title: SILAC-MS Based Characterization of LPS and Resveratrol Induced Changes in Adipocyte Proteomics - Resveratrol as Ameliorating Factor on LPS Induced Changes.
abstract: Adipose tissue inflammation is believed to play a pivotal role in the development obesity-related morbidities such as insulin resistance. However, it is not known how this (low-grade) inflammatory state develops. It has been proposed that the leakage of lipopolysaccharides (LPS), originating from the gut microbiota, through the gut epithelium could drive initiation of inflammation. To get a better understanding of which proteins and intracellular pathways are affected by LPS in adipocytes, we performed SILAC proteomic analysis and identified proteins that were altered in expression. Furthermore, we tested the anti-inflammatory compound resveratrol. A total of 927 proteins were quantified by the SILAC method and of these 57- and 64 were significantly up- and downregulated by LPS, respectively. Bioinformatic analysis (GO analysis) revealed that the upregulated proteins were especially involved in the pathways of respiratory electron transport chain and inflammation. The downregulated proteins were especially involved in protein glycosylation. One of the latter proteins, GALNT2, has previously been described to regulate the expression of liver lipases such as ANGPTL3 and apoC-III affecting lipid metabolism. Furthermore, LPS treatment reduced the protein levels of the insulin sensitizing adipokine, adiponectin, and proteins participating in the final steps of triglyceride- and cholesterol synthesis. Generally, resveratrol opposed the effect induced by LPS and, as such, functioning as an ameliorating factor in disease state. Using an unbiased proteomic approach, we present novel insight of how the proteome is altered in adipocytes in response to LPS as seen in obesity. We suggest that LPS partly exerts its detrimental effects by altering glycosylation processes of the cell, which is starting to emerge as important posttranscriptional regulators of protein expression. Furthermore, resveratrol could be a prime candidate in ameliorating dysfunctioning adipose tissue induced by inflammatory stimulation.
authors.affiliation: Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark.|Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Mark K
authors.identifier: http://orcid.org/0000-0002-5536-654X
authors.initials: MK
authors.lastname: Nøhr
authors.affiliation: Laboratory for Proteome Analysis and Protein Characterization, Department of Molecular Biology and Genetics and iNANO, Aarhus University, Aarhus, Denmark.
authors.forename: Toke P
authors.initials: TP
authors.lastname: Kroager
authors.affiliation: Laboratory for Proteome Analysis and Protein Characterization, Department of Molecular Biology and Genetics and iNANO, Aarhus University, Aarhus, Denmark.
authors.forename: Kristian W
authors.initials: KW
authors.lastname: Sanggaard
authors.affiliation: Laboratory for Proteome Analysis and Protein Characterization, Department of Molecular Biology and Genetics and iNANO, Aarhus University, Aarhus, Denmark.
authors.forename: Anders D
authors.initials: AD
authors.lastname: Knudsen
authors.affiliation: Department of Health Science and Technology, Aalborg University, Aalborg, Denmark.
authors.forename: Allan
authors.initials: A
authors.lastname: Stensballe
authors.affiliation: Laboratory for Proteome Analysis and Protein Characterization, Department of Molecular Biology and Genetics and iNANO, Aarhus University, Aarhus, Denmark.
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Enghild
authors.affiliation: Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark.|Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Jens
authors.initials: J
authors.lastname: Ølholm
authors.affiliation: Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark.|Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Bjørn
authors.initials: B
authors.lastname: Richelsen
authors.affiliation: Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark.|Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Steen B
authors.initials: SB
authors.lastname: Pedersen
mesh_terms: D017667:Adipocytes / Q000378:metabolism / Q000473:pathology; D000273:Adipose Tissue / Q000378:metabolism / Q000473:pathology; D000091303:Angiopoietin-Like Protein 3; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins / Q000096:biosynthesis; D000069196:Gastrointestinal Microbiome / Q000187:drug effects / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007249:Inflammation / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D007328:Insulin / Q000378:metabolism; D007333:Insulin Resistance / Q000235:genetics*; D050356:Lipid Metabolism; D050155:Lipogenesis / Q000187:drug effects; D008070:Lipopolysaccharides / Q000378:metabolism*; D017350:N-Acetylgalactosaminyltransferases / Q000096:biosynthesis; D009765:Obesity / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D020543:Proteome / Q000235:genetics; D040901:Proteomics; D000077185:Resveratrol; D013267:Stilbenes / Q000008:administration & dosage*; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D000091303:Angiopoietin-Like Protein 3; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins; D007328:Insulin; D008070:Lipopolysaccharides; D017350:N-Acetylgalactosaminyltransferases; D020543:Proteome; D000077185:Resveratrol; D013267:Stilbenes; D000097763:Polypeptide N-acetylgalactosaminyltransferase; C479896:ANGPTL3 protein, human