Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression in human subcutaneous adipose tissue after administration of Gastric Insulinotropic Peptide (GIP)

ABSTRACT: The intention of this clinical study was to investigate the effect of GIP administration for 240 min on gene expression in human subcutaneous adipose tissue. Three conditions have been tersted: 1. Sole infusion of GIP or NaCl as control; 2. GIP or NaCl administration under euglycemic-hyperinsulinemic clamp conditions; 3. GIP or NaCl administration under hyperglycemic-hyperinsulinemic clamp conditions to mimic the postprandial state. In each participant a complete physical examination and evaluation of medical history was performed, including an oral glucose tolerance test (oGTT) with 75 g glucose after overnight fast to ensure the metabolic state. Standard fasting laboratory and clinical chemistry evaluations were done. Synthetic human GIP (1-42) was dissolved in saline (0.9% NaCl-solution) under sterile conditions. All studies were done in the morning in the overnight fasted state (>10h since last meal). The effect of GIP administration on gene expression in subcutaneous adipose tissue was studied under 3 different conditions in a single blind design. Either the participants received only a GIP- or a saline- infusion (0.9% NaCl-isotonic solution, Fresenius, Germany) for 240 min. At different investigation days participants underwent euglycemic (EU)- and hyperglycemic (HC), hyperinsulinemic clamps combined with GIP- or placebo-infusions for 240 min at different examination days in a randomized, single-blind, crossover design. The capillary glucose concentration was 80mg/dl during EU-clamp and 140mg/dl during HC-clamp. The following numbers of treatments were performed: EU with GIP-infusion (N=9); EU with NaCl-infusion (N=9); HC with GIP-infusion (N=8), HC with NaCl-infusion (N=8); sole GIP-infusion (N=11) and sole placebo-infusion (N=11). Between examination days an intermission time of at least 7 days was maintained.

ORGANISM(S): Homo sapiens

SUBMITTER: Martin Osterhoff

PROVIDER: E-GEOD-40589 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:This study was undertaken to test the hypothesis that short term exposure (4 hours) to physiologic hyperinsulinemia in normal, healthy subjects without a family history of diabetes would induce a low grade inflammatory response, independently of glycemic status. We performed euglycemic hyperinsulinemic (80 mU/m2/min) clamps in 12 healthy, insulin sensitive subjects with no family history of diabetes followed by biopsies of the vastus lateralis muscle taken basally and after 30 and 240 minutes of insulin infusion. Gene expression profiles were generated using Affymetrix HG-U133A arrays. No probe sets had significantly altered expression at 30 minutes of the insulin clamp, but 121 probe sets (117 upregulated and 4 downregulated) were significantly altered after 240 minutes. Hyperinsulinemia in normal, healthy human subjects increased the mRNAs for a number of inflammatory genes and transcription factors. Microarray and quantitative RT-PCR revealed the upregulation of chemokine, cc motif, ligand 2 (CCL2), CCL8, thrombomodulin (THBD), ras-related associated with diabetes (RRAD), metallothionein (MT), and serum/glucocorticoid regulated kinase (SGK), and downregulation of CITED2 (a CREB-binding protein-interacting transactivator), a known coactivator of PPAR-alpha. Interestingly, SGK and CITED2 are located at chromosome 6q23, where we previously detected strong linkage to hyperinsulinemia. A control saline infusion performed on 3 normal, healthy subjects without a family history of diabetes demonstrated that the genes altered following the euglycemic-hyperinsulinemic clamp were due to insulin and independent of biopsy removal. This study demonstrates that insulin acutely regulates the expression of genes involved in inflammation and transcription, and identifies several candidate genes/pathways for further investigation. Experiment Overall Design: Twelve subjects received a vastus lateralis muscle biopsy followed by a 180-min euglycemic, hyperinsulinemic (80 mU/m2.min) clamp. Muscle biopsies from each subject were homogenized in RNAStat solution (Tel-Test Inc., Friendswood, TX), using a Polytron homogenizer (Brinkmann Instruments Westbury, NY). Total RNA was purified with RNeasy and DNase I treatment (Qiagen, Chatsworth, CA). RNA was prepared for hybridization to Affymetrix (Santa Clara, CA) HG-U133A arrays according to the manufacturerâs instructions.

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Gene expression in human subcutaneous adipose tissue after administration of Gastric Insulinotropic Peptide (GIP)

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