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Gene expression profiling in small intestine of mice administered lactoperoxidase

ABSTRACT: Lactoperoxidase (LPO) is a component of milk and other external secretions. Recently, we found antiinflammatory effect of orally administered LPO in mouse model of influenza virus-induced pneumonia. In order to study influence of bovine LPO on the digestive tract as its possible target organ, we performed comprehensive gene expression analysis of the mouse small intestine using GeneChip microarray to compare the effect of LPO with that of water or bovine serum albumin (BSA). We observed that 3 h after single ingestion of LPO the levels of 78 gene expression are upregulated and those of 9 gene expressions are downregulated. Within most changed genes, 5 upregulated and 1 downregulated genes such as FK506 binding protein 5 (FKBP5) and serum/glucocorticoid regulated kinase (SGK) are known to be transcriptionally modulated by glucocorticoid. This result suggests that ingested LPO modulates gene expressions in the small intestine in a glucocorticoid-like manner and this activity may link to its systemic antiinflammatory effects. Keywords: treatment comparison

ORGANISM(S): Mus musculus

PROVIDER: GSE5562 | GEO | 2006/09/01

SECONDARY ACCESSION(S): PRJNA95409

REPOSITORIES: GEO

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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 profiling in small intestine of mice administered lactoperoxidase

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