models. RT-PCR showed that 25HCDS significantly decreased SREBP-1/2 activities by suppressing expression of their responding genes, including ACC, FAS, and HMG-CoA reductase. Analysis of lipid profiles in the liver tissues showed that administration of 25HCDS significantly decreased cholesterol, free fatty acids, and triglycerides by 30, 25, and 20%, respectively. The results suggest that 25HCDS inhibits lipid biosynthesis via blocking SREBP signaling. We conclude that 25HCDS is a potent regulator of lipid metabolism and propose its biosynthetic pathway.
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: Shunlin
authors: S
authors: Ren
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: Jin Koung
authors: JK
authors: Kim
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: Genta
authors: G
authors: Kakiyama
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: Daniel
authors: D
authors: Rodriguez-Agudo
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: William M
authors: WM
authors: Pandak
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: Hae-Ki
authors: HK
authors: Min
authors: Department of Medicine, Veterans Affairs McGuire Medical Center/Department of Medicine, Virginia Commonwealth University, Richmond, Virginia, United States of America.
authors: Yanxia
authors: Y
authors: Ning
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000235:genetics / Q000378:metabolism; D000256:Adenoviridae / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002784:Cholesterol / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D002788:Cholesterol Esters / Q000032:analysis* / Q000138:chemical synthesis / Q000494:pharmacology; D004195:Disease Models, Animal; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism; D005260:Female; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D006888:Hydroxycholesterols / Q000032:analysis* / Q000138:chemical synthesis / Q000494:pharmacology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects; D008810:Mice, Inbred C57BL; D065626:Non-alcoholic Fatty Liver Disease / Q000378:metabolism / Q000473:pathology; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D051780:Sterol Regulatory Element Binding Protein 1 / Q000378:metabolism; D051782:Sterol Regulatory Element Binding Protein 2 / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C534977:25-hydroxycholesterol 3-sulfate; D002788:Cholesterol Esters; D006888:Hydroxycholesterols; D051780:Sterol Regulatory Element Binding Protein 1; D051782:Sterol Regulatory Element Binding Protein 2; D014764:Viral Proteins; C007997:25-hydroxycholesterol; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases; D064429:Fatty Acid Synthases; D000103:Acetyl-CoA Carboxylase