Proteomic profiling of cellular steatosis with concomitant oxidative stress in vitro.

Publisher: BioMed Central Country of Publication: England NLM ID: 101147696 Publication Model: Electronic Cited Medium: Internet ISSN: 1476-511X (Electronic) Linking ISSN: 1476511X NLM ISO Abbreviation: Lipids Health Dis Subsets: MEDLINE

Original Publication: [London] : BioMed Central, 2002-

Proteomics*; Ammonia/*pharmacology ; Caprylates/*pharmacology ; Hepatocytes/*drug effects ; Lactic Acid/*pharmacology ; Pyruvic Acid/*pharmacology; Aldehyde Reductase/genetics ; Aldehyde Reductase/metabolism ; Aldo-Keto Reductases ; Cell Line, Tumor ; Cholesterol/biosynthesis ; Fatty Acids, Nonesterified/metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; Gene Ontology ; Hepatocytes/cytology ; Hepatocytes/metabolism ; Histones/genetics ; Histones/metabolism ; Humans ; Lipid Peroxidation ; Lipogenesis/drug effects ; Lipogenesis/genetics ; Models, Biological ; Molecular Sequence Annotation ; Non-alcoholic Fatty Liver Disease/genetics ; Non-alcoholic Fatty Liver Disease/metabolism ; Non-alcoholic Fatty Liver Disease/pathology ; Oxidative Stress ; Protein Biosynthesis/drug effects

Background: Nutrient excess underpins the development of nonalcoholic fatty liver disease (NAFLD). The ensuing metabolic derangement is characterised by increased cellular respiration, oxidative stress and mitochondrial impairment. We have previously recapitulated these events in an in vitro cellular steatosis model. Here, we examined the distinct patterns of protein expression involved using a proteomics approach.
Methods: Human hepatoblastoma C3A cells were treated with a combination of energy substrates; lactate (L), pyruvate (P), octanoate (O) and ammonia (N). Proteins extracts were trypsinized and analyzed on a capillary HPLC OrbitrapXL mass spectrometer. Proteins were quantified using a label-free intensity based approach. Functional enrichment analysis was performed using ToppCluster via Gene Ontology (GO) database.
Results: Of the 1327 proteins identified, 104 were differentially expressed between LPON and untreated cells (defined as: ≥2 peptides; fold change ≥1.5; p-value <0.05). Seventy of these were upregulated with LPON. Functional enrichment analysis revealed enhanced protein biosynthesis accompanied by downregulation of histones H2A type 1-A, H1.2, H1.5 and H1.0I in LPON cells. Lipid binding annotations were also enriched as well as proteins involved in cholesterol synthesis, uptake and efflux. Increased expression of aldo-keto reductase family 1, member C1 and C3 suggests enhanced sterol metabolism and increased ROS-mediated lipid peroxidation.
Conclusions: The surge of energy substrates diverts free fatty acid metabolism towards pathways that can mitigate lipotoxicity. The histones depletion may represent an adaptation to increased protein synthesis. However, this can also expose DNA to oxidative stress thus should be explored further in the context of NAFLD progression.

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ETM/182 United Kingdom CSO_ Chief Scientist Office

Keywords: Nonalcoholic fatty liver; Oxidative stress; Steatohepatitis

0 (Caprylates)
0 (Fatty Acids, Nonesterified)
0 (Histones)
33X04XA5AT (Lactic Acid)
7664-41-7 (Ammonia)
8558G7RUTR (Pyruvic Acid)
97C5T2UQ7J (Cholesterol)
EC 1.1.1.- (Aldo-Keto Reductases)
EC 1.1.1.21 (Aldehyde Reductase)
OBL58JN025 (octanoic acid)

Date Created: 20160703 Date Completed: 20170327 Latest Revision: 20220129

20250114

PMC4930558

10.1186/s12944-016-0283-7

27368608