Effect of acute physiological hyperinsulinemia on gene expression in human skeletal muscle in vivo.

This study was undertaken to test the hypothesis that short-term exposure (4 h) to physiological hyperinsulinemia in normal, healthy subjects without a family history of diabetes would induce a low grade inflammatory response independently of glycemic status. Twelve normal glucose tolerant subjects received a 4-h euglycemic hyperinsulinemic clamp with biopsies of the vastus lateralis muscle. Microarray analysis identified 121 probe sets that were significantly altered in response to physiological hyperinsulinemia while maintaining euglycemia. In normal, healthy human subjects insulin increased the mRNAs of a number of inflammatory genes (CCL2, CXCL2 and THBD) and transcription factors (ATF3, BHLHB2, HES1, KLF10, JUNB, FOS, and FOSB). A number of other genes were upregulated in response to insulin, including RRAD, MT, and SGK. CITED2, a known coactivator of PPARalpha, was significantly downregulated. SGK and CITED2 are located at chromosome 6q23, where we previously detected strong linkage to fasting plasma insulin concentrations. We independently validated the mRNA expression changes in an additional five subjects and closely paralleled the results observed in the original 12 subjects. A saline infusion in healthy, normal glucose-tolerant subjects without family history of diabetes demonstrated that the genes altered during the euglycemic hyperinsulinemic clamp were due to hyperinsulinemia and were unrelated to the biopsy procedure per se. The results of the present study demonstrate that insulin acutely regulates the levels of mRNAs involved in inflammation and transcription and identifies several candidate genes, including HES1 and BHLHB2, for further investigation.

[1]  Ralph A. DeFronzo,et al.  Metabolic and molecular basis of insulin resistance , 2003, Journal of nuclear cardiology : official publication of the American Society of Nuclear Cardiology.

[2]  R. DeFronzo,et al.  Insulin Resistance: A Multifaceted Syndrome Responsible for NIDDM, Obesity, Hypertension, Dyslipidemia, and Atherosclerotic Cardiovascular Disease , 1991, Diabetes Care.

[3]  C. Kahn,et al.  Insulin resistance differentially affects the PI 3-kinase- and MAP kinase-mediated signaling in human muscle. , 2000, The Journal of clinical investigation.

[4]  C. Kahn,et al.  Rad: a member of the Ras family overexpressed in muscle of type II diabetic humans. , 1993, Science.

[5]  D. Auboeuf,et al.  Acute regulation by insulin of phosphatidylinositol-3-kinase, Rad, Glut 4, and lipoprotein lipase mRNA levels in human muscle. , 1996, The Journal of clinical investigation.

[6]  R. Pipek,et al.  Insulin-induced hexokinase II expression is reduced in obesity and NIDDM. , 1998, Diabetes.

[7]  L. DiPietro,et al.  Intramyocellular lipid concentrations are correlated with insulin sensitivity in humans: a 1H NMR spectroscopy study , 1999, Diabetologia.

[8]  B. Goodpaster,et al.  Skeletal muscle triglyceride: Marker or mediator of obesity-induced insulin resistance in type 2 diabetes mellitus? , 2002, Current diabetes reports.

[9]  R. DeFronzo,et al.  Regulation of hexokinase II and glycogen synthase mRNA, protein, and activity in human muscle. , 1995, The American journal of physiology.

[10]  R. DeFronzo,et al.  Glucose clamp technique: a method for quantifying insulin secretion and resistance. , 1979, The American journal of physiology.

[11]  O. Pedersen,et al.  Expression profiling of insulin action in human myotubes: induction of inflammatory and pro-angiogenic pathways in relationship with glycogen synthesis and type 2 diabetes. , 2004, Biochemical and biophysical research communications.

[12]  K. Nair,et al.  Gene expression profile in skeletal muscle of type 2 diabetes and the effect of insulin treatment. , 2002, Diabetes.

[13]  C. Haft,et al.  Activation of Serum- and Glucocorticoid-induced Protein Kinase (Sgk) by Cyclic AMP and Insulin* , 2001, The Journal of Biological Chemistry.

[14]  C. Bogardus,et al.  Microarray profiling of skeletal muscle tissues from equally obese, non-diabetic insulin-sensitive and insulin-resistant Pima Indians , 2002, Diabetologia.

[15]  R. Berria,et al.  Increased collagen content in insulin-resistant skeletal muscle. , 2006, American journal of physiology. Endocrinology and metabolism.

[16]  J. W. Davis,et al.  Identification of the CREB-binding Protein/p300-interacting Protein CITED2 as a Peroxisome Proliferator-activated Receptor α Coregulator* , 2004, Journal of Biological Chemistry.

[17]  M. Laville,et al.  Regulation by insulin of gene expression in human skeletal muscle and adipose tissue. Evidence for specific defects in type 2 diabetes. , 2001, Diabetes.

[18]  G. Reaven,et al.  Pathophysiology of insulin resistance in human disease. , 1995, Physiological reviews.

[19]  H. Vestergaard,et al.  Glycogen synthase and phosphofructokinase protein and mRNA levels in skeletal muscle from insulin-resistant patients with non-insulin-dependent diabetes mellitus. , 1993, The Journal of clinical investigation.

[20]  P. Cohen,et al.  Regulation and Physiological Roles of Serum- and Glucocorticoid-Induced Protein Kinase Isoforms , 2001, Science's STKE.

[21]  Allan Vaag,et al.  TXNIP Regulates Peripheral Glucose Metabolism in Humans , 2007, PLoS medicine.

[22]  P. O'Connell,et al.  Linkage of type 2 diabetes mellitus and of age at onset to a genetic location on chromosome 10q in Mexican Americans. , 1999, American journal of human genetics.

[23]  W. Waldhäusl.  The Glucose Clamp Technique , 1993 .

[24]  R. DeFronzo,et al.  Effect of a sustained reduction in plasma free fatty acid concentration on intramuscular long-chain fatty Acyl-CoAs and insulin action in type 2 diabetic patients. , 2005, Diabetes.

[25]  F. Dela,et al.  Metallothionein-mediated antioxidant defense system and its response to exercise training are impaired in human type 2 diabetes. Diabetes 2005;54:3089–3094 , 2005, Diabetes.

[26]  J. Eady,et al.  Variation in gene expression profiles of peripheral blood mononuclear cells from healthy volunteers. , 2005, Physiological genomics.

[27]  R. DeFronzo,et al.  Metabolic basis of obesity and noninsulin-dependent diabetes mellitus. , 1988, Diabetes/metabolism reviews.

[28]  R. DeFronzo.  Lilly lecture 1987. The triumvirate: beta-cell, muscle, liver. A collusion responsible for NIDDM. , 1988, Diabetes.

[29]  J. Beattie,et al.  Obesity and hyperleptinemia in metallothionein (-I and -II) null mice. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. DeFronzo,et al.  Effects of insulin on peripheral and splanchnic glucose metabolism in noninsulin-dependent (type II) diabetes mellitus. , 1985, The Journal of clinical investigation.

[31]  Ash A. Alizadeh,et al.  Individuality and variation in gene expression patterns in human blood , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Y. Asmann,et al.  Skeletal Muscle Mitochondrial Functions, Mitochondrial DNA Copy Numbers, and Gene Transcript Profiles in Type 2 Diabetic and Nondiabetic Subjects at Equal Levels of Low or High Insulin and Euglycemia , 2006, Diabetes.

[33]  R. Scarpulla,et al.  PGC-1-Related Coactivator, a Novel, Serum-Inducible Coactivator of Nuclear Respiratory Factor 1-Dependent Transcription in Mammalian Cells , 2001, Molecular and Cellular Biology.

[34]  Sophie Rome,et al.  Microarray Profiling of Human Skeletal Muscle Reveals That Insulin Regulates ∼800 Genes during a Hyperinsulinemic Clamp* 210 , 2003, The Journal of Biological Chemistry.

[35]  M. Daly,et al.  PGC-1α-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes , 2003, Nature Genetics.

[36]  R. DeFronzo,et al.  Lipid Infusion Decreases the Expression of Nuclear Encoded Mitochondrial Genes and Increases the Expression of Extracellular Matrix Genes in Human Skeletal Muscle* , 2005, Journal of Biological Chemistry.

[37]  Sophie Rome,et al.  Microarray profiling of human skeletal muscle reveals that insulin regulates approximately 800 genes during a hyperinsulinemic clamp. , 2003, The Journal of biological chemistry.

[38]  Peter S Linsley,et al.  Individual-specific variation of gene expression in peripheral blood leukocytes. , 2004, Genomics.

[39]  Michael Brownlee,et al.  The pathobiology of diabetic complications: a unifying mechanism. , 2005, Diabetes.

[40]  R. DeFronzo.  The Triumvirate: β-Cell, Muscle, Liver: A Collusion Responsible for NIDDM , 1988, Diabetes.

[41]  W. Hsueh,et al.  Insulin signaling in the arterial wall. , 1999, The American journal of cardiology.

[42]  B. Kahn,et al.  Glucose transporters and insulin action--implications for insulin resistance and diabetes mellitus. , 1999, The New England journal of medicine.

[43]  P. O'Connell,et al.  A major locus for fasting insulin concentrations and insulin resistance on chromosome 6q with strong pleiotropic effects on obesity-related phenotypes in nondiabetic Mexican Americans. , 2001, American journal of human genetics.

[44]  K. Polonsky,et al.  Twenty-four-hour profiles and pulsatile patterns of insulin secretion in normal and obese subjects. , 1988, The Journal of clinical investigation.

[45]  C Cobelli,et al.  Roles of glucose transport and glucose phosphorylation in muscle insulin resistance of NIDDM. , 1996, Diabetes.

[46]  A. Butte,et al.  Coordinated reduction of genes of oxidative metabolism in humans with insulin resistance and diabetes: Potential role of PGC1 and NRF1 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[47]  R. DeFronzo,et al.  Ceramide content is increased in skeletal muscle from obese insulin-resistant humans. , 2004, Diabetes.