glycogen and Diabetes-Mellitus--Type-2

Hepatalin is a hormone secreted by the liver in response to pulses of insulin after a mixed nutrient meal, but only if the liver receives two permissive synergistic feeding signals from the stomach. Hepatalin stimulates glucose uptake and storage as glycogen in skeletal muscle, heart, and kidney but not liver, intestines, or adipocytes. Insulin acts primarily on liver and fat. Reduced hepatalin action results in postprandial hyperglycemia, compensatory elevation of insulin secretion, and a resultant shift in partitioning of nutrient energy storage from glycogen in muscle, to fat. Chronic hepatalin suppression leads to a predictable chronology of dysfunctions, first diagnosable as Absence of Meal-induced Insulin Sensitization (AMIS) which progresses to prediabetes, adiposity, and type 2 diabetes. The focus on nutrient partitioning and the role of hepatalin allows AMIS to be diagnosed, prevented, and treated, including through the use of lifestyle interventions.

Topics: Blood Glucose; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin; Insulin Resistance; Muscle, Skeletal; Obesity; Prediabetic State

Topics: Animals; Diabetes Mellitus, Type 2; Glucose; Glucosides; Glycogen; Glycogen Storage Disease Type I; Hypoglycemia; Rats

The incidence of obesity, insulin resistance, and type II diabetes (T2DM) continues to rise worldwide. The liver is a central insulin-responsive metabolic organ that governs whole-body metabolic homeostasis. Therefore, defining the mechanisms underlying insulin action in the liver is essential to our understanding of the pathogenesis of insulin resistance. During periods of fasting, the liver catabolizes fatty acids and stored glycogen to meet the metabolic demands of the body. In postprandial conditions, insulin signals to the liver to store excess nutrients into triglycerides, cholesterol, and glycogen. In insulin-resistant states, such as T2DM, hepatic insulin signaling continues to promote lipid synthesis but fails to suppress glucose production, leading to hypertriglyceridemia and hyperglycemia. Insulin resistance is associated with the development of metabolic disorders such as cardiovascular and kidney disease, atherosclerosis, stroke, and cancer. Of note, nonalcoholic fatty liver disease (NAFLD), a spectrum of diseases encompassing fatty liver, inflammation, fibrosis, and cirrhosis, is linked to abnormalities in insulin-mediated lipid metabolism. Therefore, understanding the role of insulin signaling under normal and pathologic states may provide insights into preventative and therapeutic opportunities for the treatment of metabolic diseases. Here, we provide a review of the field of hepatic insulin signaling and lipid regulation, including providing historical context, detailed molecular mechanisms, and address gaps in our understanding of hepatic lipid regulation and the derangements under insulin-resistant conditions. © 2023 American Physiological Society. Compr Physiol 13:4785-4809, 2023.

Topics: Diabetes Mellitus, Type 2; Glycogen; Homeostasis; Humans; Insulin; Insulin Resistance; Lipid Metabolism; Liver; Non-alcoholic Fatty Liver Disease; Triglycerides

As the burden of type 2 diabetes mellitus (T2DM) grows in the 21st century, the need to understand glucose metabolism heightens. Increased gluconeogenesis is a major contributor to the hyperglycemia seen in T2DM. Isotope tracer experiments in humans and animals over several decades have offered insights into gluconeogenesis under euglycemic and diabetic conditions. This review focuses on the current understanding of carbon flux in gluconeogenesis, including substrate contribution of various gluconeogenic precursors to glucose production. Alterations of gluconeogenic metabolites and fluxes in T2DM are discussed. We also highlight ongoing knowledge gaps in the literature that require further investigation. A comprehensive analysis of gluconeogenesis may enable a better understanding of T2DM pathophysiology and identification of novel targets for treating hyperglycemia.

Topics: Animals; Carbon; Carbon Isotopes; Diabetes Mellitus, Type 2; Gluconeogenesis; Glucose; Glycogen; Humans; Hyperglycemia; Metabolomics

Both insulin deficiency and insulin resistance due to glucagon secretion cause fasting and postprandial hyperglycemia in patients with diabetes.. Metformin enhances insulin sensitivity, being used to prevent and treat diabetes, although its mechanism of action remains elusive.. Patients with diabetes fail to store glucose as hepatic glycogen via the direct pathway (glycogen synthesis from dietary glucose during the post-prandial period) and via the indirect pathway (glycogen synthesis from "de novo" synthesized glucose) owing to insulin deficiency and glucagoninduced insulin resistance. Depletion of the hepatic glycogen deposit activates gluconeogenesis to replenish the storage via the indirect pathway. Unlike healthy subjects, patients with diabetes experience glycogen cycling due to enhanced gluconeogenesis and failure to store glucose as glycogen. These defects raise hepatic glucose output causing both fasting and post-prandial hyperglycemia. Metformin reduces post-prandial plasma glucose, suggesting that the drug facilitates glucose storage as hepatic glycogen after meals. Replenishment of glycogen store attenuates the accelerated rate of gluconeogenesis and reduces both glycogen cycling and hepatic glucose output. Metformin also reduces fasting hyperglycemia due to declining hepatic glucose production. In addition, metformin reduces plasma insulin concentration in subjects with impaired glucose tolerance and diabetes and decreases the amount of insulin required for metabolic control in patients with diabetes, reflecting improvement of insulin activity. Accordingly, metformin preserves β-cell function in patients with type 2 diabetes.. Several mechanisms have been proposed to explain the metabolic effects of metformin, but evidence is not conclusive and the molecular basis of metformin action remains unknown.

Topics: Diabetes Mellitus, Type 2; Gluconeogenesis; Glucose; Glycogen; Humans; Hypoglycemic Agents; Insulin Resistance; Liver; Metformin

The brain contains a fairly low amount of glycogen, mostly located in astrocytes, a fact that has prompted the suggestion that glycogen does not have a significant physiological role in the brain. However, glycogen metabolism in astrocytes is essential for several key physiological processes and is adversely affected in disease. For instance, diminished ability to break down glycogen impinges on learning, and epilepsy, Alzheimer's disease, and type 2 diabetes are all associated with abnormal astrocyte glycogen metabolism. Glycogen metabolism supports astrocytic K

Topics: Alzheimer Disease; Animals; Astrocytes; Brain; Calcium; Cyclic AMP; Diabetes Mellitus, Type 2; Glutamine; Glycogen; Glycogen Phosphorylase; Humans; Isoenzymes; Learning; Memory; Neurotransmitter Agents; Potassium; Signal Transduction; Sleep

Exercise is recommended as therapeutic intervention for people at risk to develop type 2 diabetes to prevent or treat the disease. Recent studies on the influence of obesity and type 2 diabetes on the outcome of exercise programs are discussed.. Poor glycemic control before an intervention can be a risk factor of reduced therapeutic benefit from exercise. But the acute metabolic response to exercise and the transcriptional profile of the working muscle is similar in healthy controls and type 2 diabetic patients, including but not limited to intact activation of skeletal muscle AMP-activated kinase signaling, glucose uptake and expression of peroxisome proliferator-activated receptor gamma coactivator 1α. The increase in plasma acylcarnitines during exercise is not influenced by type 2 diabetes or obesity. The hepatic response to exercise is dependent on the glucagon/insulin ratio and the exercise-induced increase in hepatokines such as fibroblast growth factor 21 and follistatin is impaired in type 2 diabetes and obesity, but consequences for the benefit from exercise are unknown yet.. Severe metabolic dysregulation can reduce the benefit from exercise, but the intact response of key metabolic regulators in exercising skeletal muscle of diabetic patients demonstrates the effectiveness of exercise programs to treat the disease.

Topics: AMP-Activated Protein Kinases; Blood Glucose; Carnitine; Diabetes Mellitus, Type 2; Exercise; Fatty Acids; Glucose; Glycogen; Humans; Liver; Muscle, Skeletal; Obesity; Oxidation-Reduction; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Signal Transduction

Elevated plasma glucose leads to pancreatic β cell dysfunction and death in type 2 diabetes. Glycogen accumulation, due to impaired metabolism, contributes to this "glucotoxicity" via dysregulated biochemical pathways promoting β cell dysfunction. Here, we review emerging data, and re-examine published findings, on the role of glycogen in β cells in normoglycemia and in diabetes.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Glycogen; Glycogen Storage Disease; Humans; Insulin-Secreting Cells; Signal Transduction

Over the past decades, hypomagnesemia (serum Mg(2+) <0.7 mmol/L) has been strongly associated with type 2 diabetes mellitus (T2DM). Patients with hypomagnesemia show a more rapid disease progression and have an increased risk for diabetes complications. Clinical studies demonstrate that T2DM patients with hypomagnesemia have reduced pancreatic β-cell activity and are more insulin resistant. Moreover, dietary Mg(2+) supplementation for patients with T2DM improves glucose metabolism and insulin sensitivity. Intracellular Mg(2+) regulates glucokinase, KATP channels, and L-type Ca(2+) channels in pancreatic β-cells, preceding insulin secretion. Moreover, insulin receptor autophosphorylation is dependent on intracellular Mg(2+) concentrations, making Mg(2+) a direct factor in the development of insulin resistance. Conversely, insulin is an important regulator of Mg(2+) homeostasis. In the kidney, insulin activates the renal Mg(2+) channel transient receptor potential melastatin type 6 that determines the final urinary Mg(2+) excretion. Consequently, patients with T2DM and hypomagnesemia enter a vicious circle in which hypomagnesemia causes insulin resistance and insulin resistance reduces serum Mg(2+) concentrations. This Perspective provides a systematic overview of the molecular mechanisms underlying the effects of Mg(2+) on insulin secretion and insulin signaling. In addition to providing a review of current knowledge, we provide novel directions for future research and identify previously neglected contributors to hypomagnesemia in T2DM.

Topics: Blood Glucose; Calcium Channels, L-Type; Diabetes Mellitus, Type 2; Dietary Supplements; Disease Progression; Glucokinase; Glycogen; Glycolysis; Humans; Inflammation; Insulin; Insulin Resistance; Insulin Secretion; Insulin-Secreting Cells; KATP Channels; Liver; Magnesium; Magnesium Deficiency; Obesity; Potassium Channels, Inwardly Rectifying; Sodium Chloride Symporters; Sodium-Potassium-Exchanging ATPase; Water-Electrolyte Imbalance

Birth weight (BW) has been shown to be influenced by both fetal and maternal factors and in observational studies is reproducibly associated with future risk of adult metabolic diseases including type 2 diabetes (T2D) and cardiovascular disease. These life-course associations have often been attributed to the impact of an adverse early life environment. Here, we performed a multi-ancestry genome-wide association study (GWAS) meta-analysis of BW in 153,781 individuals, identifying 60 loci where fetal genotype was associated with BW (P < 5 × 10

Topics: Adult; Aging; Anthropometry; Birth Weight; Blood Pressure; Chromatin Assembly and Disassembly; Cohort Studies; Coronary Artery Disease; Datasets as Topic; Diabetes Mellitus, Type 2; Female; Fetus; Genetic Loci; Genetic Predisposition to Disease; Genetic Variation; Genome-Wide Association Study; Genomic Imprinting; Genotype; Glucose; Glycogen; Humans; Insulin; Male; Phenotype; Signal Transduction

Type 2 diabetes mellitus (T2D) is characterized by insulin resistance, impaired glycogen synthesis, lipid accumulation, and impaired mitochondrial function. Exercise training has received increasing recognition as a cornerstone in the prevention and treatment of T2D. Emerging research suggests that resistance training (RT) has the power to combat metabolic dysfunction in patients with T2D and seems to be an effective measure to improve overall metabolic health and reduce metabolic risk factors in diabetic patients. However, there is limited mechanistic insight into how these adaptations occur. This review provides an overview of the intervention data on the impact of RT on glucose metabolism. In addition, the molecular mechanisms that lead to adaptation in skeletal muscle in response to RT and that are associated with possible beneficial metabolic responses are discussed. Some of the beneficial adaptations exerted by RT include increased GLUT4 translocation in skeletal muscle, increased insulin sensitivity and hence restored metabolic flexibility. Increased energy expenditure and excess postexercise oxygen consumption in response to RT may be other beneficial effects. RT is increasingly establishing itself as an effective measure to improve overall metabolic health and reduce metabolic risk factors in diabetic patients.

Topics: Diabetes Mellitus, Type 2; Exercise; Glucose; Glucose Transporter Type 4; Glycogen; Humans; Insulin Resistance; Muscles; Oxygen; Resistance Training

While it has long been known that zinc (Zn) is crucial for the proper growth and maintenance of normal biological functions, Zn has also been shown to exert insulin-mimetic and anti-diabetic effects. These insulin-like properties have been demonstrated in isolated cells, tissues, and different animal models of type 1 and type 2 diabetes. Zn treatment has been found to improve carbohydrate and lipid metabolism in rodent models of diabetes. In isolated cells, it enhances glucose transport, glycogen and lipid synthesis, and inhibits gluconeogenesis and lipolysis. The molecular mechanism responsible for the insulin-like effects of Zn compounds involves the activation of several key components of the insulin signaling pathways, which include the extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphatidylinositol 3-kinase (PI3-K)/protein kinase B/Akt (PKB/Akt) pathways. However, the precise molecular mechanisms by which Zn triggers the activation of these pathways remain to be clarified. In this review, we provide a brief history of zinc, and an overview of its insulin-mimetic and anti-diabetic effects, as well as the potential mechanisms by which zinc exerts these effects.

Topics: Animals; Diabetes Mellitus; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Disease Models, Animal; ErbB Receptors; Glycogen; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Lipogenesis; Receptor, IGF Type 1; Signal Transduction; Zinc; Zinc Compounds

Proinsulin C-peptide, released in equimolar amounts with insulin by pancreatic β cells, since its discovery in 1967 has been thought to be devoid of biological functions apart from correct insulin processing and formation of disulfide bonds between A and B chains. However, in the last two decades research has brought a substantial amount of data indicating a crucial role of C-peptide in regulating various processes in different types of cells and organs. C-peptide acts presumably via either G-protein-coupled receptor or directly inside the cell, after being internalized. However, a receptor binding this peptide has not been identified yet. This peptide ameliorates pathological changes induced by type 1 diabetes mellitus, including glomerular hyperfiltration, vessel endothelium inflammation and neuron demyelinization. In diabetic patients and diabetic animal models, C-peptide substitution in physiological doses improves the functional and structural properties of peripheral neurons and protects against hyperglycemia-induced apoptosis, promoting neuronal development, regeneration and cell survival. Moreover, it affects glycogen synthesis in skeletal muscles. In vitro C-peptide promotes disaggregation of insulin oligomers, thus enhancing its bioavailability and effects on metabolism. There are controversies concerning the biological action of C-peptide, particularly with respect to its effect on Na⁺/K⁺-ATPase activity. Surprisingly, the excess of circulating peptide associated with diabetes type 2 contributes to atherosclerosis development. In view of these observations, long-term, large-scale clinical investigations using C-peptide physiological doses need to be conducted in order to determine safety and health outcomes of long-term administration of C-peptide to diabetic patients.

Topics: Animals; Apoptosis; Atherosclerosis; C-Peptide; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Diabetic Neuropathies; Disease Models, Animal; Glycogen; Humans; Hyperglycemia; Muscle, Skeletal; Peripheral Nervous System

This review deals with the role of glycogen storage in skeletal muscle for the development of insulin resistance and type 2 diabetes. Specifically, the role of the enzyme glycogen synthase, which seems to be locked in its hyperphosphorylated and inactivated state, is discussed. This defect seems to be secondary to ectopic lipid disposition in the muscle cells. These molecular defects are discussed in the context of the overall pathophysiology of hyperglycemia in type 2 diabetic subjects.

Topics: Diabetes Mellitus, Type 2; Glucose; Glycated Hemoglobin; Glycogen; Glycogen Synthase; Humans; Hyperglycemia; Insulin; Insulin Resistance; Lipid Metabolism; Muscle, Skeletal; Phosphorylation

The quest for the discovery of new antihyperglycemic agents has been more intense the last years due to the rapid increase of mortality associated with type 2 diabetes. Glycogen metabolism has been one of the major causes of the elevated blood glucose levels; hence, special attention has been drawn to the control of the enzymes implicated in the relevant pathway. To this end, the allosteric enzyme of glycogen phosphorylase, has been proposed as molecular target for the design of potential new antidiabetic agents by an interdisciplinary approach comprising organic synthesis, kinetic and X-ray crystallographic studies and physiological experiments. The results derived from the thorough investigation of the catalytic site of the enzyme with the structure-based inhibitor design approach are summarized with emphasis on the most potent inhibitors identified for different classes of compounds.

Topics: Animals; Binding Sites; Catalytic Domain; Crystallography, X-Ray; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Phosphorylase; Humans; Hypoglycemic Agents; Protein Structure, Tertiary; Rabbits

The principal modifiable risk factors for stroke are hypertension, diabetes mellitus, hypercholesterolaemia, hyperhomocysteinaemia, smoking and limited physical activity. However, it is not clear whether physical inactivity is a risk factor per se, or because it predisposes to pathological conditions that are risk factors for stroke. The limited availability of effective therapeutic approaches for stroke emphasizes the crucial role of prevention of risk factors. The global burden associated with type 2 diabetes is large and continues to grow. Convincing epidemiologic data support the role of physical activity in preventing type 2 diabetes. The increasing evidence of physical activity in preventing diabetic complications, including stroke, has generated interest in the molecular basis underlying these beneficial effects. The aim of the present review is to discuss the biological mechanisms underlying the effect of physical activity in preventing stroke in type 2 diabetes.

Topics: Animals; Diabetes Mellitus, Type 2; Endothelium; Exercise; Exercise Therapy; Glucose Transporter Type 4; Glycogen; Humans; Insulin; Lipid Metabolism; Muscle, Skeletal; Nitric Oxide; Risk Factors; Signal Transduction; Stroke

In mammals, excess carbohydrate is stored as glycogen and glycogen synthase is the enzyme that incorporates glucose units into the glycogen particle. Glycogen synthase activity is regulated by phosphorylation and allosterically activated by glucose 6-phosphate. Phosphorylation of nine serines by different kinases regulates glycogen synthase affinity for glucose 6-phosphate and its substrate UDP-glucose. Glucose 6-phosphate increases both enzyme activity and substrate affinity. Insulin and exercise increase glycogen synthase affinity for glucose 6-phosphate and activity whereas high glycogen content and adrenaline decrease affinity for glucose 6-phosphate and activity. However, insulin, exercise and adrenaline also regulate intracellular concentration of glucose 6-phosphate which will influence in vivo glycogen synthase activity. Importantly, type 2 diabetes is associated with reduced insulin-stimulated glycogen synthase activation. The nine phosphorylation sites theoretically allow 512 combinations of phosphorylation configurations of glycogen synthase with different kinetic properties. However, due to hierarchal phosphorylation, the number of configurations in vivo is most likely much lower. Unfortunately, many studies only report data on glycogen synthase activity measured with high concentration of UDP-glucose which holds back information about changes in substrate affinity. In this paper we discuss the physiological regulation of glycogen synthase phosphorylation and how the phosphorylation pattern regulates glycogen synthase kinetic properties.

Topics: Allosteric Regulation; Animals; Diabetes Mellitus, Type 2; Epinephrine; Exercise; Glycogen; Glycogen Synthase; Humans; Insulin; Insulin Resistance; Muscle, Skeletal; Phosphorylation; Uridine Diphosphate Glucose

It is now becoming evident that the liver has an important role in the control of whole body metabolism of energy nutrients. In this review, we focus on recent findings showing that AMP-activated protein kinase (AMPK) plays a major role in the control of hepatic metabolism. AMPK integrates nutritional and hormonal signals to promote energy balance by switching on catabolic pathways and switching off ATP-consuming pathways, both by short-term effects on phosphorylation of regulatory proteins and by long-term effects on gene expression. Activation of AMPK in the liver leads to the stimulation of fatty acid oxidation and inhibition of lipogenesis, glucose production and protein synthesis. Medical interest in the AMPK system has recently increased with the demonstration that AMPK could mediate some of the effects of the fat cell-derived adiponectin and the antidiabetic drugs metformin and thiazolidinediones. These findings reinforce the idea that pharmacological activation of AMPK may provide, through signalling and metabolic and gene expression effects, a new strategy for the management of metabolic hepatic disorders linked to type 2 diabetes and obesity.

Topics: AMP-Activated Protein Kinases; Animals; Diabetes Mellitus, Type 2; Enzyme Activation; Glucose; Glycogen; Humans; Liver; Liver Diseases; Multienzyme Complexes; Obesity; Protein Serine-Threonine Kinases

The brain uses glucose as a primary fuel for energy generation. Glucose gains entry into the brain by facilitated diffusion across the blood-brain barrier. Glucose transport may adapt during changes in cerebral glucose metabolism, neural activation and changes in plasma glucose levels. Within the brain, glucose is either oxidized to produce ATP or used to synthesize glycogen. To ensure the delivery of a continuous supply of glucose to maintain normal cellular function, the brain has developed a complex regulatory system to preserve its supply. Gluco-sensing neurons have been demonstrated in various regions of the brain and they appear to play an important role in not only detecting changes in brain glucose levels but also in initiating responses to maintain constant brain glucose levels. In this review, we will discuss the regulation of brain glucose metabolism (CMR(gluc)) and how it adapts to chronic changes in glycemia, like that seen in hyperglycemic patients with diabetes mellitus or patients with type 1 diabetes, recurrent hypoglycemia, and hypoglycemia unawareness. We will also consider the role of brain glycogen in providing fuel for energy under conditions of stress.

Topics: Blood-Brain Barrier; Brain; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glucose; Glucose Transporter Type 1; Glucose Transporter Type 3; Glycogen; Humans; Insulin; Neuroglia; Neurons

Type 2 diabetes is a complex metabolic disease with hyperglycemia as its recognizable hallmark. Hepatic glucose output is elevated in Type 2 diabetic patients, and evidence suggests drugs which lower hepatic glucose production are effective antihyperglycemic agents. Glycogenolysis, which is the release of monomeric glucose from its polymeric storage form called glycogen, is a key contributor to hepatic glucose output. Glycogen phosphorylase is the enzyme that catalyzes this process. This review covers advances in the design of small molecule inhibitors of this enzyme, their biological activity, and their potential as effective antihyperglycemic agents for the treatment of Type 2 diabetes.

Topics: Diabetes Mellitus, Type 2; Enzyme Inhibitors; Glucose; Glycogen; Glycogen Phosphorylase; Humans; Hyperglycemia; Hypoglycemic Agents; Liver

Topics: Acarbose; Cyclohexanes; Diabetes Mellitus, Type 2; Diet Therapy; Enzyme Inhibitors; Exercise Therapy; Glucose; Glucose Intolerance; Glycogen; Glycoside Hydrolase Inhibitors; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Insulin Secretion; Metformin; Nateglinide; Phenylalanine; Sulfonylurea Compounds; Thiazolidinediones

The etiology of type I and type II diabetes differs and so do the nutritional challenges during and after exercise. For type I diabetics, exercise may cause hypoglycemia. To avoid hypoglycemia, a carbohydrate-rich meal should be eaten 1 to 3 hours prior to exercise and the insulin dose reduced. During exercise, at least 40 g glucose per hour should be ingested; more if the insulin dose is not reduced. After exercise, it is important to rebuild the glycogen stores to reduce the risk for hypoglycemia. Carbohydrates should always be available during training and in the recovery period. Despite these difficulties, exercise is recommended for type I diabetics and competition at high level is possible. Exercise prevents development of type II diabetes and improves metabolic regulation. For type II diabetics, exercise is normally performed to improve insulin sensitivity and to reduce body weight. Carbohydrates should only be supplied to prevent hypoglycemia.

Topics: Blood Glucose; Body Weight; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Exercise; Glycemic Index; Glycogen; Homeostasis; Humans; Hypoglycemia; Muscle, Skeletal; Nutritional Physiological Phenomena; Physical Endurance

Type 2 diabetes is a complex disorder with diminished insulin secretion and insulin action contributing to the hyperglycemia and wide range of metabolic defects that underlie the disease. The contribution of glucose metabolic pathways per se in the pathogenesis of the disease remains unclear. The cellular fate of glucose begins with glucose transport and phosphorylation. Subsequent pathways of glucose utilization include aerobic and anaerobic glycolysis, glycogen formation, and conversion to other intermediates in the hexose phosphate or hexosamine biosynthesis pathways. Abnormalities in each pathway may occur in diabetic subjects; however, it is unclear whether perturbations in these may lead to diabetes or are a consequence of the multiple metabolic abnormalities found in the disease. This review is focused on the cellular fate of glucose and relevance to human type 2 diabetes.

Topics: Biological Transport; Cells; Diabetes Mellitus, Type 2; Gluconeogenesis; Glucose; Glycogen; Glycolysis; Hexosamines; Hexosephosphates; Humans; Monosaccharide Transport Proteins; Pentose Phosphate Pathway; Pentoses; Phosphorylation

Excessive hepatic glucose production is thought to be a major contributor to the type 2 diabetic state. Drug discovery efforts have yielded small synthetic inhibitors for gluconeogenic and glycogenic regulators of this pathway. The most advanced targets are outlined in this mini-review and include: the glucocorticoid receptor, 11 beta-hydroxysteroid dehydrogenase type 1, fructose 1,6-bisphosphatase, the glucagon receptor, glycogen phosphorylase, glycogen synthase kinase-3, and glucose-6-phosphatase.

Topics: Animals; Diabetes Mellitus, Type 2; Enzymes; Gluconeogenesis; Glucose; Glycogen; Humans; Hypoglycemic Agents; Liver; Models, Biological; Molecular Conformation; Receptors, Glucagon; Receptors, Glucocorticoid

Topics: Diabetes Mellitus, Type 2; Fasting; Fatty Acids, Nonesterified; Gluconeogenesis; Glycogen; Humans; Insulin; Insulin Resistance

The AMP-activated protein kinase (AMPK) system was first discovered 30 years ago. Since that time, knowledge of the diverse physiological functions of AMPK has grown rapidly and continues to evolve. Most recently, the observation that spontaneously occurring genetic mutations in the gamma regulatory subunits of AMPK give rise to a skeletal and cardiac muscle disease emphasizes the critical importance of AMPK in the maintenance of health and disease. The cardiac phenotype observed in humans harbouring genetic mutations in the gamma 2 regulatory subunit (PRKAG2) of AMPK is consistent with abnormal glycogen accumulation in the heart. The perturbation of AMPK activity induced by genetic mutations in PRKAG2 and the resultant effect on muscle cell glucose metabolism may be relevant to the issue of targeting AMPK in drug development for insulin-resistant diabetes mellitus.

Topics: AMP-Activated Protein Kinases; Animals; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Storage Disease; Humans; Multienzyme Complexes; Muscle, Skeletal; Mutation; Myocardium; Phenotype; Protein Serine-Threonine Kinases; Signal Transduction

It has recently been demonstrated that the marked increase in the systemic concentration of cytokine interleukin-6 (IL-6) seen with exercise originates from the contracting limb and that skeletal muscle cells per se are the likely source of the production. This review summarizes the possible mechanisms for activation and biological consequences of muscle-derived IL-6. It appears that intramuscular IL-6 is stimulated by complex signaling cascades initiated by both calcium (Ca2+) -dependent and -independent stimuli. It also seems likely that skeletal muscle produces IL-6 to aid in maintaining metabolic homeostasis during periods of altered metabolic demand such as muscular exercise or insulin stimulation. It may do so via local and/or systemic effects. This review also explores the efficacy that IL-6 may be used as a therapeutic drug in treating metabolic disorders such as obesity, type 2 diabetes, and atherosclerosis.

Topics: Animals; Anti-Inflammatory Agents; Diabetes Mellitus, Type 2; Exercise; Glucose; Glycogen; Humans; Insulin Resistance; Interleukin-6; Lipid Metabolism; Models, Biological; Monocytes; Muscle, Skeletal; Signal Transduction

Insulin resistance is a principal feature of type 2 diabetes and precedes the clinical development of the disease by 10 to 20 years. Insulin resistance is caused by the decreased ability of peripheral target tissues (especially muscle) to respond properly to normal circulating concentrations of insulin. Defects in muscle glycogen synthesis play a significant role in insulin resistance, and 3 potentially rate-controlling steps in muscle glucose metabolism have been implicated in its pathogenesis: glycogen synthase, hexokinase, and GLUT4 (the major insulin-stimulated glucose transporter). Results from recent studies using nuclear magnetic resonance (NMR) spectroscopy implicate intracellular defects in glucose transport as the rate-controlling step for insulin-mediated glucose uptake in muscle. These alterations in glucose transport activity are likely the result of dysregulation of intramyocellular fatty acid metabolism, whereby fatty acids cause insulin resistance by activation of a serine kinase cascade, leading to decreased insulin-stimulated insulin receptor substrate (IRS)-1 tyrosine phosphorylation and decreased IRS-1-associated phosphatidylinositol 3-kinase activity, a required step in insulin-stimulated glucose transport into muscle. The thiazolidinedione class of antidiabetic agents directly targets insulin resistance in skeletal muscle by improving glucose transport activity and insulin-stimulated muscle glycogen synthesis. Although the precise mechanism of action is not known, recent NMR studies support the hypothesis that these agents improve insulin action in skeletal muscle and liver by promoting a redistribution of fat out of these tissues and into peripheral adipocytes.

Topics: Diabetes Mellitus, Type 2; Fatty Acids; Glucose; Glycogen; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Magnetic Resonance Spectroscopy; Muscle, Skeletal; Signal Transduction; Thiazoles

Topics: Animals; Diabetes Mellitus, Type 2; Glycogen; Glycogen Synthase; Humans; Insulin; Insulin Resistance; Muscle, Skeletal; Phosphoprotein Phosphatases; Phosphorylation; Protein Kinases

Topics: Adipose Tissue; Diabetes Mellitus, Type 2; Energy Metabolism; Exercise; Exercise Therapy; Fatty Acids, Nonesterified; Glucose Transporter Type 4; Glycogen; Humans; Insulin; Insulin Resistance; Monosaccharide Transport Proteins; Muscle Proteins; Muscle, Skeletal; Receptor, Insulin

Since its discovery more than a decade ago, the Ser/Thr kinase Akt/PKB (protein kinase B) has been recognized as being remarkably well conserved across a broad range of species and involved in a diverse array of cellular processes. Among its many roles, Akt appears to be common to signaling pathways that mediate the metabolic effects of insulin in several physiologically important target tissues. Refining our understanding of those pivotal molecular components that normally coordinate insulin action throughout the body is essential for a full understanding of insulin resistance in diabetes mellitus and ultimately the successful treatment of this disease.

Topics: Adipose Tissue; Animals; Base Sequence; Diabetes Mellitus, Type 2; Disease Models, Animal; Energy Metabolism; Glucose; Glucose Transporter Type 4; Glycogen; Insulin; Lipid Metabolism; Liver; Mice; Molecular Sequence Data; Monosaccharide Transport Proteins; Muscle Proteins; Muscles; Nitric Oxide Synthase; Pancreas; Protein Biosynthesis; Protein Isoforms; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt

Skeletal muscle contains the majority of the body's glycogen stores and a similar amount of readily accessible energy as intramyocellular triglyceride (imTG). While a number of factors have been considered to contribute to the pathogenesis of insulin resistance (IR) in obesity and type 2 diabetes mellitus (DM), this review will focus on the potential role of skeletal muscle triglyceride content. In obesity and type 2 DM, there is an increased content of lipid within and around muscle fibers. Changes in muscle in fuel partitioning of lipid, between oxidation and storage of fat calories, almost certainly contribute to accumulation of imTG and to the pathogenesis of both obesity and type 2 DM. In metabolic health, skeletal muscle physiology is characterized by the capacity to utilize either lipid or carbohydrate fuels, and to effectively transition between these fuels. We will review recent findings that indicate that in type 2 DM and obesity, skeletal muscle manifests inflexibility in the transition between lipid and carbohydrate fuels. This inflexibility in fuel selection by skeletal muscle appears to be related to the accumulation of imTG and is an important aspect of IR of skeletal muscle in obesity and type 2 DM.

Topics: Adipose Tissue; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin Resistance; Leptin; Muscle, Skeletal; Obesity; Triglycerides

Insulin is the most-potent physiological anabolic agent known, promoting the synthesis and storage of carbohydrates and lipids and inhibiting their degradation and release into the circulation. This action of the hormone is due in part to the acute regulation of metabolic enzymes through changes in their phosphorylation state. In fat, liver, and muscle, insulin stimulates the dephosphorylation of a number of enzymes involved in glycogen and lipid metabolism via activation of protein phosphatases. Numerous studies have indicated that protein phosphatase-1 (PP1) is the primary phosphatase involved in insulin action. Although PP1 is a cytosolic protein, the phosphatase is compartmentalized in cells by discrete targeting subunits. These proteins confer substrate specificity to PP1 and mediate the specific regulation of intracellular pools of PP1 by a variety of extracellular signals. Four proteins have been described that target the phosphatase to the glycogen particle. G(M) and GL are expressed exclusively in striated muscle and liver, while protein targeting to glycogen (PTG) and R6 are more widely expressed. Despite a common targeting function, these four proteins are not highly conserved, suggesting profound differences in the mechanisms by which they contribute to the hormonal regulation of PP1 activity. Overexpression studies in cell lines or animals have revealed major differences among these proteins regarding basal glycogen levels and hormonal responsiveness. Furthermore, alterations in the expression or function of PP1 glycogen-targeting subunits may contribute to the onset of insulin resistance and type 2 diabetes.

Topics: Amino Acid Sequence; Animals; Aprotinin; Cell Line; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin; Insulin Resistance; Liver; Mice; Mice, Knockout; Models, Biological; Molecular Sequence Data; Muscle, Skeletal; Phosphoprotein Phosphatases; Phosphorylation; Protein Binding; Protein Phosphatase 1; Recombinant Proteins; Sequence Homology, Amino Acid; Structure-Activity Relationship

Nuclear magnetic resonance (NMR) spectroscopy has made noninvasive and repetitive measurements of human hepatic glycogen concentrations possible. Monitoring of liver glycogen in real-time mode has demonstrated that glycogen concentrations decrease linearly and that net hepatic glycogenolysis contributes only about 50 percent to glucose production during the early period of a fast. Following a mixed meal, hepatic glycogen represents approximately 20 percent of the ingested carbohydrates, while only about 10 percent of an intravenous glucose load is retained by the liver as glycogen. During mixed-meal ingestion, poorly controlled type 1 diabetic patients synthesize only about 30 percent of the glycogen stored in livers of nondiabetic humans studied under similar conditions. Reduced net glycogen synthesis can be improved but not normalized by short-term, intensified insulin treatment. A decreased increment in liver glycogen content following meals was also found in patients with maturity-onset diabetes of the young due to glucokinase mutations (MODY-2). In patients with poorly controlled type 2 diabetes, fasting hyperglycemia can be attributed mainly to increased rates of endogenous glucose production, which was found by 13C NMR to be due to increased rates of gluconeogenesis. Metformin treatment improved fasting hyperglycemia in these patients through a reduction in hepatic glucose production, which could be attributed to a decrease in gluconeogenesis. In conclusion, NMR spectroscopy has provided new insights into the pathogenesis of hyperglycemia in type 1, type 2, and MODY diabetes and offers the potential of providing new insights into the mechanism of action of novel antidabetic therapies.

Topics: Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Fasting; Glucose; Glycogen; Humans; Hyperglycemia; Liver; Liver Glycogen; Magnetic Resonance Spectroscopy; Metformin; Time Factors

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin; Insulin Resistance; Islets of Langerhans; Models, Biological; Obesity; Protein-Tyrosine Kinases; Receptor, Insulin; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Transcription Factors

Insulin resistance is defined as a clinical state in which a normal or elevated insulin level produces an attenuated biologic response. Specifically, the biologic response most studied is insulin-stimulated glucose disposal, yet the precise cellular mechanism responsible is not yet known. However, the presence of insulin resistance is observed many years before the onset of clinical hyperglycemia and the diagnosis of Type 2 diabetes. Insulin resistance at this stage appears to be significantly associated with a clustering of cardiovascular risk factors predisposing the individual to accelerated cardiovascular disease. An overview of insulin resistance and the associated clinical insulin resistant state will be discussed.

Topics: Animals; Biological Transport; Cardiovascular Diseases; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Hyperglycemia; Insulin; Insulin Resistance; Models, Chemical; Signal Transduction; Syndrome

Nuclear magnetic resonance spectroscopy provides non-invasive and real-time assessment of the metabolic fluxes in skeletal muscle during exercise, recovery from exercise and stimulation by insulin. Carbon-13 nuclear magnetic resonance spectroscopy has proved that reduced glycogen synthesis is a consistent feature of insulin-resistant type 2 diabetic patients, their offspring, and obesity. Low intracellular glucose and glucose-6-phosphate concentrations indicate that decreased glucose transport is mainly responsible for common insulin resistance. An elevation of plasma free fatty acids causes similar alterations of muscle glucose metabolism, and could play a central role in the development of impaired muscle glucose transport associated with insulin resistance.

Topics: Carbon Isotopes; Diabetes Mellitus, Type 2; Exercise; Fatty Acids, Nonesterified; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Magnetic Resonance Spectroscopy; Muscle, Skeletal; Obesity

Glycogen synthase kinase 3 (GSK3) was initially described as a key enzyme involved in glycogen metabolism, but is now known to regulate a diverse array of cell functions. The study of the substrate specificity and regulation of GSK3 activity has been important in the quest for therapeutic intervention.

Topics: Animals; Calcium-Calmodulin-Dependent Protein Kinases; Diabetes Mellitus, Type 2; Drosophila melanogaster; Embryo, Nonmammalian; Enzyme Inhibitors; Glycogen; Glycogen Synthase Kinases; Humans; Insulin; Phosphorylation; Protein Biosynthesis; Signal Transduction; Substrate Specificity

Clear cell ("sugar") tumour of the lung is a rare neoplasm, generally presenting as a discrete nodule on the chest X-ray. We report a case of clear cell tumour of the lung in a 64-year-old woman. The tumour at presentation was 1 mm in diameter.

Topics: Adenocarcinoma; Adenocarcinoma, Clear Cell; Antigens, Neoplasm; Biomarkers, Tumor; Diabetes Mellitus, Type 2; Fatal Outcome; Female; Glycogen; Hemoptysis; Humans; Lung Neoplasms; Melanoma-Specific Antigens; Middle Aged; Neoplasm Metastasis; Neoplasm Proteins; Neoplasms, Multiple Primary; Smoking; Solitary Pulmonary Nodule

Prior to the advent of nuclear magnetic resonance (NMR) spectroscopy, human glucose metabolism was studied through tracer and tissue biopsy methodology. NMR spectroscopy now provides a noninvasive means to monitor metabolic flux and intracellular metabolite concentrations continuously. 13C NMR spectroscopy has shown that muscle glycogen synthesis accounts for the majority of insulin-stimulated muscle glucose uptake in normal volunteers and that defects in this process are chiefly responsible for insulin resistance in type 1 and type 2 diabetes mellitus, as well as in other insulin resistant states (obesity, insulin-resistant offspring of type 2 diabetic parents, elevation of plasma FFA concentrations). Furthermore, using 31P NMR spectroscopy to measure intracellular glucose-6-phosphate, it has been shown that defects in insulin-stimulated glucose transport/phosphorylation activity are primarily responsible for the insulin resistance in these states.

Topics: Carbon Isotopes; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Glucose; Glucose-6-Phosphate; Glycogen; Humans; Insulin; Insulin Resistance; Magnetic Resonance Spectroscopy; Muscle, Skeletal; Obesity; Phosphorus Isotopes; Phosphorylation

Carbon nuclear magnetic resonance (13C NMR) spectroscopy and phosphorus (31p) NMR spectroscopy have been used to help define the contribution of insulin-stimulated muscle glycogen synthesis to whole-body insulin-stimulated glucose metabolism in normal individuals and the extent to which this process is defective in patients with type 2 (non-insulin-dependent) diabetes. Assessments of the response to hyperglycemic-hyperinsulinemic clamping have shown that abnormalities of muscle glycogen synthesis, apparently mediated by a defect in GLUT-4 transport and/or hexokinase activity, play a major role in causing insulin resistance in type 2 diabetes. Studies of the mechanisms by which free fatty acids (FFA) cause insulin resistance in humans indicate that increased FFA levels inhibit glucose transport, which may be a consequence of decreased insulin receptor substrate (IRS-1)-associated phosphatidylinositol 3-kinase activity. 13C NMR spectroscopy studies have documented that liver glycogen concentrations are reduced and the rate of hepatic gluconeogenesis is increased in subjects with type 2 diabetes; thus, the higher rate of glucose production in type 2 diabetes can be attributed entirely to increased rates of hepatic gluconeogenesis. These cellular mechanisms of insulin resistance can be addressed through combination therapy with agents that reverse the principal pathophysiologic defects of type 2 diabetes. The biguanide metformin appears to lower glucose by suppressing hepatic glucose production, whereas the thiazolidinedione troglitazone appears to increase glucose clearance by peripheral tissues. The two agents together have been shown to provide better glucose control than either drug alone, without stimulating insulin secretion.

Topics: Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Magnetic Resonance Spectroscopy; Muscles

Maturity onset diabetes of the young (MODY) is a genetically and clinically heterogeneous subtype of non-insulin-dependent diabetes mellitus (NIDDM) characterised by early onset, autosomal dominant inheritance and a primary defect in insulin secretion. To date, three MODY genes have been identified on chromosomes 20q (MODY1/hepatic nuclear factor (HNF)-4alpha), 7p (MODY2/glucokinase) and 12q (MODY3/HNF-1alpha). Mutations in MODY2/glucokinase result in mild chronic hyperglycaemia as a result of reduced pancreatic beta-cell responsiveness to glucose, and decreased net accumulation of hepatic glycogen and increased hepatic gluconeogenesis after meals. In contrast, MODY1 and MODY3 are characterised by severe insulin secretory defects, and by major hyperglycaemia associated with microvascular complications. The role of the three known MODY genes in susceptibility to the more common late-onset NIDDM remain uncertain. Genetic studies seem to exclude a role as major susceptibility genes, but leave unresolved whether they may have a minor role in a polygenic context or an important role in particular populations.

Topics: Adolescent; Adult; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Child; Diabetes Mellitus, Type 2; DNA-Binding Proteins; Female; Glucokinase; Glycogen; Hepatocyte Nuclear Factor 1; Hepatocyte Nuclear Factor 1-alpha; Hepatocyte Nuclear Factor 1-beta; Hepatocyte Nuclear Factor 4; Homeodomain Proteins; Humans; Liver; Male; Nuclear Proteins; Phosphoproteins; Prevalence; Trans-Activators; Transcription Factors

Insulin-mediated non-oxidative glucose metabolism is more or less identical to glycogen synthesis in skeletal muscle and that is why this pathway is specifically discussed in this paper. All three major steps in non-oxidative glucose processing--glucose transport, phosphorylation and glycogen synthesis--are found to be reduced in response to insulin in insulin-resistant type 2 diabetic subjects compared with controls. The insulin-signalling cascade from the insulin receptor to PI-3-K was also found to be abnormal, resulting in a severely reduced phosphorylation degree of the IRS-1 (IRS-2?)-PI-3-K complex, which can explain both reduced glucose transport and glycogen synthesis. The most pronounced finding in our studies is reduced glycogen synthase activation by insulin which is found in prediabetic subjects with normal glucose tolerance as well as in type 2 diabetics, but more severely. This defect was not reversible after treatment (normalization of blood glucose) and is therefore a candidate for the primary defect which is likely to be of genetic origin, but also could be caused by genetic imprinting, intrauterine malnutrition and social inheritance (obesity). Most of the abnormalities in non-oxidative glucose metabolism may be of secondary origin due to hyperglycemia itself or obesity. Both events may stimulate production of glucosamine, malonyl CoA and intramuscular triglyceride accumulation. These metabolites can theoretically induce most of the defects in glucose processing and furthermore impair insulin signalling. Whether the primary defect in activation of glycogen synthase is due to an abnormality in the enzyme complex itself or in the insulin signalling cascade still has to be investigated.

Topics: Diabetes Mellitus, Type 2; Enzyme Activation; Fatty Acids, Nonesterified; Glucose; Glucose Transporter Type 4; Glycogen; Glycogen Synthase; Glycolysis; Hexokinase; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Male; Monosaccharide Transport Proteins; Muscle Proteins; Muscle, Skeletal; Phosphorylation; Signal Transduction; Twins

Topics: Amino Acids; Diabetes Mellitus; Diabetes Mellitus, Type 2; Fructose; Gluconeogenesis; Glycerol; Glycogen; Homeostasis; Humans; Insulin; Lactates; Obesity

Topics: Blood Glucose; Diabetes Mellitus, Type 2; Glucose; Glucose Clamp Technique; Glycogen; Humans; Insulin Resistance; Muscle, Skeletal

Topics: Adenosine Triphosphate; Blood Glucose; Diabetes Mellitus, Type 2; Diet Therapy; Exercise Therapy; Female; Glycogen; Humans; Hyperglycemia; Hypoglycemic Agents; Insulin; Insulin Secretion; Male; Sulfonylurea Compounds

The activity of glycogen synthase (GSase; EC 2.4.1.11) is regulated by covalent phosphorylation. Because of this regulation, GSase has generally been considered to control the rate of glycogen synthesis. This hypothesis is examined in light of recent in vivo NMR experiments on rat and human muscle and is found to be quantitatively inconsistent with the data under conditions of glycogen synthesis. Our first experiments showed that muscle glycogen synthesis was slower in non-insulin-dependent diabetics compared to normals and that their defect was in the glucose transporter/hexokinase (GT/HK) part of the pathway. From these and other in vivo NMR results a quantitative model is proposed in which the GT/HK steps control the rate of glycogen synthesis in normal humans and rat muscle. The flux through GSase is regulated to match the proximal steps by "feed forward" to glucose 6-phosphate, which is a positive allosteric effector of all forms of GSase. Recent in vivo NMR experiments specifically designed to test the model are analyzed by metabolic control theory and it is shown quantitatively that the GT/HK step controls the rate of glycogen synthesis. Preliminary evidence favors the transporter step. Several conclusions are significant: (i) glucose transport/hexokinase controls the glycogen synthesis flux; (ii) the role of covalent phosphorylation of GSase is to adapt the activity of the enzyme to the flux and to control the metabolite levels not the flux; (iii) the quantitative data needed for inferring and testing the present model of flux control depended upon advances of in vivo NMR methods that accurately measured the concentration of glucose 6-phosphate and the rate of glycogen synthesis.

Topics: Allosteric Regulation; Animals; Diabetes Mellitus, Type 2; Gene Expression Regulation, Enzymologic; Glucose-6-Phosphate; Glucosephosphates; Glycogen; Glycogen Synthase; Humans; Models, Biological; Muscles; Phosphorylation; Rats

An increased supply of FFAs for oxidation leads to a reduced rate of glucose oxidation and interferes with the inhibitory action of insulin on hepatic glucose production. Available evidence indicates that in humans skeletal muscle is a site for such substrate competition, which involves both pyruvate oxidation and glycogen synthesis. The insulin resistance of obesity is thought to be mostly of metabolic origin, and fully reversible. A reduction in FFA supply by weight reduction can, however, reverse this defect. The insulin resistance associated with NIDDM is thought to be primary, with a strong genetic basis, and partially irreversible. Patients with NIDDM are unable to increase their glucose oxidation normally in response to insulin to meet the energy demands of the body. Increased oxidation of lipids represents a compensatory phenomenon to meet these demands. Therapeutic use of the glucose-FFA cycle to lower blood glucose levels has yielded conflicting results. Studies are in progress to develop agents that inhibit gluconeogenesis by interfering with FFA oxidation. Nicotinic acid derivatives seem to enhance glycogen synthesis acutely by activating glycogen synthetase. Whether these or similar agents can be used to restore impaired glycogen synthesis, the most characteristic genetic defect in NIDDM, cannot be answered until the effect has been proven in chronic studies. The existence of substrate competition between amino acids and glucose, and an intrinsic hypoaminoacidaemic property of amino acids, makes it possible to expand the Randel cycle into a glucose-FFA-amino acid cycle, which integrates control of substrate disposition at the whole body level.

Topics: Animals; Binding, Competitive; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Gluconeogenesis; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Lipid Metabolism; Liver; Muscles; Niacin; Obesity; Oxidation-Reduction; Rats

A metabolic hypothesis is presented for insulin resistance in obesity, in the presence or absence of Type 2 (non-insulin-dependent) diabetes mellitus. It is based on physiological mechanisms including a series of negative feed-back mechanisms, with the inhibition of the function of the glycogen cycle in skeletal muscle as a consequence of decreased glucose utilization resulting from increased lipid oxidation in the obese. It considers the inhibition of glycogen synthase activity together with inhibition of glucose storage and impaired glucose tolerance. The prolonged duration of increased lipid oxidation, considered as the initial cause, may lead to Type 2 diabetes. This hypothesis is compatible with others based on the inhibition of insulin receptor kinase and of glucose transporter activities.

Topics: Diabetes Mellitus; Diabetes Mellitus, Type 2; Glucose; Glycogen; Homeostasis; Humans; Insulin Resistance; Models, Biological; Muscles; Obesity

Topics: Diabetes Mellitus; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Insulin Secretion; Lipid Metabolism; Obesity; Oxidation-Reduction; Risk Factors

By the term "insulin resistance" we understand the attenuation of insulin-stimulated glucose uptake, which is mainly due to attenuated glycogen synthesis in skeletal muscle and is partially compensated with regard to plasma glucose homeostasis by hyperinsulinemia. Other mechanisms of insulin are either not attenuated or are less so and may contribute via hyperinsulinemia to the prevalence of hypertension, obesity, dyslipoproteinemia and type-II diabetes. At the level of insulin receptors, resistance can be due to muscle-specific, preferential expression of the low-affinity B-isoform of the insulin receptors. In rare cases of extreme resistance, it can also be due to several mutations at the insulin receptor gene or due to insulin-receptor autoantibodies. At the postreceptor level, the translocation and or expression of the insulin-responsive glucose carrier GluT-4 can be down-regulated via the hexosamine pathway by hyperglycemia plus hyperinsulinemia. Furthermore, Glut-4 can be inhibited and/or down-regulated by sustained insulin deficiency, partially via c-AMP-dependent pathways. Additionally, the insulin-induced glycogen synthesis in skeletal muscle can be attenuated by the endogenous peptides amylin and calcitonin-gene-related peptide, and by modulations of endothelial function, perfusion and capillary recruitment in the microcirculation of skeletal muscle. Epidemiological data indicate a genetic predisposition for insulin resistance. However, among the many mechanisms potentially contributing to the complex syndrome of insulin resistance, no specific localization of that predisposition can be proposed at present.

Topics: Amyloid; beta-N-Acetylhexosaminidases; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glycogen; Humans; Hyperglycemia; Hyperinsulinism; Insulin Resistance; Islet Amyloid Polypeptide; Monosaccharide Transport Proteins; Receptor, Insulin

In non-insulin-dependent diabetic subjects and in various animal models of spontaneous or experimental chronic hyperglycaemia, the secretory response of the pancreatic B-cell to a rapid rise in extracellular D-glucose concentration is characterized by a paradoxical, early and transient fall in insulin output and/or an altered anomeric specificity. These two features of B-cell glucotoxicity may be accounted for by the accumulation of glycogen in the B-cell and the interference of changes in glycogenolysis with the hexose-induced increase in glycolytic flux. The inhibitory action of D-glucose upon glycogenolysis displaying alpha-stereospecificity, the metabolic and secretory response to alpha-D-glucose is expected to be more severely affected than that evoked by the beta-anomer. Such a preferential alteration of the response to alpha-D-glucose was indeed documented in diabetic subjects, BB rats, duct-ligated rabbits, and adult rats either injected with streptozotocin during the neonatal period or rendered hyperglycaemic by the repeated administration of diazoxide. In these experimental models, the attenuation, suppression or even reversal of the anomeric preference in insulin release appeared related to the severity and duration of the hyperglycaemic state. A clear distinction ought to be made between these features of B-cell glucotoxicity and other etiopathogenic factors of B-cell dysfunction, such as the long term deleterious effect of streptozotocin upon the activity of key mitochondrial dehydrogenases.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycosylation; Humans; Insulin; Insulin Secretion; Islets of Langerhans; Sorbitol

Sulfonylureas are widely used drugs in the treatment of NIDDM when diet treatment is unsuccessful. In addition to their pancreatic effects sulfonylureas have been reported to have insulin-like and insulin-potentiating actions in vitro with respect both to glucose transport and glycogen synthase activation in isolated adipocytes and hepatocytes from rats. Glycogen synthesis in muscle accounts for the major part of non-oxidative glucose metabolism during insulin stimulation. Treatment with gliclazide of patients with NIDDM has been shown to be associated with a potentiation of both insulin-mediated glucose disposal and insulin-stimulated glycogen synthase activity in skeletal muscle. Muscle insulin receptor binding or insulin receptor kinase activity was shown not to be affected by gliclazide treatment. Whether the improved insulin sensitivity and improved insulin action on skeletal muscle glycogen synthase during gliclazide treatment is due to a direct or an indirect action of the drug is discussed.

Topics: Animals; Diabetes Mellitus, Type 2; Gliclazide; Glycogen; Humans; Insulin; Muscles

Topics: Adult; Amyloid; Animals; Diabetes Mellitus, Type 2; Finland; Glycogen; Humans; Islet Amyloid Polypeptide; Muscles; Risk Factors

Topics: Blood Glucose; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Exercise; Glucagon; Glucose; Glycogen; Hormones; Humans; Insulin; Muscles; Time Factors

Topics: Autonomic Nervous System Diseases; Blood Glucose; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Neuropathies; Energy Intake; Energy Metabolism; Glucose; Glycogen; Humans; Hypoglycemia; Hypotension, Orthostatic; Insulin; Liver; Muscles; Oxygen Consumption; Physical Exertion

In conclusion, a large body of available evidence indicates that the degree of physical conditioning is an important determinant of insulin sensitivity and overall glucose tolerance. Both acute exercise and chronic physical training are associated with enhanced disposal of a glucose load. Conversely, physical inactivity leads to a deterioration in glucose tolerance. The primary tissue responsible for accelerated glucose disposal following exercise is muscle. After an acute bout of exercise, enhanced glucose transport and augmented glycogen synthesis are largely responsible for the improvement in glucose tolerance. The beneficial effects of chronic physical training on glucose metabolism appear to be explained by multiple factors, including increased muscle mass, augmented muscle blood flow and capillary area, enhanced mitochondrial oxidative enzyme capacity, and activation of the glucose transport system. Despite these well-documented effects of training on glucose metabolism, the precise role of exercise in the treatment of diabetic patients remains to be established. In insulin-dependent (type I) diabetic individuals, acute exercise has been shown to be a helpful adjunct in establishing good glycemic control. However, the role of acute exercise in helping to smooth out glycemic control in non-insulin-dependent (type II) diabetic patients has received little attention. The role of chronic physical training in the treatment of both insulin-dependent (type I) and non-insulin-dependent (type II) diabetic individuals remains to be established.

Topics: Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Exercise Therapy; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Obesity; Physical Education and Training

Topics: Adipose Tissue; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Epinephrine; Exercise Therapy; Glucose; Glucose Tolerance Test; Glycogen; Humans; Insulin; Insulin Resistance; Lipolysis; Liver; Male; Metabolic Clearance Rate; Muscles; Obesity; Physical Conditioning, Animal; Physical Exertion

Topics: Adipose Tissue; Animals; Blood Glucose; Diabetes Mellitus; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Dietary Carbohydrates; Energy Metabolism; Exercise Therapy; Fatty Acids, Nonesterified; Food; Glucose; Glucose Tolerance Test; Glycogen; Homeostasis; Humans; Insulin; Insulin Resistance; Lipids; Liver; Muscles; Physical Exertion; Quality of Life

The degree of fasting hyperglycemia in patients with non-insulin-dependent diabetes mellitus is dependent on the rate of hepatic glucose production. The basal rate of hepatic glucose production is increased in patients with non-insulin-dependent diabetes mellitus, and there is a positive correlation between hepatic glucose production and fasting glucose levels. Diminished secretion of insulin, impaired hepatic sensitivity to insulin's effects, or a combination of these factors could contribute to the elevated hepatic glucose production in patients with non-insulin-dependent diabetes mellitus. The relationship between insulin secretion and hepatic glucose production is regulated by a closed feedback loop operating between glucose levels and pancreatic beta cells. Although fasting insulin levels are usually comparable between patients with non-insulin-dependent diabetes mellitus and normal subjects, insulin secretion is markedly impaired in non-insulin-dependent diabetes mellitus in relation to the degree of hyperglycemia present. In fact, the degree of fasting hyperglycemia in a given patient with non-insulin-dependent diabetes mellitus is closely related to the degree of impaired pancreatic beta-cell responsiveness to glucose. Such findings suggest that impaired insulin secretion leads to increased hepatic glucose production, which raises the plasma glucose level. The resulting hyperglycemia helps to maintain relatively normal basal insulin output. Chronic sulfonylurea drug therapy of patients with non-insulin-dependent diabetes mellitus enhances pancreatic islet sensitivity to glucose, leading to increased insulin secretion, suppression of hepatic glucose production, and a decline in the steady-state fasting glucose level.

Topics: Arginine; Blood Glucose; Diabetes Mellitus, Type 2; Fasting; Fatty Acids, Nonesterified; Feedback; Glucagon; Gluconeogenesis; Glucose; Glycogen; Homeostasis; Humans; Insulin; Insulin Resistance; Insulin Secretion; Islets of Langerhans; Liver; Somatostatin; Sulfonylurea Compounds

This study aimed to investigate the mechanisms known to regulate glucose homeostasis in human skeletal muscle of healthy and prediabetic subjects exercising in normobaric hypoxia. Seventeen healthy (H; 28.8 ± 2.4 yr; maximal oxygen consumption (V̇O

Topics: Adult; Anaerobic Threshold; Blood Glucose; Diabetes Mellitus, Type 2; Exercise; Glucose Tolerance Test; Glucose Transporter Type 4; Glycogen; Humans; Hypoxia; Insulin; Lipids; Male; Muscle, Skeletal; Prediabetic State

Exercise combined with adipose tissue lipolytic inhibition augments intramuscular lipid and glycogen use in type 2 diabetes patients. The present study investigates the impact of adipose tissue lipolytic inhibition during exercise on subsequent postprandial glycemic control in type 2 diabetes patients.. Sixty minutes of endurance-type exercise (at 45% W. Collectively, exercise with adipose tissue lipolytic inhibition reduces postprandial blood glucose and insulin excursions and, as such, further improves glycemic control in male type 2 diabetes patients.

Topics: Adipose Tissue; Administration, Oral; Blood Glucose; Cross-Over Studies; Diabetes Mellitus, Type 2; Double-Blind Method; Exercise; Fatty Acids; Glycogen; Humans; Hypolipidemic Agents; Insulin; Lactic Acid; Lipid Metabolism; Male; Middle Aged; Pyrazines; Triglycerides

In health, food carbohydrate is stored as glycogen in muscle and liver, preventing a deleterious rise in osmotically active plasma glucose after eating. Glycogen concentrations increase sequentially after each meal to peak in the evening, and fall to fasting levels thereafter. Skeletal muscle accounts for the larger part of this diurnal buffering capacity with liver also contributing. The effectiveness of this diurnal mechanism has not been previously studied in Type 2 diabetes. We have quantified the changes in muscle and liver glycogen concentration with 13C magnetic resonance spectroscopy at 3.0 T before and after three meals consumed at 4 h intervals. We studied 40 (25 males; 15 females) well-controlled Type 2 diabetes subjects on metformin only (HbA1c (glycated haemoglobin) 6.4±0.07% or 47±0.8 mmol/mol) and 14 (8 males; 6 females) glucose-tolerant controls matched for age, weight and body mass index (BMI). Muscle glycogen concentration increased by 17% after day-long eating in the control group (68.1±4.8 to 79.7±4.2 mmol/l; P=0.006), and this change inversely correlated with homoeostatic model assessment of insulin resistance [HOMA-IR] (r=-0.56; P=0.02). There was no change in muscle glycogen in the Type 2 diabetes group after day-long eating (68.3±2.6 to 67.1±2.0 mmol/mol; P=0.62). Liver glycogen rose similarly in normal control (325.9±25.0 to 388.1±30.3 mmol/l; P=0.005) and Type 2 diabetes groups (296.1±16.0 to 350.5±6.7 mmol/l; P<0.0001). In early Type 2 diabetes, the major physiological mechanism for skeletal muscle postprandial glycogen storage is completely inactive. This is directly related to insulin resistance, although liver glycogen storage is normal.

Topics: Carbon Isotopes; Circadian Rhythm; Diabetes Mellitus, Type 2; Female; Glycogen; Humans; Insulin Resistance; Liver; Magnetic Resonance Spectroscopy; Male; Metformin; Middle Aged; Muscle, Skeletal; Osmolar Concentration; Plasma; Postprandial Period; Statistics, Nonparametric; Triglycerides

Mitochondrial dysfunction has been implicated in the pathogenesis of type 2 diabetes. We hypothesized that any impairment in insulin-stimulated muscle ATP production could merely reflect the lower rates of muscle glucose uptake and glycogen synthesis, rather than cause it. If this is correct, muscle ATP turnover rates in type 2 diabetes could be increased if glycogen synthesis rates were normalized by the mass-action effect of hyperglycemia. Isoglycemic- and hyperglycemic-hyperinsulinemic clamps were performed on type 2 diabetic subjects and matched controls, with muscle ATP turnover and glycogen synthesis rates measured using (31)P- and (13)C-magnetic resonance spectroscopy, respectively. In diabetic subjects, hyperglycemia increased muscle glycogen synthesis rates to the level observed in controls at isoglycemia [from 19 ± 9 to 41 ± 12 μmol·l(-1)·min(-1) (P = 0.012) vs. 40 ± 7 μmol·l(-1)·min(-1) in controls]. This was accompanied by a modest increase in muscle ATP turnover rates (7.1 ± 0.5 vs. 8.6 ± 0.7 μmol·l(-1)·min(-1), P = 0.04). In controls, hyperglycemia brought about a 2.5-fold increase in glycogen synthesis rates (100 ± 24 vs. 40 ± 7 μmol·l(-1)·min(-1), P = 0.028) and a 23% increase in ATP turnover rates (8.1 ± 0.9 vs. 10.0 ± 0.9 μmol·l(-1)·min(-1), P = 0.025) from basal state. Muscle ATP turnover rates correlated positively with glycogen synthesis rates (r(s) = 0.46, P = 0.005). Changing the rate of muscle glucose metabolism in type 2 diabetic subjects alters demand for ATP synthesis at rest. In type 2 diabetes, skeletal muscle ATP turnover rates reflect the rate of glucose uptake and glycogen synthesis, rather than any primary mitochondrial defect.

Topics: Adenosine Triphosphate; Algorithms; Blood Glucose; Breath Tests; Diabetes Mellitus, Type 2; Energy Metabolism; Female; Glucose Clamp Technique; Glycogen; Humans; Hyperglycemia; Insulin; Male; Middle Aged; Muscle, Skeletal; Up-Regulation

The purpose of the study was to investigate the effect of aerobic training and type 2 diabetes on intramyocellular localization of lipids, mitochondria, and glycogen. Obese type 2 diabetic patients (n = 12) and matched obese controls (n = 12) participated in aerobic cycling training for 10 wk. Endurance-trained athletes (n = 15) were included for comparison. Insulin action was determined by euglycemic-hyperinsulinemic clamp. Intramyocellular contents of lipids, mitochondria, and glycogen at different subcellular compartments were assessed by transmission electron microscopy in biopsies obtained from vastus lateralis muscle. Type 2 diabetic patients were more insulin resistant than obese controls and had threefold higher volume of subsarcolemmal (SS) lipids compared with obese controls and endurance-trained subjects. No difference was found in intermyofibrillar lipids. Importantly, following aerobic training, this excess SS lipid volume was lowered by approximately 50%, approaching the levels observed in the nondiabetic subjects. A strong inverse association between insulin sensitivity and SS lipid volume was found (r(2)=0.62, P = 0.002). The volume density and localization of mitochondria and glycogen were the same in type 2 diabetic patients and control subjects, and showed in parallel with improved insulin sensitivity a similar increase in response to training, however, with a more pronounced increase in SS mitochondria and SS glycogen than in other localizations. In conclusion, this study, estimating intramyocellular localization of lipids, mitochondria, and glycogen, indicates that type 2 diabetic patients may be exposed to increased levels of SS lipids. Thus consideration of cell compartmentation may advance the understanding of the role of lipids in muscle function and type 2 diabetes.

Topics: Diabetes Mellitus, Type 2; Exercise Therapy; Glycogen; Humans; Lipid Metabolism; Male; Middle Aged; Mitochondria; Muscle, Skeletal; Physical Endurance; Physical Fitness; Rest; Sarcolemma; Tissue Distribution; Treatment Outcome

Activation of AMP-activated protein kinase (AMPK) by exercise induces several cellular processes in muscle. Exercise activation of AMPK is unaffected in lean (BMI approximately 25 kg/m(2)) subjects with type 2 diabetes. However, most type 2 diabetic subjects are obese (BMI >30 kg/m(2)), and exercise stimulation of AMPK is blunted in obese rodents. We examined whether obese type 2 diabetic subjects have impaired exercise stimulation of AMPK, at different signaling levels, spanning from the upstream kinase, LKB1, to the putative AMPK targets, AS160 and peroxisome proliferator-activated receptor coactivator (PGC)-1alpha, involved in glucose transport regulation and mitochondrial biogenesis, respectively. Twelve type 2 diabetic, eight obese, and eight lean subjects exercised on a cycle ergometer for 40 min. Muscle biopsies were done before, during, and after exercise. Subjects underwent this protocol on two occasions, at low (50% Vo(2max)) and moderate (70% Vo(2max)) intensities, with a 4-6 week interval. Exercise had no effect on LKB1 activity. Exercise had a time- and intensity-dependent effect to increase AMPK activity and AS160 phosphorylation. Obese and type 2 diabetic subjects had attenuated exercise-stimulated AMPK activity and AS160 phosphorylation. Type 2 diabetic subjects had reduced basal PGC-1 gene expression but normal exercise-induced increases in PGC-1 expression. Our findings suggest that obese type 2 diabetic subjects may need to exercise at higher intensity to stimulate the AMPK-AS160 axis to the same level as lean subjects.

Topics: Adult; Amino Acid Oxidoreductases; AMP-Activated Protein Kinase Kinases; AMP-Activated Protein Kinases; Blood Glucose; Diabetes Mellitus, Type 2; Exercise; Female; Glycogen; GTPase-Activating Proteins; Humans; Male; Middle Aged; Multienzyme Complexes; Muscle, Skeletal; Nuclear Respiratory Factor 1; Nucleotides; Obesity; Phosphorylation; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; Time Factors; Transcription Factors

Metabolically healthy skeletal muscle possesses the ability to switch easily between glucose and fat oxidation in response to homeostatic signals. In type 2 diabetes mellitus and obesity, the skeletal muscle shows a great reduction in this metabolic flexibility. A substrate like dodecanedioic acid (C-12), able to increase skeletal muscle glycogen stores via succinyl-CoA formation, might both postpone the fatigue and increase fatty acid utilization, since it does not affect insulin secretion. In healthy volunteers and in type 2 diabetic subjects, the effect of an oral C-12 load was compared with a glucose or water load during prolonged, moderate-intensity, physical exercise. C-12 metabolism was analyzed by a mathematical model. After C-12, diabetics were able to complete the 2 h of exercise. Nonesterified fatty acids increased both during and after the exercise in the C-12 session. C-12 oxidation provided 14% of total energy expenditure, and the sum of C-12 plus lipids oxidized after the C-12 meal was significantly greater than lipids oxidized after the glucose meal (P < 0.025). The fraction of C-12 that entered the central compartment was 47% of that ingested. During the first phase of the exercise ( approximately 60 min), the mean C-12 clearance from the central compartment toward tissues was 2.57 and 1.30 l/min during the second phase of the exercise. In conclusion, C-12 seems to be a suitable energy substrate during exercise, since it reduces muscle fatigue, is rapidly oxidized, and does not stimulate insulin secretion, which implies that lipolysis is not inhibited as reported after glucose ingestion.

Topics: Administration, Oral; Adult; Blood Glucose; Diabetes Mellitus, Type 2; Dicarboxylic Acids; Energy Metabolism; Fatty Acids, Nonesterified; Glycogen; Humans; Male; Middle Aged; Models, Biological; Muscle Fatigue; Muscle, Skeletal; Oxidation-Reduction; Physical Exertion

In the present study, we investigated the consequences of adipose tissue lipolytic inhibition on skeletal muscle substrate use in type 2 diabetic patients.. We studied ten type 2 diabetic patients under the following conditions: (1) at rest; (2) during 60 min of cycling exercise at 50% of maximal workload capacity and subsequent recovery. Studies were done under normal, fasting conditions (control trial: CON) and following administration of a nicotinic acid analogue (low plasma non-esterified fatty acid trial: LFA). Continuous [U-13C]palmitate and [6,6 -2H2]glucose infusions were applied to quantify plasma NEFA and glucose oxidation rates, and to estimate intramuscular triacylglycerol (IMTG) and glycogen use. Muscle biopsies were collected before and after exercise to determine net changes in lipid and glycogen content specific to muscle fibre type.. Following administration of the nicotinic acid analogue (Acipimox), the plasma NEFA rate of appearance was effectively reduced, resulting in lower NEFA concentrations in the LFA trial (p<0.001). Plasma NEFA oxidation rates were substantially reduced at rest, during exercise and subsequent recovery in the LFA trial. The lower plasma NEFA oxidation rates were compensated by an increase in IMTG and endogenous carbohydrate use (p<0.05). Plasma glucose disposal rates did not differ between trials. In accordance with the tracer data, a greater net decline in type I muscle fibre lipid content was observed following exercise in the LFA trial (p<0.05).. This study shows that plasma NEFA availability regulates IMTG use, and that adipose tissue lipolytic inhibition, in combination with exercise, could be an effective means of augmenting intramuscular lipid and glycogen use in type 2 diabetic patients in an overnight fasted state.

Topics: Adipose Tissue; Aged; Algorithms; Body Composition; Body Mass Index; Breath Tests; Diabetes Mellitus, Type 2; Diet; Energy Metabolism; Exercise Test; Fatty Acids, Nonesterified; Female; Glycogen; Humans; Hypolipidemic Agents; Insulin; Kinetics; Lipolysis; Male; Middle Aged; Muscle, Skeletal; Oxidation-Reduction; Palmitates; Pyrazines

Strength training represents an alternative to endurance training for patients with type 2 diabetes. Little is known about the effect on insulin action and key proteins in skeletal muscle, and the necessary volume of strength training is unknown. A total of 10 type 2 diabetic subjects and 7 healthy men (control subjects) strength-trained one leg three times per week for 6 weeks while the other leg remained untrained. Each session lasted no more than 30 min. After strength training, muscle biopsies were obtained, and an isoglycemic-hyperinsulinemic clamp combined with arterio-femoral venous catheterization of both legs was carried out. In general, qualitatively similar responses were obtained in both groups. During the clamp, leg blood flow was higher (P < 0.05) in trained versus untrained legs, but despite this, arterio-venous extraction glucose did not decrease in trained legs. Thus, leg glucose clearance was increased in trained legs (P < 0.05) and more than explained by increases in muscle mass. Strength training increased protein content of GLUT4, insulin receptor, protein kinase B-alpha/beta, glycogen synthase (GS), and GS total activity. In conclusion, we found that strength training for 30 min three times per week increases insulin action in skeletal muscle in both groups. The adaptation is attributable to local contraction-mediated mechanisms involving key proteins in the insulin signaling cascade.

Topics: Biological Transport; Capillaries; Denmark; Diabetes Mellitus, Type 2; Exercise; Glucose; Glucose Transporter Type 4; Glycogen; Humans; Insulin; Leg; Male; Monosaccharide Transport Proteins; Muscle Fibers, Skeletal; Muscle Proteins; Muscle, Skeletal; Physical Fitness; Reference Values; Tensile Strength; White People

Muscle contains the largest reservoir of glycogen (Glyc), a depot that is closely regulated and with influence on insulin sensitivity. The current study examines muscle Glyc in type 2 diabetes mellitus (T2DM) and obesity and with respect to muscle fiber type, intramyocellular lipid content (IMCL), and mitochondrial function (oxidative enzyme activity; OX-Enz). There is increasing interest in the relation of IMCL and mitochondrial dysfunction with insulin resistance (IR), yet the association with muscle Glyc has not been examined with regard to these parameters. Using a quantitative histological approach specific to muscle fiber types, we assessed muscle Glyc, IMCL, and OX-Enz in vastus lateralis obtained by percutaneous biopsy in lean nondiabetic (L; n = 16), obese nondiabetic (Ob; n = 15), and T2DM volunteers (n = 14). Insulin sensitivity was estimated using homeostasis model assessment (HOMA)-IR. Muscle Glyc was reduced in T2DM, a deficit evident for type IIa fibers, yet minor in types I and IIb fibers. Low Glyc in T2DM correlated with fasting hyperglycemia. Also, in T2DM and Ob, there was significantly higher IMCL and lower OX-Enz in all fiber types. The IMCL-to-OX-Enz ratio, especially for type I fibers, correlated strongly with IR. Similarly, a Glyc-to-OX-Enz ratio correlated with IR, particularly for type IIb fibers. This ratio tended to be higher in Ob and T2DM. In summary, there is decreased muscle Glyc in T2DM yet a disproportional Glyc-to-OX-Enz relationship that is related to IR, although not as robustly as the IMCL-to-OX-Enz ratio.

Topics: Diabetes Mellitus, Type 2; Fasting; Female; Glycogen; Homeostasis; Humans; Hyperglycemia; Insulin Resistance; Lipid Metabolism; Male; Middle Aged; Mitochondria, Muscle; Muscle Fibers, Skeletal; Muscle, Skeletal; Obesity; Reference Values

Myotube cultures from patients with type 2 diabetes mellitus (T2DM) represent an experimental in vitro model of T2DM that offers a possibility to perform gene expression studies under standardized conditions. During a time-course of insulin stimulation (1 microM) at 5.5 mM glucose for 0 (no insulin), 0.5, 1, 2, 4, 8, and 24 h, mRNA contents were analyzed in human myotubes for each time point using Affymetrix DNA chip technology. Insulin treatment induced an inflammatory and pro-angiogenic response in the myotubes, with expression of early response factors followed by inflammatory chemokines, metabolic enzymes, and finally cell cycle regulating genes. One-hundred-forty-four genes were differentially expressed in myotubes from donors with type 2 diabetes compared with control subjects, including HSP70, apolipoprotein D/E, tropomyosin, myosin, and actin previously reported from in vivo studies of diabetic skeletal muscle. We conclude, (i) that insulin induces a time-dependent inflammatory and pro-angiogenic transcriptional response in cultured human myotubes, (ii) that myotubes in vitro retain a gene expression pattern specific for type 2 diabetes and sharing five genes with that of type 2 diabetic skeletal muscle in vivo, and (iii) that insulin, despite similar metabolic effects of glucose uptake and glycogen synthesis, regulates different pools of genes in skeletal muscle during in vivo and in vitro conditions.

Topics: Adult; Cells, Cultured; Cytokines; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Gene Expression Profiling; Gene Expression Regulation; Glycogen; Humans; Inflammation; Insulin; Insulin Resistance; Kinetics; Male; Middle Aged; Muscle Fibers, Skeletal; Muscle, Skeletal; Neovascularization, Pathologic; Oligonucleotide Array Sequence Analysis; Signal Transduction

Topics: Cross-Over Studies; Diabetes Mellitus, Type 2; Energy Metabolism; Fatty Acids, Nonesterified; Food Deprivation; Gluconeogenesis; Glucose; Glycogen; Humans; Hypolipidemic Agents; Insulin Resistance; Liver; Male; Pyrazines

To investigate the effect of thalidomide on glucose turnover (glucose production and uptake), on intracellular pathways of glucose utilization (glycogen synthesis [GS], glycolysis [GLS], carbohydrate oxidation, and nonoxidative GLS), and on free fatty acid (FFA) turnover (lipolysis, FFA oxidation, and FFA reesterification).. A total of 6 patients with type 2 diabetes were studied with 4-h isoglycemic-hyperinsulinemic clamps (approximately 8 mmol/l and 500-600 pmol/l, respectively) before treatment (Prestudy), after 3 weeks of thalidomide (150 mg orally at bedtime), and after 3 weeks of placebo.. Thalidomide reduced insulin-stimulated glucose uptake by 31% (from 27.7 to 19.2 pmol x kg(-1) x min(-1), P < 0.05) compared with the prestudy and by 21% (from 24.2 to 19.2 pmol x kg(-1) x min(-1), P < 0.05) compared with placebo. Thalidomide also reduced insulin-stimulated GS by 48% (from 14.1 to 8.2 micromol x kg(-1) x min(-1), P < 0.05) compared with the prestudy and by 40% (from 13.6 to 8.2 micromol x kg(-1) x min(-1), P < 0.5) compared with placebo. Thalidomide had no effect on rates of GLS, carbohydrate oxidation, nonoxidative GLS, lipolysis, FFA oxidation, and reesterification.. We conclude that thalidomide increased insulin resistance in obese patients with type 2 diabetes by inhibiting insulin-stimulated GS and that patients taking thalidomide should be monitored for possible deterioration in their glucose tolerance.

Topics: Blood Glucose; Cross-Over Studies; Diabetes Mellitus, Type 2; Female; Glucose; Glucose Clamp Technique; Glycogen; Humans; Hyperinsulinism; Infusions, Intravenous; Insulin; Lipolysis; Male; Middle Aged; Placebos; Radioisotope Dilution Technique; Single-Blind Method; Thalidomide; Tritium

This investigation was undertaken to examine substrate utilization and glucose turnover during exercise of varying intensities in NIDDM patients.. Six male NIDDM patients (N) and six male controls (C) of similar age, body weight, % body fat, and VO2peak were studied in two experimental sessions administered in a randomized counterbalanced order. During each session the subjects cycled at a power output corresponding to 50% of VO2peak or 70% of VO2peak. Duration of exercise was adjusted so that energy expenditure (EE) was equal in both the 50% and 70% trials. Isotope infusion technique and indirect calorimetry were used to assess substrate utilization and glucose turnover during exercise.. Rates of carbohydrate (CHO) and lipid oxidation increased (P < 0.05) during both the 50% and 70% trials. Rates of CHO oxidation were greater (P < 0.05) during the 70% than during the 50% trial. However, rates of lipid oxidation were similar in the two trials. No differences in rates of CHO and lipid oxidation were observed in N and C. Rates of hepatic glucose production (Ra) and plasma glucose utilization (Rd) increased (P < 0.05) during exercise, and the increases were similar in the 50% and 70% trials. Ra did not differ between N and C. However, Rd was greater (P < 0.05) in N than in C. Plasma glucose concentration decreased (P < 0.05) in N, with the decrease being similar in the 50% and 70% trials. In contrast, plasma glucose concentration remained unchanged during both the 50% and 70% trials in C.. Exercise results in a greater increase in plasma glucose utilization in patients with NIDDM compared with that in normal individuals, and this increase mediates the decline in plasma glucose concentrations in patients with NIDDM. Under isocaloric conditions, the changes in plasma glucose utilization and plasma glucose concentrations are similar during exercise of varying intensities. Despite a greater glucose utilization, carbohydrate and fat oxidation are similar in the two groups and their relations to exercise intensity are not altered by NIDDM.

Topics: Adult; Blood Glucose; Carbohydrate Metabolism; Diabetes Mellitus, Type 2; Exercise; Fats; Glycogen; Humans; Male; Middle Aged; Muscle, Skeletal; Oxidation-Reduction

The aim of the study was to evaluate an acute decrease in NEFA levels during an oral glucose tolerance test and its effects on glucose tolerance, muscle glucose uptake and muscle indirect calorimetry in ten lean non-insulin-dependent diabetic subjects. Two 75-g oral glucose tolerance tests were performed in random order. Placebo or 250 mg acipimox (to inhibit lipolysis) were administered orally 2 h before the start of the oral glucose tolerance test. Two hours after acipimox administration (time 0), non-esterified fatty acid, glycerol and 3-hydroxybutyrate levels decreased by 84, 68 and 77% respectively, compared to basal levels. Concomitantly, muscle lipid oxidation and non-oxidative glycolysis also decreased significantly. After placebo administration, non-esterified fatty acids, glycerol and 3-hydroxybutyrate and lipid oxidation increased by 29, 28, 106 and 33%, respectively (NS vs basal levels; p < 0.001 vs acipimox). There was a negative rate of net glucose storage (interpreted as glycogenolysis) during post-absorptive conditions and at time 0 after administration of both drugs. After oral glucose tolerance test, the incremental areas of blood glucose and insulin were significantly decreased by 18 and 19% after acipimox compared to placebo. In addition, the ratio between the incremental area of forearm muscle glucose uptake and the insulin levels was significantly increased by 45% during acipimox compared to placebo administration. Glucose oxidation and non-oxidative glycolysis were significantly higher while lipid oxidation was significantly lower after acipimox than after placebo. In conclusion, our study found that in lean non-insulin-dependent diabetic subjects, an acute decrease in non-esterified fatty acid levels improves glucose tolerance, muscle glucose uptake, glucose oxidation and non-oxidative glycolysis, but is unable to normalize glucose storage.

Topics: 3-Hydroxybutyric Acid; Blood Glucose; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Forearm; Glucose; Glucose Tolerance Test; Glycerol; Glycogen; Humans; Hydroxybutyrates; Hypolipidemic Agents; Kinetics; Middle Aged; Muscle, Skeletal; Pyrazines; Regional Blood Flow; Thinness; Time Factors

To test the hypothesis that substrate utilization during mild-intensity exercise differs in non-insulin-dependent diabetes mellitus (NIDDM) compared with nondiabetic subjects, seven lean healthy subjects (L), seven obese healthy subjects (O), and seven individuals with NIDDM were studied during 40 min of mild-intensity cycling (40% of peak O2 uptake). Systemic utilization of plasma glucose (Glc Rd) was determined by using isotope dilution methods. Gas exchange was measured to determine rates of carbohydrate (CHO) and lipid oxidation. During exercise, when CHO oxidation was greater than Glc Rd, the net oxidation of glycogen was calculated as the difference: CHO oxidation - Glc Rd. During mild-intensity cycling, the respiratory exchange ratio was similar across groups (0.87 +/- 0.02, 0.85 +/- 0.02, and 0.86 +/- 0.01 in L, O, and NIDDM subjects, respectively), and CHO oxidation accounted for one-half of total energy expenditure during exercise. Glc Rd increased during exercise and was greatest in subjects with NIDDM (3.0 +/- 0.2, 2.9 +/- 0.2, and 4.5 +/- 0.4 ml.kg-1.min-1 in L, O, and NIDDM subjects, respectively, P < 0.05), yet Glc Rd was less than CHO oxidation during exercise, indicating net oxidation of glycogen. Glycogen oxidation was greater in L and O than in NIDDM subjects (3.4 +/- 1.0, 2.5 +/- 0.9, and 1.7 +/- 0.8 ml.kg-1.min-1; P < 0.05). In summary, during mild-intensity exercise, NIDDM subjects have an increased Glc Rd and a decreased oxidation of muscle glycogen.

Topics: Anaerobic Threshold; Blood Glucose; Body Composition; Diabetes Mellitus, Type 2; Energy Metabolism; Exercise; Fatty Acids, Nonesterified; Female; Glycogen; Humans; Male; Middle Aged; Obesity; Oxidation-Reduction; Pulmonary Gas Exchange

Glycogen neutrophils level was evaluated in 54 patients with non-insulin dependent diabetes mellitus (NIDDM) and 10 patients with insulin dependent diabetes mellitus (IDDM). Glycogen concentration estimated by histochemical method was lower in diabetics than in control group. Patients with NIDDM were divided in the groups according to: sex, duration of disease, a kind of complications and a way of treatment. The glycogen contents in neutrophils, defined in "score"-unit was not different in isolated groups. There was found significant correlation between glycogen contents in neutrophils and the metabolic control in patients with IDDM (r = 0.72) and less significant in patients with NIDDM (r = 0.29).

Topics: Adult; Aged; Aged, 80 and over; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glycogen; Humans; Middle Aged; Neutrophils

Finnish-specific gene variant p.P50T/AKT2 (minor allele frequency (MAF) = 1.1%) is associated with insulin resistance and increased predisposition to type 2 diabetes. Here, we have investigated in vitro the impact of the gene variant on glucose metabolism and intracellular signalling in human primary skeletal muscle cells, which were established from 14 male p.P50T/AKT2 variant carriers and 14 controls. Insulin-stimulated glucose uptake and glucose incorporation into glycogen were detected with 2-[1,2-3H]-deoxy-D-glucose and D-[14C]-glucose, respectively, and the rate of glycolysis was measured with a Seahorse XFe96 analyzer. Insulin signalling was investigated with Western blotting. The binding of variant and control AKT2-PH domains to phosphatidylinositol (3,4,5)-trisphosphate (PI(3,4,5)P3) was assayed using PIP StripsTM Membranes. Protein tyrosine kinase and serine-threonine kinase assays were performed using the PamGene® kinome profiling system. Insulin-stimulated glucose uptake and glycogen synthesis in myotubes in vitro were not significantly affected by the genotype. However, the insulin-stimulated glycolytic rate was impaired in variant myotubes. Western blot analysis showed that insulin-stimulated phosphorylation of AKT-Thr308, AS160-Thr642 and GSK3β-Ser9 was reduced in variant myotubes compared to controls. The binding of variant AKT2-PH domain to PI(3,4,5)P3 was reduced as compared to the control protein. PamGene® kinome profiling revealed multiple differentially phosphorylated kinase substrates, e.g. calmodulin, between the genotypes. Further in silico upstream kinase analysis predicted a large-scale impairment in activities of kinases participating, for example, in intracellular signal transduction, protein translation and cell cycle events. In conclusion, myotubes from p.P50T/AKT2 variant carriers show multiple signalling alterations which may contribute to predisposition to insulin resistance and T2D in the carriers of this signalling variant.

Topics: Diabetes Mellitus, Type 2; Finland; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Male; Muscle Fibers, Skeletal; Muscle, Skeletal; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction

Type 2 Diabetes Mellitus (T2DM) is characterized by hepatic insulin resistance, which results in increased glucose production and reduced glycogen storage in the liver. There is no previous study in the literature that has explored the role of Xanthosine in hepatic insulin resistance. Moreover, mechanistic explanation for the beneficial effects of Xanthosine in lowering glucose production in diabetes is yet to be determined. This study for the first time investigated the beneficial effects of Tribulus terrestris (TT) and its active constituent, Xanthosine on gluconeogenesis and glycogenesis in Free Fatty Acid (FFA)-induced CC1 hepatocytes and streptozotocin (STZ)-induced Wistar rats. Xanthosine enhanced glucose uptake and decreased glucose production through phosphorylation of AMP-activated protein kinase (AMPK) and forkhead box transcription factor O1 (FoxO1), and downregulation of two rate limiting enzymes of gluconeogenesis, phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) expression in FFA-induced CC1 cells. Xanthosine also prevented FFA-induced decreases in the phosphorylation of AKT/Protein kinase B, glycogen synthase kinase-3β (GSK3β), and increased glycogen synthase (GS) phosphorylation to increase the glycogen content in the hepatocytes. Moreover, in STZ-induced diabetic rats, oral administration of TT n-butanol fraction (TTBF) enriched with compound Xanthosine (10, 50 & 100 mg/kg body weight) improved insulin sensitivity, reduced fasting blood glucose levels, improved glucose homeostasis by reducing gluconeogenesis via AMPK/FoxO1-mediated PEPCK and G6Pase down-regulation and increasing glycogenesis via AKT/GSK3β-mediated GS activation. Overall, Xanthosine may be developed further for treating insulin resistance and hyperglycemia in T2DM.

Topics: AMP-Activated Protein Kinases; Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Gluconeogenesis; Glucose; Glycogen; Glycogen Synthase Kinase 3 beta; Glycosides; Homeostasis; Insulin Resistance; Liver; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; Xanthines

Sanhuang Xiexin Decoction (SHXXD) is a classic prescription for the treatment of diabetes. Excessive hepatic glucose production (HGP) is a major determinant of the occurrence and development of diabetes. Inhibition of HGP can significantly improve type 2 diabetes mellitus (T2DM).. To investigate the mechanism by which SHXXD inhibits HGP.. First, a mouse model of T2DM was established through high-fat diet (HFD) feeding combined with streptozotocin (STZ) injection to determine the pharmacodynamic effect of SHXXD in T2DM mice. Then, the possible pathways induced by SHXXD in the treatment of T2DM were predicted by network pharmacology combined with transcriptomics (including target prediction, network analysis and enrichment analysis). Finally, the specific mechanism of SHXXD was elucidated by in vitro experiments.. In vivo experiments showed that SHXXD reduced fasting blood glucose and alleviated weight loss in T2DM mice. Improved glucose clearance rates and insulin sensitivity improve dyslipidemia, liver tissue structural abnormalities and inflammatory cell infiltration as well as increase glycogen storage in T2DM mice. The results of network pharmacology and transcriptome analysis showed that SHXXD contained 378 compounds and 2625 targets. In total, 292 intersection targets were identified between the differentially expressed genes (DEGs) of the liver tissue insulin resistance (IR) related dataset GSE23343. KEGG enrichment analysis showed that the insulin/PI3K-Akt/FoxO signaling pathway may be related to SHXXD-mediated improvements in T2DM. In vitro experimental results showed that SHXXD increased glucose consumption by HepG2-IR cells and improved their insulin sensitivity. RT‒qPCR and Western blotting results showed that SHXXD inhibited hepatic gluconeogenesis through the insulin/PI3K-Akt/FoxO signaling pathway by promoting IGFIR, PIK3R1 and AKT2 expression and subsequently inhibiting PEPCK and FBP1 expression via phosphorylation of Foxo1. In addition, PI3K/Akt deactivated p-GSK3β through phosphorylation, thereby promoting GS expression and increasing glycogen synthesis.. SHXXD can target the liver to cooperate with the insulin/PI3K-Akt/FoxO signaling pathway to inhibit HGP to alleviate T2DM.

Topics: Animals; Diabetes Mellitus, Type 2; Glucose; Glycogen; Insulin; Insulin Resistance; Liver; Mice; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction

α-Limit dextrins (α-LDx) are slowly digestible carbohydrates that attenuate postprandial glycemic response and trigger the secretion of satiety-related hormones. In this study, more highly branched α-LDx were enzymatically synthesized to enhance the slowly digestible property by various origins of glycogen branching enzyme (GBE), which catalyzes the transglycosylation to form α-1,6 branching points after cleaving α-1,4 linkages. Results showed that the proportion of branched α-LDx in starch molecules increased around 2.2-8.1 % compared to α-LDx from starch without GBE treatment as the ratio of α-1,6 linkages increased after different types of GBE treatments. Furthermore, the enzymatic increment of branching points enhanced the slowly digestible properties of α-LDx at the mammalian α-glucosidase level by 17.3-28.5 %, although the rates of glucose generation were different depending on the source of GBE treatment. Thus, the highly branched α-LDx with a higher amount of α-1,6 linkages and a higher molecular weight can be applied as a functional ingredient to deliver glucose throughout the entire small intestine without a glycemic spike which has the potential to control metabolic diseases such as obesity and type 2 diabetes.

Topics: 1,4-alpha-Glucan Branching Enzyme; Animals; Dextrins; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Mammals; Starch

Impaired glucose regulation is one of the most important risk factors for type 2 diabetes mellitus (T2DM) and cardiovascular diseases, which have become a major public health issue worldwide. Dysregulation of carbohydrate metabolism in liver has been shown to play a critical role in the development of glucose intolerance but the molecular mechanism has not yet been fully understood. In this study, we investigated the role of hepatic LCMT1 in the regulation of glucose homeostasis using a liver-specific LCMT1 knockout mouse model. The hepatocyte-specific deletion of LCMT1 significantly upregulated the hepatic glycogen synthesis and glycogen accumulation in liver. We found that the liver-specific knockout of LCMT1 improved high fat diet-induced glucose intolerance and insulin resistance. Consistently, the high fat diet-induced downregulation of glucokinase (GCK) and other important glycogen synthesis genes were reversed in LCMT1 knockout liver. In addition, the expression of GCK was significantly upregulated in MIHA cells treated with siRNA targeting LCMT1 and improved glycogen synthesis. In this study, we provided evidences to support the role of hepatic LCMT1 in the development of glucose intolerance induced by high fat diet and demonstrated that inhibiting LCMT1 could be a novel therapeutic strategy for the treatment of glucose metabolism disorders.

Topics: Animals; Diabetes Mellitus, Type 2; Diet, High-Fat; Glucose; Glucose Intolerance; Glycogen; Insulin Resistance; Leucine; Liver; Methyltransferases; Mice; Protein O-Methyltransferase

This study examines the clinical-pathological profiles of patients with glycogenic hepatopathy in a contemporary cohort of patients at an adult acute care hospital.. Liver biopsies with glycogenic hepatopathy were retrieved from the departmental surgical pathology database, the histological findings were studied, and the clinical findings were reviewed.. Five cases of glycogenic hepatopathy were found, including cases associated with type 1 diabetes mellitus (n = 1), type 2 diabetes mellitus (n = 1), corticosteroids (n = 2), and anorexia (n = 2, including the patient with type 1 diabetes). AST and ALT were normal to mildly elevated (13-115 U/L and 7-126 U/L, respectively). Trace ascites was present in two patients. Hepatomegaly was only present in the patient with type 1 diabetes at the time of diagnosis.. Four of five cases were associated with etiologies other than type 1 diabetes, which is widely reported as the most common etiology of glycogenic hepatopathy. This study suggests that etiologies currently only rarely recognized may actually be more common causes of glycogenic hepatopathy than type 1 diabetes in a contemporary adult population. It is important not only to recognize that these rarely reported causes of glycogenic hepatopathy may be underrecognized, but that the clinical presentation may also be mild.

Topics: Adult; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glycogen; Hepatomegaly; Humans; Liver Diseases

As a multifaceted metabolic disorder, insulin resistance is accompanied by the preceding onset of type 2 diabetes mellitus, hyperinsulinemia, metabolic dysfunction-associated fatty liver disease (MAFLD) and other metabolic syndromes. Currently, the number of existing drugs and mechanism-based strategies is limited to alleviate insulin resistance in clinics. As a natural polyphenol product derivative, 1,3,6,7-tetrapropylene acyloxy-ketone (TPX) showed a significant hypoglycemic effect in our previous studies. However, whether TPX could improve hepatic insulin sensitivity was unknown.. To explore whether insulin sensitivity can be improved by the treatment with TPX and further investigate its mechanism(s) of activity.. To mimic hyperglycemia and insulin resistance in vitro, human HepG2 and HL-7702 hepatocytes were exposed to high glucose. Cellular glucose uptake, glucose consumption, glycogen synthesis, and glucose production were quantified after TPX treatment. The effects of TPX on AMP-activated protein kinase (AMPK) phosphorylation, glucose metabolism, and insulin signal transduction were evaluated by western blotting and network pharmacology analysis. The eGFP-membrane of glucose transporter type 4 (GLUT4) lentivirus transfected cells were constructed to investigate the effects of TPX on GLUT4 mobilization. Reactive oxygen species activity in high glucose-induced insulin-resistant cells was measured by DCFH-DA to show oxidative stress.. Treatment with TPX improved glycogen synthesis and inhibited gluconeogenesis by regulating GSK3β, G6Pase, and PEPCK. Furthermore, high glucose-induced inhibition of glucose consumption, glucose uptake, and GLUT4-mediated membrane translocation were reverted by TPX. Accordingly, mechanistic investigations revealed that TPX interacted with AMPK protein and activated the phosphorylation of AKT, thereby improving energy homeostasis and further ameliorating hepatic insulin resistance. Network pharmacology analysis and molecular docking further confirmed AMPK as an active target of TPX. Concordantly, the pharmacological activity of TPX was reversed by the AMPK inhibitor compound C when hepatocytes were exposed to high glucose stimulation.. In summary, our study confirmed TPX contributions to insulin resistance improvements by targeting AMPK and PI3K/AKT to restore the insulin signaling pathway, which may be an important potential treatment strategy for insulin-resistance-related diseases, including MAFLD and diabetes.

Topics: AMP-Activated Protein Kinases; Diabetes Mellitus, Type 2; Glucose; Glycogen; Hepatocytes; Humans; Insulin; Insulin Resistance; Molecular Docking Simulation; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction

Previous studies showed that high-intensity interval training (HIIT) improved fasting blood glucose and insulin resistance in type 2 diabetes mellitus (T2DM) mice. However, the effect of HIIT on the kidneys of mice with T2DM has not been examined. This study aimed to investigate the impact of HIIT on the kidneys of T2DM mice.. T2DM mice were induced with a high-fat diet (HFD) and one-time 100 mg/kg streptozotocin intraperitoneal injection, and then T2DM mice were treated with 8 weeks of HIIT. Renal function and glycogen deposition were observed by serum creatinine levels and PAS staining, respectively. Sirius red staining, hematoxylin-eosin staining, and Oil red O staining were used to detect fibrosis and lipid deposition. Western blotting was performed to detect the protein levels.. HIIT significantly ameliorated the body composition, fasting blood glucose, and serum insulin of the T2DM mice. HIIT also improved glucose tolerance, insulin tolerance, and renal lipid deposition of T2DM mice. However, we found that HIIT increased serum creatinine and glycogen accumulation in the kidneys of T2DM mice. Western blot analysis showed that the PI3K/AKT/mTOR signaling pathway was activated after HIIT. The expression of fibrosis-related proteins (TGF-β1, CTGF, collagen-III, α-SMA) increased, while the expression of klotho (sklotho) and MMP13 decreased in the kidneys of HIIT mice.. This study concluded that HIIT induced renal injury and fibrosis, although it also improved glucose homeostasis in T2DM mice. The current study reminds us that patients with T2DM should be cautious when participating in HIIT.

Topics: Animals; Blood Glucose; Creatinine; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Fibrosis; Glycogen; High-Intensity Interval Training; Insulins; Kidney; Lipids; Mice; Phosphatidylinositol 3-Kinases

Objectives: manganese (Mn) is closely related to type 2 diabetes mellitus and insulin resistance (IR), but the exact mechanism is unclear. This study aimed to explore the regulatory effects and mechanism of Mn on IR using hepatocyte IR model induced by high palmitate (PA), high glucose (HG) or insulin. Methods: HepG2 cells were exposed to PA (200 μM), HG (25 mM) or insulin (100 nM) respectively, alone or with 5 μM Mn for 24 hours. The expression of key proteins in insulin signaling pathway, intracellular glycogen content and glucose accumulation, reactive oxygen species (ROS) level and Mn superoxide dismutase (MnSOD) activity were detected. Results: compared with control group, the expression of phosphorylated protein kinase B (Akt), glycogen synthase kinase-3β (GSK-3β) and forkhead box O1 (FOXO1) in the three IR groups was declined, and this decrease was reversed by Mn. The reduction of intracellular glycogen content and increase in glucose accumulation in IR groups were also inhibited by Mn. Additionally, the production of ROS was increased in IR models, compared with normal control group, while Mn reduced the excessive production of ROS induced by PA, HG or insulin. However, Mn did not alter the activity of MnSOD in the three IR models. Conclusion: this study demonstrated that Mn treatment can improve IR in hepatocytes. The mechanism is probably by reducing the level of intracellular oxidative stress, enhancing the activity of Akt/GSK-3β/FOXO1 signal pathway, promoting glycogen synthesis, and inhibiting gluconeogenesis.. Objetivos: el manganeso (Mn) está estrechamente relacionado con la diabetes mellitus tipo 2 y la resistencia a la insulina (RI), pero el mecanismo exacto aún no está claro. Este estudio tuvo como objetivo explorar los efectos reguladores y el mecanismo del Mn sobre la RI utilizando un modelo de RI en hepatocitos inducido por palmitato alto (PA), glucosa alta (HG) o insulina. Métodos: las células HepG2 se expusieron a PA (200 μM), HG (25 mM) o insulina (100 nM), solas o junto con 5 μM de Mn durante 24 horas. Se evaluó la expresión de proteínas clave en la vía de señalización de la insulina, el contenido intracelular de glucógeno y la acumulación de glucosa, el nivel de especies reactivas de oxígeno (ROS) y la actividad superóxido dismutasa del manganeso (MnSOD). Resultados: en comparación con el grupo de control, la expresión de proteína quinasa B fosforilada (Akt), la glucógeno sintasa quinasa-3β (GSK-3β) y la proteína forkhead box O1 (FOXO1) en los tres grupos de RI se redujo, y esta disminución fue revertida por el Mn. La reducción del contenido de glucógeno intracelular y el aumento de la acumulación de glucosa en los grupos de RI también fueron inhibidos por el Mn. Además, la producción de ROS aumentó en los modelos de RI en comparación con el grupo de control normal. Mientras que el Mn redujo la producción excesiva de ROS inducida por PA, HG o insulina. Sin embargo, el Mn no alteró la actividad de la MnSOD en los tres modelos de RI. Conclusión: este estudio demostró que el tratamiento con Mn puede mejorar la RI en hepatocitos. El mecanismo probablemente sea mediante la reducción del nivel de estrés oxidativo intracelular, mejorando la actividad de la vía de señalización Akt/GSK-3β/FOXO1, promoviendo la síntesis de glucógeno e inhibiendo la gluconeogénesis.

Topics: Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Synthase Kinase 3 beta; Hepatocytes; Humans; Insulin; Insulin Resistance; Manganese; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species

Diabetes is one of the major global public health problems. Our previous results found that oat β-D-glucan exhibited ameliorative effects on diabetic mice, but the underlying mechanism is unclear. The present study indicates that oat β-D-glucan increased glycogen content, decreased glycogen synthase (GS) phosphorylation and increased hepatic glycogen synthase kinase 3β (GSK3β) phosphorylation for glycogen synthesis via PI3K/AKT/GSK3-mediated GS activation. Moreover, oat β-D-glucan inhibited gluconeogenesis through the PI3K/AKT/Foxo1-mediated phosphoenolpyruvate carboxykinase (PEPCK) decrease. In addition, oat β-D-glucan enhanced glucose catabolism through elevated protein levels of COQ9, UQCRC2, COXIV and ATP5F complexes involved in oxidative phosphorylation, as well as that of TFAM, a key regulator of mitochondrial gene expression. Importantly, our results showed that oat β-D-glucan maintained hepatic glucose balance via TLR4-mediated intracellular signal. After TLR4 blocking with anti-TLR4 antibody, oat β-D-glucan had almost no effect on high glucose-induced HepG2 cells. These data revealed that oat β-D-glucan maintains glucose balance by regulating the TLR4/PI3K/AKT signal pathway.

Topics: Animals; Avena; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glucans; Glucose; Glycogen; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Insulin Resistance; Mice; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Toll-Like Receptor 4

The antidiabetic activity and underlying mechanisms of Fucus vesiculosus polysaccharide (FVP) were studied in type 2 diabetic rats. Our results exhibited that FVP intervention reversed body weight loss, alleviated hyperglycemia and insulin resistance in diabetic rats. FVP also had the potential to ameliorate dyslipidemia, liver and kidney dysfunction, decrease oxidative stress, promote glycogen synthesis, and boost short-chain fatty acid production and total bile acid excretion. 16S rRNA gene sequencing analysis suggested that FVP interfered with the gut microbiota in a beneficial manner. Moreover, RT-qPCR results demonstrated that the antidiabetic activity of FVP in connection with the acceleration of blood glucose absorption and glycogen synthesis, the inhibition of gluconeogenesis, and the regulation of lipid metabolism in the liver. These findings suggested that FVP had antidiabetic effects on high-fat diet and STZ-induced diabetic rats and could be a potential resource for treating type 2 diabetes mellitus.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Fucus; Gastrointestinal Microbiome; Gene Expression; Glycogen; Glycolipids; Hypoglycemic Agents; Lipid Metabolism; Polysaccharides; Rats; RNA, Ribosomal, 16S

A prediabetic population has an increased risk of cognitive decline and type 2 diabetes mellitus (T2DM). This study investigated whether the progression of memory dysfunction and dysregulated brain glycogen metabolism is prevented with 4 mo of exercise intervention from the presymptomatic stage in a T2DM rat model. Memory function and biochemical and molecular profiles were assessed in the presymptomatic stage of Otsuka-Long-Evans-Tokushima fatty (OLETF) rats, a T2DM model, with Long-Evans Tokushima (LETO) rats as genetic control. These rats were subjected to light- or moderate-intensity treadmill running for 4 mo with repetition of the same experiments. Significant hippocampal-dependent memory dysfunction was observed in the presymptomatic stage of OLETF rats, accompanied by downregulated levels of hippocampal monocarboxylate transporter 2 (MCT2), a neuronal lactate-transporter, without alteration in hippocampal glycogen levels. Four months of light or moderate exercise from the presymptomatic stage of T2DM normalized glycemic parameters and hippocampal molecular normalization through MCT2, glycogen, and brain-derived neurotrophic factor (BDNF) levels with the improvement of memory dysfunction in OLETF rats. A 4-mo exercise regimen from the presymptomatic stage of T2DM at a light and moderate intensities contributed to the prevention of the development of T2DM and the progression of cognitive decline with hippocampal lactate-transport and BDNF improvement.

Topics: Animals; Blood Glucose; Brain-Derived Neurotrophic Factor; Cognitive Dysfunction; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glycogen; Hippocampus; Humans; Lactates; Physical Conditioning, Animal; Prediabetic State; Rats; Rats, Inbred OLETF; Rats, Long-Evans

Drosophila melanogaster has been used as the most successful invertebrate model for studying metabolic diseases such as type 2 diabetes (T2D). We induced T2D by feeding Drosophila larvae on a high-sugar diet (HSD). The glucose and trehalose, glycogen, lipid, triglyceride, and protein levels were determined in HSD-fed larvae. Moreover, larval food intake, water content, size, and weight in addition to the development until pupation were observed. Levels of Drosophila insulin-like peptides (DILPs 2, 3, and 5), as well as adipokinetic hormone (AKH), were also determined in HSD-fed larvae by quantitative real-time polymerase chain reaction. The results demonstrated that HSD could induce elevated levels of glucose, trehalose, glycogen, and proteins in larvae. The larvae consumed less food intake and were smaller, lighter, and less developed on HSD than those on the control diet. Moreover, the water content of larvae fed HSD was similar to that fed the control diet. HSD induced higher expression of DILP3 and AKH, confirming hyperglycemia with insulin resistance. In sum, Drosophila offers an appropriate model for quick and inexpensive in vivo experimentation on human metabolic diseases.

Topics: Animals; Diabetes Mellitus, Type 2; Drosophila; Drosophila melanogaster; Glucose; Glycogen; Larva; Trehalose; Water

In this study, a novel peptide GPPGPA was screened from the collagen hydrolysates of Chinese giant salamander (

Topics: alpha-Glucosidases; Animals; Collagen; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycoside Hydrolase Inhibitors; Hep G2 Cells; Humans; Hypoglycemic Agents; Insulin Resistance; Kinetics; Malondialdehyde; Molecular Docking Simulation; Network Pharmacology; Oxidative Stress; Peptide Fragments; Protein Hydrolysates; Skin; Superoxide Dismutase; Urodela

Major stores of glucose are found as glycogen in skeletal muscle and liver. Skeletal muscle is a heterogenous tissue, with cellular metabolic and contractile distinctions dependent on whether the cell (fibre) is slow-twitch (Type I) or fast-twitch (Type II). We hypothesised that proteins important for glycogen metabolism would be differentially abundant between these diverse fibres. We further hypothesised that the cellular location of these proteins would be different in muscle samples between control (CON) and individuals with type 2 diabetes (T2D). We dissected individual muscle fibre segments from vastus lateralis skeletal muscle biopsy samples from CON and T2D and used cell-type-specific approaches to address muscle heterogeneity. We measured glycogen and glycogen-related proteins by immunoblotting techniques. A lower proportion of Type I fibres was found in muscle in T2D compared with CON. AMPK-β2, glycogen branching enzyme (GBE), glycogen debranching enzyme (GDE), and glycogen phosphorylase (GP) were differentially localized between fibre types and in fibres from CON and T2D individuals. A key novel finding was that the majority of glycogen is loosely bound or cytosolic in location in human skeletal muscle. The proportion of this diffusible pool of glycogen was significantly lower in Type I fibres in T2D compared to CON. A hyperinsulinaemic, euglycaemic clamp in people with type 2 diabetes had no effect on the proportion of diffusible glycogen. We identify cell-type as an important consideration when assessing glycogen metabolism in muscle. Our findings demonstrate varying glucose handling abilities in specific muscle fibre types in type 2 diabetes. A model is presented to provide an overview of the cell-specific differences in glycogen metabolism in type 2 diabetes.

Topics: Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Muscle Fibers, Skeletal; Muscle, Skeletal

This study aims to explore the mechanism of Astragali Radix-Puerariae Lobatae Radix(AP) combination in the treatment of type 2 diabetes mellitus(T2 DM) based on network pharmacology and experiment. The effective components and targets of the pair were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and targets of T2 DM from each disease database. On this basis, the common targets of the medicinals and the disease were screened out. The protein-protein interaction(PPI) network was established based on STRING. Then Cytoscape 3.7.1 was employed for visualization of the common targets and the network topology analysis of key targets, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of core targets by DAVID. Thereby, the possible molecular mechanism was unveiled. High-fat diet was combined with streptozotocin(STZ, injected into tail vein) for T2 DM rat modeling. Rats were classified into the normal group, model group, positive control group(metformin hydrochloride), AP high-dose, medium-dose, and low-dose groups. After 4 weeks of intragastric administration, serum fasting blood glucose(FBG), fasting insulin(FINS), aspartate aminotransferase(AST), alanine aminotransferase(ALT), triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), interleukin(IL)-6, and tumor necrosis factor(TNF)-α of rats in each group were measured. The expression of insulin receptor substrate-2(IRS-2), adenosine monophosphate-activated protein kinase(AMPK), phosphorylated AMPK(p-AMPK), glucose 6 phosphatase(G6 Pase), and phosphoenolpyruvate carboxy kinase(Pepck) in rat liver was detected by Western blot. A total of 131 core targets of the combination in the treatment of T2 DM were screened out, among which protein kinase B(AKT) 1, mitogen-activated protein kinase(MAPK) 1, TNF-α, IL-6 were more critical. KEGG enrichment analysis suggested that the combination decreased blood glucose mainly through PI3 K/AKT signaling pathway, AMPK signaling pathway, TNF signaling pathway, and MAPK signaling pathway. The levels of FBG and FINS were lower and the glycogen level was higher in the AP high-dose and medium-dose groups than in the model group. The levels of AST, ALT, TG, and LDL-C in the three AP groups and the level of TC in AP high-dose and low-dose groups decreased compared with those in the model g

Topics: AMP-Activated Protein Kinases; Animals; Astragalus Plant; Blood Glucose; Cholesterol, LDL; Diabetes Mellitus, Type 2; Drugs, Chinese Herbal; Glycogen; Interleukin-6; Network Pharmacology; Proto-Oncogene Proteins c-akt; Pueraria; Rats; Signal Transduction; Streptozocin; Tumor Necrosis Factor-alpha

Dysfunction of glucokinase (GCK) caused by mutations in the GCK gene is the main cause of maturity-onset diabetes of the young type-2 (MODY2, also known as GCK-MODY), which is usually present in adolescence or young adulthood. MODY2 is characterized by mild, stable fasting hyperglycemia that presents at birth, usually 5.4-8.3 mmol L

Topics: Adolescent; Child, Preschool; Diabetes Mellitus, Type 2; Female; Glucokinase; Glycogen; Humans; Hyperglycemia; Kinetics; Male; Mutation; Young Adult

Liver glycogen is a highly branched glucose polymer found as β particles (~20 nm in diameter), which can bind together into larger composite α particles. Hepatic α particles have been shown to be structurally fragile (breaking up into smaller particles in certain solvents) in mouse models of diabetes; if occurring in vivo, the resulting small glycogen particles could exacerbate the poor blood-sugar homeostasis characteristic of the disease. Here we tested if this α-particle fragility also occurred in liver glycogen obtained from humans with diabetes. It was found that liver glycogen from diabetic humans was indeed more fragile than from non-diabetic humans, which was also seen in the mouse experiments we ran in parallel. Proteomic analysis revealed three candidate proteins from differentially expressed glycogen proteins (Diabetes/ Non-diabetes) in both human and mouse groups. Identifying these proteins may give clues to the binding mechanism that holds together α particles together, which, being different in diabetic glycogen, is relevant to diabetes prevention and management.

Topics: Animals; Diabetes Mellitus, Type 2; Glycogen; Humans; Liver; Liver Glycogen; Mice; Pilot Projects; Proteomics

KCNH6 has been proven to affect glucose metabolism and insulin secretion both in humans and mice. Further study revealed that Kcnh6 knockout (KO) mice showed impaired glucose tolerance. However, the precise function of KCNH6 in the liver remains unknown. Mitochondria have been suggested to maintain intracellular Ca

Topics: Animals; Calcium; Diabetes Mellitus, Type 2; Ether-A-Go-Go Potassium Channels; Glucose; Glycogen; Humans; Insulin; JNK Mitogen-Activated Protein Kinases; Liver; Mice; Mice, Knockout; Mitochondria; NADPH Oxidases; Oxidative Stress; Reactive Oxygen Species; Superoxides

Acarbose, as an alpha-glucosidase inhibitor, is widely used clinically to treat type II diabetes. In its industrial production, Actinoplanes sp. SE50/110 is used as the production strain. Lack of research on its regulatory mechanisms and unexplored gene targets are major obstacles to rational strain design. Here, transcriptome sequencing was applied to uncover more gene targets and rational genetic engineering was performed to increase acarbose production.. In this study, with the help of transcriptome information, a TetR family regulator (TetR1) was identified and confirmed to have a positive effect on the synthesis of acarbose by promoting the expression of acbB and acbD. Some genes with low expression levels in the acarbose biosynthesis gene cluster were overexpressed and this resulted in a significant increase in acarbose yield. In addition, the regulation of metabolic pathways was performed to retain more glucose-1-phosphate for acarbose synthesis by weakening the glycogen synthesis pathway and strengthening the glycogen degradation pathway. Eventually, with a combination of multiple strategies and fed-batch fermentation, the yield of acarbose in the engineered strain increased 58% compared to the parent strain, reaching 8.04 g/L, which is the highest fermentation titer reported.. In our research, acarbose production had been effectively and steadily improved through genetic engineering based on transcriptome analysis and fed-batch culture strategy.

Topics: Acarbose; Actinoplanes; Diabetes Mellitus, Type 2; Fermentation; Genetic Engineering; Glycogen; Humans

Type 2 diabetes mellitus (T2DM) is a systemic metabolic disorder characterized by insulin deficiency and insulin resistance. Recently, it has become a significant threat to public health.

Topics: Animals; Blood Glucose; Cholesterol; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glycogen; Glycogen Synthase Kinase 3 beta; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Mice; Polygonatum

Type 2 diabetes mellitus (T2DM) is a disease characterized by hyperglycemia resulting from insulin resistance. In recent years, the incidence of T2DM has been increasing. Women with T2DM often suffer from infertility and early miscarriage; however, the underlying mechanisms remain unclear. Insulin is the most important regulatory hormone of glycogen metabolism. In addition, 5' adenosine monophosphate-activated protein kinase (AMPK) is an important regulator of glycogen metabolism. Patients with T2DM have inhibited AMPK expression in the liver, which leads to impaired glucose metabolism. However, the role of AMPK in endometrial glycogen metabolism has not been reported. In this study, a mouse model of T2DM was established to investigate whether altered endometrial glucose metabolism affects early embryo implantation. Metformin and insulin were used for therapy; the resulting changes to glycogen metabolism and embryo implantation were examined. The results indicate that the concentrations of glycogen decreased significantly in T2DM mice, resulting in insufficient energy supplies for proper endometrial function, and thereby impeding embryonic implantation. Interestingly, endometrial AMPK was not found to be overactivated. Insulin treatment was found to partially resolve the embryo implantation defects in T2DM mice. Metformin improved blood glucose but did not have a significant effect on local endometrial glucose metabolism. This study explored the changes in endometrial glucose metabolism in T2DM mouse, and the effects of these changes on embryo implantation. We found that insulin, but not metformin, significantly resolved embryo implantation problems. These findings will help to increase our understanding of the pathomechanisms of infertility and early miscarriage in women with T2DM.

Topics: AMP-Activated Protein Kinases; Animals; Biomarkers; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Embryo Implantation; Endometrium; Female; Glycogen; Homeostasis; Hypoglycemic Agents; Infertility, Female; Insulin; Metformin; Mice, Inbred ICR; Pregnancy

Environmental exposure to arsenic can cause a variety of health problems. Epidemiological and experimental studies have established a diabetogenic role for arsenic, but the mechanisms responsible for arsenic-induced impairment of insulin action are unclear. MicroRNAs (miRNAs) are involved in various metabolic disorders, particularly in the development of insulin resistance. The present study investigated whether arsenite, an active form of arsenic, induces hepatic insulin resistance and the mechanisms underlying it. After male C57BL/6J mice were exposed to arsenite (0 or 20 ppm) in drinking water for 12 months, intraperitoneal glucose tolerance tests (IPGTTs) and insulin tolerance tests (ITTs) revealed an arsenite-induced glucose metabolism disorder. Hepatic glycogen levels were lower in arsenite-exposed mice. Further, for livers of mice exposed to arsenite, miR-191 levels were higher, and protein levels of insulin receptor substrate 1 (IRS1), p-IRS1, and phospho-protein kinase B (p-AKT) were lower. Further, glucose transporter 4 (GLUT4) had lower levels on the plasma membrane. For insulin-treated L-02 cells, arsenite decreased glucose consumption and glycogen levels, increased miR-191 levels, and inhibited the IRS1/AKT pathway and the translocation of GLUT4 from the cytoplasm to the plasma membrane. For insulin-treated L-02 cells, the decreases of glucose consumption, glycogen levels, GLUT4 on the plasma membrane, and p-AKT levels induced by arsenite were reversed by SC79 (agonist of AKT) and an miR-191 inhibitor; these effects caused by miR-191 inhibitor were restored by IRS1 siRNA. In insulin-treated L-02 cells, miR-191, via IRS1, was involved in the arsenite-induced decreases of glucose consumption and glycogen levels and in inhibition of the translocation of GLUT4. Thus, miR-191 blocking the translocation of GLUT4 was involved in arsenite-induced hepatic insulin resistance through inhibiting the IRS1/AKT pathway. Our study reveals a mechanism for arsenite-induced hepatic insulin resistance, which provides clues for discovering biomarkers for the development of type 2 diabetes and for prevention and treatment of arsenic poisoning.

Topics: Animals; Arsenites; Diabetes Mellitus, Type 2; Glucose; Glucose Transporter Type 4; Glycogen; Insulin Receptor Substrate Proteins; Insulin Resistance; Liver; Male; Mice; Mice, Inbred C57BL; MicroRNAs; Proto-Oncogene Proteins c-akt; Signal Transduction

The hypoglycemic effects and potential mechanism of sweet potato leaf polyphenols (SPLP) on type 2 diabetes mellitus (T2DM) were investigated. Results showed that oral administration of SPLP to mice could alleviate body weight loss, decrease fasting blood glucose levels (by 64.78%) and improve oral glucose tolerance compared with those of untreated diabetic mice. Furthermore, increased fasting serum insulin levels (by 100.11%), ameliorated insulin resistance and improved hepatic glycogen (by 126.78%) and muscle glycogen (increased by 135.85%) were observed in the SPLP treatment group. SPLP also could reverse dyslipidemia, as indicated by decreased total cholesterol, triglycerides, low density lipoprotein-cholesterol and promoted high density lipoprotein-cholesterol. Histopathological analysis revealed that SPLP could relieve liver inflammation and maintain the islet structure to inhibit β-cell apoptosis. A quantitative real-time polymerase chain reaction confirmed that SPLP could up-regulate the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3β signaling pathway to improve glucose metabolism and up-regulate the phosphatidylinositol 3-kinase/protein kinase B/glucose transporter 4 signaling pathway in the skeletal muscle to enhance glucose transport. This study provides useful information to support the application of SPLP as a natural product for the treatment of T2DM.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Eating; Glucose Transporter Type 4; Glycogen; Glycogen Synthase Kinase 3 beta; Hyperglycemia; Hypoglycemic Agents; Insulin; Ipomoea batatas; Islets of Langerhans; Lipids; Liver; Liver Glycogen; Male; Mice; Pancreas; Phosphatidylinositol 3-Kinase; Plant Leaves; Polyphenols; Proto-Oncogene Proteins c-akt; Signal Transduction

Diosmetin (Dios), a flavonoid compound with multiple pharmacological activities. However, fewer studies have reported its effects on type 2 diabetic mellitus (T2DM). Here, we address the effect of Dios on glucose metabolism and gut microbiota in KK-Ay diabetic mice.. Wild type C57BL/6 J mice or diabetic KK-Ay mice were treated with vehicle or Dios for one month. The ELISA kit and fluorescence microscope system were respectively employed to the evaluation of serum biochemical indicators and histopathological changes. Liver RNA-Seq and western blot were used to reveal the key signaling pathway. The effects of Dios on gut microbiota was investigated by the 16S rRNA gene sequencing, as well as the relationship between Dios and C. glu on glucose metabolism was explored with the C. glu transplantation.. Dios treatment significantly decreased blood glucose and increased serum insulin concentrations. RNA-Seq analysis found that the underlying action mechanism of Dios on T2DM was via modulating glucose metabolism, which was proved by up-regulating IRS/PI3K/AKT signaling pathway to promote glycogen synthesis and GLUT4 translocation. Besides, Dios treatment reshaped the unbalanced gut microbiota by suppressing the ratio of Firmicutes/Bacteroidetes and markedly increasing the richness of C. glu. Moreover, treatment with C. glu and Dios together could markedly ameliorate glucose metabolism by up-regulating IRS/PI3K/AKT signaling pathway to promote glycogen synthesis and GLUT4 translocation.. Dios treatment remarkably ameliorated glucose metabolism in KK-Ay diabetic mice by the regulation of C. glu via IRS/PI3K/AKT signaling pathway and reshaped the unbalanced gut microbiota. Our study provided evidence for the application of Dios to the treatment of T2DM.

Topics: Animals; Blood Glucose; Corynebacterium glutamicum; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; DNA-Binding Proteins; Flavonoids; Gastrointestinal Microbiome; Glycogen; Hypoglycemic Agents; Insulin; Male; Mice, Inbred C57BL; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; RNA, Ribosomal, 16S; Transcription Factors

Diabetic patients have an increased predisposition to thromboembolic events, in most cases originating from thrombi in the left atrial appendage (LAA). Remodeling of the LAA, which predisposes to thrombi formation, has been previously described in diabetic patients with atrial fibrillation, but whether remodeling of the LAA occurs in diabetics also in the absence of atrial fibrillation is unknown. To investigate the contribution of diabetes, as opposed to atrial fibrillation, to remodeling of the LAA, we went from humans to the animal model.. We studied by echocardiography the structure and function of the heart over multiple time points during the evolution of diabetes in the Cohen diabetic sensitive rat (CDs/y) provided diabetogenic diet over a period of 4 months; CDs/y provided regular diet and the Cohen diabetic resistant (CDr/y), which do not develop diabetes, served as controls. All animals were in sinus rhythm throughout the study period.. Compared to controls, CDs/y developed during the evolution of diabetes a greater heart mass, larger left atrial diameter, wider LAA orifice, increased LAA depth, greater end-diastolic and end-systolic diameter, and lower E/A ratio-all indicative of remodeling of the LAA and left atrium (LA), as well as the development of left ventricular diastolic dysfunction. To investigate the pathophysiology involved, we studied the histology of the hearts at the end of the study. We found in diabetic CDs/y, but not in any of the other groups, abundance of glycogen granules in the atrial appendages , atria  and ventricles, which may be of significance as glycogen granules have previously been associated with cell and organ dysfunction in the diabetic heart.. We conclude that our rodent model of diabetes, which was in sinus rhythm, reproduced structural and functional alterations previously observed in hearts of human diabetics with atrial fibrillation. Remodeling of the LAA and of the LA in our model was unrelated to atrial fibrillation and associated with accumulation of glycogen granules. We suggest that myocardial accumulation of glycogen granules is related to the development of diabetes and may play a pathophysiological role in remodeling of the LAA and LA, which predisposes to atrial fibrillation, thromboembolic events and left ventricular diastolic dysfunction in the diabetic heart.

Topics: Animals; Atrial Appendage; Atrial Function, Left; Atrial Remodeling; Diabetes Mellitus, Type 2; Diabetic Cardiomyopathies; Disease Models, Animal; Disease Progression; Echocardiography, Doppler, Color; Glycogen; Heart Rate; Male; Rats, Inbred Strains; Time Factors; Ventricular Function, Left

Previous study suggests that

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Diet, High-Fat; Dietary Fats; DNA, Bacterial; Gastrointestinal Microbiome; Glucagon; Glycogen; Hypoglycemic Agents; Insulin; Lacticaseibacillus casei; Liver; Male; Metformin; Mice; Probiotics; Random Allocation; RNA, Bacterial; RNA, Ribosomal, 16S

Spexin (SPX) is a 14 aa peptide discovered in 2007 using bioinformatics methods. SPX inhibits food intake and regulates lipid, and carbohydrate metabolism. Here, we evaluate the ability of SPX at improving metabolic control and liver function in obese and type 2 diabetic animals. The effects of 30 days SPX treatment of mice with experimentally induced obesity (DIO) or type 2 diabetes (T2DM) on serum glucose and lipid levels, insulin sensitivity and hormonal profile (insulin, glucagon, adiponectin, leptin, TNF alpha, IL-6 and IL-1β) are characterized. In addition, alterations of hepatic lipid and glycogen contents are evaluated. We report that SPX decreases body weight in healthy and DIO mice, and reduces lipid content in all three animal groups. SPX improves insulin sensitivity in DIO and T2DM animals. In addition, SPX modulates hormonal and metabolic profile by regulating the concentration of adiponectin (concentration increase) and leptin (concentration decrease) in the serum blood of DIO and T2DM mice. Lastly, SPX decreases lipid content as well as IL-6 and TNF-α protein levels in liver of DIO and T2DM mice, and reduces IL-6 and TNF-alpha concentrations in the serum derived from T2DM mice. Based on our results, we conclude that SPX could be involved in the development of obesity and type 2 diabetes mellitus and it can be further evaluated as a potential target for therapy of DIO and T2DM.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Diet, High-Fat; Disease Models, Animal; Female; Glycogen; Insulin Resistance; Lipid Metabolism; Lipids; Liver Function Tests; Mice; Obesity; Peptide Hormones

Dendrobium officinale polysaccharide (DOP) is the main active ingredient of Dendrobium officinale Kimura & Migo, which is a precious traditional Chinese medicine and often used in treatment of hepatitis, diabetes, obesity and rheumatoid arthritis.. DOP exhibits significant hypoglycemic activity, while its mechanism remains unclear. The present study aims to investigate the hypoglycemic mechanisms of DOP based on the glucagon-mediated signaling pathways and the liver glycogen structure, which catalyze hepatic glucose metabolism, and provide new knowledge about the antidiabetic mechanism of DOP and further evidence for its clinical use for diabetes.. DOP were obtained from the dry stems of Dendrobium officinale by water extraction and alcohol precipitation method. T2DM mice model was established by high-fat diet combined with streptozotocin. Liver histopathological changes were observed by H&E and PAS straining. Pancreatic histology was studied by H&E staining and immunofluorescence analysis. The levels of glucagon and insulin were detected by Elisa Kit and the hepatic glycogen content was detected by GOPOD. The expressions of the hepatic glycogen-related metabolism enzymes, hepatic gluconeogenesis enzymes, and the related protein in cAMP-PKA and Akt/FoxO1 signaling pathways were detected by western blots. Liver glycogen was extracted from the liver tissues by sucrose density gradient centrifugation, and size exclusion chromatography (SEC) was used to analyze the structure of liver glycogen.. DOP could significantly affect the glucagon-mediated signaling pathways, cAMP-PKA and Akt/FoxO1, to further promote hepatic glycogen synthesis, inhibit hepatic glycogen degradation and hepatic gluconeogenesis. Moreover, DOP could reverse the instability of the liver glycogen structure and thus probably suppressed glycogen degradation. Thus, DOP finally would ameliorate hepatic glucose metabolism via glucagon-mediated signaling pathways and modifying liver-glycogen structure in diabetic mice.. The hypoglycemic mechanism of DOP might be associated with the regulation of glucagon-mediated hepatic glycogen metabolism and gluconeogenesis, and of liver glycogen structure, contributing to improved hepatic glucose metabolism in diabetic mice.

Topics: Animals; Dendrobium; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Glucagon; Glucose; Glycogen; Hypoglycemic Agents; Liver; Male; Mice, Inbred C57BL; Molecular Structure; Plant Extracts; Polysaccharides; Signal Transduction; Streptozocin

Flowering tops of Trifolium pratense L. (Fabaceae) are known for its traditional medicinal values. In present study, our aim was to investigate effect of standardized aqueous extract of flowering tops of Trifolium pratense L. on insulin resistance and SIRT1 expression in type 2 diabetic rats. Type 2 diabetes was induced by feeding high fat diet and administering low dose of streptozotocin. Diabetic animals were treated with standardized aqueous extract at three different doses. Parameters such as blood glucose, lipid profile, glycohemoglobin, insulin sensitivity, HOMA-IR and liver glycogen content were measured. Changes in morphology and expression of SIRT1 in pancreatic tissue were measured in histopathological and immunohistological studies. Aqueous extract treatment showed reduction in hyperglycemia and improved insulin sensitivity. Extract treatment also showed reduction in formation of glycated hemoglobin and improved liver glycogen level. Histopathological study revealed protecting effect of extract in pancreatic tissue against hyperglycemia induced damage. Treatment increased expression of SIRT1 in rat pancreatic tissue. Results indicate that the aqueous extract of Trifolium pratense had beneficial role in improving insulin sensitivity and SIRT1 expression.

Topics: Animals; Blood Glucose; Body Weight; Chromatography, High Pressure Liquid; Diabetes Mellitus, Type 2; Diet, High-Fat; Disease Models, Animal; Flowers; Glycogen; Hyperglycemia; Isoflavones; Male; Pancreas; Plant Extracts; Protective Agents; Rats; Rats, Sprague-Dawley; Sirtuin 1; Streptozocin; Trifolium

Non-alcoholic fatty liver disease and steatohepatitis are highly associated with obesity and type 2 diabetes mellitus. Cotadutide, a GLP-1R/GcgR agonist, was shown to reduce blood glycemia, body weight and hepatic steatosis in patients with T2DM. Here, we demonstrate that the effects of Cotadutide to reduce body weight, food intake and improve glucose control are predominantly mediated through the GLP-1 signaling, while, its action on the liver to reduce lipid content, drive glycogen flux and improve mitochondrial turnover and function are directly mediated through Gcg signaling. This was confirmed by the identification of phosphorylation sites on key lipogenic and glucose metabolism enzymes in liver of mice treated with Cotadutide. Complementary metabolomic and transcriptomic analyses implicated lipogenic, fibrotic and inflammatory pathways, which are consistent with a unique therapeutic contribution of GcgR agonism by Cotadutide

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Glucagon-Like Peptide-1 Receptor; Glycogen; Lipogenesis; Liver; Liver Cirrhosis; Male; Mice; Mice, Knockout; Mitochondria; Non-alcoholic Fatty Liver Disease; Peptides; Proteomics

SDM and T2DM rat models were established. During this time, PET/CT imaging was used to measure the %ID/g value of skeletal muscle and liver to evaluate glucose uptake. The pancreatic, skeletal muscle and liver were analyzed by immunohistochemistry.. SDM rats showed increased fasting blood glucose and insulin levels, hyperplasia of islet α and β cells, increased FDG uptake in skeletal muscle accompanied by an up-regulation of PI3Kp85α, IRS-1, and GLUT4, no significant changes in liver uptake, and that glycogen storage in the liver and skeletal muscle increased. T2DM rats showed atrophy of pancreatic islet β cells and decreased insulin levels, significantly reduced FDG uptake and glycogen storage in skeletal muscle and liver.. The pathogenesis of SDM is different from that of T2DM. The increased glucose metabolism of skeletal muscle may be related to the increased compensatory secretion of insulin. Glucocorticoids promote the proliferation of islet α cells and cause an increase in gluconeogenesis in the liver, which may cause increased blood glucose.

Topics: Animals; Blood Glucose; Diabetes Mellitus; Diabetes Mellitus, Type 2; Fasting; Fluorodeoxyglucose F18; Glucocorticoids; Glucose Transporter Type 4; Glycogen; Humans; Insulin; Insulin Receptor Substrate Proteins; Liver; Male; Muscle, Skeletal; Positron Emission Tomography Computed Tomography; Rats; Rats, Wistar

Understanding the mechanism by which the exogenous biomolecule modulates the GLUT-4 signalling cascade along with the information on glucose metabolism is essential for finding solutions to increasing cases of diabetes and metabolic disease. This study aimed at investigating the effect of hamamelitannin on glycogen synthesis in an insulin resistance model using L6 myotubes. Glucose uptake was determined using 2-deoxy-D-[1-

Topics: Animals; Biological Transport; Carbohydrate Metabolism; Cell Survival; Diabetes Mellitus, Type 2; Gallic Acid; Genistein; Glucose; Glucose Transporter Type 4; Glycogen; Glycogen Synthase Kinase 3 beta; Hexoses; Insulin; Insulin Antagonists; Insulin Resistance; Muscle Fibers, Skeletal; Myoblasts; Phosphatidylinositol 3-Kinases; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Rats; Signal Transduction; Wortmannin

The aim of this study was to evaluate the hypoglycemic effects of Pea oligopeptide on the glycemic and lipidemic status of mice with type 2 diabetes (T2D) induced by a high-fat diet and streptozotocin (STZ). Using HPLC-MS/MS spectra processing, 70 significant peptide (2-3 amino acids) sequences were identified, noting four peptides from Pea oligopeptide with a proline residue at the C-terminus, which might have dipeptidase-IV (DPP-IV) inhibitory activity for the treatment of T2D. After a 4-week administration of Pea oligopeptide and metformin, various blood biochemical indexes and organic histopathologies were detected to aid the discussion regarding potential mechanisms. The results showed a significant reduction in the levels of blood glucose, lipid profiles, and liver fat deposition in diabetic mice. Furthermore, Pea oligopeptide and metformin improved glucose tolerance, promoted glycogen synthesis, and protected the liver and kidney structures in diabetic mice. The results indicated that Pea oligopeptide played an essential role in the hypoglycemic effect in the T2D mice model. Practical applications This paper examined the preliminary hypoglycemic activities of Pea oligopeptide in a high-fat diet and STZ-induced T2D mice. Furthermore, four kinds of dipeptides and tripeptides that might exhibit antidiabetic functions were detected using HPLC-MS/MS. The results provided practical knowledge regarding the hypoglycemic effects of Pea oligopeptide and established the foundation of its structure-function relationships.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Glycogen; Hypoglycemic Agents; Insulin; Lipids; Metformin; Mice; Oligopeptides; Pisum sativum; Streptozocin

Type 2 diabetes (T2D) has become a major health problem worldwide. Skeletal muscle (SKM) is the key tissue for whole-body glucose disposal and utilization. New drugs aimed at improving insulin sensitivity of SKM would greatly expand available therapeutic options. β-arrestin-1 and -2 (Barr1 and Barr2, respectively) are two intracellular proteins best known for their ability to mediate the desensitization and internalization of G protein-coupled receptors (GPCRs). Recent studies suggest that Barr1 and Barr2 regulate several important metabolic functions including insulin release and hepatic glucose production. Since SKM expresses many GPCRs, including the metabolically important β2-adrenergic receptor, the goal of this study was to examine the potential roles of Barr1 and Barr2 in regulating SKM and whole-body glucose metabolism. Using SKM-specific knockout (KO) mouse lines, we showed that the loss of SKM Barr2, but not of SKM Barr1, resulted in mild improvements in glucose tolerance in diet-induced obese mice. SKM-specific Barr1- and Barr2-KO mice did not show any significant differences in exercise performance. However, lack of SKM Barr2 led to increased glycogen breakdown following a treadmill exercise challenge. Interestingly, mice that lacked both Barr1 and Barr2 in SKM showed no significant metabolic phenotypes. Thus, somewhat surprisingly, our data indicate that SKM β-arrestins play only rather subtle roles (SKM Barr2) in regulating whole-body glucose homeostasis and SKM insulin sensitivity.

Topics: Animals; beta-Arrestin 1; beta-Arrestin 2; Diabetes Mellitus, Type 2; Diet, High-Fat; Disease Models, Animal; Glucose; Glucose Clamp Technique; Glycogen; Humans; Insulin; Insulin Resistance; Male; Mice; Mice, Knockout; Muscle, Skeletal; Obesity; Signal Transduction

Cognitive dysfunction is one of the comorbidities of diabetes mellitus, but hippocampus-dependent learning and memory, a component of cognitive function, shows particular decline in type 2 diabetes, suggesting an increased risk for dementia and Alzheimer's disease. Cognitive function is related to dysregulated glucose metabolism, which is the typical cause of type 2 diabetes; however, hippocampal glycogen and its metabolite lactate are also crucial for hippocampus-dependent memory function. Type 2 diabetes induced hippocampus-dependent learning and memory dysfunction can be improved by chronic exercise and this improvement may possibly mediate through an adaptation of the astrocyte-neuron lactate shuttle (ANLS). This chapter focuses on the dysregulation of hippocampal glycometabolism in type 2 diabetes examining both existing evidence as well as the potential underlying pathophysiological mechanism responsible for memory dysfunction in type 2 diabetes, and showing for the first time that chronic exercise could be an effective therapy for type-2-diabetes-induced hippocampal memory decline.

Topics: Diabetes Mellitus, Type 2; Exercise; Exercise Therapy; Glycogen; Hippocampus; Humans; Lactic Acid; Neurons; Spatial Memory

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Diet, High-Fat; Enzyme Inhibitors; Fructose; Glycogen; Kidney; Lipase; Lipids; Liver; Male; Mice; Obesity; Phytoestrogens

Imbalance of iron homeostasis has been involved in clinical courses of metabolic diseases such as type 2 diabetes mellitus, obesity, and nonalcoholic fatty liver, through mechanisms not yet fully elucidated. Herein, we evaluated the effect of dietary iron on the development of diabetic syndromes in genetically obese db/db mice. Mice (aged 7 weeks) were fed with high-iron (HI) diets (1000 mg/kg chow) or low-iron (LI) diets (12 mg/kg) for 9 weeks. HI diets increased hepatic iron threefold and led to fourfold higher mRNA levels of hepcidin. HI also induced a 60% increase in fasting glucose due to insulin resistance, as confirmed by decreased hepatic glycogen deposition eightfold and a 21% decrease of serum adiponectin level. HI-fed mice had lower visceral adipose tissue mass estimated by epididymal and inguinal fat pad, associated with iron accumulation and smaller size of adipocytes. Gene expression analysis of liver showed that HI diet upregulated gluconeogenesis and downregulated lipogenesis. These results suggested that excess dietary iron leads to reduced mass, increased fasting glucose, decreased adiponectin level, and enhancement of insulin resistance, which indicated a multifactorial role of excess iron in the development of diabetes in the setting of obesity.

Topics: Adipose Tissue; Animals; Diabetes Mellitus, Type 2; Glucose; Glycogen; Hepcidins; Iron, Dietary; Lipid Metabolism; Lipogenesis; Liver; Male; Mice; Obesity; Real-Time Polymerase Chain Reaction

Pumpkin polysaccharides (PPe) have various biological activities. This research was to optimize the acid hydrolysis process of PPe with OH scavenging ability based on central composite design (CCD), and to explore the antioxidant and antidiabetic activities of the acid hydrolysates (PPe-H). A rat model of type 2 diabetes mellitus (T2DM) using high-fat diet and low-dose streptozotocin were established to assess the bioactivities. Both PPe and PPe-H could distinctly reduce fasting blood glucose level, prevent the weight loss in T2DM rats, and exhibited the remarkable ability to enhance the activities of the antioxidant enzymes (CAT and GR, p < 0.01, p < 0.01) and the level of GSH (p < 0.05, p < 0.01). Besides, PPe-H could significantly decrease the level of MDA (p < 0.05). Furthermore, PPe-H could cause an evident improvement to glucose stimulated GLP-1 secretion from 0 min to 30 min (p < 0.05). PPe and PPe-H were both the heteropolysaccharide and composed of rhamnose, arabinose, glucose and galactose, their molecular weight were 104.27 kDa and 37.58 kDa, respectively. The potential antidiabetic mechanism of PPe-H might be related to stimulating the secretion of endogenous GLP-1, decreasing oxidative damages, and then slowing down the process of diabetes.

Topics: Animals; Antioxidants; Cucurbita; Diabetes Mellitus, Type 2; Diet, High-Fat; Glycogen; Hydrogen-Ion Concentration; Hydrolysis; Hypoglycemic Agents; Insulin; Male; Molecular Weight; Polysaccharides; Rats; Rats, Wistar

Gold nanoparticles are known for their many applications in the fields of therapeutics and diagnosis.. This article focuses mainly on the green method of synthesizing gold nanoparticles by using the leaf powder extract of the insulin plant Chamaecostus cuspidatus and on the characterization of developed plant-mediated synthesis of gold nanoparticles. Furthermore, we investigated the free-radical scavenging activity of green-synthesized gold nanoparticles.. The free radicals were exhibited in a dose-dependent manner. The 50% inhibition of free radicals by gold nanoparticles showed that it was similar to that of the standard inhibition. Toxicity studies generally examine changes in blood serum chemistry and cell populations in tissue morphology through histologic analysis without inducing any lethal effects in the mouse model, thereby accomplishing sustained control over the progression of diabetes mellitus, which plays a leading role in vascular complications in patients. The treatment by gold nanoparticles of the mice with diabetes for a period of 21 days restored their blood glucose, glycogen and insulin levels.. The use of gold nanoparticles as antidiabetes materials has been achieved. Further studies are required before gold nanoparticle-based drugs are more widely used.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Free Radical Scavengers; Free Radicals; Glycogen; Gold; Hypoglycemic Agents; Insulin; Male; Metal Nanoparticles; Mice; Rats, Wistar; Wound Healing

The probiotic Lactobacillus gasseri SBT2055 (LG2055) has a protective effect against metabolic syndrome in rats and humans. Metabolic syndrome increases the risk of type 2 diabetes mellitus. In this study, Goto-Kakizaki rats were used as a diabetic model and fed diets containing LG2055-fermented or nonfermented skim milk for 4 wk. Indices of diabetes such as blood glucose levels, serum glucagon levels, plasma levels of insulin, C-peptide, and glucagon-like peptide-1, tissue glycogen contents, and pancreatic mRNA levels were measured. The plasma C-peptide levels and pancreatic mRNA levels of insulin genes (Ins1 and Ins2) and Pdx1 (a transcriptional factor of insulin genes) were increased in LG2055 diet-fed rats. The increase in insulin secretion corresponded to an improvement in serum and pancreatic inflammatory status, associated with decreases in serum levels of serum amyloid P and pancreatic levels of granulocyte colony-stimulating factor. Insulin resistance in Goto-Kakizaki rats was ameliorated by increased glycogen storage in the liver and quadriceps femoris muscles and decreased serum free fatty acid levels. This improvement may be related to the increased cecal production of short-chain fatty acids. In conclusion, dietary LG2055 improved insulin secretion in diabetic rats by improving the inflammatory status in the pancreas and serum.

Topics: Animals; Blood Glucose; Cecum; Diabetes Mellitus, Type 2; Diet; Fatty Acids, Volatile; Glucagon-Like Peptide 1; Glycogen; Humans; Insulin; Insulin Resistance; Insulin Secretion; Lactobacillus gasseri; Liver; Male; Muscle, Skeletal; Probiotics; Rats

The miR-181 family plays an important role in the regulation of various cellular functions. However, whether miR-181b-5p mediates hepatic insulin resistance remains unknown. In this study, we investigated the effect of miR-181b-5p on the regulation of hepatic glycogen synthesis.. The miR-181b-5p levels in the livers of diabetic mice were detected by real-time PCR. The glycogen levels and AKT/GSK pathway activation were examined in human hepatic L02 cells and HepG2 cells transfected with miR-181b-5p mimic or inhibitor. The potential target genes of miR-181b-5p were evaluated using a luciferase reporter assay and Western blot analysis. EGR1-specific siRNA and pCMV-EGR1 were used to further determine the role of miR-181b-5p in hepatic glycogen synthesis in vitro. Hepatic inhibition of miR-181b-5p in mice was performed using adeno-associated virus 8 (AAV8) vectors by tail intravenous injection.. The miR-181b-5p levels were significantly decreased in the serum and livers of diabetic mice as well as the serum of type 2 diabetes patients. Importantly, inhibition of miR-181b-5p expression impaired the AKT/GSK pathway and reduced glycogenesis in hepatocytes. Moreover, upregulation of miR-181b-5p reversed high-glucose-induced suppression of glycogenesis. Further analysis revealed that early growth response 1 (EGR1) was a downstream target of miR-181b-5p. Silencing of EGR1 expression rescued miR-181b-5p inhibition-reduced AKT/GSK pathway activation and glycogenesis in hepatocytes. Hepatic inhibition of miR-181b-5p led to insulin resistance in C57BL/6 J mice.. We demonstrated that miR-181b-5p contributes to glycogen synthesis by targeting EGR1, thereby regulating PTEN expression to mediate hepatic insulin resistance.

Topics: Adult; Animals; Diabetes Mellitus, Type 2; Disease Models, Animal; Early Growth Response Protein 1; Female; Glycogen; Hep G2 Cells; Humans; Insulin Resistance; Liver; Male; Mice, Inbred C57BL; MicroRNAs; Middle Aged; PTEN Phosphohydrolase; Signal Transduction

Dysregulation of glucose and glycogen metabolism are crucial mechanisms implicated in type 2 diabetes mellitus (T2DM). Centella asiatica (L.) Urban (Apiaceae) has been utilized as a traditional medicine in Africa and Asia for centuries and is commercially available as a dietary supplement.. We explored for the first time, the possible efficacy of Centella asiatica (CA) extract in ameliorating T2DM-induced changes in key enzymes involved in glucose and glycogen metabolism in the rat skeletal muscle.. Diabetic rats were orally treated with vehicle, CA (500 and 1000 mg/kg) or metformin (300 mg/kg) daily for 14 days. Skeletal muscle activities of hexokinase (HK), phosphofructokinase (PFK) and fructose 1,6-bisphosphatase (FBPase) were determined by spectrophotometric assays while those of glycogen synthase (GS) and glycogen phosphorylase (GP) were assayed radio-chemically. Histological examination of skeletal muscle was also performed.. Rats with induced T2DM had reduced activities of HK (25%), PFK (88%), and GS (38%) when compared to non-diabetic rats. Treatment of diabetic rats with CA500 increased the activities of PFK (7-fold), and FBPase (23%). Further, treatment of diabetic rats with CA1000 also increased the activities of GS (27%) and GP (50%) with little change in these parameters for diabetic rats treated with CA500. These effects probably led to the reduced blood glucose level and elevated skeletal muscle glycogen content observed in CA-treated rats relative to diabetic controls. Furthermore, CA treated rats had reduced the morphological damage of skeletal muscle fibres compared to the non-treated diabetic control rats.. Our findings strongly suggest that the anti-diabetic effects of CA in part target muscle glucose and glycogen metabolism and hence supporting its folkloric medical use as an anti-diabetic remedy.

Topics: Animals; Blood Glucose; Centella; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Phosphorylase; Glycogen Synthase; Hypoglycemic Agents; Male; Medicine, Traditional; Muscle, Skeletal; Plant Extracts; Plant Leaves; Rats, Sprague-Dawley; Streptozocin; Triterpenes

Protein tyrosine phosphatase 1B (PTP1B) is a widely confirmed target of the type 2 diabetes mellitus (T2DM) treatment. Herein, we reported a highly specific PTP1B inhibitor 2,2',3,3'-tetrabromo-4,4',5,5'-tetrahydroxydiphenylmethane (compound 1), which showed promising hypoglycemic activity in diabetic BKS db mice. With the IC

Topics: Administration, Oral; Animals; Benzhydryl Compounds; Catalytic Domain; Cell Line; Diabetes Mellitus, Type 2; Glycogen; Hydrogen Bonding; Hypoglycemic Agents; Inhibitory Concentration 50; Insulin; Insulin Resistance; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Molecular Docking Simulation; Myoblasts; Plant Extracts; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Rhodophyta; Signal Transduction

Korean pine nut protein (PNP) has a variety of biological activities, which are good for human health, but its ability to preventing diabetes has not been reported. This study evaluated the effects of water-soluble proteins of Korean pine nut obtained from a dilute alkali extract on carbohydrate metabolism of type 2 diabetic mice on a model of diabetes induced using a high fat diet combined with streptozotocin. The results showed that the hypoglycemic effect of PNP at a middle dose was the most significant, which was 38.7% lower than that of control. The extract significantly improved the oral glucose tolerance and liver indexes, increased the activity of the carbohydrate metabolism enzymes, and regulated the expression of the function of key genes for carbohydrate metabolism. It had a positive effect on both insulin resistance and glycolytic/gluconeogenesis signaling. In conclusion, PNP can regulate fasting blood glucose, improve insulin resistance, correct the glucose metabolism disorder in diabetic mice, and have a positive regulatory role. As the functional food, it has the potential to be beneficial in the treatment of type 2 diabetes mellitus as a new hypoglycemic functional food.

Topics: Administration, Oral; Amino Acids; Animals; Blood Glucose; Body Weight; Carbohydrate Metabolism; Diabetes Mellitus, Type 2; Diet; Drinking Behavior; Fasting; Feeding Behavior; Gene Expression Regulation; Glucose Tolerance Test; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Male; Mice; Muscles; Nut Proteins; Pinus; Solubility; Water

During the development of type 2 diabetes mellitus (T2DM), hyperinsulinemia is the earliest symptom. It is believed that long-term high insulin stimulation might facilitate insulin resistance in the liver, but the underlying mechanism remains unknown. Herein, we report that hyperinsulinemia could induce persistent early growth response gene-1 (Egr-1) activation in hepatocytes, which provides negative feedback inhibition of insulin sensitivity by inducing the expression of protein tyrosine phosphatase-1B (PTP1B). Deletion of Egr-1 in the liver remarkably decreases glucose production, thus improving systemic glucose tolerance and insulin sensitivity. Mechanistic analysis indicates that Egr-1 inhibits insulin receptor phosphorylation by directly activating PTP1B transcription in the liver. Our results reveal the molecular mechanism by which hyperinsulinemia accelerates insulin resistance in hepatocytes during the progression of T2DM.

Topics: Animals; Cells, Cultured; Diabetes Mellitus, Type 2; Disease Models, Animal; Early Growth Response Protein 1; Gene Knockout Techniques; Glucose; Glycogen; Hyperinsulinism; Insulin; Insulin Resistance; Liver; Male; Mice; Phosphorylation; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Receptor, Insulin; Transcriptional Activation

The compound 2-deoxy-2-fluoro-α-D-glucopyranosyl fluoride (F

Topics: Animals; Deoxyglucose; Diabetes Mellitus, Type 2; Glycogen; Glycogen Synthase; Hepatocytes; Male; Protein Transport; Rats; Rats, Sprague-Dawley

Cigarette smoke causes insulin resistance, which is associated with type 2 diabetes mellitus (T2DM). However, the mechanism by which this occurs remains poorly understood. Because the involvement of microRNAs (miRNAs) in the development of insulin resistance is largely unknown, we investigated, in hepatocytes, the roles of miR-191 in cigarette smoke extract (CSE)-induced insulin resistance. In L-02 cells, CSE not only decreased glucose uptake and glycogen levels but also reduced levels of insulin receptor substrate-1 (IRS-1) and Akt activation, effects that were blocked by SC79, an activator of Akt. CSE also increased miR-191 levels in L-02 cells. Furthermore, the inhibition of miR-191 blocked the decreases of IRS-1 and p-Akt levels, which antagonized the decreases of glucose uptake and glycogen levels in L-02 cells induced by CSE. These results reveal a mechanism by which miR-191 is involved in CSE-induced hepatic insulin resistance via the IRS-1/Akt signaling pathway, which helps to elucidate the mechanism for cigarette smoke-induced T2DM.

Topics: Cell Line; Cigarette Smoking; Diabetes Mellitus, Type 2; Glycogen; Hepatocytes; Humans; Insulin Receptor Substrate Proteins; Insulin Resistance; MicroRNAs; Proto-Oncogene Proteins c-akt; Signal Transduction

Type 2 diabetes mellitus contributes to an increased risk of metabolic and morphological changes in key organs, such as the liver. We aimed to assess the effect of the açaí seed extract (ASE) associated with exercise training on hepatic steatosis induced by high-fat (HF) diet plus streptozotocin (STZ) in rats. Type 2 diabetes was induced by feeding rats with HF diet (55% fat) for 5 weeks, followed by a single low dose of STZ (35 mg/kg i.p.). Control and diabetic groups were subdivided into four groups that were fed with standard chow diet for 4 weeks. Control (C) group was subdivided into Sedentary C, Training C, ASE Sedentary C and ASE Training C. Diabetic (D) group was subdivided into Sedentary D, Training D, ASE Sedentary D and ASE Training D. ASE (200 mg/kg/day) was administered by intragastric gavage, and the exercise training was performed on a treadmill (30 min/day; 5 days/week). Treatment with ASE associated with exercise training reduced the blood glucose (70.2%), total cholesterol (81.2%), aspartate aminotransferase (51.7%) and hepatic triglyceride levels (66.8%) and steatosis (72%) in ASE Training D group compared with the Sedentary D group. ASE associated with exercise training reduced the hepatic lipogenic proteins' expression (77.3%) and increased the antioxidant defense (63.1%), pAMPK expression (70.2%), cholesterol transporters (71.1%) and the pLKB1/LKB1 ratio (57.1%) in type 2 diabetic rats. In conclusion, ASE treatment associated with exercise training protects against hepatic steatosis in diabetic rats by reducing hepatic lipogenesis and increasing antioxidant defense and cholesterol excretion.

Topics: Animals; Antioxidants; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Enzymes; Euterpe; Glycogen; Lipid Metabolism; Liver; Male; Non-alcoholic Fatty Liver Disease; Physical Conditioning, Animal; Plant Extracts; Protein Carbonylation; Proteins; Rats, Wistar; Seeds

In this study, we have investigated whether silymarin intake influences lipid and glycogen content in conjunction with sirtuin1 (SIRT1) and sterol regulatory element-binding protein 1c (SREBP-1c) expressions in liver of type 2 diabetic rat.. Thirty-six male Wistar rats were randomly divided into six groups: control groups (C) and diabetic groups (D); the control groups received 60 or 120 mg/kg silymarin (C+S60 or C+S120), and the diabetic groups received 60 or 120 mg/kg silymarin (D+S60 or D+S120) daily for 8 weeks. Serum biochemical parameters, as well as glycogen, lipid and oxidative stress biomarkers, in the liver tissue were measured by spectrophotometric methods. Additionally, SIRT1 and SREBP-1c messenger RNA (mRNA) expressions were evaluated by quantitative polymerase chain reaction.. Diabetes caused a significantly increased fasting blood sugar, homeostasis model assessment for insulin resistance, liver total cholesterol and triglyceride (TG) content, which were attenuated after the administration of silymarin. Dietary silymarin caused the improvement of lipid content in the liver of diabetic rats. Moreover, silymarin administration promoted SIRT1, suppressed SREBP-1c mRNA expression, reduced liver nitric oxide and protein carbonyl content, and increased liver glycogen, catalase and glutathione peroxidase activity. Furthermore, histopathological changes were improved in the treated groups.. Silymarin administration considerably restored hepatic changes induced by streptozotocin and nicotinamide. The upregulation of SIRT1 mRNA expression by silymarin may be associated with decreased lipid, increased glycogen content and downregulation of the SREBP-1c gene in the liver.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Glycogen; Lipid Metabolism; Liver; Male; Niacinamide; Protective Agents; Rats; Rats, Wistar; RNA, Messenger; Silymarin; Sirtuin 1; Sterol Regulatory Element Binding Protein 1; Streptozocin; Up-Regulation

AMPK activated protein kinase (AMPK), a master regulator of energy homeostasis, is activated in response to an energy shortage imposed by physical activity and caloric restriction. We here report on the identification of PAN-AMPK activator O304, which - in diet-induced obese mice - increased glucose uptake in skeletal muscle, reduced β cell stress, and promoted β cell rest. Accordingly, O304 reduced fasting plasma glucose levels and homeostasis model assessment of insulin resistance (HOMA-IR) in a proof-of-concept phase IIa clinical trial in type 2 diabetes (T2D) patients on Metformin. T2D is associated with devastating micro- and macrovascular complications, and O304 improved peripheral microvascular perfusion and reduced blood pressure both in animals and T2D patients. Moreover, like exercise, O304 activated AMPK in the heart, increased cardiac glucose uptake, reduced cardiac glycogen levels, and improved left ventricular stroke volume in mice, but it did not increase heart weight in mice or rats. Thus, O304 exhibits a great potential as a novel drug to treat T2D and associated cardiovascular complications.

Topics: AMP-Activated Protein Kinases; Animals; Blood Glucose; Blood Pressure; Cardiomegaly; Cardiovascular Diseases; Diabetes Mellitus, Type 2; Glucose; Glycogen; Heart; Heterocyclic Compounds; Holoprosencephaly; Homeostasis; Humans; Insulin Resistance; Insulin-Secreting Cells; Jaw Abnormalities; Metformin; Mice; Mice, Obese; Muscle, Skeletal; Rats; Stroke Volume

Insulin resistance (IR) is considered as a major factor of type 2 diabetes (T2D), which is seriously detrimental to human health. In our present study, we found that the expression of miR-138-5p was increased in the insulin-resistant HepG2 cells induced by TNF-α. Therefore, we hypothesized that miR-138-5p might play a regulatory role in the IR. To examine this hypothesis, HepG2 cells were transfected with miR-138-5p inhibitor. Silencing of miR-138-5p increased glucose uptake and glycogen synthesis of TNF-α-stimulated HepG2 cells and decreased glucose concentration in medium, suggesting that downregulation of miR-138-5p suppressed IR in HepG2 cells. Besides that, we found that sirtuin 1 (SIRT1) was the target gene of the miR-138-5p. Moreover, co-transfection with SIRT1-siRNA and miR-138-5p inhibitor suppressed glucose uptake and glycogen synthesis of HepG2 cells compared with miR-138-5p inhibitor-transfected group, indicating that downregulation of SIRT1 weakened the inhibitory effect of miR-138-5p inhibitor on IR. In addition, overexpressed SIRT1 increased Beclin1, LC3 II/I level, and the number of GFP-LC3 dots and decreased p62 level, whereas downregulation of SIRT1 had the opposite effects. Our results demonstrated that overexpressed SIRT1 activated autophagy in HepG2 cells. Moreover, we observed that 3-methyladenine (an inhibitor of autophagy) treatment decreased the high glucose uptake and glycogen synthesis of miR-138-5p inhibitor-transfected HepG2 cells, suggesting that the inhibition of autophagy abolished the inhibitory effect of miR-138-5p inhibitor on IR in HepG2 cells. Taken together, this study suggested that miR-138-5p contributed to the TNF-α-induced IR, possibly through inducing autophagy in HepG2 cells by regulating SIRT1. MiR-138-5p might be a potential and promising target for the treatment of IR.

Topics: Adenine; Autophagy; Diabetes Mellitus, Type 2; Disease Progression; Down-Regulation; Glucose; Glycogen; Hep G2 Cells; Humans; Insulin; Insulin Resistance; MicroRNAs; Sirtuin 1; Tumor Necrosis Factor-alpha

Astrocyte-neuron lactate shuttle (ANLS) is a pathway that supplies glycogen-derived lactate to active neurons via monocarboxylate transporter 2 (MCT2), and is important for maintaining brain functions. Our study revealed alterations of ANLS with hippocampal hyper-glycogen levels and downregulated MCT2 protein levels underlying hippocampal dysfunctions as a complication in type 2 diabetic (T2DM) animals. Since T2DM rats exhibit brain dysfunctions involving several brain regions, we examined whether there might also be T2DM effects on ANLS's disturbances in other brain loci. OLETF rats exhibited significantly higher glycogen levels in the hippocampus, hypothalamus, and cerebral cortex than did LETO rats. MCT2 protein levels in OLETF rats decreased significantly in the hippocampus and hypothalamus compared to their controls, but a significant correlation with glycogen levels was only observed in the hippocampus. This suggests that the hippocampus may be more vulnerable to T2DM compared to other brain regions in the context of ANLS disruption.

Topics: Animals; Astrocytes; Cerebral Cortex; Diabetes Mellitus, Type 2; Glycogen; Hippocampus; Male; Monocarboxylic Acid Transporters; Neurons; Rats; Rats, Inbred OLETF

Metformin has been introduced for treatment of type 2 diabetes but may also have ergogenic properties at high altitude by improving muscle glycogen repletion. However, very little information is available on potential risks associated with the (mis)use of metformin by healthy people.

Topics: Altitude; Diabetes Mellitus, Type 2; Glycogen; Humans; Metformin; Performance-Enhancing Substances

[6]-Gingerol, a major component of Zingiber officinale, was previously reported to ameliorate hyperglycemia in type 2 diabetic mice. Endocrine signaling is involved in insulin secretion and is perturbed in db/db Type-2 diabetic mice. [6]-Gingerol was reported to restore the disrupted endocrine signaling in rodents. In this current study on Lepr. 4-weeks treatment of [6]-Gingerol dramatically increased glucose-stimulated insulin secretion and improved glucose tolerance. Plasma GLP-1 was found to be significantly elevated in the treated mice. Pharmacological intervention of GLP-1 levels regulated the effect of [6]-Gingerol on insulin secretion. Mechanistically, [6]-Gingerol treatment upregulated and activated cAMP, PKA, and CREB in the pancreatic islets, which are critical components of GLP-1-mediated insulin secretion pathway. [6]-Gingerol upregulated both Rab27a GTPase and its effector protein Slp4-a expression in isolated islets, which regulates the exocytosis of insulin-containing dense-core granules. [6]-Gingerol treatment improved skeletal glycogen storage by increased glycogen synthase 1 activity. Additionally, GLUT4 transporters were highly abundant in the membrane of the skeletal myocytes, which could be explained by the increased expression of Rab8 and Rab10 GTPases that are responsible for GLUT4 vesicle fusion to the membrane.. Collectively, our study reports that GLP-1 mediates the insulinotropic activity of [6]-Gingerol, and [6]-Gingerol treatment facilitates glucose disposal in skeletal muscles through increased activity of glycogen synthase 1 and enhanced cell surface presentation of GLUT4 transporters.

Topics: Animals; Blood Glucose; Catechols; Diabetes Mellitus, Type 2; Fatty Alcohols; Glucagon-Like Peptide 1; Glucose Transporter Type 4; Glycogen; Glycogen Synthase; Hyperglycemia; Insulin; Insulin Secretion; Insulin-Secreting Cells; Membrane Proteins; Mice; Mice, Inbred NOD; Mice, Knockout; Muscle, Skeletal; Phytotherapy; Plant Extracts; rab GTP-Binding Proteins; Secretory Pathway; Vesicular Transport Proteins; Zingiber officinale

Topics: Animal Feed; Animals; Animals, Newborn; Birth Weight; Blood Glucose; Diabetes Mellitus, Type 2; Diet; Female; Fetal Growth Retardation; Glucose-6-Phosphatase; Glycogen; Hyperglycemia; Insulin Resistance; Liver; Male; Methionine; Muscles; Phosphoenolpyruvate Carboxykinase (GTP); Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction; Swine; Weaning

Recent studies have shown that rambutan peel phenolic (RPP) extract demonstrate high antioxidant and antiglycation activities in vitro and in vivo. This study further evaluated the anti-diabetic activity of RPP in a mouse model of Type II diabetes induced by streptozotocin combined with high-fat diet. Results showed that RPP increased the body weight and reduced the fasting blood glucose level of the diabetic mice. RPP significantly reduced the serum levels of total cholesterol, triglyceride, creatinine, and glycated serum protein in diabetic mice in a dose-dependent manner. Glycogen content in mice liver was recovered by RPP, which further increased the activity of superoxide dismutase and glutathione peroxidase and reduced lipid peroxidation in diabetic mice. Histological analysis showed that RPP effectively protected the tissue structure of the liver, kidney, and pancreas. In addition, RPP decreased the mesangial index and inhibited the expression of TGF-β in the kidney of diabetic mice.

Topics: Animals; Biomarkers; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Dose-Response Relationship, Drug; Fruit; Glutathione Peroxidase; Glycated Hemoglobin; Glycogen; Hypoglycemic Agents; Kidney; Lipid Peroxidation; Lipids; Liver; Male; Mice, Inbred ICR; Pancreas; Phenols; Phytotherapy; Plant Extracts; Plants, Medicinal; Sapindaceae; Streptozocin; Superoxide Dismutase; Time Factors; Transforming Growth Factor beta1

Type 2 diabetes is likely to be an independent risk factor for hippocampal-based memory dysfunction, although this complication has yet to be investigated in detail. As dysregulated glycometabolism in peripheral tissues is a key symptom of type 2 diabetes, it is hypothesised that diabetes-mediated memory dysfunction is also caused by hippocampal glycometabolic dysfunction. If so, such dysfunction should also be ameliorated with moderate exercise by normalising hippocampal glycometabolism, since 4 weeks of moderate exercise enhances memory function and local hippocampal glycogen levels in normal animals.. The hippocampal glycometabolism in OLETF rats (model of human type 2 diabetes) was assessed and, subsequently, the effects of exercise on memory function and hippocampal glycometabolism were investigated.. OLETF rats, which have memory dysfunction, exhibited higher levels of glycogen in the hippocampus than did control rats, and breakdown of hippocampal glycogen with a single bout of exercise remained unimpaired. However, OLETF rats expressed lower levels of hippocampal monocarboxylate transporter 2 (MCT2, a transporter for lactate to neurons). Four weeks of moderate exercise improved spatial memory accompanied by further increase in hippocampal glycogen levels and restoration of MCT2 expression independent of neurotrophic factor and clinical symptoms in OLETF rats.. Our findings are the first to describe detailed profiles of glycometabolism in the type 2 diabetic hippocampus and to show that 4 weeks of moderate exercise improves memory dysfunction in type 2 diabetes via amelioration of dysregulated hippocampal glycometabolism. Dysregulated hippocampal lactate-transport-related glycometabolism is a possible aetiology of type-2-diabetes-mediated memory dysfunction.

Topics: Animals; Blood Glucose; Blotting, Western; Body Weight; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Eating; Glycogen; Hippocampus; Male; Memory; Physical Conditioning, Animal; Rats; Rats, Inbred OLETF

DNA methylation is altered by environmental factors. We hypothesized that DNA methylation is altered in skeletal muscle in response to either insulin or glucose exposure. We performed a genome-wide DNA methylation analysis in muscle from healthy men before and after insulin exposure. DNA methylation of selected genes was determined in muscle from healthy men and men with type 2 diabetes before and after a glucose tolerance test. Insulin altered DNA methylation in the 3' untranslated region of the calcium pump

Topics: Biopsy; Blood Glucose; Case-Control Studies; Death-Associated Protein Kinases; Diabetes Mellitus, Type 2; DNA Methylation; Glucose; Glucose Tolerance Test; Glycogen; Humans; Hypoglycemic Agents; In Vitro Techniques; Insulin; Male; Middle Aged; Muscle, Skeletal; Real-Time Polymerase Chain Reaction; RNA, Messenger; Sarcoplasmic Reticulum Calcium-Transporting ATPases

Infusion of mesenchymal stem cells (MSCs) has been identified in the rapid alleviation in hyperglycemia of diabetic individuals, but the mechanism involved has not been adequately explained by these cells' potential role in modulating system insulin sensitivity and islet regeneration. In this study, we demonstrated adipose-derived mesenchymal stem cells (ASCs) produced significantly lower blood glucose via promoting hepatic glycogen synthesis and inhibiting hepatic glucose production within 24 h after infusion in T2DM rats. In vitro, HepG2 cells treated with palmitate (PA) were used as a model of hepatic glucose metabolism disorder to confirm that ASCs stimulates the phosphorylation of hepatic AMP-activated protein kinase (AMPK) to restores hepatic glucose metabolism in type 2 diabetes. In summary, this study indicated that ASCs improve hyperglycemia via regulating hepatic glucose metabolism. Additionally, the effect of ASCs on hepatic glucose metabolism depended on the AMPK signaling pathway. Thus, this is the new research of the molecular mechanisms of MSCs administration to improve glucose metabolism, and it may indicate a new treatment target of MSCs in T2DM.

Topics: Adipose Tissue; Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Enzymes; Glucose; Glycogen; Hep G2 Cells; Humans; Hyperglycemia; Infusions, Intravenous; Liver; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Palmitates; Rats, Sprague-Dawley

Diabetes mellitus is chronic metabolic disorder, resulting from insulin deficiency, characterized by hyperglycemia altered metabolism of carbohydrates, proteins and lipids and an increased risk of vascular complications. There are different classes of anti-diabetic drugs in allopathic system of medicine. Metformin (dimethyl biguanide) is a blood glucose lowering agent used in the treatment of non-insulin dependent diabetes mellitus. Almost in all diseases the blood serves as the primary metabolic transport system in the body. Its composition is the preferred indicator with respect to the pathophysiological condition of the patient. Instead of analyzing blood to diagnose diabetes, hair could be used to detect diabetes using FTIR-ATR technique. The most important components of hair are fibrous proteins (keratins), melanins, glycogen, and lipids. Hair follicles are located 3-4mm below the surface of the skin and are surrounded by rich blood capillary system. In the present study, ten diabetic subjects were considered to evaluate the efficacy of metformin hydrochloride for the treatment of diabetes mellitus using FTIR-ATR spectroscopy. The spectra of diabetic hair fibre samples have been recorded in the mid infrared region of 4000-450cm

Topics: Biomarkers; Data Interpretation, Statistical; Diabetes Mellitus, Type 2; Female; Glucose; Glycogen; Hair; Humans; Hypoglycemic Agents; Keratins; Lipoproteins, HDL; Lipoproteins, LDL; Male; Metformin; Middle Aged; Spectroscopy, Fourier Transform Infrared; Treatment Outcome

Topics: Adrenal Medulla; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Disease Models, Animal; Fasting; Glucose; Glucose Tolerance Test; Glycogen; Hypoxia; Insulin; Insulin Resistance; Liver; Male; Mice; Mice, Inbred C57BL; Oxygen; Oxyhemoglobins; Sleep Apnea, Obstructive

The decoctions of Ficus carica Linn. (Moraceae) leaves are used in the folklore treatment of diabetes.. To evaluate the effect of F. carica on glucose and lipids levels, carbohydrate metabolism enzymes and β-cells protective effects in type 2 diabetes.. Diabetes was induced in 15 days high-fat diet (HFD)-fed Wistar rats by intraperitoneal injection of streptozotocin (STZ) (40 mg/kg). The ethyl acetate extract (250 and 500 mg/kg) of F. carica leaves was administered for 28 days. Oral glucose tolerance (OGTT) and intraperitoneal insulin tolerance tests (ITT) were evaluated on 15th and 25th days, respectively.. The ethyl acetate extract (250 and 500 mg/kg) of n F. carica leaves showed significant effect (p < 0.005) in the levels of blood glucose, total cholesterol (TC), triglycerides (TG), body weight and hepatic glycogen. In OGTT, F. carica (250 and 500 mg/kg) significantly (p < 0.005) detained the increase in blood glucose levels at 60 and 120 min and in ITT, F. carica enhanced the glucose utilization significantly (p < 0.005) over 30 and 60 min compared to diabetic control. Further, the altered activities of key carbohydrate metabolizing enzymes such as glucose-6-phosphatase, fructose-1,6-bisphosphatase and hexokinase in the liver tissue of diabetic rats were significantly (p < 0.005) reverted to near normal levels upon treatment with F. carica. Immumohistochemical studies of islets substantiated the cytoprotective effect on pancreatic β-cells.. F. carica leaves exerted significant effect on carbohydrate metabolism enzymes with promising hypoglycemic and hypolipidemic activities in type 2 diabetic rats.

Topics: Acetates; Animals; Biomarkers; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Dose-Response Relationship, Drug; Ficus; Glucose Tolerance Test; Glyburide; Glycogen; Hypoglycemic Agents; Hypolipidemic Agents; Insulin; Insulin Resistance; Insulin-Secreting Cells; Lipids; Liver; Male; Phytotherapy; Plant Extracts; Plant Leaves; Plants, Medicinal; Rats, Wistar; Solvents; Streptozocin; Time Factors

The impact of diet-induced weight loss and weight loss due to RYGB in patients with (T2DM, N = 16) and without (OB, N = 27) type 2 diabetes was studied.. At inclusion (A), after diet-induced weight loss (B), 4 months post-surgery (C) and 18 months post-surgery (D) body composition, hepatic glucose production (HGP), insulin-mediated glucose uptake (GIR), respiratory exchange ratio, hepatic insulin sensitivity and clearance were determined. GLUT4, intramuscular triglycerides (IMTG) and glycogen content were measured in skeletal muscle.. Weight loss was 35-40 kg, and approximately one-third of the total improvement in GIR in T2DM was observed after the diet-induced weight loss of only ~6 kg (B). Insulin clearance, visceral fat and fasting plasma insulin also improved significantly after the diet (P < 0.05). Throughout the study, HGP, GLUT4 and glycogen content did not change significantly, but IMTG decreased significantly consistent with significant increases in GIR. Metabolic flexibility and hepatic insulin sensitivity improved after RYGB.. Metabolic improvements of RYGB are present already after the diet-induced weight loss prior to surgery. GLUT4 content in skeletal muscle cannot and IMTG content can only partly explain increases in GIR after RYGB.

Topics: Adult; Blood Glucose; Diabetes Mellitus, Type 2; Diet, Reducing; Female; Gastric Bypass; Glycogen; Humans; Insulin; Insulin Resistance; Male; Obesity; Triglycerides; Weight Loss

Glycogen is a vital highly branched polymer of glucose that is essential for blood glucose homeostasis. In this article, the structure of liver glycogen from mice is investigated with respect to size distributions, degradation kinetics, and branching structure, complemented by a comparison of normal and diabetic liver glycogen. This is done to screen for differences that may result from disease. Glycogen α-particle (diameter ∼ 150 nm) and β-particle (diameter ∼ 25 nm) size distributions are reported, along with in vitro γ-amylase degradation experiments, and a small angle X-ray scattering analysis of mouse β-particles. Type 2 diabetic liver glycogen upon extraction was found to be present as large loosely bound, aggregates, not present in normal livers. Liver glycogen was found to aggregate in vitro over a period of 20 h, and particle size is shown to be related to rate of glucose release, allowing a structure-function relationship to be inferred for the tissue specific distribution of particle types. Application of branching theories to small angle X-ray scattering data for mouse β-particles revealed these particles to be randomly branched polymers, not fractal polymers. Together, this article shows that type 2 diabetic liver glycogen is present as large aggregates in mice, which may contribute to the inflexibility of interconversion between glucose and glycogen in type 2 diabetes, and further that glycogen particles are randomly branched with a size that is related to the rate of glucose release.

Topics: Animals; Diabetes Mellitus, Type 2; Glucose; Glycogen; Liver; Mice

Adaptation to hypoxia makes the heart more oxygen efficient, by metabolising more glucose. In contrast, type 2 diabetes makes the heart metabolise more fatty acids. Diabetes increases the chances of the heart being exposed to hypoxia, but whether the diabetic heart can adapt and respond is unknown. In this study we show that diabetic hearts retain the ability to adapt their metabolism in response to hypoxia, with functional hypoxia signalling pathways. However, the hypoxia-induced changes in metabolism are additive to abnormal baseline metabolism, resulting in hypoxic diabetic hearts metabolising more fat and less glucose than controls. This stops the diabetic heart being able to recover its function when stressed. These results demonstrate that the diabetic heart retains metabolic flexibility to adapt to hypoxia, but is hindered by the baseline effects of the disease. This increases our understanding of how the diabetic heart is affected by hypoxia-associated complications of the disease.. Hypoxia activates the hypoxia-inducible factor (HIF), promoting glycolysis and suppressing mitochondrial respiration. In the type 2 diabetic heart, glycolysis is suppressed whereas fatty acid metabolism is promoted. The diabetic heart experiences chronic hypoxia as a consequence of increased obstructive sleep apnoea and cardiovascular disease. Given the opposing metabolic effects of hypoxia and diabetes, we questioned whether diabetes affects cardiac metabolic adaptation to hypoxia. Control and type 2 diabetic rats were housed for 3 weeks in normoxia or 11% oxygen. Metabolism and function were measured in the isolated perfused heart using radiolabelled substrates. Following chronic hypoxia, both control and diabetic hearts upregulated glycolysis, lactate efflux and glycogen content and decreased fatty acid oxidation rates, with similar activation of HIF signalling pathways. However, hypoxia-induced changes were superimposed on diabetic hearts that were metabolically abnormal in normoxia, resulting in glycolytic rates 30% lower, and fatty acid oxidation 36% higher, in hypoxic diabetic hearts than hypoxic controls. Peroxisome proliferator-activated receptor α target proteins were suppressed by hypoxia, but activated by diabetes. Mitochondrial respiration in diabetic hearts was divergently activated following hypoxia compared with controls. These differences in metabolism were associated with decreased contractile recovery of the hypoxic diabetic heart following an acute hypoxic insult. In conclusion, type 2 diabetic hearts retain metabolic flexibility to adapt to hypoxia, with normal HIF signalling pathways. However, they are more dependent on oxidative metabolism following hypoxia due to abnormal normoxic metabolism, which was associated with a functional deficit in response to stress.

Topics: Adaptation, Physiological; Animals; Cell Hypoxia; Diabetes Mellitus, Type 2; Diabetic Cardiomyopathies; Glycogen; Glycolysis; Lactic Acid; Male; Mitochondria, Muscle; Myocardium; Oxidative Stress; Oxygen; PPAR gamma; Rats; Rats, Wistar; Signal Transduction

The present study was to evaluate the beneficial effect of polysaccharide isolated from Ganoderma atrum (PSG-1) on liver function in type 2 diabetic rats. Results showed that PSG-1 decreased the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), while increasing hepatic glycogen levels. PSG-1 also exerted strong antioxidant activities, together with upregulated mRNA expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), glucose transporter-4 (GLUT4), phosphoinositide 3-kinase (PI3K), and phosphorylated-Akt (p-Akt) in the liver of diabetic rats. Moreover, the concentrations of short-chain fatty acids (SCFA) were significantly higher in the liver, serum, and faeces of diabetic rats after treating with PSG-1 for 4 weeks. These results suggest that the improvement of PSG-1 on liver function in type 2 diabetic rats may be due to its antioxidant effects, SCFA excretion in the colon from PSG-1, and regulation of hepatic glucose uptake by inducing GLUT4 translocation through PI3K/Akt signaling pathways.

Topics: Alanine Transaminase; Animals; Antioxidants; Aspartate Aminotransferases; Diabetes Mellitus, Type 2; Fatty Acids, Volatile; Feces; Ganoderma; Gene Expression; Glucose Transporter Type 4; Glycogen; Liver; Male; Phosphatidylinositol 3-Kinases; Polysaccharides; PPAR gamma; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; RNA, Messenger; Signal Transduction

The regulation of microRNAs (miRNAs) at different stages of the progression of type 2 diabetes mellitus (T2DM) and their role in glucose homeostasis was investigated.. Microarrays were used to assess miRNA expression in skeletal muscle biopsies taken from healthy individuals and patients with pre-diabetes or T2DM, and insulin resistant offspring of rat dams fed a high fat diet during pregnancy.. Twenty-three miRNAs were differentially expressed in patients with T2DM, and 7 in the insulin resistant rat offspring compared to their controls. Among these, only one miRNA was similarly regulated: miR-194 expression was significantly reduced by 25 to 50% in both the rat model and in human with pre-diabetes and established diabetes. Knockdown of miR-194 in L6 skeletal muscle cells induced an increase in basal and insulin-stimulated glucose uptake and glycogen synthesis. This occurred in conjunction with an increased glycolysis, indicated by elevated lactate production. Moreover, oxidative capacity was also increased as we found an enhanced glucose oxidation in presence of the mitochondrial uncoupler FCCP. When miR-194 was down-regulated in vitro, western blot analysis showed an increased phosphorylation of AKT and GSK3β in response to insulin, and an increase in expression of proteins controlling mitochondrial oxidative phosphorylation.. Type 2 diabetes mellitus is associated with regulation of several miRNAs in skeletal muscle. Interestingly, miR-194 was a unique miRNA that appeared regulated across different stages of the disease progression, from the early stages of insulin resistance to the development of T2DM. We have shown miR-194 is involved in multiple aspects of skeletal muscle glucose metabolism from uptake, through to glycolysis, glycogenesis and glucose oxidation, potentially via mechanisms involving AKT, GSK3 and oxidative phosphorylation. MiR-194 could be down-regulated in patients with early features of diabetes as an adaptive response to facilitate tissue glucose uptake and metabolism in the face of insulin resistance.

Topics: Animals; Cell Line; Diabetes Mellitus, Type 2; Diet, High-Fat; Disease Models, Animal; Female; Gene Expression Regulation; Glucose; Glycogen; Glycogen Synthase Kinase 3; Humans; Insulin; Insulin Resistance; Male; Mice, Inbred C57BL; MicroRNAs; Mitochondria; Muscle, Skeletal; Myoblasts; Oxidative Phosphorylation; Prediabetic State; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction

There are several claims about the beneficial effects of supplementing L-glutamine to both type 1 and type 2 diabetes. The purpose of the present study was to provide detailed knowledge about the fate of this amino acid in the liver, the first organ that receives the compound when ingested orally. The study was done using the isolated perfused rat liver, an experimental system that preserves the microcirculation of the organ and that allows to measured several parameters during steady-state and pre steady-state conditions. L-Glutamine was infused in the portal vein (5 mM) and several parameters were monitored. Livers from type 1 diabetic rats showed an accelerated response to L-glutamine infusion. In consequence of this accelerated response livers from type 1 diabetic rats presented higher rates of ammonia, urea, glucose and lactate output during the first 25-30 minutes following L-glutamine infusion. As steady-state conditions approached, however, the difference between type 1 diabetes and control livers tended to disappear. Measurement of the glycogen content over a period of 100 minutes revealed that, excepting the initial phase of the L-glutamine infusion, the increased glucose output in livers from type 1 diabetic rats was mainly due to accelerated glycogenolysis. Livers from type 2 diabetic rats behaved similarly to control livers with no accelerated glucose output but with increased L-alanine production. L-Alanine is important for the pancreatic β-cells and from this point of view the oral intake of L-glutamine can be regarded as beneficial. Furthermore, the lack of increased glucose output in livers from type 2 diabetic rats is consistent with observations that even daily L-glutamine doses of 30 g do not increase the glycemic levels in well controlled type 2 diabetes patients.

Topics: Alanine; Ammonia; Animals; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Gluconeogenesis; Glucose; Glutamine; Glycogen; Lactic Acid; Liver; Male; Oxygen; Rats; Rats, Wistar; Urea

This study aims to establish the corneal nerve fiber (CNF) morphological alterations in a large cohort of type 2 diabetic patients and to investigate the association between the bead size, a novel parameter representing composite of accumulated mitochondria, glycogen particles, and vesicles in CNF, and the neurophysiological dysfunctions of the peripheral nerves. 162 type 2 diabetic patients and 45 healthy control subjects were studied in detail with a battery of clinical and neurological examinations and corneal confocal microscopy. Compared with controls, patients had abnormal CNF parameters. In particular the patients had reduced density and length of CNF and beading frequency and increased bead size. Alterations in CNF parameters were significant even in patients without neuropathy. The HbA1c levels were tightly associated with the bead size, which was inversely related to the motor and sensory nerve conduction velocity (NCV) and to the distal latency period of the median nerve positively. The CNF density and length positively correlated with the NCV and amplitude. The hyperglycemia-induced expansion of beads in CNF might be a predictor of slow NCV in peripheral nerves in type 2 diabetic patients.

Topics: Case-Control Studies; Cornea; Diabetes Mellitus, Type 2; Diabetic Neuropathies; Female; Glycated Hemoglobin; Glycogen; Humans; Intravital Microscopy; Male; Median Nerve; Microscopy, Confocal; Middle Aged; Mitochondria; Nerve Fibers, Unmyelinated; Neural Conduction; Peripheral Nerves; Sensory Thresholds; Synaptic Vesicles

In the postprandial state, the liver regulates glucose homeostasis by glucose uptake and conversion to glycogen and lipids. Glucose and insulin signalling finely regulate glycogen synthesis through several mechanisms. Glucose uptake in hepatocytes is favoured by the insulin receptor isoform A (IRA), rather than isoform B (IRB). Thus, we hypothesised that, in hepatocytes, IRA would increase glycogen synthesis by promoting glucose uptake and glycogen storage.. We addressed the role of insulin receptor isoforms on glycogen metabolism in vitro in immortalised neonatal hepatocytes. In vivo, IRA or IRB were specifically expressed in the liver using adeno-associated virus vectors in inducible liver insulin receptor knockout (iLIRKO) mice, a model of type 2 diabetes. The role of IR isoforms in glycogen synthesis and storage in iLIRKO was subsequently investigated.. In immortalised hepatocytes, IRA, but not IRB expression induced an increase in insulin signalling that was associated with elevated glycogen synthesis, glycogen synthase activity and glycogen storage. Similarly, elevated IRA, but not IRB expression in the livers of iLIRKO mice induced an increase in glycogen content.. We provide new insight into the role of IRA in the regulation of glycogen metabolism in cultured hepatocytes and in the livers of a mouse model of type 2 diabetes. Our data strongly suggest that IRA is more efficient than IRB at promoting glycogen synthesis and storage. Therefore, we suggest that IRA expression in the liver could provide an interesting therapeutic approach for the regulation of hepatic glucose content and glycogen storage.

Topics: Animals; Blotting, Western; Cell Line; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Phosphorylase; Glycogen Synthase; Glycogenolysis; Hepatocytes; Liver; Liver Glycogen; Mice; Mice, Knockout; Protein Isoforms; Receptor, Insulin

AMPK dysregulation contributes to the onset and development of type 2 diabetes (T2DM). AMPK is known to be activated by reactive oxygen species (ROS) and antioxidant interference. However the mechanism by which redox state mediates such contradictory result remains largely unknown. Here we used streptozotocin-high fat diet （STZ-HFD） induced-type 2 diabetic rats and cells lines (L02 and HEK 293) to explore the mechanism of redox-mediated AMPK activation. We show glutaredoxins (Grxs) concomitant with optimal ROS act as an essential mediator for AMPK activation. ROS level results in different mechanisms for AMPK activation. Under low ROS microenvironment, Grxs-mediated S-glutathionylation on AMPK-α catalytic subunit activates AMPK to improve glucose transportation and degradation while inhibiting glycogen synthesis and keeping redox balance. While, under high ROS microenvironment, AMPK is activated by an AMP-dependent mechanism, however sustained high level ROS also causes loss of AMPK protein. This finding provides evidence for a new approach to diabetes treatment by individual doses of ROS or antioxidant calibrated against the actual redox level in vivo. Moreover, the novel function of Grxs in promoting glucose metabolism may provide new target for T2DM treatment.

Topics: AMP-Activated Protein Kinases; Animals; Cell Line; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Epithelial Cells; Gene Expression Regulation; Glucose; Glutaredoxins; Glycogen; HEK293 Cells; Hepatocytes; Humans; Liver; Male; Oxidation-Reduction; Protein Subunits; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Signal Transduction; Streptozocin

Nonalcoholic fatty liver disease (NAFLD) is a risk factor for type 2 diabetes (T2D), but whether NAFLD plays a causal role in the pathogenesis of T2D is uncertain. One proposed mechanism linking NAFLD to hepatic insulin resistance involves diacylglycerol-mediated (DAG-mediated) activation of protein kinase C-ε (PKCε) and the consequent inhibition of insulin receptor (INSR) kinase activity. However, the molecular mechanism underlying PKCε inhibition of INSR kinase activity is unknown. Here, we used mass spectrometry to identify the phosphorylation site Thr1160 as a PKCε substrate in the functionally critical INSR kinase activation loop. We hypothesized that Thr1160 phosphorylation impairs INSR kinase activity by destabilizing the active configuration of the INSR kinase, and our results confirmed this prediction by demonstrating severely impaired INSR kinase activity in phosphomimetic T1160E mutants. Conversely, the INSR T1160A mutant was not inhibited by PKCε in vitro. Furthermore, mice with a threonine-to-alanine mutation at the homologous residue Thr1150 (InsrT1150A mice) were protected from high fat diet-induced hepatic insulin resistance. InsrT1150A mice also displayed increased insulin signaling, suppression of hepatic glucose production, and increased hepatic glycogen synthesis compared with WT controls during hyperinsulinemic clamp studies. These data reveal a critical pathophysiological role for INSR Thr1160 phosphorylation and provide further mechanistic links between PKCε and INSR in mediating NAFLD-induced hepatic insulin resistance.

Topics: Amino Acid Substitution; Animals; Diabetes Mellitus, Type 2; Dietary Fats; Glycogen; Insulin Resistance; Liver; Mice; Mice, Mutant Strains; Mutation, Missense; Non-alcoholic Fatty Liver Disease; Phosphorylation; Protein Kinase C-epsilon; Receptor, Insulin; Signal Transduction

Sang-Tong-Jian (STJ), a novel formula composed of flavonoids and alkaloids derived from mulberry leaf, has been found to reduce blood glucose levels in rats with type 2 diabetes mellitus (T2DM) in our previous studies. However, the precise mechanisms remain unknown. Insulin resistance is the main characteristic of T2DM, which may be due to impairment of the PI3K/AKT signaling pathway. In this study, we investigated the effects of STJ on glycometabolism and insulin resistance in KKAy mice.. A total of 50 KKAy male mice were randomly divided into five groups: model, metformin at 260mg/kg, and STJ at 105, 210 and 420mg/kg. C57BL/6J mice were used as the control group. Random blood glucose levels in KKAy mice were determined every 10days after treatments. At the 10th and 13th week, oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were conducted after a 12h overnight fast, respectively. After 13-week treatments, glycosylated hemoglobin (GHb) and serum insulin were measured using a colorimetric method and ELISA kits. Liver glycogen and muscle glycogen levels were analyzed using a colorimetric method. The morphology of pancreas, liver, skeletal muscle and epididymal fat were visualized by haematoxylin and eosin staining. The gene level of GLUT2 (liver) and GLUT4 (skeletal muscle, epididymal fat) were detected by real-time PCR. The proteins of GLUT2, GLUT4, IRS1, PI3K, AKT and their phosphorylation were assayed by Western blot analyses.. STJ significantly decreased the random blood glucose and GHb levels, and increased liver and muscle glycogen levels. The results of OGTT and ITT and measurement of serum insulin indicated that STJ ameliorated insulin resistance in KKAy mice. STJ treatments also ameliorated the histopathological alterations in pancreas, liver, skeletal muscle and epididymal fat in KKAy mice. Furthermore, STJ upregulated the gene and protein expression of GLUT2 (liver) and GLUT4 (skeletal muscle, epididymal fat). Meanwhile, GLUT4 translocation and phosphorylation of IRS1, p85-PI3K and AKT were significantly increased by STJ treatments.. Our results indicated that STJ ameliorated glycometabolism and insulin resistance in KKAy mice, which might be due to activation of PI3K/AKT pathway.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Drugs, Chinese Herbal; Gene Expression Regulation; Glucose; Glucose Tolerance Test; Glucose Transporter Type 2; Glucose Transporter Type 4; Glycated Hemoglobin; Glycogen; Insulin; Insulin Resistance; Male; Mice; Mice, Inbred C57BL; Organ Specificity; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; RNA, Messenger; Signal Transduction

Diabetic nephropathy is derived from long-term effects of high blood glucose on kidney function in type 2 diabetic patients. Several antidiabetic drugs and herbal medications have failed to prevent episodes of DN. Hence, this study aimed to further investigate the renal injury-reducing effect of antidiabetic CmNo1, a novel combination of powders of fruiting bodies and mycelia of Cordyceps militaris. After being administered with streptozotocin-nicotinamide and high-fat-diet, the diabetic nephropathy mouse model displayed elevated blood glucose and renal dysfunction markers including serum creatinine and kidney-to-body weight ratio. These elevated markers were significantly mitigated following 8 weeks CmNo1 treatment. Moreover, the chronic hyperglycemia-induced pathological alteration in renal tissue were also ameliorated. Besides, immunohistochemical study demonstrated a substantial reduction in elevated levels of carboxymethyl lysine, an advanced glycation end product. Elevated collagenous deposition in DN group was also attenuated through CmNo1 administration. Moreover, the enhanced levels of transforming growth factor-β1, a fibrosis-inducing protein in glomerulus were also markedly dampened. Furthermore, auxiliary risk factors in DN like serum triglycerides and cholesterol were found to be increased but were decreased by CmNo1 treatment. Conclusively, the results suggests that CmNo1 exhibit potent and efficacious renoprotective action against hyperglycemia-induced DN.

Topics: Animals; Biological Products; Collagen; Cordyceps; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Fruiting Bodies, Fungal; Glycation End Products, Advanced; Glycogen; Hypoglycemic Agents; Kidney; Kidney Function Tests; Mice; Mice, Inbred C57BL; Mycelium; Streptozocin; Transforming Growth Factor beta1

Insulin secretion from pancreatic β-cells is impaired in all forms of diabetes. The resultant hyperglycaemia has deleterious effects on many tissues, including β-cells. Here we show that chronic hyperglycaemia impairs glucose metabolism and alters expression of metabolic genes in pancreatic islets. In a mouse model of human neonatal diabetes, hyperglycaemia results in marked glycogen accumulation, and increased apoptosis in β-cells. Sulphonylurea therapy rapidly normalizes blood glucose levels, dissipates glycogen stores, increases autophagy and restores β-cell metabolism. Insulin therapy has the same effect but with slower kinetics. Similar changes are observed in mice expressing an activating glucokinase mutation, in in vitro models of hyperglycaemia, and in islets from type-2 diabetic patients. Altered β-cell metabolism may underlie both the progressive impairment of insulin secretion and reduced β-cell mass in diabetes.

Topics: Animals; Apoptosis; Autophagy; Blood Glucose; Cell Line; Diabetes Mellitus, Type 2; Disease Models, Animal; Glucokinase; Glycogen; Humans; Hyperglycemia; Hypoglycemic Agents; In Vitro Techniques; Infant, Newborn; Infant, Newborn, Diseases; Insulin; Insulin-Secreting Cells; Mice; Mutation; Rats; Sulfonylurea Compounds

The IV anesthetic, propofol, when administered as fat emulsion-based formulation (Diprivan) promotes insulin resistance, but the direct effects of propofol and its solvent, Intralipid, on cardiac insulin resistance are unknown.. Hearts of healthy and type-2 diabetic rats (generated by fructose feeding) were aerobically perfused for 60 minutes with 10 μM propofol in the formulation of Diprivan or an equivalent concentration of its solvent Intralipid (25 μM) ± insulin (100 mU•L). Glucose uptake, glycolysis, and glycogen metabolism were measured using [H]glucose. Activation of Akt, GSK3β, AMPK, ERK1/2, p38MAPK, S6K1, JNK, protein kinase Cθ (PKCθ), and protein kinase CCβII (PKCβII) was determined using immunoblotting. GLUT4 trafficking and phosphorylations of insulin receptor substrate-1 (IRS-1) at Ser307(h312), Ser1100(h1101), and Tyr608(hTyr612) were measured. Mass spectrometry was used to determine acylcarnitines, phospholipids, and sphingolipids.. Diprivan and Intralipid reduced insulin-induced glucose uptake and redirected glucose to glycogen stores in diabetic hearts. Reduced glucose uptake was accompanied by lower GLUT4 trafficking to the sarcolemma. Diprivan and Intralipid inactivated GSK3β but activated AMPK and ERK1/2 in diabetic hearts. Only Diprivan increased phosphorylation of Akt(Ser473/Thr308) and translocated PKCθ and PKCβII to the sarcolemma in healthy hearts, whereas it activated S6K1 and p38MAPK and translocated PKCβII in diabetic hearts. Furthermore, only Diprivan phosphorylated IRS-1 at Ser1100(h1101) in healthy and diabetic hearts. JNK expression, phosphorylation of Ser307(h312) of IRS-1, and PKCθ expression and translocation were increased, whereas GLUT4 expression was reduced in insulin-treated diabetic hearts. Phosphatidylglycerol, phosphatidylethanolamine, and C18-sphingolipids accumulated in Diprivan-perfused and Intralipid-perfused diabetic hearts.. Propofol and Intralipid promote insulin resistance predominantly in type-2 diabetic hearts.

Topics: Anesthetics, Intravenous; Animals; Citrate (si)-Synthase; Diabetes Mellitus, Type 2; Emulsions; Fat Emulsions, Intravenous; Fructose; Glucose; Glucose Transporter Type 4; Glycogen; Glycolysis; Heart; Insulin Resistance; Male; Phospholipids; Propofol; Rats; Rats, Sprague-Dawley; Soybean Oil

Nymphaea stellata (Willd.) has been used in traditional medicine for centuries to treat several illnesses, including diabetes. However, scientific evidence supporting its mechanism of action is lacking. Here, we showed that an N. stellata flower chloroform extract (NSFCExt) has significant plasma glucose lowering ability. Furthermore, an active compound was identified and purified by column chromatography, and the structure of this compound, nymphayol, was determined by X-ray crystallographic analysis. Nymphayol was tested for its effects on insulin secretion by RIN-5F cells cultured in low or high glucose medium; we found that nymphayol treatment improved glucose-stimulated insulin secretion in vitro. Additionally, insulin sensitization and glucose uptake were increased in L6 myotubes. Nymphayol was administered to type 2 diabetic male Wistar rats at several doses (5, 10 or 20 mg/kg/day) for 45 days. After nymphayol administration, the plasma glucose concentration was significantly (p⩽0.05) lower (60.33%) than in control diabetic rats, and the plasma insulin level increased in a dose-dependent manner. In addition, the cellular insulin response was analyzed in type 2 diabetic rats; oral administration of nymphayol increased IRS1 phosphorylation and GLUT4 protein expression in liver and muscle. Nymphayol significantly (p⩽0.05) restored the levels of HbA1c, hepatic glycogen and hepatic glucose-metabolizing enzyme (hexokinase, glucose-6-phosphate dehydrogenase, glucose-6-phosphatase, fructose-1, 6-bisphosphatase, glycogen synthase and glycogen phosphorylase) activity in diabetic rats. The administration of glibenclamide, a reference drug (600 μg/kg), also produced a significant (p⩽0.05) reduction in blood glucose in STZ-nicotinamide induced diabetic rats. The results suggest that nymphayol may be a useful therapy for diabetes because it stimulates insulin secretion and promotes glucose absorption.

Topics: Animals; Biological Transport; Blood Glucose; Body Weight; Cell Line; Cell Proliferation; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Drinking; Gene Expression Regulation; Glucose; Glucose Transporter Type 4; Glycated Hemoglobin; Glycogen; Insulin; Insulin Receptor Substrate Proteins; Insulin Resistance; Insulin Secretion; Insulin-Secreting Cells; Liver; Male; Muscle Fibers, Skeletal; Phytosterols; Rats; Rats, Wistar; Reactive Oxygen Species

The present study was designed to evaluate the potential hypoglycemic and hypolipidemic effects of Angelica sinensis polysaccharide (ASP), purified from the fresh roots of Angelica sinensis (AS), in prediabetic and streptozotocin (STZ)-induced diabetic BALB/c mice. It was observed that fasting blood glucose (FBG) levels in both models were reduced after a 4-week oral administration of ASP or metformin, and abnormal fasting serum insulin (FINS) concentrations were ameliorated as well. Moreover, the homeostasis model assessment-insulin resistance (HOMA-IR) index was decreased strikingly and body weight (BW) was reduced significantly in prediabetic mice after treatment with ASP. In addition, ASP also contributed to improving the dyslipidemia conditions. Elevated serum total cholesterol (TC) or triglyceride (TG) concentrations were reduced after treatment with ASP in prediabetic mice or STZ-induced diabetic mice. Meanwhile, hepatic glycogen (HG) and muscle glycogen (MG) concentrations were increased while insulin resistance (IR)-related inflammatory factors IL-6 and TNF-α in serum were reduced in STZ-induced diabetic mice. Histopathological examination indicated that the impaired pancreatic/hepatic tissues or adipose tissues were effectively restored in STZ-induced diabetic mice or prediabetic mice after the ASP treatment. Taken together, these results revealed that ASP efficiently exerted hypoglycemic and hypolipidemic benefits, and its potential effect was associated with the amelioration of IR. ASP can be applied in the prevention and treatment of diabetes.

Topics: Adipose Tissue, White; Angelica sinensis; Animals; Anti-Inflammatory Agents, Non-Steroidal; Diabetes Mellitus, Type 2; Glycogen; Hyperglycemia; Hyperinsulinism; Hyperlipidemias; Hypoglycemic Agents; Hypolipidemic Agents; Insulin Resistance; Lipid Metabolism Disorders; Liver; Male; Mice, Inbred BALB C; Muscle, Skeletal; Pancreas; Plant Roots; Polysaccharides; Prediabetic State; Random Allocation

TWIST proteins are important for development of embryonic skeletal muscle and play a role in the metabolism of tumor and white adipose tissue. The impact of TWIST on metabolism in skeletal muscle is incompletely studied. Our aim was to assess the impact of TWIST1 and TWIST2 overexpression on glucose and lipid metabolism. In intact mouse muscle, overexpression of Twist reduced total glycogen content without altering glucose uptake. Expression of TWIST1 or TWIST2 reduced Pdk4 mRNA, while increasing mRNA levels of Il6, Tnfα, and Il1β. Phosphorylation of AKT was increased and protein abundance of acetyl CoA carboxylase (ACC) was decreased in skeletal muscle overexpressing TWIST1 or TWIST2. Glycogen synthesis and fatty acid oxidation remained stable in C2C12 cells overexpressing TWIST1 or TWIST2. Finally, skeletal muscle mRNA levels remain unaltered in ob/ob mice, type 2 diabetic patients, or in healthy subjects before and after 3 months of exercise training. Collectively, our results indicate that TWIST1 and TWIST2 are expressed in skeletal muscle. Overexpression of these proteins impacts proteins in metabolic pathways and mRNA level of cytokines. However, skeletal muscle levels of TWIST transcripts are unaltered in metabolic diseases.

Topics: Animals; Case-Control Studies; Cells, Cultured; Diabetes Mellitus, Type 2; Female; Gene Expression Regulation; Glycogen; Humans; Inflammation; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Mice, Transgenic; Middle Aged; Muscle, Skeletal; Nuclear Proteins; Repressor Proteins; Twist-Related Protein 1

Circulating redox state changes, determined by the ratio of reduced/oxidized pairs of different metabolites, have been associated with metabolic diseases. However, the pathogenic contribution of these changes and whether they modulate normal tissue function is unclear. As alterations in hepatic gluconeogenesis and glycogen metabolism are hallmarks that characterize insulin resistance and type 2 diabetes, we tested whether imposed changes in the extracellular redox state could modulate these processes. Thus, primary hepatocytes were treated with different ratios of the following physiological extracellular redox couples: β-hydroxybutyrate (βOHB)/acetoacetate (Acoc), reduced glutathione (GSH)/oxidized glutathione (GSSG), and cysteine/cystine. Exposure to a more oxidized ratio via extracellular βOHB/Acoc, GSH/GSSG, and cysteine/cystine in hepatocytes from fed mice increased intracellular hydrogen peroxide without causing oxidative damage. On the other hand, addition of more reduced ratios of extracellular βOHB/Acoc led to increased NAD(P)H and maximal mitochondrial respiratory capacity in hepatocytes. Greater βOHB/Acoc ratios were also associated with decreased β-oxidation, as expected with enhanced lipogenesis. In hepatocytes from fasted mice, a more extracellular reduced state of βOHB/Acoc led to increased alanine-stimulated gluconeogenesis and enhanced glycogen synthesis capacity from added glucose. Thus, we demonstrated for the first time that the extracellular redox state regulates the major metabolic functions of the liver and involves changes in intracellular NADH, hydrogen peroxide, and mitochondrial respiration. Because redox state in the blood can be communicated to all metabolically sensitive tissues, this work confirms the hypothesis that circulating redox state may be an important regulator of whole body metabolism and contribute to alterations associated with metabolic diseases.

Topics: 3-Hydroxybutyric Acid; Acetoacetates; Animals; Cysteine; Cystine; Diabetes Mellitus, Type 2; Gluconeogenesis; Glutathione; Glutathione Disulfide; Glycogen; Hepatocytes; Humans; Hydrogen Peroxide; Insulin Resistance; Mice; Mitochondria; NAD; Oxidation-Reduction; Respiration

A decrease in skeletal muscle lipolysis and hormone sensitive-lipase (HSL) expression has been linked to insulin resistance in obesity. The purpose of this study was to identify potential intrinsic defects in lipid turnover and lipolysis in myotubes established from obese and type 2 diabetic subjects. Lipid trafficking and lipolysis were measured by pulse-chase assay with radiolabeled substrates in myotubes from non-obese/non-diabetic (lean), obese/non-diabetic (obese) and obese/diabetic (T2D) subjects. Lipolytic protein content and level of Akt phosphorylation were measured by Western blot. HSL was overexpressed by adenovirus-mediated gene delivery. Myotubes established from obese and T2D subjects had lower lipolysis (-30-40%) when compared to lean, using oleic acid as precursor. Similar observations were also seen for labelled glycerol. Incorporation of oleic acid into diacylglycerol (DAG) and free fatty acid (FFA) level was lower in T2D myotubes, and acetate incorporation into FFA and complex lipids was also lower in obese and/or T2D subjects. Both protein expression of HSL (but not ATGL) and changes in DAG during lipolysis were markedly lower in cells from obese and T2D when compared to lean subjects. Insulin-stimulated glycogen synthesis (-60%) and Akt phosphorylation (-90%) were lower in myotubes from T2D, however, overexpression of HSL in T2D myotubes did not rescue the diabetic phenotype. In conclusion, intrinsic defects in lipolysis and HSL expression co-exist with reduced insulin action in myotubes from obese T2D subjects. Despite reductions in intramyocellular lipolysis and HSL expression, overexpression of HSL did not rescue defects in insulin action in skeletal myotubes from obese T2D subjects.

Topics: Biological Transport; Carbon Radioisotopes; Diabetes Mellitus, Type 2; Diglycerides; Female; Gene Expression Regulation; Glycerol; Glycogen; Humans; Insulin; Lipolysis; Male; Middle Aged; Muscle Fibers, Skeletal; Obesity; Oleic Acid; Primary Cell Culture; Proto-Oncogene Proteins c-akt; Signal Transduction; Sterol Esterase

Melicope lunu-ankenda leaves are used to treat diabetes in folklore medicinal practices in India and Malaysia. Here we report the isolation of an O-prenylated flavonoid (3,5,4'-trihydroxy-8,3'-dimethoxy-7-(3-methylbut-2-enoxy)flavone; OPF) from the leaves of M. lunu-ankenda and its antidiabetes activity against type-2 diabetes mellitus (T2DM).. OPF was isolated from M. lunu-ankenda leaves by extraction and repeated column chromatography and its structure was elucidated by IR, UV-vis, 1D-, 2D-NMR and mass spectral analyses. Blood glucose lowering activity of OPF was tested in normal rats by oral glucose tolerance test and its efficacy was tested in STZ-induced type-2 diabetic rats. SGOT, SGPT, ALP, serum urea, total triglycerides, total cholesterol and reduction in HDLC, protein and serum insulin levels in normal rats and STZ-induced type-2 diabetic rats were measured. Acute toxicity of OPF was tested at 500 mg/kg dose. Mechanism of antidiabetes action of OPF was elucidated by insulin release from RIN 5F cells.. OPF isolated from M. lunu-ankenda showed significant blood glucose lowering activity in oral glucose tolerance test on overnight fasted, glucose loaded normal rats and the optimum activity was observed at a dose of 10mg/kg body weight. In neonatal streptozotocin (STZ) induced diabetic rats, the OPF treatment for 20 days significantly ameliorated the derailed blood glucose levels, liver glycogen and serum biological parameters including insulin to normal levels. OPF on acute toxicity evaluation did not show any conspicuous toxic symptoms even at a higher dose of 500 mg/kg body weight in mice. On evaluating the mechanism of antidiabetes action, it was observed that, OPF induced insulin release from cultured RIN 5F cells in vitro from which it was evident that the OPF acts on pancreatic β-cells for insulin release thereby correcting the derailed blood glucose levels, serum biochemical parameters and ameliorate various diabetic complications in STZ-induced diabetic rats.. This study shows the potent antidiabetes activity of OPF and describes its mechanism of action. OPF is a promising candidate for the development of new generation anti-DM drugs. Isolation of the O-prenylated flavonoid justifies the use of M. lunu-ankenda for diabetic treatments in folklore practices.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Flavonoids; Glycogen; Hypoglycemic Agents; Liver; Male; Mice; Plant Leaves; Prenylation; Rats, Wistar; Rutaceae; Toxicity Tests, Acute

Insulin and exercise stimulate skeletal muscle glycogen synthase (GS) activity by dephosphorylation and changes in kinetic properties. The aim of this study was to investigate the effects of insulin, exercise and post-exercise insulin stimulation on GS phosphorylation, activity and substrate affinity in obesity and type 2 diabetes.. Obese men with type 2 diabetes (n = 13) and weight-matched controls (n = 14) underwent euglycaemic-hyperinsulinaemic clamps in the rested state and 3 h after 60 min of cycling (70% maximal pulmonary oxygen uptake [VO2max]). Biopsies from vastus lateralis muscle were obtained before and after clamps, and before and immediately after exercise.. Insulin-stimulated glucose uptake was lower in diabetic patients vs obese controls with or without prior exercise. Post exercise, glucose partitioning shifted away from oxidation and towards storage in both groups. Insulin and, more potently, exercise increased GS activity (fractional velocity [FV]) and substrate affinity in both groups. Both stimuli caused dephosphorylation of GS at sites 3a + 3b, with exercise additionally decreasing phosphorylation at sites 2 + 2a. In both groups, changes in GS activity, substrate affinity and dephosphorylation at sites 3a + 3b by exercise were sustained 3 h post exercise and further enhanced by insulin. Post exercise, reduced GS activity and substrate affinity as well as increased phosphorylation at sites 2 + 2a were found in diabetic patients vs obese controls.. Exercise-induced activation of muscle GS in obesity and type 2 diabetes involves dephosphorylation of GS at sites 3a + 3b and 2 + 2a and enhanced substrate affinity, which is likely to facilitate glucose partitioning towards storage. Lower GS activity and increased phosphorylation at sites 2 + 2a in type 2 diabetes in the recovery period imply an impaired response to exercise.

Topics: Bicycling; Biopsy; Cohort Studies; Diabetes Mellitus, Type 2; Exercise; Glucose Clamp Technique; Glycogen; Glycogen Synthase; Humans; Hypoglycemic Agents; Insulin; Kinetics; Male; Middle Aged; Muscle, Skeletal; Obesity; Phosphorylation; Uridine Diphosphate Glucose

Type 2 diabetes, obesity, and sex difference affect myocardial glucose uptake and utilization. However, their effect on the intramyocellular fate of glucose in humans has been unknown. How the heart uses glucose is important, because it affects energy production and oxygen efficiency, which in turn affect heart function and adaptability. We hypothesized that type 2 diabetes, sex difference, and obesity affect myocardial glucose oxidation, glycolysis, and glycogen production. In a first-in-human study, we measured intramyocardiocellular glucose metabolism from time-activity curves generated from previously obtained positron emission tomography scans of 110 subjects in 3 groups: nonobese, obese, and diabetes. Group and sex difference interacted in the prediction of all glucose uptake, utilization, and metabolism rates. Group independently predicted fractional glucose uptake and its components: glycolysis, glycogen deposition, and glucose oxidation rates. Sex difference predicted glycolysis rates. However, there were fewer differences in glucose metabolism between diabetic patients and others when plasma glucose levels were included in the modeling. The potentially detrimental effects of obesity and diabetes on myocardial glucose metabolism are more pronounced in men than women. This sex difference dimorphism needs to be taken into account in the design, trials, and application of metabolic modulator therapy. Slightly higher plasma glucose levels improve depressed glucose oxidation and glycogen deposition rates in diabetic patients.

Topics: Adult; Aged; Blood Glucose; Diabetes Mellitus, Type 2; Energy Metabolism; Female; Glycogen; Glycolysis; Hemodynamics; Humans; Kinetics; Male; Middle Aged; Myocardium; Obesity; Oxidation-Reduction; Positron-Emission Tomography; Sex Factors; Young Adult

Our previous study showed that chromium malate improved the regulation of blood glucose in mice with alloxan-induced diabetes. The present study was designed to evaluate the effect of chromium malate on glycometabolism, glycometabolism-related enzymes and lipid metabolism in type 2 diabetic rats. Our results showed that fasting blood glucose, serum insulin level, insulin resistance index and C-peptide level in the high dose group had a significant downward trend when compared with the model group, chromium picolinate group and chromium trichloride group. The hepatic glycogen, glucose-6-phosphate dehydrogenase, glucokinase, Glut4, phosphor-AMPKβ1 and Akt levels in the high dose group were significantly higher than those of the model, chromium picolinate and chromium trichloride groups. Chromium malate in a high dose group can significantly increase high density lipoprotein cholesterol level while decreasing the total cholesterol, low density lipoprotein cholesterol and triglyceride levels when compared with chromium picolinate and chromium trichloride. The serum chromium content in chromium malate and chromium picolinate group is significantly higher than that of the chromium trichloride group. The results indicated that the curative effects of chromium malate on glycometabolism, glycometabolism-related enzymes and lipid metabolism changes are better than those of chromium picolinate and chromium trichloride. Chromium malate contributes to glucose uptake and transport in order to improved glycometabolism and glycometabolism-related enzymes.

Topics: Animals; Biological Transport; Blood Glucose; Body Weight; Chromium; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dietary Supplements; Disease Models, Animal; Fasting; Gastrointestinal Microbiome; Glucose; Glycogen; Insulin; Lipid Metabolism; Liver; Malates; Male; Maze Learning; Rats

The incidence of type 2 diabetes mellitus and its prodromal stage, pre-diabetes, is rapidly increasing among young men, leading to disturbances in testosterone synthesis. However, the impact of testosterone deficiency induced by these progressive stages of diabetes on the metabolic behavior of Sertoli cells remains unknown. We evaluated the effects of testosterone deficiency associated with pre-diabetes and type 2 diabetes on Sertoli cells metabolism, by measuring (1) the expression and/or activities of glycolysis and glycogen metabolism-related proteins and (2) the metabolite secretion/consumption in Sertoli cells obtained from rat models of different development stages of the disease, to unveil the mechanisms by which testosterone deregulation may affect spermatogenesis. Glucose and pyruvate uptake were decreased in cells exposed to the testosterone concentration found in pre-diabetic rats (600nM), whereas the decreased testosterone concentrations found in type 2 diabetic rats (7nM) reversed this profile. Lactate production was not altered, although the expression and/or activity of lactate dehydrogenase and monocarboxylate transporter 4 were affected by progressive testosterone-deficiency. Sertoli cells exposed to type 2 diabetic conditions exhibited intracellular glycogen accumulation. These results illustrate that gradually reduced levels of testosterone, induced by progressive stages of diabetes mellitus, favor a metabolic reprogramming toward glycogen synthesis. Our data highlights a pivotal role for testosterone in the regulation of spermatogenesis metabolic support by Sertoli cells, particularly in individuals suffering from metabolic diseases. Such alterations may be in the basis of male subfertility/infertility associated with the progression of diabetes mellitus.

Topics: Animals; Blotting, Western; Cells, Cultured; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Estradiol; Gene Expression; Glucose; Glycogen; Inhibins; Male; Prediabetic State; Rats; Rats, Wistar; Receptors, Androgen; Reverse Transcriptase Polymerase Chain Reaction; Sertoli Cells; Testis; Testosterone

Increased glucose production and reduced hepatic glycogen storage contribute to metabolic abnormalities in diabetes. Irisin, a newly identified myokine, induces the browning of white adipose tissue, but its effects on gluconeogenesis and glycogenesis are unknown. In the present study, we investigated the effects and underlying mechanisms of irisin on gluconeogenesis and glycogenesis in hepatocytes with insulin resistance, and its therapeutic role in type 2 diabetic mice. Insulin resistance was induced by glucosamine (GlcN) or palmitate in human hepatocellular carcinoma (HepG2) cells and mouse primary hepatocytes. Type 2 diabetes was induced by streptozotocin/high-fat diet (STZ/HFD) in mice. In HepG2 cells, irisin ameliorated the GlcN-induced increases in glucose production, phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) expression, and glycogen synthase (GS) phosphorylation; it prevented GlcN-induced decreases in glycogen content and the phosphoinositide 3-kinase (PI3K) p110α subunit level, and the phosphorylation of Akt/protein kinase B, forkhead box transcription factor O1 (FOXO1) and glycogen synthase kinase-3 (GSK3). These effects of irisin were abolished by the inhibition of PI3K or Akt. The effects of irisin were confirmed in mouse primary hepatocytes with GlcN-induced insulin resistance and in human HepG2 cells with palmitate-induced insulin resistance. In diabetic mice, persistent subcutaneous perfusion of irisin improved the insulin sensitivity, reduced fasting blood glucose, increased GSK3 and Akt phosphorylation, glycogen content and irisin level, and suppressed GS phosphorylation and PEPCK and G6Pase expression in the liver. Irisin improves glucose homoeostasis by reducing gluconeogenesis via PI3K/Akt/FOXO1-mediated PEPCK and G6Pase down-regulation and increasing glycogenesis via PI3K/Akt/GSK3-mediated GS activation. Irisin may be regarded as a novel therapeutic strategy for insulin resistance and type 2 diabetes.

Topics: Animals; Blotting, Western; Cells, Cultured; Chromones; Class I Phosphatidylinositol 3-Kinases; Diabetes Mellitus, Type 2; Fibronectins; Gluconeogenesis; Glucose; Glucose-6-Phosphatase; Glycogen; Glycogen Synthase; Hep G2 Cells; Hepatocytes; Heterocyclic Compounds, 3-Ring; Humans; Insulin Resistance; Liver; Male; Mice, Inbred C57BL; Morpholines; Phosphatidylinositol 3-Kinases; Phosphoenolpyruvate Carboxykinase (ATP); Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Proto-Oncogene Proteins c-akt; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction

Recently, there has been a growing interest in alternative therapies and in the therapeutic use of natural products for the treatment of diabetes. Therefore, in this study, we investigated the hypoglycemic and hypolipidemic effects of brown algae, Padina arborescens, in an animal model of type 2 diabetes. For 6 weeks, male C57BL/KsJ-db/db mice were administrated either control diet with no treatment or were treated with rosiglitazone (RG; 0.005%, w/w) or P. arborescens extract (PAE; 0.5%, w/w). At the end of the experimental period, the blood glucose levels, glycosylated hemoglobin levels, and plasma insulin levels were significantly lower in the RG and PAE groups compared with the control group. In addition, glucose tolerance was significantly improved in the RG and PAE groups. The homeostatic index of insulin resistance was lower in the RG and PAE groups than the diabetic control group. Also, the total cholesterol, LDL-cholesterol, triglyceride, and free fatty acid levels were lower in the PAE group than in the control group, whereas the HDL-C level was higher in the PAE group. Supplementation with PAE significantly lowered hepatic glucose-6-phosphatase and phosphoenolpyruvate carboxykinase activities, and increased glucokinase activity in the liver. Consequently, these results suggest that PAE may be beneficial in improving insulin resistance, hyperglycemia, and dyslipidemia in type 2 diabetics.

Topics: Adiponectin; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Diet; Disease Models, Animal; Dyslipidemias; Fasting; Glucose Tolerance Test; Glycated Hemoglobin; Glycogen; Hyperglycemia; Hypoglycemic Agents; Insulin; Insulin Resistance; Lipids; Male; Mice; Mice, Inbred C57BL; Phaeophyceae; Rosiglitazone; Thiazolidinediones

In West Africa, various preparations of the fruit, seed and leaf of Aframomum melegueta K. Schum. are reputably used for the management of diabetes mellitus (DM) and other metabolic disorders. The present study evaluated the anti-diabetic effects of A. melegueta ethyl acetate fraction (AMEF) from fruit ethanolic extract in a type 2 diabetes (T2D) model of rats.. T2D was induced in rats by feeding a 10% fructose solution ad libitum for two weeks followed by a single intraperitoneal injection of streptozotocin (40 mg/kg body weight) and the animals were orally treated with 150 or 300 mg/kg body weight (bw) of the AMEF once daily for four weeks.. At the end of the intervention, diabetic untreated animals showed significantly higher serum glucose, serum fructosamine, LDH, CK-MB, serum lipids, liver glycogen, insulin resistance (HOMA-IR), AI, CRI and lower serum insulin, pancreatic β-cell function (HOMA- β) and glucose tolerance ability compared to the normal animals. Histopathological examination of their pancreas revealed corresponding pathological changes in the islets and β-cells. These alterations were reverted to near-normal after the treatment of AMEF at 150 and 300 mg/kg bw when, the effects were more pronounced at 300 mg/kg bw compared to the 150 mg/kg bw.. The results of our study suggest that AMEF treatment at 300 mg/kg bw showed potent anti-diabetic effect in a T2D model of rats.

Topics: Acetates; alpha-Amylases; alpha-Glucosidases; Animals; Blood Glucose; Creatine Kinase, MB Form; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Ethanol; Fructosamine; Fruit; Glycogen; Glycoside Hydrolase Inhibitors; Insulin; Insulin-Secreting Cells; L-Lactate Dehydrogenase; Lipids; Liver; Male; Plant Extracts; Rats, Sprague-Dawley; Solvents; Zingiberaceae

Thiazolidinediones constitute a family of antidiabetic drugs, and rosiglitasone (RSG) has an extensive usage in treating the complications of type 2 diabetes mellitus. Carvacrol (CVL), a monoterpenic phenol that occurs in many essential oils of the family Labiatae including Origanum, Satureja, Thymbra, Thymus, and Corydothymus species, possess a wide variety of pharmacological properties including antioxidant potential. We hypothesized that carvacrol in combination with RSG would prove beneficial to ameliorate the dysregulated carbohydrate metabolism in high-fat diet (HFD)-induced type 2 diabetic C57BL/6J mice. Mice were divided into six groups and fed HFD, for 10 weeks. CVL (20 mg/kg BW) and RSG (4 mg/kg BW) were administered post-orally, daily for 35 days. HFD mice showed an elevation in plasma glucose, insulin, glycosylated hemoglobin and a decrease in hemoglobin. The activities of carbohydrate metabolic enzymes such as glucose-6-phosphatase and fructose-1,6-bisphosphatase increased whereas glucokinase and glucose-6-phosphate dehydrogenase activities decreased in the liver of HFD mice. The activities of hepatic marker enzymes such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and gamma-glutamyl transpeptidase increased in HFD mice. Combination of CVL and RSG prevented the above changes toward normalcy. Histopathological analysis of H&E stained pancreas was also in agreement with the biochemical findings. These major findings provide evidence that combination of CVL with RSG has better antidiabetic properties.

Topics: Animals; Biomarkers; Blood Glucose; Body Weight; Cymenes; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Dose-Response Relationship, Drug; Drug Therapy, Combination; Glycated Hemoglobin; Glycogen; Hypoglycemic Agents; Liver; Male; Mice; Mice, Inbred C57BL; Monoterpenes; Rosiglitazone; Thiazolidinediones

Several studies showed that the orphan Bombesin Receptor Subtype-3 (BRS-3) - member of the bombesin receptor family - has an important role in glucose homeostasis (v.g.: BRS-3-KO mice developed mild obesity, and decreased levels of BRS-3 mRNA/protein have been described in muscle from obese (OB) and type 2 diabetic (T2D) patients). In this work, to gain insight into BRS-3 receptor cell signaling pathways, and its implication on glucose metabolism, primary cultured myocytes from normal subjects, OB or T2D patients were tested using high affinity ligand - [d-Tyr(6),β-Ala(11),Phe(13),Nle(14)]bombesin6-14. In muscle cells from all metabolic conditions, the compound significantly increased not only MAPKs, p90RSK1, PKB and p70s6K phosphorylation levels, but also PI3K activity; moreover, it produced a dose-response stimulation of glycogen synthase a activity and glycogen synthesis. Myocytes from OB and T2D patients were more sensitive to the ligand than normal, and T2D cells even more than obese myocytes. These results widen the knowledge of human BRS-3 cell signaling pathways induced by a BRS-3 agonist, described its insulin-mimetic effects on glucose metabolism, showed the role of BRS-3 receptor in glucose homeostasis, and also propose the employing of BRS-3/ligand system, as participant in the obese and diabetic therapies.

Topics: Adult; Aged; Bombesin; Cells, Cultured; Diabetes Mellitus, Type 2; Female; Glucose; Glycogen; Glycogen Synthase; Homeostasis; Humans; Male; Middle Aged; Mitogen-Activated Protein Kinases; Muscle Fibers, Skeletal; Obesity; Peptide Fragments; Phosphatidylinositol 3-Kinases; Phosphorylation; Protein Processing, Post-Translational; Proto-Oncogene Proteins c-akt; Receptors, Bombesin; Ribosomal Protein S6 Kinases, 70-kDa; Ribosomal Protein S6 Kinases, 90-kDa; Signal Transduction

We found that the 50% aqueous EtOH extract of clove (Syzygium aromaticum) had potent dose-dependent inhibitory activity toward glycogen phosphorylase b and glucagon-stimulated glucose production in primary rat hepatocytes. Among the components, eugeniin inhibited glycogen phosphorylase b and glucagon-stimulated glucose production in primary rat hepatocytes, with IC50 values of 0.14 and 4.7 μM, respectively. In sharp contrast, eugenol showed no significant inhibition toward glycogen phosphorylase b, even at a concentration of 400 μM. Eugenol-reduced clove extracts (erCE) were prepared and when fed to a db/db mouse they clearly suppressed the blood glucose and HbA1c levels. Furthermore, plasma triglyceride and non-esterified fatty acid levels in 5% and 10% erCE-fed db/db mice were significantly lowered, compared with control db/db mice without erCE supplementation. These results suggested that dietary supplementation with the erCE could beneficially modify glucose and lipid metabolism and contribute to the prevention of the progress of hyperglycemia and metabolic syndrome.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Disease Models, Animal; Eugenol; Flowers; Glycated Hemoglobin; Glycogen; Glycogen Phosphorylase; Hepatocytes; Humans; Male; Mice; Mice, Inbred C57BL; Plant Extracts; Rats; Syzygium

Sterol regulatory element-binding protein-1 (SREBP-1) is a key transcription factor that regulates genes in the de novo lipogenesis and glycolysis pathways. The levels of SREBP-1 are significantly elevated in obese patients and in animal models of obesity and type 2 diabetes, and a vast number of studies have implicated this transcription factor as a contributor to hepatic lipid accumulation and insulin resistance. However, its role in regulating carbohydrate metabolism is poorly understood. Here we have addressed whether SREBP-1 is needed for regulating glucose homeostasis. Using RNAi and a new generation of adenoviral vector, we have silenced hepatic SREBP-1 in normal and obese mice. In normal animals, SREBP-1 deficiency increased Pck1 and reduced glycogen deposition during fed conditions, providing evidence that SREBP-1 is necessary to regulate carbohydrate metabolism during the fed state. Knocking SREBP-1 down in db/db mice resulted in a significant reduction in triglyceride accumulation, as anticipated. However, mice remained hyperglycemic, which was associated with up-regulation of gluconeogenesis gene expression as well as decreased glycolysis and glycogen synthesis gene expression. Furthermore, glycogen synthase activity and glycogen accumulation were significantly reduced. In conclusion, silencing both isoforms of SREBP-1 leads to significant changes in carbohydrate metabolism and does not improve insulin resistance despite reducing steatosis in an animal model of obesity and type 2 diabetes.

Topics: Animals; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Gene Expression Regulation; Gene Knockdown Techniques; Gluconeogenesis; Glycogen; Liver; Male; Mice; Obesity; Sterol Regulatory Element Binding Protein 1

Cucumis prophetarum (L.) is used in traditional Indian medicine for the treatment of inflammation related problems.. The present investigation was designed to study the effect of N-Trisaccharide (a new compound isolated from the fruit of C. prophetarum (L.)) on hyperglycemia in streptozotocin (STZ)-nicotinamide (NA) induced type 2 diabetic rats.. Different doses of N-Trisaccharide (25 and 50 mg/kgb.w.) were administered once daily for 28 days to STZ-NA induced diabetic rats. Plasma insulin and glycogen levels were measured. The activities of hexokinase, glucose-6-phosphatase, fructose-1,6-bisphosphatase, glucose-6-phosphate dehydrogenase, glycogen synthase and glycogen phosphorylase were measured. Further, histological studies on pancreas were also carried out.. The active compound at doses of 25 and 50 mg/kgb.w. given orally for 14 days showed 47.7% and 69.3% antihyperglycemic activity, respectively. Treatment at the same doses for 28 days provided complete protection against STZ-NA challenge (65 and 230 mg/kgb.w., respectively), intraperitoneally. N-Trisaccharide significantly (p≤0.05) increased the plasma insulin and liver glycogen levels in diabetic rats. The altered enzyme activities of carbohydrate metabolism in the liver and kidney of the diabetic rats were significantly (p≤0.05) improved. Additionally, N-Trisaccharide increased glycogen synthase and decreased glycogen phosphorylase activity in diabetic rats. Histological studies confirmed an increase in insulin level is due to stimulation of injured pancreatic β-cells.. The results of the study suggested that N-Trisaccharide possesses propitious effect on STZ-NA induced type 2 diabetes, indicating its usefulness in diabetes management.

Topics: Animals; Blood Glucose; Cucumis; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Drug Evaluation, Preclinical; Female; Fruit; Glycogen; Hypoglycemic Agents; Liver; Male; Pancreas; Phytotherapy; Plant Extracts; Plants, Medicinal; Rats, Wistar; Trisaccharides

Efficacy of Kalpaamruthaa on the activities of lipid and carbohydrate metabolic enzymes, electron transport chain complexes and mitochondrial ATPases were studied in heart and liver of experimental rats. Cardiovascular damage (CVD) was developed in 8 weeks after type 2 diabetes mellitus induction with high fat diet (2 weeks) and low dose of streptozotocin (2 × 35 mg/kg b.w. i.p. in 24 hr interval). In CVD-induced rats, the activities of total lipase, cholesterol ester hydrolase and cholesterol ester synthetase were increased, while lipoprotein lipase and lecithin-cholesterol acyltransferase activities were decreased. The activities of lipid-metabolizing enzymes were altered by Kalpaamruthaa in CVD-induced rats towards normal. Kalpaamruthaa modulated the activities of glycolytic enzymes (hexokinase, phosphogluco-isomerase, aldolase and glucose-6-phosphate dehydrogenase), gluconeogenic enzymes (glucose-6-phosphatase and fructose-1, 6-bisphosphatase) and glycogenolytic enzyme (glycogen phosphorylase) along with increased glycogen content in the liver of CVD-induced rats. The activities of isocitrate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, α-ketoglutarate dehydrogenase, Complexes and ATPases (Na(+)/K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase) were decreased in CVD-induced rats, which were ameliorated by the treatment with Kalpaamruthaa. This study ascertained the efficacy of Kalpaamruthaa for the treatment of CVD in diabetes through the modulation of metabolizing enzymes and mitochondrial dysfunction.

Topics: Adenosine Triphosphatases; Animals; Blood Glucose; Cardiovascular Diseases; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Glycogen; Heart; Lipid Metabolism; Liver; Magnoliopsida; Male; Medicine, Ayurvedic; Mitochondria; Phytotherapy; Plant Extracts; Rats, Sprague-Dawley

Khaya senegalensis A. Juss (Meliaceae) is commonly exploited for the traditional treatment of diabetes mellitus in Nigeria and Togo. The present study was conducted to examine the anti-diabetic activity of Khaya senegalensis butanol fraction (KSBF) of root ethanolic extract in a type 2 diabetes (T2D) model of rats.. T2D was induced in rats by feeding a 10% fructose solution ad libitum for two weeks followed by a single intraperitoneal injection of streptozotocin (40 mg/kg body weight) and the animals were treated with 150 and 300 mg/kg body weight (BW) of the fraction for five days in a week. Relevant diabetes-related parameters were analyzed in all experimental animals.. The KSBF treatment, at 300 mg/kg BW, significantly (p<0.05) reduced blood glucose level, improved oral glucose tolerance ability and β-cell function (HOMA-β), decreased insulin resistance (HOMA-IR), stimulated hepatic glycogen synthesis, ameliorated serum lipids alterations and prevented hepatic and renal damages compared to untreated diabetic rats. Additionally, the fraction insignificantly (p>0.05) improved weight gain, decreased food and fluid intake, stimulated insulin secretion and lowered serum fructosamine concentrations compared to untreated diabetic rats.. Data from this study suggests that orally administered KSBF, at 300 mg/kg BW, possess remarkable anti-type 2 diabetic activity and could ameliorate some diabetes-associated complications and hence can be considered as a source of potential anti-type 2 diabetic medicine.

Topics: Animals; Blood Glucose; Butanols; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Disease Models, Animal; Drinking; Drugs, Chinese Herbal; Eating; Glycogen; Hypoglycemic Agents; Insulin; Insulin Secretion; Insulin-Secreting Cells; Male; Meliaceae; Plant Roots; Rats; Rats, Sprague-Dawley; Streptozocin

The aim of this study was to evaluate the effects of Bifidobacterium lactis HY8101 on insulin resistance induced using tumour necrosis factor-α (TNF-α) in rat L6 skeletal muscle cells and on the KK-A(Y) mouse noninsulin-dependent diabetes mellitus (NIDDM) model.. The treatment using HY8101 improved the insulin-stimulated glucose uptake and translocation of GLUT4 via the insulin signalling pathways AKT and IRS-1(Tyr) in TNF-α-treated L6 cells. HY8101 increased the mRNA levels of GLUT4 and several insulin sensitivity-related genes (PPAR-γ) in TNF-α-treated L6 cells. In KK-A(Y) mice, HY8101 decreased fasting insulin and blood glucose and significantly improved insulin tolerance. HY8101 improved diabetes-induced plasma total cholesterol and triglyceride (TG) levels and increased the muscle glycogen content. We observed concurrent transcriptional changes in the skeletal muscle tissue and the liver. In the skeletal muscle tissue, the glycogen synthesis-related gene pp-1 and GLUT4 were up-regulated in mice receiving HY8101 treatment. In the liver, the hepatic gluconeogenesis-regulated genes (PCK1 and G6PC) were down-regulated in mice receiving HY8101 treatment.. Bifidobacterium lactis HY8101 can be used to moderate glucose metabolism, lipid metabolism and insulin sensitivity in mice and in cells.. Bifidobacterium lactis HY8101 might have potential as a probiotic candidate for alleviating metabolic syndromes such as diabetes.

Topics: Animals; Bifidobacterium; Blood Glucose; Cell Line; Diabetes Mellitus, Type 2; Glucose; Glucose Transporter Type 4; Glycogen; Insulin; Insulin Receptor Substrate Proteins; Insulin Resistance; Liver; Male; Mice; Mice, Inbred C57BL; Muscle, Skeletal; Probiotics; Rats; Signal Transduction; Tumor Necrosis Factor-alpha

Ursolic acid (UA) is a pentacyclic triterpenoid compound that naturally occurs in fruits, leaves and flowers of medicinal herbs. This study investigated the dose-response efficacy of UA (0.01 and 0.05%) on glucose metabolism, the polyol pathway and dyslipidemia in streptozotocin/nicotinamide-induced diabetic mice. Supplement with both UA doses reduced fasting blood glucose and plasma triglyceride levels in non-obese type 2 diabetic mice. High-dose UA significantly lowered plasma free fatty acid, total cholesterol and VLDL-cholesterol levels compared with the diabetic control mice, while LDL-cholesterol levels were reduced with both doses. UA supplement effectively decreased hepatic glucose-6-phosphatase activity and increased glucokinase activity, the glucokinase/glucose-6-phosphatase ratio, GLUT2 mRNA levels and glycogen content compared with the diabetic control mice. UA supplement attenuated hyperglycemia-induced renal hypertrophy and histological changes. Renal aldose reductase activity was higher, whereas sorbitol dehydrogenase activity was lower in the diabetic control group than in the non-diabetic group. However, UA supplement reversed the biochemical changes in polyol pathway to normal values. These results demonstrated that low-dose UA had preventive potency for diabetic renal complications, which could be mediated by changes in hepatic glucose metabolism and the renal polyol pathway. High-dose UA was more effective anti-dyslipidemia therapy in non-obese type 2 diabetic mice.

Topics: Animals; Antineoplastic Agents, Phytogenic; Blotting, Western; Diabetes Complications; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dyslipidemias; Glucokinase; Glucose; Glucose Transporter Type 2; Glucose-6-Phosphatase; Glycogen; Hyperglycemia; Kidney Diseases; Male; Mice; Mice, Inbred ICR; Mice, Inbred NOD; Polymers; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Triterpenes; Ursolic Acid

This study was aimed at assessing the potential of essential oil from the leaf of Hoslundia opposita in the treatment of diabetes. Forty-eight rats (Rattus norvegicus) were randomized into two groups; nondiabetic and diabetic groups, each with four subgroups. Animals in the diabetic group were induced with diabetes using a single dose of alloxan monohydrate, 160 mg/kg body weight (b. wt.). The rats were treated with 110 and 220 mg/kg b. wt. of the essential oil. All treatments were administered, intraperitoneally, once a day for 4 days. In the nondiabetic condition, there was no effect of the oil on fasting blood glucose (FBG) levels in rats. In diabetic rats, the oil caused a significant reduction in FBG levels. Treatment with 110 mg/kg b. wt. of the oil reduced FBG almost to the normoglycemic level by day 4 and the overall glucose excursion during a 3-h intraperitoneal glucose tolerance test approached the baseline level at 120 min. Also, hepatic glycogen was significantly higher, while the glucose concentrations were lower in the diabetic-treated group when compared with the diabetic untreated group. Histological examinations revealed a mildly distorted architecture of the pancreatic islets β-cells of diabetic rats treated with the oil, while those of the untreated rats were severely degenerated. Overall, the in vivo antihyperglycemic activity of the essential oil may prove to be of clinical importance in the management of type 2 diabetes.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Female; Glycogen; Humans; Hypoglycemic Agents; Lamiaceae; Liver; Male; Oils, Volatile; Plant Extracts; Plant Leaves; Rats

Two-dimensional difference gel electrophoresis (2-D DIGE)-based proteome analysis has revealed intrinsic insulin resistance in myotubes derived from type 2 diabetic patients. Using 2-D DIGE-based proteome analysis, we identified a subset of insulin-resistant proteins involved in protein turnover in skeletal muscle of type 2 diabetic patients, suggesting aberrant regulation of the protein homeostasis maintenance system underlying metabolic disease. We then validated the role of the ubiquitin-proteasome system (UPS) in myotubes to investigate whether impaired proteasome function may lead to metabolic arrest or insulin resistance. Myotubes derived from muscle biopsies obtained from people with normal glucose tolerance (NGT) or type 2 diabetes were exposed to the proteasome inhibitor bortezomib (BZ; Velcade) without or with insulin. BZ exposure increased protein carbonylation and lactate production yet impaired protein synthesis and UPS function in myotubes from type 2 diabetic patients, marking the existence of an insulin-resistant signature that was retained in cultured myotubes. In conclusion, BZ treatment further exacerbates insulin resistance and unmasks intrinsic features of metabolic disease in myotubes derived from type 2 diabetic patients. Our results highlight the existence of a confounding inherent abnormality in cellular protein dynamics in metabolic disease, which is uncovered through concurrent inhibition of the proteasome system.

Topics: Boronic Acids; Bortezomib; Cells, Cultured; Diabetes Mellitus, Type 2; Enzyme Inhibitors; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Lipid Metabolism; Male; Muscle, Skeletal; Oxidative Stress; Proteasome Endopeptidase Complex; Proteasome Inhibitors; Protein Carbonylation; Proteome; Pyrazines; RNA Interference; Signal Transduction

Benzopyrones are proven antidiabetic drug candidate in diabetic drug discovery. In this view novel synthetic benzopyrone analogues were selected for testing in experimental diabetes. Type 2 diabetes (T2D) was induced in Wistar rats by streptozotocin (60 mg/kg, i.p.) followed by nicotinamide (120 mg/kg i.p.). Rats having fasting blood glucose (FBG)>200 mg/dL, 7 days after T2D-induction, are selected for the study. Test compounds and standard treatment were continued for 15 days. FBG, oral glucose tolerance test (OGTT), and insulin tolerance test (ITT) were determined on 21st day after induction of T2D. Plasma lipids and serum insulin were estimated. Homeostatic model assessment (HOMA-IR) was then calculated from serum insulin. Rats were sacrificed and pancreas was isolated for histopathological observations. Oxidative stress markers were estimated in liver homogenate. Quercetin, a natural product with benzopyrone ring, showed significant hypoglycemic activity comparable to glibenclamide. Treatment with test compounds lowered the FBG and insulin resistance was significant alleviated as determined by OGTT, HOMA-IR, and ITT. There was significant normalisation of liver antioxidant enzymes compared to diabetic rats indicating that all the synthesised benzopyrone analogues are beneficial in reducing oxidative stress and are on par with the standard quercetin and glibenclamide in experimental T2D.

Topics: Animals; Biomarkers; Blood Glucose; Coumarins; Creatinine; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Fasting; Glucose Tolerance Test; Glycation End Products, Advanced; Glycogen; Hyperglycemia; Hypoglycemic Agents; Insulin; Insulin Resistance; Lipids; Liver; Male; Niacinamide; Oxidative Stress; Pancreas; Rats, Wistar; Streptozocin

Failure of white adipose tissue to appropriately store excess metabolic substrate seems to underpin obesity-associated type 2 diabetes. Encouraging "browning" of white adipose has been suggested as a therapeutic strategy to help dispose of excess stored lipid and ameliorate the resulting insulin resistance. Genetic variation at the DNA locus encoding the novel proteolipid neuronatin has been associated with obesity, and we recently observed that neuronatin expression is reduced in subcutaneous adipose tissue from obese humans. Thus, to explore the function of neuronatin further, we used RNAi to silence its expression in murine primary adipocyte cultures and examined the effects on adipocyte phenotype. We found that primary adipocytes express only the longer isoform of neuronatin. Loss of neuronatin led to increased mitochondrial biogenesis, indicated by greater intensity of MitoTracker Green staining. This was accompanied by increased expression of UCP1 and the key genes in mitochondrial oxidative phosphorylation, PGC-1α, Cox8b, and Cox4 in primary subcutaneous white adipocytes, indicative of a "browning" effect. In addition, phosphorylation of AMPK and ACC was increased, suggestive of increased fatty acid utilization. Similar, but less pronounced, effects of neuronatin silencing were also noted in primary brown adipocytes. In contrast, loss of neuronatin caused a reduction in both basal and insulin-stimulated glucose uptake and glycogen synthesis, likely mediated by a reduction in Glut1 protein upon silencing of neuronatin. In contrast, loss of neuronatin had no effect on insulin signaling. In conclusion, neuronatin appears to be a novel regulator of browning and metabolic substrate disposal in white adipocytes.

Topics: Adipocytes, White; Adipogenesis; Adipose Tissue, Brown; Adult; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Glucose Transporter Type 1; Glycogen; Humans; Membrane Proteins; Mice; Mice, 129 Strain; Mice, Knockout; Middle Aged; Mitochondria; Nerve Tissue Proteins; Obesity; Phenotype; Primary Cell Culture

Plasma glucose levels are maintained within a narrow range in normal individuals. Both insulin-dependent and insulin-independent processes contribute to fasting and postprandial plasma glucose regulation. The brain and nervous system are insulin independent. Muscle and adipose tissue are responsive to insulin and can use either glucose or ketones and free fatty acids as their primary metabolic fuel. The essential components of metabolic syndrome are obesity, glucose intolerance, insulin resistance, lipid disturbances, and hypertension. The risk of type 2 diabetes increases exponentially as body mass index increases above about 25 kg/m2. The links between obesity and type 2 diabetes include proinflammatory cytokines, insulin resistance, deranged fatty acid metabolism, and cellular processes. Modest weight reduction can improve glycaemic control and reduce diabetes risk. Obesity also leads to hyperinsulinaemia and insulin resistance, with a progressive decrease in insulin secretory function. Ageing is another important risk factor for metabolic disorders, including obesity, impaired glucose tolerance, and type 2 diabetes.

Topics: Aging; Body Mass Index; Diabetes Mellitus, Type 2; Glucagon; Glucose; Glucose Intolerance; Glucose Transport Proteins, Facilitative; Glycogen; Homeostasis; Humans; Inflammation Mediators; Insulin; Insulin Resistance; Metabolic Syndrome; Obesity; Obesity, Abdominal; Sodium-Glucose Transport Proteins

This study explores the hypoglycemic effects of borapetoside A, the most active principle among three major diterpenoids (borapetosides A, B, and C) isolated from ethanol extract of Tinospora crispa vines.. We employed mouse mitogenic C2C12 and hepatocellular carcinoma Hep3B cells in this study. Furthermore, the mice were divided into three groups, including streptozotocin-induced type 1 diabetes mellitus, diet-induced type 2 diabetes mellitus, and normal control. The mice in each group were treated with assigned vehicle control, borapetoside A, or other active agents.. Borapetoside A was shown to increase the glycogen content and decrease the plasma glucose concentration in a concentration or dose-dependent manner in vitro and in vivo. The hypoglycemic effects in the normal mice and the mice with type 2 diabetes mellitus were associated with the increases of the plasma insulin levels; whereas, the insulin levels remained unchanged in the mice with type 1 diabetes mellitus. Borapetoside A not only attenuated the elevation of plasma glucose induced by an intraperitoneal glucose tolerance test, but also increased the glycogen synthesis of IL-6 treated C2C12 cells. Moreover, the elevated protein expression levels of phosphoenolpyruvate carboxykinase were reversed after borapetoside A treatment twice a day for 7 days.. The hypoglycemic effects of borapetoside A were mediated through both the insulin-dependent and the insulin-independent pathways. Furthermore, borapetoside A was shown to increase the glucose utilization in peripheral tissues, to reduce the hepatic gluconeogenesis, and to activate the insulin signaling pathway; they thereby contributed to the lowering of the plasma glucose. Comparison of the structures of three borapetosides suggests clearly that the C-8 stereochemistry plays a key role in hypoglycemic effect since the active borapetoside A and C possess 8R-chirality but the inactive borapetoside B possess 8S-chirality. The location of glycoside at C-3 for borapetoside A but C-6 for borapetoside C and the formation of lactone between C-4 and C-6 for borapetoside A, could account for the different potency in hypoglycemic action for these two compounds.

Topics: Animals; Blood Glucose; Cell Line; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Disease Models, Animal; Diterpenes; Gluconeogenesis; Glucose; Glucosides; Glycogen; Hypoglycemic Agents; Insulin; Interleukin-6; Liver; Male; Mice; Mice, Inbred ICR; Muscles; Phosphoenolpyruvate Carboxykinase (GTP); Phytotherapy; Plant Extracts; Plants, Medicinal; Signal Transduction; Tinospora

The measurement of tissue lipid and glycogen contents and the establishment of normal levels of variability are important when assessing changes caused by pathology or treatment. We measured hepatic and skeletal muscle lipid and glycogen levels using (1)H and (13)C MRS at 3 T in groups of subjects with and without type 2 diabetes. Within-visit reproducibility, due to repositioning and instrument errors was determined from repeat measurements made over 1 h. Natural variability was assessed from separate measurements made on three occasions over 1 month. Hepatic lipid content was greater in subjects with diabetes relative to healthy subjects (p = 0.03), whereas levels of hepatic and skeletal muscle glycogen, and of intra- and extra-myocellular lipid, were similar. The single-session reproducibility values (coefficient of variation, CV) for hepatic lipid content were 12% and 7% in groups of subjects with and without diabetes, respectively. The variability of hepatic lipid content over 1 month was greater than the reproducibility, with CV = 22% (p = 0.08) and CV = 44% (p = 0.004) in subjects with and without diabetes, respectively. Similarly, levels of variation in basal hepatic glycogen concentrations (subjects with diabetes, CV = 38%; healthy volunteers, CV = 35%) were significantly larger than single-session reproducibility values (CV = 17%, p = 0.02 and CV = 13%, p = 0.05, respectively), indicating substantial biological changes in basal concentrations over 1 month. There was a decreasing correlation in measurements of both hepatic lipid and glycogen content with increasing time between scans. Levels of variability in intra- and extra-myocellular lipid in the soleus muscle, and glycogen concentrations in the gastrocnemius muscle, tended to be larger than expected from single-session reproducibility, although these did not reach significance.

Topics: Carbon Isotopes; Diabetes Mellitus, Type 2; Fasting; Female; Glycogen; Humans; Lipid Metabolism; Liver; Liver Glycogen; Longitudinal Studies; Magnetic Resonance Spectroscopy; Male; Middle Aged; Muscle, Skeletal; Protons; Reproducibility of Results

Genome-wide association (GWA) studies have described a large number of new candidate genes that contribute to of Type 2 Diabetes (T2D). In some cases, small clusters of genes are implicated, rather than a single gene, and in all cases, the genetic contribution is not defined through the effects on a specific organ, such as the pancreas or liver. There is a significant need to develop and use human cell-based models to examine the effects these genes may have on glucose regulation. We describe the development of a primary human hepatocyte model that adjusts glucose disposition according to hormonal signals. This model was used to determine whether candidate genes identified in GWA studies regulate hepatic glucose disposition through siRNAs corresponding to the list of identified genes. We find that several genes affect the storage of glucose as glycogen (glycolytic response) and/or affect the utilization of pyruvate, the critical step in gluconeogenesis. Of the genes that affect both of these processes, CAMK1D, TSPAN8 and KIF11 affect the localization of a mediator of both gluconeogenesis and glycolysis regulation, CRTC2, to the nucleus in response to glucagon. In addition, the gene CDKAL1 was observed to affect glycogen storage, and molecular experiments using mutant forms of CDK5, a putative target of CDKAL1, in HepG2 cells show that this is mediated by coordinate regulation of CDK5 and PKA on MEK, which ultimately regulates the phosphorylation of ribosomal protein S6, a critical step in the insulin signaling pathway.

Topics: Calcium-Calmodulin-Dependent Protein Kinase Type 1; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinase 5; Diabetes Mellitus, Type 2; Extracellular Signal-Regulated MAP Kinases; Gene Knockdown Techniques; Genome-Wide Association Study; Genome, Human; Glucagon; Glucose; Glycogen; Hep G2 Cells; Hepatocytes; Homeostasis; Humans; Intracellular Signaling Peptides and Proteins; Phosphoenolpyruvate Carboxykinase (GTP); Phosphorylation; Polymorphism, Single Nucleotide; Primary Cell Culture; Protein Processing, Post-Translational; Protein Transport; Pyruvic Acid; Ribosomal Protein S6 Kinases; RNA Interference; RNA, Small Interfering; Signal Transduction; Transcription Factors; tRNA Methyltransferases

CCR2 inhibition has produced promising experimental and clinical anti-hyperglycemic effects. These results support the thesis that insulin resistance and Type 2 diabetes (T2D) are associated with chronic unresolved inflammation. The aim of this study was to provide a broad analysis of the various physiological changes occurring in mouse models of T2D in connection with pharmacological CCR2 inhibition.. A mouse-active chemical analogue of the clinical candidate CCX140-B was tested in diet-induced obese (DIO) mice and db/db mice. Measurements included: adipose tissue inflammatory macrophage counts; peripheral blood glucose levels at steady-state and after glucose and insulin challenges; peripheral blood insulin and adiponectin levels; 24-h urine output and urinary glucose levels; pancreatic islet number and size; hepatic triglyceride and glycogen content; and hepatic glucose-6-phosphatase levels.. In DIO mice, the CCR2 antagonist completely blocked the recruitment of inflammatory macrophages to visceral adipose tissue. The mice exhibited reduced hyperglycemia and insulinemia, improved insulin sensitivity, increased circulating adiponectin levels, decreased pancreatic islet size and increased islet number. It also reduced urine output, glucose excretion, hepatic glycogen and triglyceride content and glucose 6-phosphatase levels. Similar effects were observed in the db/db diabetic mice.. These data indicate that pharmacological inhibition of CCR2 in models of T2D can reduce inflammation in adipose tissue, alter hepatic metabolism and ameliorate multiple diabetic parameters. These mechanisms may contribute to the promising anti-diabetic effects seen in humans with at least one CCR2 antagonist.

Topics: Adiponectin; Adipose Tissue; Animals; Biomarkers; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Dose-Response Relationship, Drug; Glucose-6-Phosphatase; Glycogen; Glycosuria; Hypoglycemic Agents; Inflammation; Insulin; Insulin Resistance; Insulin-Secreting Cells; Liver; Macrophages; Male; Mice; Mice, Inbred C57BL; Obesity; Receptors, CCR2; Triglycerides

To study the pathogenesis of diabetic cardiomyopathy, reliable animal models of type 2 diabetes are required. Physiologically relevant rodent models are needed, which not only replicate the human pathology but also mimic the disease process. Here we characterised cardiac metabolic abnormalities, and investigated the optimal experimental approach for inducing disease, in a new model of type 2 diabetes.. Male Wistar rats were fed a high-fat diet for three weeks, with a single intraperitoneal injection of low dose streptozotocin (STZ) after fourteen days at 15, 20, 25 or 30 mg/kg body weight. Compared with chow-fed or high-fat diet fed control rats, a high-fat diet in combination with doses of 15-25 mg/kg STZ did not change insulin concentrations and rats maintained body weight. In contrast, 30 mg/kg STZ induced hypoinsulinaemia, hyperketonaemia and weight loss. There was a dose-dependent increase in blood glucose and plasma lipids with increasing concentrations of STZ. Cardiac and hepatic triglycerides were increased by all doses of STZ, in contrast, cardiac glycogen concentrations increased in a dose-dependent manner with increasing STZ concentrations. Cardiac glucose transporter 4 protein levels were decreased, whereas fatty acid metabolism-regulated proteins, including uncoupling protein 3 and pyruvate dehydrogenase (PDH) kinase 4, were increased with increasing doses of STZ. Cardiac PDH activity displayed a dose-dependent relationship between enzyme activity and STZ concentration. Cardiac insulin-stimulated glycolytic rates were decreased by 17% in 15 mg/kg STZ high-fat fed diabetic rats compared with control rats, with no effect on cardiac contractile function.. High-fat feeding in combination with a low dose of STZ induced cardiac metabolic changes that mirror the decrease in glucose metabolism and increase in fat metabolism in diabetic patients. While low doses of 15-25 mg/kg STZ induced a type 2 diabetic phenotype, higher doses more closely recapitulated type 1 diabetes, demonstrating that the severity of diabetes can be modified according to the requirements of the study.

Topics: Animals; Biomarkers; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Cardiomyopathies; Diet, High-Fat; Energy Metabolism; Glycogen; Glycolysis; Lipid Metabolism; Lipids; Male; Myocardium; Phenotype; Rats; Rats, Wistar; Time Factors

The beneficial effects of green tea polyphenols (GTP) against metabolic syndrome and type 2 diabetes by suppressing appetite and nutrient absorption have been well reported. However the direct effects and mechanisms of GTP on glucose and lipid metabolism remain to be elucidated. Since the liver is an important organ involved in glucose and lipid metabolism, we examined the effects and mechanisms of GTP on glycogen synthesis and lipogenesis in HepG2 cells. Concentrations of GTP containing 68% naturally occurring (-)-epigallocatechin-3-gallate (EGCG) were incubated in HepG2 cells with high glucose (30 mM) under 100 nM of insulin stimulation for 24 h. GTP enhanced glycogen synthesis in a dose-dependent manner. 10  μM of EGCG significantly increased glycogen synthesis by 2fold (P < 0.05) compared with insulin alone. Western blotting revealed that phosphorylation of Ser9 glycogen synthase kinase 3 β and Ser641 glycogen synthase was significantly increased in GTP-treated HepG2 cells compared with nontreated cells. 10  μM of EGCG also significantly inhibited lipogenesis (P < 0.01). We further demonstrated that this mechanism involves enhanced expression of phosphorylated AMP-activated protein kinase α and acetyl-CoA carboxylase in HepG2 cells. Our results showed that GTP is capable of enhancing insulin-mediated glucose and lipid metabolism by regulating enzymes involved in glycogen synthesis and lipogenesis.

Topics: Antioxidants; Catechin; Diabetes Mellitus, Type 2; Glycogen; Hep G2 Cells; Hepatocytes; Humans; Lipid Metabolism; Lipogenesis; Metabolic Syndrome; Phosphorylation; Proto-Oncogene Proteins c-akt; Tea

The dried succulent stem of Cistanche tubulosa (Schenk) R. Wight is one component of traditional Chinese medicine prescriptions for diabetes. However, there have been no modern scientific reports to confirm this traditional claim for the Cistanche species until now. Thus, we investigated the effects of Cistanche tubulosa on glucose homeostasis and serum lipids in male BKS.Cg-Dock7(m) +/+ Lepr(db)/J (db/db) mice, a model of type 2 diabetes.. The verbascoside and echinacoside contents of Cistanche tubulosa powder were evaluated using HPLC. The total phenolic content, polysaccharide content and antioxidant activity of Cistanche tubulosa powder were also evaluated. Then, different doses of Cistanche tubulosa (equivalent to 120.9, 72.6 or 24.2mg verbascoside/kg) were administered orally once daily for 45 days to male db/db mice. Age matched db/+ mice were used as normal controls. Body weight, fasting blood glucose, postprandial blood glucose and insulin tolerance test were measured during the experiment. At the time of sacrifice, blood was collected for measurement of insulin level, the homeostatic model assessment of insulin resistance (HOMA-IR), and total cholesterol, triglyceride, HDL-c, LDL-c and VLDL-c levels; liver and muscle were harvested for measurement of glycogen levels.. Cistanche tubulosa significantly suppressed the elevated fasting blood glucose and postprandial blood glucose levels, improved insulin resistance and dyslipidemia, and suppressed body weight loss in db/db mice. However, Cistanche tubulosa did not significantly affect serum insulin levels or hepatic and muscle glycogen levels.. This study provides scientific evidence for the traditional use of Cistanche tubulosa to treat diabetes, suggesting that Cistanche tubulosa has the potential for development into a functional food ingredient or drug to prevent hyperglycemia and treat hyperlipidemia.

Topics: Animals; Blood Glucose; Cistanche; Diabetes Mellitus, Type 2; Glycogen; Hypoglycemic Agents; Hypolipidemic Agents; Insulin Resistance; Lipids; Liver; Male; Mice; Mice, Mutant Strains; Muscle, Skeletal; Phytotherapy; Plant Extracts; Plant Stems

The objective of this study is to fully investigate the therapeutic effect and mechanisms of action of Gegen Qinlian decoction (GD) on type 2 diabetes mellitus (type 2 DM). A rat model of type 2 DM was established with the combination of high-fat diet and multiple low doses of streptozotocin (STZ). Biochemical indicators related to glucose metabolism disorders, insulin resistance, oxidative stress were observed. The type 2 DM rats were administrated with GD for 80 days, the above-mentioned indexes were detected. The results indicated that the hepatic glycogen synthesis level was promoted, fasting blood glucose level and fasting blood insulin level were significantly reduced, insulin sensitivity index was significantly improved; the level of superoxide dismutase (SOD) was increased and the level of malondialdehyde (MDA) was reduced; pathologic morphology of pancreas and kidney was ameliorated in the GD group. It was indicated that the therapeutic mechanisms of action of GD on type 2 DM might be related to its effect of ameliorating glucose metabolism disorders, relieving insulin resistance, increasing the tissues' sensitivity to insulin, improving the antioxidative ability of living system, GD has therapeutic effect on type 2 DM and protective effects against damaged pancreatic function.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Drugs, Chinese Herbal; Female; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Kidney; Liver; Male; Malondialdehyde; Pancreas; Phytotherapy; Plants, Medicinal; Random Allocation; Rats; Rats, Sprague-Dawley; Streptozocin; Superoxide Dismutase

Insulin resistance in Type 2 diabetes leads to hepatic steatosis that can accompanied by progressive inflammation of the liver. Citrus unshiu peel is a rich source of citrus flavonoids that possess anti-inflammatory, anti-diabetic and lipid-lowering effects. However, the ability of citrus unshiu peel ethanol extract (CPE) to improve hyperglycemia, adiposity and hepatic steatosis in Type 2 diabetes is unknown. Thus, we evaluated the effects of CPE on markers for glucose, lipid metabolism and inflammation in Type 2 diabetic mice. Male C57BL/KsJ-db/db mice were fed a normal diet with CPE (2 g/100 g diet) or rosiglitazone (0.001 g/100 g diet) for 6 weeks. Mice supplemented with the CPE showed a significant decrease in body weight gain, body fat mass and blood glucose level. The antihyperglycemic effect of CPE appeared to be partially mediated through the inhibition of hepatic gluconeogenic phosphoenolpyruvate carboxykinase mRNA expression and its activity and through the induction of insulin/glucagon secretion. CPE also ameliorated hepatic steatosis and hypertriglyceridemia via the inhibition of gene expression and activities of the lipogenic enzymes and the activation of fatty acid oxidation in the liver. These beneficial effects of CPE may be related to increased levels of anti-inflammatory adiponectin and interleukin (IL)-10, and decreased levels of pro-inflammatory markers (IL-6, monocyte chemotactic protein-1, interferon-γ and tumor necrosis factor-α) in the plasma or liver. Taken together, we suggest that CPE has the potential to improve both hyperglycemia and hepatic steatosis in Type 2 diabetes.

Topics: Adipose Tissue; Animals; Blood Glucose; Citrus; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Enzymes; Fatty Liver; Gene Expression Regulation; Glycogen; Hyperglycemia; Inflammation; Lipid Metabolism; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Phosphoenolpyruvate Carboxykinase (ATP); Plant Extracts; Weight Gain

Emerging evidences demonstrate that excess aldosterone and insulin interact at target tissues. It has been shown that increased levels of aldosterone contribute to the development of insulin resistance and thus act as a risk factor for the development of type-2 diabetes mellitus. However, the molecular mechanisms involved in this scenario are yet to be identified. This study was designed to assess the dose-dependent effects of aldosterone on insulin signal transduction and glucose oxidation in the skeletal muscle (gastrocnemius) of adult male rat. Healthy adult male albino rats of Wistar strain (Rattus norvegicus) weighing 180-200 g were used in this study. Rats were divided into four groups. Group I: control (treated with 1 % ethanol only), group II: aldosterone treated (10 μg /kg body weight, twice daily for 15 days), group III: aldosterone treated (20 μg /kg body weight, twice daily for 15 days), and group IV: aldosterone treated (40 μg/kg body weight, twice daily for 15 days). Excess aldosterone caused glucose intolerance in a dose-dependent manner. Serum insulin and aldosterone were significantly increased, whereas serum testosterone was decreased. Aldosterone treatment impaired the rate of glucose uptake, oxidation, and insulin signal transduction in the gastrocnemius muscle through defective expression of IR, IRS-1, Akt, AS160, and GLUT4 genes. Phosphorylation of IRS-1, β-arrestin-2, and Akt was also reduced in a dose-dependent manner. Excess aldosterone results in glucose intolerance as a result of impaired insulin signal transduction leading to decreased glucose uptake and oxidation in skeletal muscle. In addition to this, it is inferred that excess aldosterone may act as one of the causative factors for the onset of insulin resistance and thus increased incidence of type-2 diabetes.

Topics: Aldosterone; Animals; Arrestins; beta-Arrestin 2; beta-Arrestins; Blood Glucose; CSK Tyrosine-Protein Kinase; Diabetes Mellitus, Type 2; Glucose Intolerance; Glucose Transporter Type 4; Glycogen; GTPase-Activating Proteins; Insulin; Insulin Receptor Substrate Proteins; Lipid Peroxidation; Male; Muscle, Skeletal; Oxidation-Reduction; Phosphorylation; Protein Processing, Post-Translational; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; Reactive Oxygen Species; Receptor, Insulin; RNA, Messenger; Signal Transduction; src-Family Kinases; Testosterone

Interleukin-6 (IL-6) has a dual role in modulating insulin sensitivity, with evidence for this cytokine as both an enhancer and inhibitor of insulin action. We determined the effect of IL-6 exposure on glucose and lipid metabolism in cultured myotubes established from people with normal glucose tolerance or type 2 diabetes. Acute IL-6 exposure increased glycogen synthesis, glucose uptake, and signal transducer and activator of transcription 3 (STAT3) phosphorylation in cultured myotubes from normal glucose tolerant subjects. However, in type 2 diabetic patients, IL-6 was without effect on glucose metabolism and STAT3 signaling, concomitant with increased suppressor of cytokine signaling 3 (SOCS3) expression. IL-6 increased fatty acid oxidation in myotubes from type 2 diabetic and normal glucose tolerant subjects. Expression of IL-6, IL-6 receptor (IL-6R), or glycoprotein 130, as well as IL-6 secretion, was unaltered between cultured myotubes from normal glucose tolerant or type 2 diabetic subjects. Circulating serum IL-6 concentration was unaltered between normal glucose tolerant and type 2 diabetic subjects. In summary, skeletal muscle cells from type 2 diabetic patients display selective IL-6 resistance for glucose rather than lipid metabolism. In conclusion, IL-6 appears to play a differential role in regulating metabolism in type 2 diabetic patients compared with normal glucose tolerant subjects.

Topics: Animals; Cells, Cultured; Diabetes Mellitus, Type 2; Fatty Acids; Female; Glucose; Glycogen; Glycoproteins; Humans; Insulin Resistance; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Middle Aged; Muscle Fibers, Skeletal; Muscle, Skeletal; Phosphorylation; Receptors, Interleukin-6; STAT3 Transcription Factor; Suppressor of Cytokine Signaling 3 Protein; Suppressor of Cytokine Signaling Proteins

Lower quantity and quality of high density lipoprotein (HDL) are important characteristics of type 2 diabetes mellitus. Acute HDL infusion results in a greater fall of plasma glucose in diabetes patients. Here, we aim to investigate the influence of long-term HDL infusion on metabolic phenotypes of diabetic db/db mice.. High density lipoprotein was introduced to db/db mice twice a week for 4 weeks. The phenotypes of the mice were monitored by analyzing metabolic parameters. Glycogen analysis was performed with amyloglucosidase. The corresponding signaling molecules were detected by western blot.. Long-term introduction of HDL decreased plasma glucose levels of db/db mice. Glycogen deposition was enhanced in gastrocnemius muscle, paralleling the elevated glycogen synthase kinase-3 phosphorylation. Meanwhile, increased Akt-Ser473 and adenosine monophosphate-activated protein kinase phosphorylations were detected in the muscle. Moreover, HDL reduced blood glucose and free fatty acids and improved pancreatic islet structure and function with increased C-peptide. Furthermore, decreased interleukin-6, C-reactive protein, monocyte chemoattractant protein-1, resistin, and malondialdehyde, as well as enhanced leptin levels were detected in HDL-treated mice.. Results of the present study suggest that long-term HDL infusion has positive therapeutic effects on the metabolic disturbances of db/db diabetic mice.

Topics: Adipose Tissue; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Glycogen; Insulin Resistance; Islets of Langerhans; Lipid Metabolism; Lipid Peroxidation; Lipoproteins, HDL; Male; Mice; Mice, Inbred C57BL; Muscle, Skeletal; Phenotype

Phycocyanin (PC) has been proven to have many therapeutic properties, but its effects on diabetes have not been investigated.. Antidiabetic activity of PC isolated from Spirulina platensis was evaluated in this study.. Oral administration of PC (100 mg/kg, once per day for 3 weeks) on KKAy mice were investigated by monitoring the changes in body weight, food intake, fasting plasma glucose level, 24 h random blood glucose levels, oral glucose tolerance tests (OGTTs), glycosylated serum protein (GSP), fasting serum insulin (FINS), glycogen, triglyceride (TG), total cholesterol (TC), total antioxidative capability (T-AOC) and malondialdehyde (MDA). Histopathological changes in the pancreas were also examined with hematoxylin-eosin staining.. Administration of PC significantly decreased the body weight, fasting plasma glucose, 24 h random blood glucose levels, FINS and GSP levels, TG and TC content in serum and livers, MDA content in livers (p < 0.05 or p < 0.01). On the other hand, glucose tolerance to glucose administration, T-AOC, and the content of glycogen in liver and muscle were enhanced following PC treatment (p < 0.05 or p < 0.01). Histopathological results showed that PC administration suppressed the abnormal enlargement of islets observed in the pancreas of KKAy mice.. The antidiabetic effect of PC on KKAy mice is most likely due to its ability to enhance insulin sensitivity, amelioration of insulin resistance of peripheral target tissues and regulation of glucolipide metabolism. Therefore, PC may have a potential clinical utility in combating type-2 diabetes.

Topics: Administration, Oral; Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Female; Glucose Tolerance Test; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Islets of Langerhans; Liver; Mice; Phycocyanin; Spirulina

The present study was designed to systematically investigate the antidiabetic potential of Amaranthus spinosus leaves which are traditionally known to have various medicinal properties and used for the treatment of diabetes mellitus. The ethanolic extract of leaves of Amaranthus spinosus was administered (150, 300 and 450 mg/kg bw) to type-1 and type-2 diabetic rats. Standard drugs, glibenclamide and metformin were used as a positive control for comparison. Changes in carbohydrate and lipid metabolism and antioxidants were assessed and compared with control and standard drug treated animals. Among the standardized extract doses tested (150, 300 and 450 mg/kg bw), higher doses significantly decreased plasma glucose levels (p<0.01 and p<0.001), hepatic glucose-6-phophatase activity (p<0.01 and p<0.001) and increased the hepatic glycogen content (p<0.01) with a concurrent increase in hexokinase activity in both type 1 and 2 diabetic rats (p<0.01 and p<0.001). Besides, the higher doses also significantly lowered the plasma and hepatic lipids, urea, creatinine levels (p<0.001) and lipid peroxidation with an improvement in the antioxidant profiles (p<0.001) of both type-1 and type-2 diabetic rats. It is concluded that Amaranthus spinosus has potential antidiabetic activity and significantly improves disrupted metabolisms and antioxidant defense in type-1 and type-2 diabetic rats.

Topics: Amaranthus; Animals; Blood Glucose; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glucose-6-Phosphatase; Glycogen; Hypoglycemic Agents; Lipid Metabolism; Liver; Male; Plant Leaves; Plant Preparations; Rats

Diabetes mellitus is one of the three most common modem diseases. A number of animal models is used in investigations of the mechanisms of development of the disease. Most of these models replicate the symptoms of type 1 diabetes mellitus. The development of type 2 diabetes is caused by the insulin resistance, hyperglycemia, structural and functional disorders of the pancreatic cells. Investigation of pathogenesis of type 2 diabetes is complicated by the lack of adequate models of this disease. In this work, based on existing hyperglycemia model, we propose the model of metabolic syndrome as a precursor of type 2 diabetes. The development of metabolic syndrome symptoms was caused by 28 days long intramuscular injection of protamine sulfate to guinea pigs at a dose of 15 mg/kg along with keeping of animals on a high glucose diet. Increased blood glucose and cholesterol levels, reduction of glycogen in liver, the structural and functional damage and reduce in the number of functionally active beta-cells in the pancreas of the experimental animals were observed. The results confirm the development of the metabolic syndrome symptoms in experimental animals, which makes it possible to use such methodical approach in creation of promising type 2 diabetes model.

Topics: Animals; Diabetes Mellitus, Type 2; Diet; Disease Models, Animal; Glucose; Glycogen; Guinea Pigs; Hyperglycemia; Injections, Intramuscular; Insulin Resistance; Insulin-Secreting Cells; Liver; Male; Metabolic Syndrome; Protamines

Hepatic insulin resistance is the major contributor to fasting hyperglycemia in type 2 diabetes. The protein kinase Akt plays a central role in the suppression of gluconeogenesis involving forkhead box O1 (Foxo1) and peroxisome proliferator-activated receptor gamma co-activator 1 alpha (PGC-1α), and in the control of glycogen synthesis involving the glycogen synthase kinase beta (GSK3β) in the liver. It has been demonstrated that endosomal adaptor protein APPL1 interacts with Akt and blocks the association of Akt with its endogenous inhibitor, tribbles-related protein 3 (TRB3), improving the action of insulin in the liver. Here, we demonstrated that chronic exercise increased the basal levels and insulin-induced Akt serine phosphorylation in the liver of diet-induced obese mice. Endurance training was able to increase APPL1 expression and the interaction between APPL1 and Akt. Conversely, training reduced both TRB3 expression and TRB3 and Akt association. The positive effects of exercise on insulin action are reinforced by our findings that showed that trained mice presented an increase in Foxo1 phosphorylation and Foxo1/PGC-1α association, which was accompanied by a reduction in gluconeogenic gene expressions (PEPCK and G6Pase). Finally, exercised animals demonstrated increased at basal and insulin-induced GSK3β phosphorylation levels and glycogen content at 24 h after the last session of exercise. Our findings demonstrate that exercise increases insulin action, at least in part, through the enhancement of APPL1 and the reduction of TRB3 expression in the liver of obese mice, independently of weight loss.

Topics: Adaptor Proteins, Signal Transducing; Animals; Cell Cycle Proteins; Diabetes Mellitus, Type 2; Diet; Forkhead Box Protein O1; Forkhead Transcription Factors; Gluconeogenesis; Glucose-6-Phosphatase; Glycogen; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Insulin; Liver; Male; Mice; Mice, Obese; Obesity; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Phosphorylation; Physical Conditioning, Animal; Physical Endurance; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; Trans-Activators; Transcription Factors; Weight Loss

The number of people suffering from diabetes is hastily increasing and the condition is associated with altered brain glucose homeostasis. Brain glycogen is located in astrocytes and being a carbohydrate reservoir it contributes to glucose homeostasis. Furthermore, glycogen has been indicated to be important for proper neurotransmission under normal conditions. Previous findings from our laboratory suggested that glucose metabolism was reduced in type 2 diabetes, and thus we wanted to investigate more specifically how brain glycogen metabolism contributes to maintain energy status in the type 2 diabetic state. Also, our objective was to elucidate the contribution of glycogen to support neurotransmitter glutamate and GABA homeostasis. A glycogen phosphorylase (GP) inhibitor was administered to Sprague-Dawley (SprD) and Zucker Diabetic Fatty (ZDF) rats in vivo and after one day of treatment [1-¹³C]glucose was used to monitor metabolism. Brain levels of ¹³C labeling in glucose, lactate, alanine, glutamate, GABA, glutamine and aspartate were determined. Our results show that inhibition of brain glycogen metabolism reduced the amounts of glutamate in both the control and type 2 diabetes models. The reduction in glutamate was associated with a decrease in the pyruvate carboxylase/pyruvate dehydrogenase ratio in the control but not the type 2 diabetes model. In the type 2 diabetes model GABA levels were increased suggesting that brain glycogen serves a role in maintaining a proper ratio between excitatory and inhibitory neurotransmitters in type 2 diabetes. Both the control and the type 2 diabetic states had a compensatory increase in glucose-derived ¹³C processed through the TCA cycle following inhibition of glycogen degradation. Finally, it was indicated that the type 2 diabetes model might have an augmented necessity for compensatory upregulation at the glycolytic level.

Topics: Animals; Aspartic Acid; Brain Chemistry; Cerebral Cortex; Diabetes Mellitus, Type 2; gamma-Aminobutyric Acid; Glucose; Glutamic Acid; Glycogen; Glycogen Phosphorylase; Homeostasis; Indoles; Lactic Acid; Magnetic Resonance Spectroscopy; Male; Phenylbutyrates; Pyruvate Carboxylase; Pyruvate Dehydrogenase Complex; Rats; Rats, Sprague-Dawley; Rats, Zucker; Synaptic Transmission

The tissue-specific role of mitochondrial respiratory capacity in the development of insulin resistance and type 2 diabetes is unclear. We determined mitochondrial function in glycolytic and oxidative skeletal muscle and liver from lean (+/?) and obese diabetic (db/db) mice. In lean mice, the mitochondrial respiration pattern differed between tissues. Tissue-specific mitochondrial profiles were then compared between lean and db/db mice. In liver, mitochondrial respiratory capacity and protein expression, including peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), was decreased in db/db mice, consistent with increased mitochondrial fission. In glycolytic muscle, mitochondrial respiration, as well as protein and mRNA expression of mitochondrial markers, was increased in db/db mice, suggesting increased mitochondrial content and fatty acid oxidation capacity. In oxidative muscle, mitochondrial complex I function and PGC-1α and mitochondrial transcription factor A (TFAM) protein levels were decreased in db/db mice, along with increased level of proteins related to mitochondrial dynamics. In conclusion, mitochondrial respiratory performance is under the control of tissue-specific mechanisms and is not uniformly altered in response to obesity. Furthermore, insulin resistance in glycolytic skeletal muscle can be maintained by a mechanism independent of mitochondrial dysfunction. Conversely, insulin resistance in liver and oxidative skeletal muscle from db/db mice is coincident with mitochondrial dysfunction.

Topics: Animal Nutritional Physiological Phenomena; Animals; Blood Glucose; Blotting, Western; Cell Lineage; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; DNA-Binding Proteins; Glycogen; Glycolysis; High Mobility Group Proteins; Insulin Resistance; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Mice, Obese; Mitochondria; Mitochondria, Liver; Mitochondria, Muscle; Obesity; Oxygen Consumption; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Real-Time Polymerase Chain Reaction; Trans-Activators; Transcription Factors; Triglycerides

Insulin stimulates glycogen synthase (GS) through dephosphorylation of serine residues, and this effect is impaired in skeletal muscle from insulin-resistant [obese and type 2 diabetic (T2DM)] subjects. Exercise also increases GS activity, yet it is not known whether the ability of exercise to affect GS is impaired in insulin-resistant subjects. The objective of this study was to examine the effect of acute exercise on GS phosphorylation and enzyme kinetic properties in muscle from insulin-resistant individuals. Lean normal glucose-tolerant (NGT), obese NGT, and obese T2DM subjects performed 40 min of moderate-intensity cycle exercise (70% of Vo(2max)). GS kinetic properties and phosphorylation were measured in vastus lateralis muscle before exercise, immediately after exercise, and 3.5 h postexercise. In lean subjects, GS fractional activity increased twofold after 40 min of exercise, and it remained elevated after the 3.5-h rest period. Importantly, exercise also decreased GS K(m) for UDP-glucose from ≈0.5 to ≈0.2 mM. In lean subjects, exercise caused significant dephosphorylation of GS by 50-70% (Ser(641), Ser(645), and Ser(645,649,653,657)), and phosphorylation of these sites remained decreased after 3.5 h; Ser⁷ phosphorylation was not regulated by exercise. In obese NGT and T2DM subjects, exercise increased GS fractional activity, decreased K(m) for UDP-glucose, and decreased GS phosphorylation as effectively as in lean NGT subjects. We conclude that the molecular regulatory process by which exercise promotes glycogen synthesis in muscle is preserved in insulin-resistant subjects.

Topics: Adult; Bicycling; Biopsy; Body Mass Index; Diabetes Mellitus, Type 2; Female; Glycogen; Glycogen Synthase; Humans; Insulin Resistance; Kinetics; Male; Middle Aged; Motor Activity; Obesity; Oxygen Consumption; Phosphorylation; Protein Processing, Post-Translational; Quadriceps Muscle; Uridine Diphosphate Glucose

Diabetes mellitus (DM) often leads to disability from vascular complications and neurological complications. Tinospora crispa has been widely used in Asia and Africa as a remedy for diabetes and other diseases. In this study, we investigated the hypoglycemic actions of borapetoside C isolated from T. crispa, and the mechanisms underlying its actions. Acute treatment with borapetoside C (5mg/kg, i.p.) attenuated the elevated plasma glucose induced by oral glucose in normal and type 2 DM (T2DM) mice. Compared to the effect of injected insulin (0.5 IU/kg), borapetoside C caused a more prominent increase of glycogen content in skeletal muscle of T2DM mice, but a less increase in type 1 DM (T1DM) mice. Combined treatment of a low dose borapetoside C (0.1mg/kg, i.p.) plus insulin enhanced insulin-induced lowering of the plasma glucose level and insulin-induced increase of muscle glycogen content. Continuous treatment with 5mg/kg borapetoside C (twice daily) for 7 days increased phosphorylation of insulin receptor (IR) and protein kinase B (Akt) as well as the expression of glucose transporter-2 (GLUT2) in T1DM mice. Combined treatment of a low dose borapetoside C (0.1mg/kg, twice daily) plus insulin for 7 days enhanced insulin-induced IR and Akt phosphorylation and GLUT2 expression in the liver of T1DM mice. This study proved that borapetoside C can increase glucose utilization, delayed the development of insulin resistance and enhanced insulin sensitivity. The activation of IR-Akt-GLUT2 expression and the enhancement of insulin sensitivity may contribute to the hypoglycemic action of borapetoside C in diabetic mice.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Diterpenes; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Drug Therapy, Combination; Glucose Tolerance Test; Glucose Transporter Type 2; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Male; Mice; Mice, Inbred ICR; Muscle, Skeletal; Phosphorylation; Plants, Medicinal; Proto-Oncogene Proteins c-akt; Receptor, Insulin; Signal Transduction; Tinospora

The purpose of this study was to investigate the influence of exercise training on intramyocellular lipid (IMCL) content and test the hypothesis that the effect of endurance-oriented exercise training on IMCL is dependent on characteristics of the population studied. Lean (N = 11, body mass index (BMI) = 22.2 ± 0.7 kg·m⁻²), obese (N = 14, BMI = 38.8 ± 1.7 kg·m⁻²), and type 2 diabetic (N = 9, BMI = 35.5 ± 2.5 kg·m⁻²) participants were examined before and after 10 consecutive days of endurance-oriented (60 min·day⁻¹ at ~70% [Formula: see text]O(2peak)) exercise training. IMCL and muscle glycogen were measured by Oil-Red-O and periodic acid - Schiff staining, respectively. The results indicated that IMCL was elevated (p < 0.05) in the obese and diabetic groups compared with the lean subjects prior to training. After training, IMCL content decreased (-35%) in the participants with type 2 diabetes; there were no changes in IMCL in the lean or obese groups. Muscle glycogen content was lower in the diabetic subjects than in the lean subjects both before and after training. These data indicate that changes in IMCL with exercise training do not exhibit a universal response but rather depend on the metabolic status of the population studied.

Topics: Adult; Biopsy, Needle; Body Mass Index; Diabetes Mellitus, Type 2; Exercise; Female; Glycogen; Humans; Lipid Metabolism; Male; Middle Aged; Muscle Fibers, Fast-Twitch; Muscle Fibers, Slow-Twitch; Obesity; Oxygen Consumption; Physical Exertion; Quadriceps Muscle; Sedentary Behavior

This study investigated the effects of resveratrol (RV) on diabetes-related metabolic changes in a spontaneous model of type 2 diabetes, as well as activation of AMP-activated protein kinase (AMPK) and downstream targets.. C57BL/KsJ-db/db mice were fed a normal diet with RV (0.005% and 0.02%, w/w) or rosiglitazone (RG, 0.001%, w/w) for 6 weeks. Both doses of RV significantly decreased blood glucose, plasma free fatty acid, triglyceride, apo B/apo AІ levels and increased plasma adiponectin levels. RV activated AMPK and downstream targets leading to decreased blood HbA1c levels, hepatic gluconeogenic enzyme activity, and hepatic glycogen, while plasma insulin levels, pancreatic insulin protein, and skeletal muscle GLUT4 protein were higher after RV supplementation. The high RV dose also significantly increased hepatic glycolytic gene expression and enzyme activity, along with skeletal muscle glycogen synthase protein expression, similar to RG. Furthermore, RV dose dependently decreased hepatic triglyceride content and phosphorylated I kappa B kinase (p-IKK) protein expression, while hepatic uncoupling protein (UCP) and skeletal muscle UCP expression were increased.. RV potentiates improving glycemic control, glucose uptake, and dyslipidemia, as well as protecting against pancreatic β-cell failure in a spontaneous type 2 diabetes model. Dietary RV has potential as an antidiabetic agent via activation of AMPK and its downstream targets.

Topics: Adiponectin; AMP-Activated Protein Kinases; Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Dietary Supplements; Dyslipidemias; Glucose Transporter Type 4; Glycated Hemoglobin; Glycogen; Insulin; Insulin Secretion; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Muscle, Skeletal; Resveratrol; Rosiglitazone; Stilbenes; Thiazolidinediones; Triglycerides

Hepatitis C virus (HCV) infection may cause liver diseases of various severities ranging from primary acute infection to life-threatening diseases, such as cirrhosis or hepatocellular carcinoma with poor prognosis. According to clinical findings, HCV infection may also lead to some extra-hepatic symptoms, including type 2 diabetes mellitus (DM). Since insulin resistance is the major etiology for type 2 DM and numerous evidences showed that HCV infection associated with insulin resistance, the involvement of E2 in the pathogenesis of type 2 DM and underlying mechanisms were investigated in this study.. Reverse transcription and real-time PCR, Western blot assay, Immunoprecipitation, Glucose uptake assay and analysis of cellular glycogen content.. Results showed that E2 influenced on protein levels of insulin receptor substrate-1 (IRS-1) and impaired insulin-induced Ser308 phosphorylation of Akt/PKB and Ser9 phosphorylation of GSK3β in Huh7 cells, leading to an inhibition of glucose uptake and glycogen synthesis, respectively, and eventually insulin resistance.. Therefore, HCV E2 protein indeed involved in the pathogenesis of type 2 DM by inducing insulin resistance.

Topics: Cell Line; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Hepacivirus; Hepatocytes; Humans; Insulin Receptor Substrate Proteins; Insulin Resistance; Phosphorylation; Proto-Oncogene Proteins c-akt; Serine; Signal Transduction; Transfection; Viral Envelope Proteins

Mutations in Wnt receptor LRP5/6 and polymorphism in Wnt-regulated transcription factor TCF7L2 are associated with dysregulation of glucose metabolism. However, it is not clear whether Wnt antagonist Dickkopf (Dkk) has a significant role in the regulation of glucose metabolism. Here, we identified small-molecule inhibitors of Wnt antagonist Dkk through molecular modeling, computation-based virtual screens, and biological assays. One of the Dkk inhibitors reduced basal blood-glucose concentrations and improved glucose tolerance in mice. This Dkk inhibitor appeared to act through DKK2 because the inhibitor exerted no additional effects on glucose metabolism in the Dkk2(-/-) mice. Our study of Dkk2(-/-) mice showed that DKK2 deficiency was associated with increased hepatic glycogen accumulation and decreased hepatic glucose output. DKK2 deficiency did not cause in increase in insulin production but resulted in increased Wnt activity and GLP1 production in the intestines. Given that the Dkk inhibitor improved glucose tolerance in a murine model of type 2 diabetes (db/db), we suggest that DKK2 may be a potential therapeutic target for treating type 2 diabetes.

Topics: Animals; Diabetes Mellitus, Type 2; Gene Expression Regulation; Glucose; Glucose Tolerance Test; Glycogen; Intercellular Signaling Peptides and Proteins; Kinetics; Liver; Mice; Mice, Transgenic; Models, Genetic; Polymorphism, Genetic; Signal Transduction; Software; Transcription Factor 7-Like 2 Protein; Wnt Proteins

American ginseng (Panax quinquefolius L.) root health benefits include treatment of type 2 diabetes and this study evaluated the hypoglycemic effects of American red ginseng (ARG). ARG roots have increased bioactive phenolic contents, such as cinnamic acid and ferulic acid during the steaming process. The antihyperglycemic effects of methanol fraction extract of ARG, ferulic acid, and cinnamic acid were examined using a type 2 diabetic mouse model. The ARG treated group presented relatively lower blood glucose levels than the control group (P < 0.05). In addition, the glycogen and high density lipoprotein (HDL) contents were significantly increased while levels of plasma cholesterol and low density lipoprotein (LDL) concentration were significantly decreased in the ARG treated group. The groups treated with ferulic and cinnamic acids showed similar effects as those found in the ARG treated group. Thus, it is suggested that ARG roots, ferulic acid, and cinnamic acid have hypoglycemic effects in an animal model.. This study was conducted to elucidate the hypoglycemic effects of American red ginseng (ARG) using a type 2 diabetic mouse model. ARG showed an enhanced antioxidant capacity and higher antihyperglycemia effect. The glycogen and high density lipoprotein (HDL) contents were significantly increased while levels of plasma cholesterol and low density lipoprotein (LDL) concentration were significantly decreased in the ARG treated group. It is suggested that ARG has a potential to be used for human diabetic treatment.

Topics: Animals; Blood Glucose; Cinnamates; Coumaric Acids; Diabetes Mellitus, Type 2; Disease Models, Animal; Glutathione Peroxidase; Glycogen; Hypoglycemic Agents; Insulin; Lipid Peroxidation; Lipoproteins, HDL; Lipoproteins, LDL; Male; Malondialdehyde; Mice; Mice, Inbred Strains; Panax; Phytotherapy; Plant Extracts; Plant Roots; Superoxide Dismutase

The aim of the present study is to investigate the antidiabetic properties of oligosaccharides of Ophiopogonis japonicus (OOJ) in experimental type 2 diabetic rats. OOJ was administered orally in doses of 225 and 450 mg/kg body weight to high-fat diet and low-dose streptozotocin (STZ)-induced type 2 diabetic rats for 3 weeks. The results showed that OOJ treatment could increase body weight, decrease organ related weights of liver and kidney, reduce fasting blood glucose level, and improve oral glucose tolerance in diabetic rats. Moreover, increased glycogen content in liver and skeletal muscle, reduced urinary protein excretion, higher hepatic GCK enzyme activity, lower hepatic PEPCK enzyme activity, enhanced GLP-1 level, decreased glucagon level and alleviated histopathological changes of pancreas occurred in OOJ-treated diabetic rats by comparison with untreated diabetic rats. This study demonstrates, for the first time to our knowledge, that OOJ exerts remarkable antidiabetic effect in experimental type 2 diabetes mellitus, thus justifying its traditional usage.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Fasting; Glucagon; Glucagon-Like Peptide 1; Glucose Tolerance Test; Glycogen; Hypoglycemic Agents; Liver; Male; Muscle, Skeletal; Oligosaccharides; Ophiopogon; Organ Size; Proteinuria; Rats; Rats, Sprague-Dawley

Acute hyperglycaemia rapidly suppresses endogenous glucose production (EGP) in non-diabetic individuals, mainly by inhibiting glycogenolysis. Loss of this 'glucose effectiveness' contributes to fasting hyperglycaemia in type 2 diabetes. Elevated NEFA levels characteristic of type 2 diabetes impair glucose effectiveness, although the mechanism is not fully understood. Therefore we examined the impact of increasing NEFA levels on the ability of hyperglycaemia to regulate pathways of EGP.. We performed 4 h 'pancreatic clamp' studies (somatostatin; basal glucagon/growth hormone/insulin) in seven non-diabetic individuals. Glucose fluxes (D-[6,6-(2)H(2)]glucose) and hepatic glycogen concentrations ((13)C magnetic resonance spectroscopy) were quantified under three conditions: euglycaemia, hyperglycaemia and hyperglycaemia with elevated NEFA (HY-NEFA).. EGP was suppressed by hyperglycaemia, but not by HY-NEFA. Hepatic glycogen concentration decreased ~14% with prolonged fasting during euglycaemia and increased by ~12% with hyperglycaemia. In contrast, raising NEFA levels in HY-NEFA caused a substantial ~23% reduction in hepatic glycogen concentration. Moreover, rates of gluconeogenesis were decreased with hyperglycaemia, but increased with HY-NEFA.. Increased NEFA appear to profoundly blunt the ability of hyperglycaemia to inhibit net glycogenolysis under basal hormonal conditions.

Topics: Adult; Blood Glucose; Diabetes Mellitus, Type 2; Fasting; Fatty Acids, Nonesterified; Glucagon; Glucose Clamp Technique; Glycogen; Glycogenolysis; Human Growth Hormone; Humans; Hyperglycemia; Insulin; Liver; Male; Somatostatin

Recent studies have suggested resveratrol (RSV) as a new natural therapeutic agent to treat type 2 diabetes and lipid-induced insulin resistance. Here, we investigated whether RSV could reverse palmitate-induced insulin resistance in human primary muscle cells.. Myotubes obtained from six healthy men (54 ± 3 years (mean ± SE), BMI 25.0 ± 1.7 kg/m(2), fasting plasma glucose concentration (fP-glucose) 5.47 ± 0.09 mmol/l) were treated for 4 h with 100 μmol/l RSV and/or 0.2 mmol/l palmitate, and stimulated with or without 100 nmol/l insulin. Assays of glucose uptake, glycogen synthesis, palmitate oxidation, intracellular signalling and AMP-activated protein kinase (AMPK) activity were performed.. RSV did not reverse palmitate-induced impairment of glucose metabolism. Surprisingly, RSV decreased glucose uptake and glycogen synthesis in human skeletal muscle cells. Palmitate oxidation and phosphorylation of AMPK and its downstream target acetyl-CoA carboxylase β (ACCβ) were inhibited by RSV, and RSV completely blocked the activity of AMPK isoform complexes α1/β2/γ1 and α2/β2/γ1 in in-vitro kinase activity assays. Endoplasmic reticulum (ER) stress was increased in response to RSV, as indicated by increased phosphorylation of eukaryotic initiation factor 2α (eIF2α) and increased expression of CCAAT/enhancer binding protein homologous protein (CHOP).. Acute exposure to RSV inhibits AMPK activity, fatty-acid oxidation and glucose metabolism in human myotubes.

Topics: AMP-Activated Protein Kinases; Cell Differentiation; Diabetes Mellitus, Type 2; Drug Interactions; Enzyme Inhibitors; Glucose; Glycogen; Humans; Insulin Resistance; Male; Middle Aged; Muscle Fibers, Skeletal; Palmitates; Phosphorylation; Primary Cell Culture; Resveratrol; Signal Transduction; Stilbenes

A major problem in the insulin therapy of patients with diabetes type 2 (T2DM) is the increased occurrence of hypoglycemic events which, if left untreated, may cause confusion or fainting and in severe cases seizures, coma, and even death. To elucidate the potential contribution of the liver to hypoglycemia in T2DM we applied a detailed kinetic model of human hepatic glucose metabolism to simulate changes in glycolysis, gluconeogenesis, and glycogen metabolism induced by deviations of the hormones insulin, glucagon, and epinephrine from their normal plasma profiles. Our simulations reveal in line with experimental and clinical data from a multitude of studies in T2DM, (i) significant changes in the relative contribution of glycolysis, gluconeogenesis, and glycogen metabolism to hepatic glucose production and hepatic glucose utilization; (ii) decreased postprandial glycogen storage as well as increased glycogen depletion in overnight fasting and short term fasting; and (iii) a shift of the set point defining the switch between hepatic glucose production and hepatic glucose utilization to elevated plasma glucose levels, respectively, in T2DM relative to normal, healthy subjects. Intriguingly, our model simulations predict a restricted gluconeogenic response of the liver under impaired hormonal signals observed in T2DM, resulting in an increased risk of hypoglycemia. The inability of hepatic glucose metabolism to effectively counterbalance a decline of the blood glucose level becomes even more pronounced in case of tightly controlled insulin treatment. Given this Janus face mode of action of insulin, our model simulations underline the great potential that normalization of the plasma glucagon profile may have for the treatment of T2DM.

Topics: Algorithms; Blood Glucose; Carbohydrate Metabolism; Computer Simulation; Diabetes Mellitus, Type 2; Epinephrine; Glucagon; Glucose; Glycogen; Hepatocytes; Humans; Hypoglycemia; Hypoglycemic Agents; Insulin; Insulin Resistance; Kinetics; Liver; Models, Biological; Phosphorylation; Postprandial Period; Protein Processing, Post-Translational

Persimmon Leaf (PL), commonly consumed as herbal tea and traditional medicines, contains a variety of compounds that exert antioxidant, α-amylase and α-glucosidase inhibitory activity. However, little is known about the in vivo effects and underlying mechanisms of PL on hyperglycemia, hyperlipidemia and hepatic steatosis in type 2 diabetes. Powered PL (5%, w/w) was supplemented with a normal diet to C57BL/KsJ-db/db mice for 5 weeks. PL decreased blood glucose, HOMA-IR, plasma triglyceride and total cholesterol levels, as well as liver weight, hepatic lipid droplets, triglycerides and cholesterol contents, while increasing plasma HDL-cholesterol and adiponectin levels. The anti-hyperglycemic effect was linked to decreased activity of gluconeogenic enzymes as well as increased glycogen content, glucokinase activity and its mRNA level in the liver. PL also led to a decrease in lipogenic transcriptional factor PPARγ as well as gene expression and activity of enzymes involved in lipogenesis, with a simultaneous increase in fecal lipids, which are seemingly attributable to the improved hyperlipidemia and hepatic steatosis and decreased hepatic fatty acid oxidation. Furthermore, PL ameliorated plasma and hepatic oxidative stress. Supplementation with PL may be an effective dietary strategy to improve type 2 diabetes accompanied by dyslipidemia and hepatic steatosis by partly modulating the activity or gene expression of enzymes related to antioxidant, glucose and lipid homeostasis.

Topics: Adiponectin; Animals; Antioxidants; Blood Glucose; Cholesterol; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dietary Supplements; Diospyros; Dyslipidemias; Fatty Liver; Gene Expression; Glucokinase; Glycogen; Hyperglycemia; Hypoglycemic Agents; Insulin Resistance; Lipid Metabolism; Lipogenesis; Liver; Male; Mice; Mice, Inbred C57BL; Oxidative Stress; Phytotherapy; Plant Leaves; Plant Preparations; PPAR gamma; RNA, Messenger; Triglycerides

Type 2 diabetes is caused by relative deficiency of insulin secretion and is associated with dysregulation of glucagon secretion during the late stage of diabetes development. Like insulin secretion from beta cells, glucagon secretion is dependent on calcium signals and a calcium sensing protein, synaptotagmin-7. In this study, we tested the relative contribution of dysregulated glucagon secretion and reduced insulin release in the development of hyperglycaemia and type 2 diabetes by using synaptotagmin-7 knockout (KO) mice, which exhibit glucose intolerance, reduced insulin secretion and nearly abolished Ca(2+)-stimulated glucagon secretion.. We fed the synaptotagmin-7 KO and control mice with a high-fat diet (HFD) for 14 weeks, and compared their body weight, glucose levels, glucose and insulin tolerance, and insulin and glucagon secretion.. On the HFD, synaptotagmin-7 KO mice showed progressive impairment of glucose tolerance and insulin secretion, along with continued maintenance of a low glucagon level. The control mice were less affected in terms of glucose intolerance, and showed enhanced insulin secretion with a concurrent increase in glucagon levels. Unexpectedly, after 14 weeks of HFD feeding, only the control mice displayed resting hyperglycaemia, whereas in synaptotagmin-7 KO mice defective insulin secretion and reduced insulin sensitivity were not sufficient to cause hyperglycaemia in the absence of enhanced glucagon secretion.. Our data uncover a previously overlooked role of dysregulated glucagon secretion in promoting hyperglycaemia and the ensuing diabetes, and strongly suggest maintenance of adequate regulation of glucagon secretion as an important therapeutic target in addition to the preservation of beta cell function and mass in the prevention and treatment of diabetes.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Dietary Fats; Glucagon; Glycogen; Hyperglycemia; Male; Mice; Mice, Knockout; Synaptotagmins

Sulfonylureas (SUs) may impair outcome in patients with acute coronary syndrome. Most experimental studies of the myocardial effects of SU treatment are performed in non-diabetic models. We compared the effect of two widely used SUs, glibenclamide (gb) and gliclazide (gc), with high and low myocardial K(ATP) channel affinity, respectively, at therapeutic concentrations on infarct size, left ventricular (LV) function and myocardial glycogen, lactate and alanine content before and after ischaemia/reperfusion (I/R).. Non-diabetic Wistar and diabetic Goto-Kakizaki rat hearts were investigated in a Langendorff preparation. Gb (0.1 μmol/l) and gc (1.0 μmol/l) were administrated throughout the study. Infarct size was evaluated after 120 min of reperfusion. Myocardial metabolite content was measured before and after ischaemia.. Infarct size was smaller in diabetic hearts than in non-diabetic hearts (0.33 ± 0.03 vs 0.51 ± 0.05, p < 0.05). Gb increased infarct size (0.54 ± 0.04 vs 0.33 ± 0.03, p < 0.05) and reduced post-ischaemic LV developed pressure (60 ± 3 vs 76 ± 3 mmHg, p < 0.05) and coronary flow (4.9 ± 0.5 vs 7.1 ± 0.4 ml min(-1) g(-1), p < 0.05) in gb-treated diabetic rats compared with untreated diabetic rats. On comparing gb-treated diabetic rats with untreated diabetic rats, glycogen content was reduced before (9.1 ± 0.6 vs 13.6 ± 1.0 nmol/mg wet weight, p < 0.01) and after ischaemia (0.9 ± 0.2 vs 1.8 ± 0.2 nmol/mg wet weight, p < 0.05), and lactate (4.8 ± 0.4 vs 3.2 ± 0.3 nmol/mg wet weight, p < 0.01) and alanine (1.38 ± 0.12 vs 0.96 ± 0.09 nmol/mg wet weight, p < 0.05) contents were increased during reperfusion. Gc-treatment of diabetic and non-diabetic rats did not affect any of the measured variables.. Gb, but not gc, exacerbates I/R injury and deteriorates LV function in diabetic hearts. These effects of gb on diabetic hearts may be due to detrimental effects on myocardial carbohydrate metabolism.

Topics: Animals; Diabetes Mellitus, Type 2; Gliclazide; Glyburide; Glycogen; Lactic Acid; Male; Myocardial Infarction; Myocardium; Potassium Channels; Rats; Rats, Wistar; Sulfonylurea Compounds

Symplocos cochinchinensis (Lour.) S. Moore. is used in Indian system of traditional medicine to treat diabetes mellitus. The present study aims to investigate the antidiabetic efficacy of the hexane extract of Symplocos cochinchinensis leaves in high fat diet-low streptozotocin (STZ) induced type 2 diabetic rats.. The doses for the study were fixed based on Irwin test. The hypoglycemic effect of the hexane extract of Symplocos cochinchinensis leaves were studied in normal rats. Oral glucose and insulin tolerance tests were carried out. The antihyperglycemic effect of the hexane extract at 250 and 500 mg/kg was studied in high fat diet-low STZ induced type 2 diabetic rats for 28 days.. The extracts showed no adverse effects up to 5 g/kg concentration. In hypoglycemic study, after treatment with hexane extract at 250 and 500 mg/kg the blood glucose was mildly reduced. In oral glucose tolerance test, the treatment with the hexane extract at 250 and 500 mg/kg showed a highly significant reduction of 12.07% and 23.58% in plasma glucose levels, respectively 30 min after glucose load. The insulin tolerance test also showed improved insulin sensitivity after 60 min of insulin treatment. In high fat diet-low STZ induced type 2 diabetic rats, after 28 days treatment with the hexane extract at 250 and 500 mg/kg reduced the plasma glucose level by 17.04% and 42.10%, respectively. A significant reduction in plasma insulin, plasma and hepatic total cholesterol (TC), triglycerides (TG) and free fatty acids (FFA) and a significant increase in liver glycogen were observed in treated diabetic rats.. This study demonstrated the potential antidiabetic property of hexane extract of Symplocos cochinchinensis leaves on type 2 diabetes mellitus, thus justifying its traditional usage.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Dietary Fats; Glucose Tolerance Test; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Lipids; Liver; Male; Phytotherapy; Plant Extracts; Plant Leaves; Rats; Rats, Wistar; Streptozocin

Recent extensive studies have revealed that molecular hydrogen (H(2)) has great potential for improving oxidative stress-related diseases by inhaling H(2) gas, injecting saline with dissolved H(2), or drinking water with dissolved H(2) (H(2)-water); however, little is known about the dynamic movement of H(2) in a body. First, we show that hepatic glycogen accumulates H(2) after oral administration of H(2)-water, explaining why consumption of even a small amount of H(2) over a short span time efficiently improves various disease models. This finding was supported by an in vitro experiment in which glycogen solution maintained H(2). Next, we examined the benefit of ad libitum drinking H(2)-water to type 2 diabetes using db/db obesity model mice lacking the functional leptin receptor. Drinking H(2)-water reduced hepatic oxidative stress, and significantly alleviated fatty liver in db/db mice as well as high fat-diet-induced fatty liver in wild-type mice. Long-term drinking H(2)-water significantly controlled fat and body weights, despite no increase in consumption of diet and water. Moreover, drinking H(2)-water decreased levels of plasma glucose, insulin, and triglyceride, the effect of which on hyperglycemia was similar to diet restriction. To examine how drinking H(2)-water improves obesity and metabolic parameters at the molecular level, we examined gene-expression profiles, and found enhanced expression of a hepatic hormone, fibroblast growth factor 21 (FGF21), which functions to enhance fatty acid and glucose expenditure. Indeed, H(2) stimulated energy metabolism as measured by oxygen consumption. The present results suggest the potential benefit of H(2) in improving obesity, diabetes, and metabolic syndrome.

Topics: Animals; Diabetes Mellitus, Type 2; Energy Metabolism; Fatty Liver; Fibroblast Growth Factors; Glycogen; Hydrogen; Hyperglycemia; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Obesity; Oxidative Stress; Random Allocation; Rats; Rats, Sprague-Dawley; RNA, Messenger; Water

Substrate imbalance is a well-recognized feature of diabetic cardiomyopathy. Insulin resistance effectively limits carbohydrate oxidation, resulting in abnormal cardiac glycogen accumulation. Aims of the present study were to 1) characterize the role of glycogen-associated proteins involved in excessive glycogen accumulation in type 2 diabetic hearts and 2) determine if exercise training can attenuate abnormal cardiac glycogen accumulation. Control (db(+)) and genetically diabetic (db/db) C57BL/KsJ-lepr(db)/lepr(db) mice were subjected to sedentary or treadmill exercise regimens. Exercise training consisted of high-intensity/short-duration (10 days) and low-intensity/long-duration (6 wk) protocols. Glycogen levels were elevated by 35-50% in db/db hearts. Exercise training further increased (2- to 3-fold) glycogen levels in db/db hearts. Analysis of soluble and insoluble glycogen pools revealed no differential accumulation of one glycogen subspecies. Phosphorylation (Ser(640)) of glycogen synthase, an indicator of enzymatic fractional activity, was greater in db/db mice subjected to sedentary and exercise regimens. Elevated glycogen levels were accompanied by decreased phosphorylation (Thr(172)) of 5'-AMP-activated kinase and phosphorylation (Ser(79)) of its downstream substrate acetyl-CoA carboxylase. Glycogen concentration was not associated with increases in other glycogen-associated proteins, including malin and laforin. Novel observations show that exercise training does not correct diabetes-induced elevations in cardiac glycogen but, rather, precipitates further accumulation.

Topics: Animals; Body Weight; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Cardiomyopathies; Exercise Therapy; Glycogen; Glycogen Storage Disease Type IIb; Mice; Mice, Inbred C57BL; Mice, Transgenic; Myocardium; Physical Conditioning, Animal; Receptors, Leptin

Suppression of lipolysis by acipimox is known to improve insulin-stimulated glucose disposal, and this is an important phenomenon. The mechanism has been assumed to be an enhancement of glucose storage as glycogen, but no direct measurement has tested this concept or its possible relationship to the reported impairment in insulin-stimulated muscle ATP production. Isoglycaemic-hyperinsulinaemic clamps with [13C]glucose infusion were performed on Type 2 diabetic subjects and matched controls with measurement of glycogen synthesis by 13C MRS (magnetic resonance spectroscopy) of muscle. 31P saturation transfer MRS was used to quantify muscle ATP turnover rates. Glucose disposal rates were restored to near normal in diabetic subjects after acipimox (6.2 ± 0.8 compared with 4.8 ± 0.6 mg·kgffm⁻¹·min⁻¹; P<0.01; control 6.6 ± 0.5 mg·kgffm⁻¹·min⁻¹; where ffm, is fat-free mass). The increment in muscle glycogen concentration was 2-fold higher in controls compared with the diabetic group, and acipimox administration to the diabetic group did not increase this (2.0 ± 0.8 compared with 1.9 ± 1.1 mmol/l; P<0.05; control, 4.0 ± 0.8 mmol/l). ATP turnover rates did not increase during insulin stimulation in any group, but a modest decrease in the diabetes group was prevented by lowering plasma NEFAs (non-esterified fatty acids; 8.4 ± 0.7 compared with 7.1 ± 0.5 μmol·g⁻¹·min⁻¹; P<0.05; controls 8.6 ± 0.8 μmol·g⁻¹·min⁻¹). Suppression of lipolysis increases whole-body glucose uptake with no increase in the rate of glucose storage as glycogen but with increase in whole-body glucose oxidation rate. ATP turnover rate in muscle exhibits no relationship to the acute metabolic effect of insulin.

Topics: Adenosine Triphosphate; Blood Glucose; Breath Tests; Diabetes Mellitus, Type 2; Double-Blind Method; Fatty Acids, Nonesterified; Female; Glucose Clamp Technique; Glycogen; Humans; Hypolipidemic Agents; Insulin; Lipolysis; Magnetic Resonance Spectroscopy; Male; Middle Aged; Muscles; Pyrazines

Diabetes is a chronic metabolic disorder affecting a vast number of people worldwide. Oxidative stress is the causative agent amplifying diabetic complications in various organs by generating noxious amount of free radicals. A huge interest always exists in exploring nutraceuticals from plant materials to replace synthetic drugs in order to overcome their adverse effects and also for economic reasons. The anti-diabetic efficiency of a medicinal plant, Tinospora cordifolia (TC) was studied in experimentally induced type 2 diabetes in Sprague-Dawley rats. Diabetes was induced by a combination of high fat diet (HFD) for a period of 10 weeks followed by intraperitoneal injection of streptozotocin (STZ, 35 mg/kg of body weight). Oral treatment of TC (100 and 200 mg/kg body weight) for 14 days regulated blood glucose, provoked insulin secretion and also suppressed oxidative stress marker, thiobarbituric acid reactive substances (TBARS), formation and restored cellular defence anti-oxidant markers including superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione (GSH), in liver. Treatment with TC (100 and 200 mg/kg) also inhibited glucose 6-phosphatase and fructose 1,6-diphosphatase (p < 0.001); and restored glycogen content in liver (p < 0.005), which was also studied by histopathological staining with periodic acid-Schiff stain. In conclusion, the traditional plant Tinospora cordifolia mediates its anti-diabetic potential through mitigating oxidative stress, promoting insulin secretion and also by inhibiting gluconeogenesis and glycogenolysis, thereby regulating blood glucose.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Type 2; Gluconeogenesis; Glutaredoxins; Glycogen; Hypoglycemic Agents; Insulin; Male; Oxidative Stress; Plant Extracts; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Thioredoxins; Tinospora

Amylin is co-secreted with insulin, responds to the same stimuli, is anorectic, lowers body weight by reducing fat mass, and is proposed for diabetes treatment. We examined the effect of a 3-day constant infusion of close to physiological doses of amylin in Wistar rats, on glucotransporter expression, glycogen content (G), glycogen synthase a activity (GSa) and glucose transport (GT), in liver, muscle and fat from insulin resistant (IR) and type 2 diabetic (T2D) models, compared to normal (N) animals; plasma glucose and insulin were measured. Plasma insulin in IR was higher than in N or T2D, and amylin normalized the value. In both, IR and T2D, liver G was lower than normal, accompanied by GLUT-2, mRNA and protein, higher and lower, respectively, than in N; amylin normalized G in both groups, without changes in GLUT-2, except for an mRNA increase in T2D. In IR and T2D, muscle GSa was reduced, together with respective over- and under-GLUT-4 expression; amylin induced only a trend toward GSa normalization in both groups. In isolated adipocytes, GT and GLUT-4 in IR and T2D were lower and higher, respectively, than in N; after amylin, not only GT was normalized in both groups but also the response to insulin was much more pronounced, including that in N, without major changes in GLUT-4. This suggests that the beneficial effect of amylin in states running with altered glucose homeostasis could occur by partially acting on the hexose metabolism of the liver and mainly on that of the adipose tissue.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glucose Transporter Type 2; Glucose Transporter Type 4; Glycogen; Glycogen Synthase; Homeostasis; Humans; Insulin; Insulin Resistance; Islet Amyloid Polypeptide; Male; Rats; Rats, Wistar

To investigate the relationship between liver glucose, glycogen, and plasma glucose in diabetic patients, in vivo liver carbon-13 magnetic resonance spectroscopy ((13)C MRS) with a clinical 3.0T MR system was performed. Subjects were healthy male volunteers (n=5) and male type-2 diabetic patients (n=5). Pre- and during oral glucose tolerance tests (OGTT), (13)C MR spectra without proton decoupling were acquired in a monitoring period of over 6h, and in total seven spectra were obtained from each subject. For OGTT, 75g of glucose, including 5g of [1-(13)C]glucose, was administered. The MR signals of liver [1-(13)C]glucose and glycogen were detected and their time-course changes were assessed in comparison with the plasma data obtained at screening. The correlations between the fasting plasma glucose level and liver glycogen/glucose rate (Spearman: rho=-0.68, p<0.05, n=10) and the fasting plasma glucose level and liver glycogen peak/fasting rate (Spearman: rho=-0.67, p<0.05, n=10) indicated that (13)C MRS can perform noninvasive measurement of glycogen storage/degradation ability in the liver individually and can assist in tailor-made therapy for diabetes. In conclusion, (13)C MRS has a potential to become a powerful tool in diagnosing diabetes multilaterally.

Topics: Carbon Isotopes; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Liver; Magnetic Resonance Spectroscopy; Male; Metabolic Clearance Rate

Caveolin, a member of the membrane-anchoring protein family, accumulates various growth receptors in caveolae and inhibits their function. Upregulation of caveolin attenuates cellular proliferation and growth. However, the role of caveolin in regulating insulin signals remains controversial. Here, we demonstrate that caveolin potently enhances insulin receptor (IR) signaling when overexpressed in the liver in vivo. Adenovirus-mediated gene transfer was used to overexpress caveolin specifically in the liver of diabetic obese mice, which were generated with a high-fat diet. Expression of molecules involved in IR signaling, such as IR or Akt, remained unchanged after gene transfer. However, hepatic glycogen synthesis was markedly increased with a decrease in phosphoenolpyruvate carboxykinase protein expression. Insulin sensitivity was increased after caveolin gene transfer as determined by decreased blood glucose levels in response to insulin injection and fasting blood glucose levels. Glucose tolerant test performance was also improved. Similar improvements were obtained in KKA(y) genetically diabetic mice. Adenovirus-mediated overexpression of caveolin-3 in hepatic cells also enhanced IR signaling, as shown by increased phosphorylation of IR in response to insulin stimulation and higher glycogen synthesis at baseline. These effects were attributed mostly to increased insulin receptor activity and caveolin-mediated, direct inhibition of protein tyrosine phosphatase 1B, which was increased in obese mouse livers. In conclusion, our results suggest that caveolin is an important regulator of glucose metabolism that can enhance insulin signals.

Topics: Adenoviridae; Age Factors; Aging; Animals; Blood Glucose; Caveolin 3; Diabetes Mellitus, Type 2; Dietary Fats; Disease Models, Animal; Gene Transfer Techniques; Genetic Vectors; Glucose Tolerance Test; Glycogen; Hep G2 Cells; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Mice; Obesity; Phosphoenolpyruvate Carboxykinase (GTP); Phosphorylation; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Rats; Receptor, Insulin; Signal Transduction

The present study focuses on identifying and developing an anti-diabetic molecule from plant sources that would effectively combat insulin resistance through proper channeling of glucose metabolism involving glucose transport and storage.. Insulin-stimulated glucose uptake formed the basis for isolation of a bioactive molecule through column chromatography followed by its characterization using NMR and mass spectroscopic analysis. Mechanism of glucose transport and storage was evaluated based on the expression profiling of signaling molecules involved in the process.. The study reports (i) the isolation of a bioactive compound 3beta-taraxerol from the ethyl acetate extract (EAE) of the leaves of Mangifera indica (ii) the bioactive compound exhibited insulin-stimulated glucose uptake through translocation and activation of the glucose transporter (GLUT4) in an IRTK and PI3K dependent fashion. (iii) the fate of glucose following insulin-stimulated glucose uptake was ascertained through glycogen synthesis assay that involved the activation of PKB and suppression of GSK3beta.. This study demonstrates the dual activity of 3beta-taraxerol and the ethyl acetate extract of Mangifera indica as a glucose transport activator and stimulator of glycogen synthesis. 3beta-taraxerol can be validated as a potent candidate for managing the hyperglycemic state.

Topics: 3T3 Cells; Adipocytes; Animals; Blood Glucose; Cell Survival; Deoxyglucose; Diabetes Mellitus, Type 2; Enzyme Activation; Glucose; Glycogen; Humans; Insulin Resistance; Mangifera; Mice; Oleanolic Acid; Phosphatidylinositol 3-Kinases; Plant Extracts

Topics: Adenoviridae; Animals; Blood Glucose; Caveolin 3; Diabetes Mellitus, Type 2; Gene Transfer Techniques; Genetic Vectors; Glycogen; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Mice; Phosphoenolpyruvate Carboxykinase (GTP); Phosphorylation; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Receptor, Insulin; Signal Transduction

Studies using chemical inhibitors have suggested that the Ca(2+)-sensitive serine/threonine kinase Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is a key regulator of both insulin- and contraction-stimulated glucose uptake in skeletal muscle. However, due to nonspecificity of these inhibitors, the specific role that CaMKII may play in the regulation of glucose uptake is not known. We sought to determine whether specific inhibition of CaMKII impairs insulin- and/or contraction-induced glucose uptake in mouse skeletal muscle. Expression vectors containing green fluorescent protein conjugated to a CaMKII inhibitory (KKALHRQEAVDCL) or control (KKALHAQERVDCL) peptide were transfected into tibialis anterior muscles by in vivo electroporation. After 1 wk, muscles were assessed for peptide expression, CaMK activity, insulin- and contraction-induced 2-[(3)H]deoxyglucose uptake, glycogen concentrations, and changes in intracellular signaling proteins. Expression of the CaMKII inhibitory peptide decreased muscle CaMK activity approximately 35% compared with control peptide. Insulin-induced glucose uptake was not changed in muscles expressing the inhibitory peptide. In contrast, expression of the inhibitory peptide significantly decreased contraction-induced muscle glucose uptake (approximately 30%). Contraction-induced decreases in muscle glycogen were not altered by the inhibitory peptide. The CaMKII inhibitory peptide did not alter expression of the glucose transporter GLUT4 and did not impair contraction-induced increases in the phosphorylation of AMP-activated protein kinase (Thr(172)) or TBC1D1/TBC1D4 on phospho-Akt substrate sites. These results demonstrate that CaMKII does not regulate insulin-stimulated glucose uptake in skeletal muscle. However, CaMKII plays a critical role in the regulation of contraction-induced glucose uptake in mouse skeletal muscle.

Topics: Animals; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Diabetes Mellitus, Type 2; Female; Glucose; Glucose Transporter Type 4; Glycogen; Immunoblotting; In Vitro Techniques; Insulin; Mice; Mice, Inbred ICR; Muscle Contraction; Muscle, Skeletal; Peptide Fragments; Transfection

Obesity and type 2 diabetes have reached epidemic proportions; however, scarce information about how these metabolic syndromes influence brain energy and neurotransmitter homeostasis exist. The objective of this study was to elucidate how brain glycogen and neurotransmitter homeostasis are affected by these conditions. [1-(13)C]glucose was administered to Zucker obese (ZO) and Zucker diabetic fatty (ZDF) rats. Sprague-Dawley (SprD), Zucker lean (ZL), and ZDF lean rats were used as controls. Several brain regions were analyzed for glycogen levels along with (13)C-labeling and content of glutamate, glutamine, GABA, aspartate, and alanine. Blood glucose concentrations and (13)C enrichment were determined. (13)C-labeling in glutamate was lower in ZO and ZDF rats in comparison with the controls. The molecular carbon labeling (MCL) ratio between alanine and glutamate was higher in the ZDF rats. The MCL ratios of glutamine and glutamate were decreased in the cerebellum of the ZO and the ZDF rats. Glycogen levels were also lower in this region. These results suggest that the obese and type 2 diabetic models were associated with lower brain glucose metabolism. Glucose metabolism through the TCA cycle was more decreased than glycolytic activity. Furthermore, reduced glutamate-glutamine cycling was also observed in the obese and type 2 diabetic states.

Topics: Alanine; Amino Acids; Animals; Aspartic Acid; Blood Glucose; Brain; Diabetes Mellitus, Type 2; gamma-Aminobutyric Acid; Glutamic Acid; Glutamine; Glycogen; Male; Obesity; Rats; Rats, Sprague-Dawley; Rats, Zucker

Type 2 diabetes mellitus (T2DM) and aging are characterized by insulin resistance and impaired mitochondrial energetics. In lower organisms, remodeling by the protease pcp1 (PARL ortholog) maintains the function and lifecycle of mitochondria. We examined whether variation in PARL protein content is associated with mitochondrial abnormalities and insulin resistance. PARL mRNA and mitochondrial mass were both reduced in elderly subjects and in subjects with T2DM. Muscle knockdown of PARL in mice resulted in malformed mitochondrial cristae, lower mitochondrial content, decreased PGC1alpha protein levels, and impaired insulin signaling. Suppression of PARL protein in healthy myotubes lowered mitochondrial mass and insulin-stimulated glycogen synthesis and increased reactive oxygen species production. We propose that lower PARL expression may contribute to the mitochondrial abnormalities seen in aging and T2DM.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Animals; Cells, Cultured; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin; Metalloproteases; Mice; Mice, Knockout; Middle Aged; Mitochondria; Mitochondrial Proteins; Muscle, Skeletal; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Reactive Oxygen Species; Signal Transduction; Trans-Activators; Transcription Factors

In view of multi-dimensional activity of plant drugs beneficial to complex disorders like diabetes, the present study has been undertaken to evaluate the effect of aqueous extract of C. peltata roots on serum glucose, lipid profile, insulin, inflammatory marker namely tumour necrosis factor (TNF)-alpha and muscle glycogen in type 2 diabetic rats. Aqueous extract of C. peltata at 40 and 60 mg/kg dose significantly decreased both the fasting and postprandial blood glucose of type 2 diabetic rats; 60 mg/kg dose having more pronounced effect on hyperglycemia. An enhanced insulin levels by the aqueous extract is primary for its glucose and lipid lowering activity. The extract significantly decreased the elevated TNF-alpha in type 2 diabetic rats. The extract at 40 and 60 mg/kg dose increased the glycogen levels in skeletal muscle by 58 and 60% respectively. Improved glycogen in peripheral tissue such as skeletal muscle indicates the ability of plant drug to combat insulin resistance of type 2 diabetes.

Topics: Animals; Blood Glucose; Cyclea; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Drug Evaluation, Preclinical; Female; Glyburide; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Lipids; Male; Medicine, Ayurvedic; Metformin; Mice; Muscle, Skeletal; Phytotherapy; Plant Extracts; Plant Roots; Plants, Medicinal; Random Allocation; Rats; Rats, Wistar; Solvents; Tumor Necrosis Factor-alpha; Water

Exendin-4, like GLP-1, is insulinotropic, antidiabetic and glucoregulatory among other properties, which are thought to be exerted through the pancreatic GLP-1 receptor; exendin-4 is also an agonist of the GLP-1 stimulatory action upon liver and muscle glucose metabolism, where GLP-1 receptor is distinct from that in the pancreas. We investigated the action of prolonged treatment with exendin-4 upon glucose transport parameters in skeletal muscle and liver of normal rats and streptozotocin-induced type 2 diabetic rats (T2D). Muscle of T2D showed lower than normal glucose transport; exendin-4 did not modify the value in normal but normalized that in the T2D; unlike previously detected with GLP-1, no apparent modification was observed in GLUT-4 expression in either group after exendin-4, except for an increased GLUT-4 protein in normal rats. Yet, exendin-4 significantly stimulated liver GLUT-2-mRNA and -protein in T2D and normal rats, the effect upon GLUT-2-protein in T2D being higher than that in normal animals; this was accompanied by a normalizing action of exendin-4 upon the lower than normal liver glycogen in T2D rats. These data suggest that the liver may represent at least one of the major target organs for exendin-4 to exert its plasma lowering effect in diabetic state.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Exenatide; Glucagon-Like Peptide-1 Receptor; Glucose; Glycogen; Liver; Male; Muscle, Skeletal; Peptides; Rats; Rats, Wistar; Receptors, Glucagon; Venoms

We examined the intracellular metabolic fate of plasma glucose during a hyperglycemic clamp in impaired glucose-tolerant (IGT; n = 21) and normal glucose-tolerant subjects (n = 10) using a combination of [3-(3)H]glucose infusion with measurement of [(3)H]water formation and indirect calorimetry. IGT was associated with approximately 35% reduced first-phase insulin responses, normal second-phase insulin response, and 25-30% reduced insulin sensitivity, resulting in approximately 35% reduced plasma glucose disposal. This was coupled with approximately 55% reduced storage of plasma glucose (P < 0.01) and approximately 15-20% reduced glycolysis of plasma glucose (P < 0.03), accounting for approximately 75 and 25% of the reduction in glucose disposal, respectively. Decreased glucose oxidation accounted for virtually all the decrease in glycolysis. Therefore, nonoxidative glycolysis of plasma glucose in IGT was similar to that in NGT (P > 0.9) and accounted for an increased proportion of systemic glucose disposal (P < 0.05). We conclude that, in IGT, decreased disposal of plasma glucose involves mainly decreased glycogen synthesis and to a lesser extent decreased glycolysis, which is accounted for by decreased glucose oxidation. An increased proportion of plasma glucose hence undergoes nonoxidative glycolysis, representing a novel early abnormality in the pathogenesis of T2DM.

Topics: Alanine; Blood Glucose; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Female; Glucagon; Glucose Clamp Technique; Glucose Intolerance; Glycogen; Glycolysis; Humans; Hyperglycemia; Lactic Acid; Male; Middle Aged; Oxidation-Reduction

Obesity is a risk factor for type 2 diabetes in cats. The risk of developing diabetes is severalfold greater for male cats than for females, even after having been neutered early in life. The purpose of this study was to investigate the role of different metabolic pathways in the regulation of endogenous glucose production (EGP) during the fasted state considering these risk factors. A triple tracer protocol using (2)H(2)O, [U-(13)C(3)]propionate, and [3,4-(13)C(2)]glucose was applied in overnight-fasted cats (12 lean and 12 obese; equal sex distribution) fed three different diets. Compared with lean cats, obese cats had higher insulin (P < 0.001) but similar blood glucose concentrations. EGP was lower in obese cats (P < 0.001) due to lower glycogenolysis and gluconeogenesis (GNG; P < 0.03). Insulin, body mass index, and girth correlated negatively with EGP (P < 0.003). Female obese cats had approximately 1.5 times higher fluxes through phosphoenolpyruvate carboxykinase (P < 0.02) and citrate synthase (P < 0.05) than male obese cats. However, GNG was not higher because pyruvate cycling was increased 1.5-fold (P < 0.03). These results support the notion that fasted obese cats have lower hepatic EGP compared with lean cats and are still capable of maintaining fasting euglycemia, despite the well-documented existence of peripheral insulin resistance in obese cats. Our data further suggest that sex-related differences exist in the regulation of hepatic glucose metabolism in obese cats, suggesting that pyruvate cycling acts as a controlling mechanism to modulate EGP. Increased pyruvate cycling could therefore be an important factor in modulating the diabetes risk in female cats.

Topics: Animals; Blood Glucose; Body Mass Index; Body Weight; Carbon Isotopes; Cats; Citrate (si)-Synthase; Citric Acid Cycle; Diabetes Mellitus, Type 2; Diet; Disease Models, Animal; Eating; Fasting; Female; Gluconeogenesis; Glycerol; Glycogen; Glycogenolysis; Indicator Dilution Techniques; Insulin; Liver; Magnetic Resonance Spectroscopy; Male; Obesity; Phosphoenolpyruvate Carboxykinase (GTP); Pyruvic Acid; Sex Factors

Major urinary protein (MUP) 1 is a lipocalin family member abundantly secreted into the circulation by the liver. MUP1 binds to lipophilic pheromones and is excreted in urine. Urinary MUP1/pheromone complexes mediate chemical communication in rodents. However, it is unclear whether circulatory MUP1 has additional physiological functions. Here we show that MUP1 regulates glucose and lipid metabolism. MUP1 expression was markedly reduced in both genetic and dietary fat-induced obesity and diabetes. Mice were infected with MUP1 adenoviruses via tail vein injection, and recombinant MUP1 was overexpressed in the liver and secreted into the bloodstream. Recombinant MUP1 markedly attenuated hyperglycemia and glucose intolerance in genetic (db/db) and dietary fat-induced type 2 diabetic mice as well as in streptozotocin-induced type 1 diabetic mice. MUP1 inhibited the expression of both gluconeogenic genes and lipogenic genes in the liver. Moreover, recombinant MUP1 directly decreased glucose production in primary hepatocyte cultures by inhibiting the expression of gluconeogenic genes. These data suggest that MUP1 regulates systemic glucose and/or lipid metabolism through the paracrine/autocrine regulation of the hepatic gluconeogenic and/or lipogenic programs, respectively.

Topics: Adenoviridae; Animals; Diabetes Mellitus, Type 2; Gene Expression Regulation; Gluconeogenesis; Glucose; Glucose Tolerance Test; Glycogen; Hepatocytes; Lipid Metabolism; Mice; Mice, Inbred C57BL; Proteins; Recombinant Proteins; Triglycerides

To examine whether escitalopram enhances net hepatic glucose uptake during a hyperinsulinemic hyperglycemic clamp, studies were performed in conscious 42-h-fasted dogs. The experimental period was divided into P1 (0-90 min) and P2 (90-270 min). During P1 and P2 somatostatin (to inhibit insulin and glucagon secretion), 4x basal intraportal insulin, basal intraportal glucagon, and peripheral glucose (2x hepatic glucose load) were infused. Saline was infused intraportally during P1 in all groups. In one group saline infusion was continued in P2 (SAL, n = 11), while escitalopram was infused intraportally at 2 microg/kg/min (L-ESC, n = 6) or 8 microg/kg/min (H-ESC, n = 7) during P2 in two other groups. The arterial insulin concentrations rose approximately four fold (to 123 +/- 8, 146 +/- 13 and 148 +/- 15 pmol/L) while glucagon concentrations remained basal (41 +/- 3, 44 +/- 9 and 40 +/- 3 ng/L) in all groups. The hepatic glucose load averaged 216 +/- 13, 223 +/- 19 and 202 +/- 12 micromol/kg/min during the entire experimental period (P1 and P2) in the SAL, L-ESC and H-ESC groups, respectively. Net hepatic glucose uptake was 11.6 +/- 1.4, 10.1 +/- 0.9 and 10.4 +/- 2.3 micromol/kg/min in P1 and averaged 16.9 +/- 1.5, 15.7 +/- 1.3 and 22.6 +/- 3.7 (P < 0.05) in the SAL, L-ESC and H-ESC groups, respectively during the last hour of P2 (210-270 min). Net hepatic carbon retention (glycogen storage) was 15.4 +/- 1.3, 14.9 +/- 0.6 and 20.9 +/- 2.6 (P < 0.05) micromol/kg/min in SAL, L-ESC and H-ESC respectively during the last hour of P2. Escitalopram enhanced net hepatic glucose uptake and hepatic glycogen deposition, showing that it can improve hepatic glucose clearance under hyperinsulinemic hyperglycemic conditions. Its use in individuals with diabetes may, therefore, result in improved glycemic control.

Topics: Animals; Antidepressive Agents, Second-Generation; Carbon; Citalopram; Diabetes Mellitus, Type 2; Dogs; Glucose; Glucose Clamp Technique; Glycogen; Hyperinsulinism; Infusions, Intravenous; Liver; Portal Vein; Somatostatin; Time Factors

Obesity is an important risk factor for the development of type 2 diabetes, but not all obese individuals develop this complication. The clinical signs of type 2 diabetes can often be reversed with weight loss; however, it is unknown whether the skeletal muscle oxidative stress associated with type 2 diabetes remains after weight loss. We hypothesised that chronic exposure to high glucose and insulin would re-elicit impaired metabolism in primary myotubes from patients with a history of type 2 diabetes.. Obese participants with or without type 2 diabetes completed a standardised weight loss protocol, following which all participants were euglycaemic and had similar indices of insulin sensitivity. Satellite cells were isolated from muscle biopsies and differentiated under low or high glucose and insulin conditions (HGI).. Cells from participants with no history of type 2 diabetes showed robust increases in mitochondrial content, citrate synthase and cytochrome c oxidase activities when exposed to HGI. This increase in oxidative capacity was absent in cells from patients with a history of type 2 diabetes. High glucose and insulin caused increased oxidative damage in cells from the latter, despite higher superoxide dismutase expression. Cells from patients with a history of type 2 diabetes were unable to decrease mitochondrial membrane potential in response to HGI, potentially due to lower levels of uncoupling protein-3.. This is the first report to note that primary myotubes from patients with a history of type 2 diabetes are unable to adapt to a hyperglycaemic-hyperinsulinaemic challenge. We have demonstrated that impaired mitochondrial biogenesis and an inability to manage oxidative stress define a muscle phenotype at risk of obesity-associated type 2 diabetes.

Topics: Adult; Body Composition; Body Mass Index; Diabetes Complications; Diabetes Mellitus, Type 2; Female; Glucose Clamp Technique; Glycated Hemoglobin; Glycogen; Humans; Hyperinsulinism; Hypoglycemic Agents; Insulin; Ion Channels; Male; Middle Aged; Mitochondrial Proteins; Muscle Fibers, Skeletal; Muscle, Skeletal; Obesity; Oxidative Stress; Triglycerides; Uncoupling Protein 3; Weight Loss

Moutan Cortex is a well-known herb in traditional Korean, Chinese, and Japanese anti-diabetic formulae. In the current study, we investigated the metabolic effects of isolated triterpenes (1-7) in HepG2 cells under high glucose conditions. These compounds remakably stimulated AMP-activated protein kinase (AMPK), GSK-3beta, and ACC phosphorylation. The compounds also increased glucose uptake and enhanced glycogen synthesis. Among these, compound 1 displayed the greatest potential anti-diabetic activity though the AMPK activation pathway. Compound 1 significantly increased the levels of phospho-AMPK, phospho-ACC, and phospho-GSK-3beta and stimulated glucose uptake and glycogen synthesis in a dose-dependent manner. In conclusion, our results suggest that these compounds, especially compound 1, may have beneficial roles in glucose metabolism via the AMPK pathway.

Topics: AMP-Activated Protein Kinases; Cell Line, Tumor; Diabetes Mellitus, Type 2; Drugs, Chinese Herbal; Glucose; Glycogen; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Hypoglycemic Agents; Insulin Resistance; Paeonia; Phosphorylation; Terpenes; Triterpenes

1-[4-[2-(4-Bromobenzenesulfonamino)ethyl]phenylsulfonyl]-3-(trans-4-methylcyclohexyl)urea (I4, CAS 865483-06-3), a totally synthetic new sulfonylurea compound, incorporating part of the hypoglycemic structure of glimepiride (CAS 93479-97-1) and having anti-TXA2 receptor properties, was designed and synthesized. Its hypoglycemic property had not been reported yet.. To study the hypoglycemic effects of I4 and its primary mechanisms of action.. A rat model of type 2 diabetes was established by intraperitoneal injection of small doses of streptozotocin combined with high calorie feeding. Normal fasted mice and type 2 diabetic rats were used to assay the hypoglycemic actions of I4. Blood glucose and immunoreactive insulin concentrations were measured and the effects of I4 on insulin release from rat isolated pancreatic islets were examined. A liver cell line, Hep G2, was used to examine effects on glucose consumption, glycogen synthesis and glucokinase activity.. Oral administration of I4 (1-10 mg/kg) produced a marked, dose-dependent reduction in blood glucose in normal mice and type 2 diabetic rats and improved oral glucose tolerance. Plasma insulin concentrations were increased, and 14 increased insulin release from rat isolated pancreatic islets and from the isolated perfused rat pancreas. I4 (1-100 micromol x L(-1)) also produced an insulin-independent increase in glucose consumption by Hep G2 cells, increased glycogen synthesis and glucokinase activity of these cells. Glimepiride produced similar effects on glucose consumption and glycogen synthesis but did not facilitate glucokinase activity of Hep G2 cells.. I4 markedly improved glucose metabolism in normal animals and type 2 diabetic rats, probably by increasing insulin secretion and stimulating hepatic glucose uptake and glycogen synthesis.

Topics: Animals; Cell Line; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dietary Fats; Glucokinase; Glucose; Glucose Tolerance Test; Glycogen; Hypoglycemic Agents; In Vitro Techniques; Indicators and Reagents; Insulin; Islets of Langerhans; Male; Mice; Mice, Inbred C57BL; Pancreas; Rats; Rats, Sprague-Dawley; Sulfhydryl Compounds; Urea

An alcoholic extract of Artemisia dracunculus L (PMI 5011) has been shown to decrease glucose and improve insulin levels in animal models, suggesting an ability to enhance insulin sensitivity. We sought to assess the cellular mechanism by which this botanical affects carbohydrate metabolism in primary human skeletal muscle culture. We measured basal and insulin-stimulated glucose uptake, glycogen accumulation, phosphoinositide 3 (PI-3) kinase activity, and Akt phosphorylation in primary skeletal muscle culture from subjects with type 2 diabetes mellitus incubated with or without various concentrations of PMI 5011. We also analyzed the abundance of insulin receptor signaling proteins, for example, IRS-1, IRS-2, and PI-3 kinase. Glucose uptake was significantly increased in the presence of increasing concentrations of PMI 5011. In addition, glycogen accumulation, observed to be decreased with increasing free fatty acid levels, was partially restored with PMI 5011. PMI 5011 treatment did not appear to significantly affect protein abundance for IRS-1, IRS-2, PI-3 kinase, Akt, insulin receptor, or Glut-4. However, PMI 5011 significantly decreased levels of a specific protein tyrosine phosphatase, that is, PTP1B. Time course studies confirmed that protein abundance of PTP1B decreases in the presence of PMI 5011. The cellular mechanism of action to explain the effects by which an alcoholic extract of A dracunculus L improves carbohydrate metabolism on a clinical level may be secondary to enhancing insulin receptor signaling and modulating levels of a specific protein tyrosine phosphatase, that is, PTP1B.

Topics: Artemisia; Cell Culture Techniques; Cells, Cultured; Diabetes Mellitus, Type 2; Drug Evaluation, Preclinical; Gene Expression Regulation, Enzymologic; Glucose; Glycogen; Glycogen Synthase; Humans; Insulin; Male; Middle Aged; Models, Biological; Muscle, Skeletal; Obesity; Phosphatidylinositol 3-Kinases; Plant Extracts; Signal Transduction

Insulin-resistant type 2 diabetic patients have been reported to have impaired skeletal muscle mitochondrial respiratory function. A key question is whether decreased mitochondrial respiration contributes directly to the decreased insulin action. To address this, a model of impaired cellular respiratory function was established by incubating human skeletal muscle cell cultures with the mitochondrial inhibitor sodium azide and examining the effects on insulin action. Incubation of human skeletal muscle cells with 50 and 75 microM azide resulted in 48 +/- 3% and 56 +/- 1% decreases, respectively, in respiration compared with untreated cells mimicking the level of impairment seen in type 2 diabetes. Under conditions of decreased respiratory chain function, insulin-independent (basal) glucose uptake was significantly increased. Basal glucose uptake was 325 +/- 39 pmol/min/mg (mean +/- SE) in untreated cells. This increased to 669 +/- 69 and 823 +/- 83 pmol/min/mg in cells treated with 50 and 75 microM azide, respectively (vs. untreated, both P < 0.0001). Azide treatment was also accompanied by an increase in basal glycogen synthesis and phosphorylation of AMP-activated protein kinase. However, there was no decrease in glucose uptake following insulin exposure, and insulin-stimulated phosphorylation of Akt was normal under these conditions. GLUT1 mRNA expression remained unchanged, whereas GLUT4 mRNA expression increased following azide treatment. In conclusion, under conditions of impaired mitochondrial respiration there was no evidence of impaired insulin signaling or glucose uptake following insulin exposure in this model system.

Topics: AMP-Activated Protein Kinases; Cells, Cultured; Diabetes Mellitus, Type 2; Electron Transport; Electron Transport Complex IV; Enzyme Inhibitors; Glucose; Glucose Transporter Type 1; Glucose Transporter Type 4; Glycogen; Humans; Insulin; Mitochondria; Multienzyme Complexes; Muscle Fibers, Skeletal; Muscle, Skeletal; Phosphorylation; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; RNA, Messenger; Signal Transduction; Sodium Azide

Insulin promotes dephosphorylation and activation of glycogen synthase (GS) by inactivating glycogen synthase kinase (GSK) 3 through phosphorylation. Insulin also promotes glucose uptake and glucose 6-phosphate (G-6-P) production, which allosterically activates GS. The relative importance of these two regulatory mechanisms in the activation of GS in vivo is unknown. The aim of this study was to investigate if dephosphorylation of GS mediated via GSK3 is required for normal glycogen synthesis in skeletal muscle with insulin. We employed GSK3 knockin mice in which wild-type GSK3 alpha and -beta genes are replaced with mutant forms (GSK3 alpha/beta S21A/S21A/S9A/S9A), which are nonresponsive to insulin. Although insulin failed to promote dephosphorylation and activation of GS in GSK3 alpha/beta S21A/S21A/S9A/S9A mice, glycogen content in different muscles from these mice was similar compared with wild-type mice. Basal and epinephrine-stimulated activity of muscle glycogen phosphorylase was comparable between wild-type and GSK3 knockin mice. Incubation of isolated soleus muscle in Krebs buffer containing 5.5 mM glucose in the presence or absence of insulin revealed that the levels of G-6-P, the rate of [14C]glucose incorporation into glycogen, and an increase in total glycogen content were similar between wild-type and GSK3 knockin mice. Injection of glucose containing 2-deoxy-[3H]glucose and [14C]glucose also resulted in similar rates of muscle glucose uptake and glycogen synthesis in vivo between wild-type and GSK3 knockin mice. These results suggest that insulin-mediated inhibition of GSK3 is not a rate-limiting step in muscle glycogen synthesis in mice. This suggests that allosteric regulation of GS by G-6-P may play a key role in insulin-stimulated muscle glycogen synthesis in vivo.

Topics: Animals; Blood Glucose; Carbon Radioisotopes; Diabetes Mellitus, Type 2; Enzyme Activation; Epinephrine; Glucose-6-Phosphate; Glycogen; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Hypoglycemic Agents; Insulin; Mice; Mice, Mutant Strains; Muscle, Skeletal; Phosphorylation; Signal Transduction; Sympathomimetics; Tritium

Topics: Animals; Cardiomyopathies; Codon, Nonsense; Diabetes Mellitus, Type 2; Exercise; Frameshift Mutation; Gene Frequency; Glycogen; Glycogen Phosphorylase; Glycogen Synthase; Heart; Humans; Infant, Newborn; Insulin; Insulin Resistance; Mice; Muscle, Skeletal; Myocardium; Organ Specificity; Phosphoprotein Phosphatases; United Kingdom; White People

Stored glycogen is an important source of energy for skeletal muscle. Human genetic disorders primarily affecting skeletal muscle glycogen turnover are well-recognised, but rare. We previously reported that a frameshift/premature stop mutation in PPP1R3A, the gene encoding RGL, a key regulator of muscle glycogen metabolism, was present in 1.36% of participants from a population of white individuals in the UK. However, the functional implications of the mutation were not known. The objective of this study was to characterise the molecular and physiological consequences of this genetic variant.. In this study we found a similar prevalence of the variant in an independent UK white population of 744 participants (1.46%) and, using in vivo (13)C magnetic resonance spectroscopy studies, demonstrate that human carriers (n = 6) of the variant have low basal (65% lower, p = 0.002) and postprandial muscle glycogen levels. Mice engineered to express the equivalent mutation had similarly decreased muscle glycogen levels (40% lower in heterozygous knock-in mice, p < 0.05). In muscle tissue from these mice, failure of the truncated mutant to bind glycogen and colocalize with glycogen synthase (GS) decreased GS and increased glycogen phosphorylase activity states, which account for the decreased glycogen content.. Thus, PPP1R3A C1984DeltaAG (stop codon 668) is, to our knowledge, the first prevalent mutation described that directly impairs glycogen synthesis and decreases glycogen levels in human skeletal muscle. The fact that it is present in approximately 1 in 70 UK whites increases the potential biomedical relevance of these observations.

Topics: Adult; Animals; Codon, Nonsense; Diabetes Mellitus, Type 2; Female; Frameshift Mutation; Gene Frequency; Glycogen; Glycogen Phosphorylase; Glycogen Synthase; Humans; Male; Mice; Mice, Transgenic; Middle Aged; Molecular Sequence Data; Muscle, Skeletal; Phosphoprotein Phosphatases; Postprandial Period; Structure-Activity Relationship; United Kingdom; White People

In Type 2 diabetes, increased glycogenolysis contributes to the hyperglycaemic state, therefore the inhibition of GP (glycogen phosphorylase), a key glycogenolytic enzyme, is one of the possibilities to lower plasma glucose levels. Following this strategy, a number of GPis (GP inhibitors) have been described. However, certain critical issues are associated with their mode of action, e.g. an impairment of muscle function. The interaction between GP and the liver glycogen targeting subunit (termed G(L)) of PP1 (protein phosphatase 1) has emerged as a new potential anti-diabetic target, as the disruption of this interaction should increase glycogen synthesis, potentially providing an alternative approach to counteract the enhanced glycogenolysis without inhibiting GP activity. We identified an inhibitor of the G(L)-GP interaction (termed G(L)-GPi) and characterized its mechanism of action in comparison with direct GPis. In primary rat hepatocytes, at elevated glucose levels, the G(L)-GPi increased glycogen synthesis similarly to direct GPis. Direct GPis significantly reduced the cellular GP activity, caused a dephosphorylation of the enzyme and decreased the amounts of GP in the glycogen-enriched fraction; the G(L)-GPi did not influence any of these parameters. Both mechanisms increased glycogen accumulation at elevated glucose levels. However, at low glucose levels, only direct GPis led to increased glycogen amounts, whereas the G(L)-GPi allowed the mobilization of glycogen because it did not block the activity of GP. Due to this characteristic, G(L)-GPi in comparison with GPis could offer an advantageous risk/benefit profile circumventing the potential downsides of a complete prevention of glycogen breakdown while retaining glucose-lowering efficacy, suggesting that inhibition of the G(L)-GP interaction may provide an attractive novel approach for rebalancing the disturbed glycogen metabolism in diabetic patients.

Topics: Animals; Carrier Proteins; Cells, Cultured; Diabetes Mellitus, Type 2; Glycogen; Glycogen Phosphorylase; Glycogen Synthase; Hepatocytes; Humans; Male; Phosphoprotein Phosphatases; Protein Phosphatase 1; Protein Subunits; Rats; Rats, Wistar

We investigated the effect of low-fat (2.5%) dahi containing probiotic Lactobacillus acidophilus and Lactobacillus casei on progression of high fructose-induced type 2 diabetes in rats.. Diabetes was induced in male albino Wistar rats by feeding 21% fructose in water. The body weight, food and water intakes, fasting blood glucose, glycosylated hemoglobin, oral glucose tolerance test, plasma insulin, liver glycogen content, and blood lipid profile were recorded. The oxidative status in terms of thiobarbituric acid-reactive substances and reduced glutathione contents in liver and pancreatic tissues were also measured.. Values for blood glucose, glycosylated hemoglobin, glucose intolerance, plasma insulin, liver glycogen, plasma total cholesterol, triacylglycerol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, and blood free fatty acids were increased significantly after 8 wk of high fructose feeding; however, the dahi-supplemented diet restricted the elevation of these parameters in comparison with the high fructose-fed control group. In contrast, high-density lipoprotein cholesterol decreased slightly and was retained in the dahi-fed group. The dahi-fed group also exhibited lower values of thiobarbituric acid-reactive substances and higher values of reduced glutathione in liver and pancreatic tissues compared with the high fructose-fed control group.. The probiotic dahi-supplemented diet significantly delayed the onset of glucose intolerance, hyperglycemia, hyperinsulinemia, dyslipidemia, and oxidative stress in high fructose-induced diabetic rats, indicating a lower risk of diabetes and its complications.

Topics: Animals; Area Under Curve; Blood Glucose; Diabetes Mellitus, Type 2; Fructose; Glucose Tolerance Test; Glutathione; Glycated Hemoglobin; Glycogen; Insulin; Lacticaseibacillus casei; Lactobacillus acidophilus; Lipid Metabolism; Male; Oxidative Stress; Probiotics; Rats; Rats, Wistar; Risk Factors; Thiobarbituric Acid Reactive Substances; Yogurt

Disturbances in substrate source metabolism and, more particularly, in fatty acid metabolism, play an important role in the aetiology and progression of type 2 diabetes. However, data on substrate source utilisation in type 2 diabetes are inconclusive.. [U-(13)C]palmitate and [6,6-(2)H(2)]glucose tracers were used to assess plasma NEFA and glucose oxidation rates and to estimate the use of muscle- and/or lipoprotein-derived triacylglycerol and muscle glycogen. Subjects were ten male patients who had a long-term (7 +/- 1 years) diagnosis of type 2 diabetes and were overweight, and ten matched healthy, male control subjects. Muscle biopsy samples were collected before and after exercise to assess muscle fibre type-specific intramyocellular lipid and glycogen content.. At rest and during exercise, the diabetes patients had greater values than the controls for palmitate rate of appearance (Ra) (rest, 2.46 +/- 0.18 and 1.85 +/- 0.20 respectively; exercise, 3.71 +/- 0.36 and 2.84 +/- 0.20 micromol kg(-1) min(-1)) and rate of disappearance (Rd) (rest, 2.45 +/- 0.18 and 1.83 +/- 0.20; exercise, 3.64 +/- 0.35 and 2.80 +/- 0.20 micromol kg(-1) min(-1) respectively). This was accompanied by significantly higher fat oxidation rates at rest and during recovery in the diabetes patients (rest, 0.11 +/- 0.01 in diabetes patients and 0.09 +/- 0.01 in controls; recovery, 0.13 +/- 0.01 and 0.11 +/- 0.01 g/min respectively), despite significantly greater plasma glucose Ra, Rd and circulating plasma glucose concentrations. Furthermore, exercise significantly lowered plasma glucose concentrations in the diabetes patients, as a result of increased blood glucose disposal.. This study demonstrates that substrate source utilisation in long-term-diagnosed type 2 diabetes patients, in whom compensatory hyperinsulinaemia is no longer present, shifts towards an increase in whole-body fat oxidation rate and is accompanied by disturbances in fat and carbohydrate handling.

Topics: Biopsy; Blood Glucose; Case-Control Studies; Diabetes Mellitus, Type 2; Energy Metabolism; Exercise; Fatty Acids, Nonesterified; Glycerol; Glycogen; Humans; Insulin; Lipid Metabolism; Male; Middle Aged; Muscle, Skeletal; Obesity; Rest

Topics: Adaptor Proteins, Signal Transducing; Animals; Blood Glucose; Carrier Proteins; Diabetes Mellitus, Type 2; Fasting; Glucagon; Glucokinase; Glucose; Glucose-6-Phosphatase; Glycogen; Insulin; Liver; Phosphorylation; Rats; Rats, Zucker; Sorbitol; Time Factors

Effect of stimulation of glucokinase (GK) export from the nucleus by small amounts of sorbitol on hepatic glucose flux in response to elevated plasma glucose was examined in 6-h fasted Zucker diabetic fatty rats at 10 wk of age. Under basal conditions, plasma glucose, insulin, and glucagon were approximately 8 mM, 2,000 pmol/l, and 60 ng/l, respectively. Endogenous glucose production (EGP) was 44 +/- 4 micromol x kg(-1) x min(-1). When plasma glucose was raised to approximately 17 mM, GK was still predominantly localized with its inhibitory protein in the nucleus. EGP was not suppressed. When sorbitol was infused at 5.6 and 16.7 micromol x kg(-1) x min(-1), along with the increase in plasma glucose, GK was exported to the cytoplasm. EGP (23 +/- 19 and 12 +/- 5 micromol x kg(-1) x min(-1)) was suppressed without a decrease in glucose 6-phosphatase flux (145 +/- 23 and 126 +/- 16 vs. 122 +/- 10 micromol x kg(-1) x min(-1) without sorbitol) but increased in glucose phosphorylation as indicated by increases in glucose recycling (122 +/- 17 and 114 +/- 19 vs. 71 +/- 11 microl x kg(-1) x min(-1)), glucose-6-phosphate content (254 +/- 32 and 260 +/- 35 vs. 188 +/- 20 nmol/g liver), fractional contribution of plasma glucose to uridine 5'-diphosphate-glucose flux (43 +/- 8 and 42 +/- 8 vs. 27 +/- 6%), and glycogen synthesis from plasma glucose (20 +/- 4 and 22 +/- 5 vs. 9 +/- 4 mumol glucose/g liver). The decreased glucose effectiveness to suppress EGP and stimulate hepatic glucose uptake may result from failure of the sugar to activate GK by stimulating the translocation of the enzyme.

Topics: Animals; Blood Glucose; Carrier Proteins; Diabetes Mellitus, Type 2; Fasting; Glucagon; Glucokinase; Glucose; Glucose-6-Phosphatase; Glycogen; Insulin; Liver; Liver Glycogen; Male; Phosphorylation; Rats; Rats, Zucker; Sorbitol; Time Factors

High-fat diets and oxidative damage may contribute to the development of type 2 diabetes. Hypolipidaemic drugs and antioxidants were supposed to prevent the development of the disease. In this study, we investigated preventive effects of fenofibrate (200 mg kg(-1)), vitamin C (30 mg kg(-1)), combination of both in mice induced by streptozotocin (35 mg kg(-1)) and soluble lard (15 ml kg(-1)). The results showed the mice demonstrated hyperglycaemia and hypercholesterolaemia, visceral fat accumulation, and a slight increase in liver glycogen/triglyceride and oxidative stress within 60 days of treatment. Fenofibrate enhanced insulin sensitivity, improved glycaemic control, lowered serum triglycerides, reduced body and visceral fat weights, and decreased liver glycogen/lipid levels but showed hepatotoxicity in the mice. Vitamin C neither itself prevented nor enhanced preventive effects of fenofibrate on glucose and lipid metabolism but partly attenuated the hepatotoxicity of fenofibrate. These results suggest that fenofibrate inhibit development of type 2 diabetes induced by high-fat diets and oxidative stress. However, here, vitamin C just can serve as an adjunct to fenofibrate therapy against its hepatotoxicity. In the future study, we should investigate if higher dosage of vitamin C or other antioxidants would enhance preventive effects of fenofibrate in type 2 diabetes.

Topics: Animals; Antioxidants; Ascorbic Acid; Blood Glucose; Chemical and Drug Induced Liver Injury; Cholesterol; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dietary Fats; Drug Therapy, Combination; Fenofibrate; Glycogen; Hypercholesterolemia; Hyperglycemia; Hypoglycemic Agents; Hypolipidemic Agents; Insulin Resistance; Intra-Abdominal Fat; Liver; Liver Diseases; Male; Matrix Metalloproteinase 9; Mice; Oxidative Stress; Streptozocin; Triglycerides

D-Glucose-6-phosphatase is a key regulator of endogenous glucose production, and its inhibition may improve glucose control in type 2 diabetes. Herein, 2'-O-(2-methoxy)ethyl-modified phosphorothioate antisense oligonucleotides (ASOs) specific to the glucose 6-phosphate transporter-1 (G6PT1) enabled reduction of hepatic D-Glu-6-phosphatase activity in diabetic ob/ob mice. Treatment with G6PT1 ASOs decreased G6PT1 expression, reduced G6PT1 activity, blunted glucagon-stimulated glucose production, and lowered plasma glucose concentration in a dose-dependent manner. In contrast to G6PT1 knock-out mice and patients with glycogen storage disease, excess hepatic and renal glycogen accumulation, hyperlipidemia, neutropenia, and elevations in plasma lactate and uric acid did not occur. In addition, hypoglycemia was not observed in animals during extended periods of fasting, and the ability of G6PT1 ASO-treated mice to recover from an exogenous insulin challenge was not impaired. Together, these results demonstrate that effective glucose lowering by G6PT1 inhibitors can be achieved without adversely affecting carbohydrate and lipid metabolism.

Topics: Acidosis, Lactic; Animals; Antiporters; Blood Glucose; Diabetes Complications; Diabetes Mellitus, Type 2; Glucagon; Glucose-6-Phosphatase; Glycogen; Glycogen Storage Disease; Hyperlipidemias; Hyperuricemia; Hypoglycemia; Kidney; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Obese; Monosaccharide Transport Proteins; Oligoribonucleotides, Antisense; RNA, Messenger

Hypersecretion of glucagon contributes to abnormally increased hepatic glucose output in type 2 diabetes. Somatostatin (SST) inhibits murine glucagon secretion from isolated pancreatic islets via somatostatin receptor subtype-2 (sst2). Here, we characterize the role of sst2 in controlling glucose homeostasis in mice with diet-induced obesity. Sst2-deficient (sst2(-/-)) and control mice were fed high-fat diet for 14 wk, and the parameters of glucose homeostasis were monitored. Hepatic glycogen and lipid contents were quantified enzymatically and visualized histomorphologically. Enzymes regulating glycogen and lipid synthesis and breakdown were measured by real-time PCR and/or Western blot. Gluconeogenesis and glycogenolysis were determined from isolated primary hepatocytes and glucagon or insulin secretion from isolated pancreatic islets. Nonfasting glucose, glucagon, and fasting nonesterified fatty acids of sst2(-/-) mice were increased. Inhibition of glucagon secretion from sst2-deficient pancreatic islets by glucose or somatostatin was impaired. Insulin less potently reduced blood glucose concentration in sst2-deficient mice as compared with wild-type mice. Sst2-deficient mice had decreased nonfasting hepatic glycogen and lipid content. The activity/expression of enzymes controlling hepatic glycogen synthesis of sst2(-/-) mice was decreased, whereas enzymes facilitating glycogenolysis and lipolysis were increased. Somatostatin and an sst2-selective agonist decreased glucagon-induced glycogenolysis, without influencing de novo glucose production using cultured primary hepatocytes. This study demonstrates that ablation of sst2 leads to hyperglucagonemia. Increased glucagon concentration is associated with impaired glucose control in sst2(-/-) mice, resulting from decreased hepatic glucose storage, increased glycogen breakdown, and reduced lipid accumulation. Sst2 may constitute a therapeutic target to lower hyperglucagonemia in type 2 diabetes.

Topics: Animal Feed; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Disease Models, Animal; Fasting; Fatty Acids, Nonesterified; Female; Glucagon; Gluconeogenesis; Glycogen; Glycogen Synthase; Glycogenolysis; Homeostasis; Hyperglycemia; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Mice, Obese; Obesity; Receptors, Somatostatin; Triglycerides

Intraportal infusion of serotonin (5-hydroxytryptamine, 5-HT) or inhibitors of its cellular uptake stimulate hepatic glucose uptake in vivo by either direct or indirect mechanisms. The aims of this study were to determine the direct effects of 5-HT in hepatocytes and to test the hypothesis that atypical antipsychotic drugs that predispose to type 2 diabetes counter-regulate the effects of 5-HT.. Rat hepatocytes were studied in short-term primary culture.. Serotonin (5-HT) stimulated glycogen synthesis at nanomolar concentrations but inhibited it at micromolar concentrations. The stimulatory effect was mimicked by alpha-methyl-5-HT, a mixed 5-HT1/5-HT2 receptor agonist, whereas the inhibition was counteracted by a 5-HT2B/2C receptor antagonist. alpha-Methyl-5-HT stimulated glycogen synthesis additively with insulin, but unlike insulin, did not stimulate glucose phosphorylation and glycolysis, nor did it cause Akt (protein kinase B) phosphorylation. Stimulation of glycogen synthesis by alpha-methyl-5-HT correlated with depletion of phosphorylase a. This effect could not be explained by elevated levels of glucose 6-phosphate, which causes inactivation of phosphorylase, but was explained, at least in part, by decreased phosphorylase kinase activity in situ. The antipsychotic drugs clozapine and olanzapine, which bind to 5-HT receptors, counteracted the effect of alpha-methyl-5-HT on phosphorylase inactivation.. This study provides evidence for both stimulation and inhibition of glycogen synthesis in hepatocytes by serotonergic mechanisms. The former effects are associated with the inactivation of phosphorylase and are counteracted by atypical antipsychotic drugs that cause hepatic insulin resistance. Antagonism of hepatic serotonergic mechanisms may be a component of the hepatic dysregulation caused by antipsychotic drugs that predispose to type 2 diabetes.

Topics: Amides; Animals; Antipsychotic Agents; Benzodiazepines; Blotting, Western; Cells, Cultured; Clozapine; Diabetes Mellitus, Type 2; Enzyme Activation; Glycogen; Hepatocytes; Immunoblotting; Indoles; Male; Olanzapine; Phosphorylases; Rats; Rats, Wistar; Serotonin; Serotonin 5-HT1 Receptor Agonists; Serotonin 5-HT2 Receptor Agonists; Serotonin 5-HT2 Receptor Antagonists

In the present study, we investigated triacylglycerol (TAG) accumulation, glucose and fatty acid (FA) uptake, and glycogen synthesis (GS) in human myotubes from healthy, lean, and obese subjects with and without type 2 diabetes (T2D), exposed to increasing palmitate (PA) and oleate (OA) concentrations with/without high glucose and/or high insulin concentrations for 4 days. We showed that these myotubes expressed an increased TAG accumulation (P<0.001) without differences between groups. Chronically high insulin, but not high glucose concentrations, increases TAG accumulation by 25% (P<0.001). Inhibition of oxidative phosphorylation by antimycin A and oligomyin was followed by a reduced lipid oxidation (P<0.05) and increased TAG accumulation (P<0.05), but only in the presence of FAs. Both chronic PA and OA exposure reduced the insulin-mediated PA and OA uptake (fold change) (P<0.001), but could not induce insulin resistance at the level of glucose uptake, whereas high insulin concentrations induced insulin resistance (P<0.001). Chronic, high PA, but not OA, induced insulin resistance at the GS level in control subjects (P<0.05). The TAG content correlated negatively with insulin-stimulated FA uptake (P<0.001), but did not correlate with insulin-stimulated glucose uptake for PA or OA (P>0.05). These results indicate that (1) TAG accumulation is not primarily affected in skeletal muscle tissue of obese and T2D; (2) induced inhibition of oxidative phosphorylation is followed by TAG accumulation; (3) increasing FA and insulin availability, and reduced oxidative phosphorylation, and to a lesser extent glucose, are determinants for differences in intramyocellular TAG accumulation; (4) quantitative TAG content may not be the best marker for insulin resistance. Thus, increased TAG content in skeletal muscle of obese and T2D subjects is adaptive.

Topics: Cells, Cultured; Diabetes Complications; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Middle Aged; Muscle Fibers, Skeletal; Obesity; Reference Values; Triglycerides

Hepatic insulin resistance in the leptin-receptor defective Zucker fa/fa rat is associated with impaired glycogen synthesis and increased activity of phosphorylase-a. We investigated the coupling between phosphorylase-a and glycogen synthesis in hepatocytes from fa/fa rats by modulating the concentration of phosphorylase-a. Treatment of hepatocytes from fa/fa rats and Fa/? controls with a selective phosphorylase inhibitor caused depletion of phosphorylase-a, activation of glycogen synthase and stimulation of glycogen synthesis. The flux-control coefficient of phosphorylase on glycogen synthesis was glucose dependent and at 10 mm glucose was higher in fa/fa than Fa/? hepatocytes. There was an inverse correlation between the activities of glycogen synthase and phosphorylase-a in both fa/fa and Fa/? hepatocytes. However, fa/fa hepatocytes had a higher activity of phosphorylase-a, for a corresponding activity of glycogen synthase. This defect was, in part, normalized by expression of the glycogen-targeting protein, PTG. Hepatocytes from fa/fa rats had normal expression of the glycogen-targeting proteins G(L) and PTG but markedly reduced expression of R6. Expression of R6 protein was increased in hepatocytes from Wistar rats after incubation with leptin and insulin. Diminished hepatic R6 expression in the leptin-receptor defective fa/fa rat may be a contributing factor to the elevated phosphorylase activity and/or its high control strength on glycogen synthesis.

Topics: Animals; Carrier Proteins; Cells, Cultured; Diabetes Mellitus, Type 2; Disease Models, Animal; Female; Glycogen; Hepatocytes; Insulin; Insulin Resistance; Intracellular Signaling Peptides and Proteins; Leptin; Male; Obesity; Phosphoprotein Phosphatases; Phosphorylase a; Protein Subunits; Rats; Rats, Wistar; Rats, Zucker; Receptors, Cell Surface; Receptors, Leptin

A study was undertaken to evaluate the antihyperglycemic activity of aqueous extract of bark of Garuga pinnata Roxb. (Burseraceae). The various parameters studied included fasting blood sugar levels, serum lipid levels, liver glycogen content, serum insulin level and glycated hemoglobin in diabetic and normal rats. Streptozotocin-nicotinamide was used to induce type-II diabetes mellitus. Treatment with the extract at two dose levels showed a significant increase in the liver glycogen and serum insulin level and a significant decrease in fasting blood glucose and glycated hemoglobin levels. The total cholesterol and serum triglycerides levels were also significantly reduced and the HDL cholesterol levels were significantly increased upon treatment with the extract thus proving the potent antidiabetic property of the plant.

Topics: Animals; Blood Glucose; Burseraceae; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glycated Hemoglobin; Glycogen; Hypoglycemic Agents; Lipids; Male; Pancreas; Plant Bark; Plant Extracts; Rats; Rats, Wistar; Toxicity Tests, Acute; Water

Creatine supplementation may exert beneficial effects on muscle performance and facilitate peripheral glucose disposal in both rats and human subjects. The present study was undertaken to explore the effects of creatine supplementation on the ATP, creatine, phosphocreatine and glycogen content of white and red gastrocnemius and soleus muscles and on blood D-glucose and plasma insulin concentrations before and during an intravenous glucose tolerance test in Goto-Kakizaki rats, a current animal model of inherited type 2 diabetes mellitus. Creatine supplementation increased muscle creatine content, especially in the soleus muscle of young rats (+35.5-/+15.8%; d.f.=10; p<0.05), and lowered the insulinogenic index, i.e. the paired ratio between plasma insulin and blood D-glucose concentrations. The latter change was mainly attributable to a lowering of plasma insulin concentration. It is proposed, therefore, that creatine supplementation may improve the sensitivity to insulin in extrapancreatic sites in the present animal model of type 2 diabetes.

Topics: Adenosine Triphosphate; Animals; Blood Glucose; Body Weight; Creatine; Diabetes Mellitus, Type 2; Disease Models, Animal; Eating; Glucose Tolerance Test; Glycogen; Insulin; Male; Muscle, Skeletal; Rats; Rats, Inbred Strains

Glycogen phosphorylase inhibition represents a promising strategy to suppress inappropriate hepatic glucose output, while muscle glycogen is a major source of fuel during contraction. Glycogen phosphorylase inhibitors (GPi) currently being investigated for the treatment of type 2 diabetes do not demonstrate hepatic versus muscle glycogen phosphorylase isoform selectivity and may therefore impair patient aerobic exercise capabilities. Skeletal muscle energy metabolism and function are not impaired by GPi during high-intensity contraction in rat skeletal muscle; however, it is unknown whether glycogen phosphorylase inhibitors would impair function during prolonged lower-intensity contraction. Utilizing a novel red cell-perfused rodent gastrocnemius-plantaris-soleus system, muscle was pretreated for 60 min with either 3 micromol/l free drug GPi (n=8) or vehicle control (n=7). During 60 min of aerobic contraction, GPi treatment resulted in approximately 35% greater fatigue. Muscle glycogen phosphorylase a form (P<0.01) and maximal activity (P<0.01) were reduced in the GPi group, and postcontraction glycogen (121.8 +/- 16.1 vs. 168.3 +/- 8.5 mmol/kg dry muscle, P<0.05) was greater. Furthermore, lower muscle lactate efflux and glucose uptake (P<0.01), yet higher muscle Vo(2), support the conclusion that carbohydrate utilization was impaired during contraction. Our data provide new confirmation that muscle glycogen plays an essential role during submaximal contraction. Given the critical role of exercise prescription in the treatment of type 2 diabetes, it will be important to monitor endurance capacity during the clinical evaluation of nonselective GPi. Alternatively, greater effort should be devoted toward the discovery of hepatic-selective GPi, hepatic-specific drug delivery strategies, and/or alternative strategies for controlling excess hepatic glucose production in type 2 diabetes.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Enzyme Inhibitors; Female; Glucose; Glycogen; Glycogen Phosphorylase; Glycogen Phosphorylase, Liver Form; Glycogen Phosphorylase, Muscle Form; Lactic Acid; Liver Glycogen; Muscle Contraction; Muscle, Skeletal; Rats; Rats, Wistar

Ethanolic extracts of Ananas comosus L. leaves (AC) enriched with phenols have hypoglycemic activity in diabetic rats. Here, we investigated the effect of AC on insulin sensitivity in rats and HepG2. In high-fat diet-fed and low-dose streptozotozin-treated diabetic Wistar rats subjected to challenge with exogenous human insulin, AC treatment at an oral dose of 0.40 g/kg could significantly improve sensitivity to exogenous insulin. After a sub-acute treatment, AC also could inhibit the development of insulin resistance in high-fat diet-fed and low-dose streptozotozin-treated diabetic rats following the test of loss of tolbutamide-induced blood glucose lowering action. For intravenous insulin/glucose infusion test, high-fat diet-fed and low-dose alloxan-treated Wistar rats were associated with insulin resistance, which was improved after AC or fenofibrate treatment. AC application inhibited the development of insulin resistance in HepG2 cells. The above animal models were well developed to simulate type 2 diabetes. Taken together, our results suggest that AC may improve insulin sensitivity in type 2 diabetes and could be developed into a new potential natural product for handling of insulin resistance in diabetic patients.

Topics: Ananas; Animals; Blood Glucose; Cell Line, Tumor; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Ethanol; Glucose; Glycogen; Humans; Insulin Resistance; Male; Phytotherapy; Plant Extracts; Plant Leaves; Rats; Rats, Wistar

Accumulation of intramuscular long-chain acyl-CoA esters (LCACoA) has previously in animal and human models been suggested to play an important role in lipid induced insulin resistance. The aim of this study was to examine whether myotubes established from type 2 diabetic (T2D) subjects and lean controls express differences in long-chain acyl-CoA esters (LCACoA) precultured under physiological conditions and during chronic exposure to palmitate (PA) and oleic acids (OA) with/without acute insulin stimulation. No significant differences were found between diabetic and control myotubes, neither in the total amount nor among individual LCA-CoA species during basal and acute insulin stimulation. LCA-CoA accumulated during exposure to palmitic acid but not during exposure to oleic acid. During PA and OA exposure, only palmitoyl-CoA, oleoyl-CoA and total LCA-CoA change. PA exposure increased the palmitoyl-CoA, whereas oleoyl-CoA was reduced and vice versa during OA exposure. No differences were found in the LCA-CoA level between T2D and control subjects, neither in the total amount nor in the individual specific LCA-CoA species during fatty acid exposure. Chronic (24 h), high PA, but not OA exposure induced insulin resistance at the level of glycogen synthesis in control subjects. These results indicate that (1) no primary defects are responsible for LCA-CoA accumulation in diabetic subjects; (2) LCA-CoA changes in vivo are partly adaptive to changes in the PA level and possibly other saturated fatty acids; and (3) PA induced insulin resistance may be mediated through an increased level of palmitoyl-CoA.

Topics: Acyl Coenzyme A; Blood Glucose; Body Mass Index; Diabetes Mellitus, Type 2; Glycated Hemoglobin; Glycogen; Humans; Insulin; Middle Aged; Muscle Fibers, Skeletal; Obesity; Reference Values

Defective fatty acid oxidation in skeletal muscle is one of the possible causes of insulin resistance. Peroxisome proliferator-activated receptor beta activators are strong inducers of fatty acid oxidation. The aim of this study was to verify whether activation of fatty acid oxidation by PPARbeta agonists in human skeletal muscle cells prepared from type 2 diabetic patients could improve the reduced responses to insulin that characterized this cell model. GW0742 (10 nM) significantly increased fatty acid oxidation and oxidative gene expression in myotubes prepared from both healthy subjects and type 2 diabetic patients. In cells from control subjects, incubation with the agonist for 48 h affected neither insulin-induced rate of glycogen synthesis nor the phosphorylation state of protein kinase B (PKB serine 473). Myotubes from type 2 diabetic patients displayed marked reduction in the effects of insulin on glycogen synthesis and on PKB phosphorylation. However, treatment with PPARbeta agonists did not restore these defects. Therefore, these results indicate that induction of fatty acid oxidation with PPARbeta activators during short-term exposition is not sufficient to correct for insulin resistance in muscle cells from type 2 diabetic patients. This suggests that additional studies are needed to better characterize the link between fatty acid oxidation and insulin sensitivity in humans.

Topics: Cells, Cultured; Diabetes Mellitus, Type 2; Female; Glycogen; Humans; Insulin; Male; Middle Aged; Muscle Cells; Oxidation-Reduction; Palmitic Acid; Phosphorylation; PPAR delta; PPAR-beta; Proto-Oncogene Proteins c-akt; Thiazoles; Time Factors

The effect of restoration of normoglycemia by a novel sodium-dependent glucose transporter inhibitor (T-1095) on impaired hepatic glucose uptake was examined in 14-week-old Zucker diabetic fatty (ZDF) rats. The nontreated group exhibited persistent endogenous glucose production (EGP) despite marked hyperglycemia. Gluconeogenesis and glucose cycling (GC) were responsible for 46 and 51% of glucose-6-phosphatase (G6Pase) flux, respectively. Net incorporation of plasma glucose into hepatic glycogen was negligible. Glucokinase (GK) and its inhibitory protein, GK regulatory protein (GKRP), were colocalized in the cytoplasm of hepatocytes. At day 7 of drug administration, EGP was slightly reduced, but G6Pase flux and GC were markedly lower compared with the nontreated group. In this case, GK and GKRP were colocalized in the nuclei of hepatocytes. When plasma glucose and insulin levels were raised during a clamp, EGP was completely suppressed and GC, glycogen synthesis from plasma glucose, and the fractional contribution of plasma glucose to uridine diphosphoglucose flux were markedly increased. GK, but not GKRP, was translocated from the nucleus to the cytoplasm. Glucotoxicity may result in the blunted response of hepatic glucose flux to elevated plasma glucose and/or insulin associated with impaired regulation of GK by GKRP in ZDF rats.

Topics: Animals; Blood Glucose; Carbonates; Carrier Proteins; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Glucagon; Glucokinase; Glucose; Glucose-6-Phosphatase; Glucosides; Glycogen; Glycogen Synthase; Hyperglycemia; Insulin; Liver; Liver Glycogen; Male; Muscle, Skeletal; Phosphorylases; Rats; Rats, Zucker

The present study tested the hypothesis that the phosphorylation and regulation of metabolic proteins implicated in glucose homeostasis were impaired in the heart of the type 2 diabetic Zucker-diabetic-fatty (ZDF) rat model. The onset of hyperglycaemia in ZDF rats was not uniform, instead it either progressed rapidly (3-4 weeks) or was delayed (6-8 weeks). In both the early and late onset hyperglycaemic ZDF rats, AMPKalpha Thr172 phosphorylation in the heart was significantly decreased. In the early onset hyperglycaemic ZDF rats, PKB Ser473 phosphorylation was reduced, whereas Thr308 phosphorylation was significantly increased. In the late onset hyperglycaemic ZDF rats, PKB Ser473 phosphorylation was unchanged, but Thr308 phosphorylation remained elevated. Cardiac GLUT4 protein and mRNA expression were significantly reduced in the early onset hyperglycaemic ZDF rats, whereas increased protein expression was observed in the late onset hyperglycaemic ZDF rats. In conclusion, the present study has demonstrated that following a more rapid onset of hyperglycaemia, the type 2 diabetic heart is more prone to alterations in the signaling proteins implicated in glucose metabolism.

Topics: AMP-Activated Protein Kinases; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Disease Models, Animal; Disease Progression; Glucose; Glucose Transporter Type 4; Glycogen; Glycogen Synthase Kinase 3; Hyperglycemia; Insulin; Male; Multienzyme Complexes; Myocardium; Phosphorylation; Protein Kinases; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Zucker; RNA, Messenger; Time Factors

D-mannose is an essential monosaccharide constituent of glycoproteins and glycolipids. However, it is unknown how plasma mannose is supplied. The aim of this study was to explore the source of plasma mannose. Oral administration of glucose resulted in a significant decrease of plasma mannose concentration after 20 min in fasted normal rats. However, in fasted type 2 diabetes model rats, plasma mannose concentrations that were higher compared with normal rats did not change after the administration of glucose. When insulin was administered intravenously to fed rats, it took longer for plasma mannose concentrations to decrease significantly in diabetic rats than in normal rats (20 and 5 min, respectively). Intravenous administration of epinephrine to fed normal rats increased the plasma mannose concentration, but this effect was negated by fasting or by administration of a glycogen phosphorylase inhibitor. Epinephrine increased mannose output from the perfused liver of fed rats, but this effect was negated in the presence of a glucose-6-phosphatase inhibitor. Epinephrine also increased the hepatic levels of hexose 6-phosphates, including mannose 6-phosphate. When either lactate alone or lactate plus alanine were administered as gluconeogenic substrates to fasted rats, the concentration of plasma mannose did not increase. When lactate was used to perfuse the liver of fasted rats, a decrease, rather than an increase, in mannose output was observed. These findings indicate that hepatic glycogen is a source of plasma mannose.

Topics: Administration, Oral; Alanine; Animals; Arabinose; Blood Glucose; Chlorogenic Acid; Diabetes Mellitus, Type 2; Disease Models, Animal; Epinephrine; Glucose; Glucose-6-Phosphatase; Glycogen; Glycogen Phosphorylase; Hexosephosphates; Imino Furanoses; Injections, Intravenous; Insulin; Lactic Acid; Liver; Male; Mannose; Models, Biological; Rats; Rats, Inbred Strains; Rats, Wistar; Sugar Alcohols

The metabolic mechanism of hepatic glucose overproduction was investigated in 3,3'-5-triiodo-l-thyronine (T3)-treated rats and Zucker diabetic fatty (ZDF) rats (fa/fa) after a 24-h fast. 2H2O and [U-13C3]propionate were administered intraperitoneally, and [3,4-13C2]glucose was administered as a primed infusion for 90 min under ketamine-xylazine anesthesia. 13C NMR analysis of monoacetone glucose derived from plasma glucose indicated that hepatic glucose production was twofold higher in both T3-treated rats and ZDF rats compared with controls, yet the sources of glucose overproduction differed significantly in the two models by 2H NMR analysis. In T3-treated rats, the hepatic glycogen content and hence the contribution of glycogenolysis to glucose production was essentially zero; in this case, excess glucose production was due to a dramatic increase in gluconeogenesis from TCA cycle intermediates. 13C NMR analysis also revealed increased phosphoenolpyruvate carboxykinase flux (4x), increased pyruvate cycling flux (4x), and increased TCA flux (5x) in T3-treated animals. ZDF rats had substantial glycogen stores after a 24-h fast, and consequently nearly 50% of plasma glucose originated from glycogenolysis; other fluxes related to the TCA cycle were not different from controls. The differing mechanisms of excess glucose production in these models were easily distinguished by integrated 2H and 13C NMR analysis of plasma glucose.

Topics: Animals; Blood Glucose; Citric Acid Cycle; Diabetes Mellitus, Type 2; Glucose; Glycerol; Glycogen; Kinetics; Liver; Male; Nuclear Magnetic Resonance, Biomolecular; Phosphoenolpyruvate; Rats; Rats, Sprague-Dawley; Rats, Zucker; Triiodothyronine

Liver X receptors (LXRs) are important regulators of cholesterol and lipid metabolism and are also involved in glucose metabolism. However, the functional role of LXRs in human skeletal muscle is at present unknown. This study demonstrates that chronic ligand activation of LXRs by a synthetic LXR agonist increases the uptake, distribution into complex cellular lipids, and oxidation of palmitate as well as the uptake and oxidation of glucose in cultured human skeletal muscle cells. Furthermore, the effect of the LXR agonist was additive to acute effects of insulin on palmitate uptake and metabolism. Consistently, activation of LXRs induced the expression of relevant genes: fatty acid translocase (CD36/FAT), glucose transporters (GLUT1 and -4), sterol regulatory element-binding protein-1c, peroxisome proliferator-activated receptor-gamma, carnitine palmitoyltransferase-1, and uncoupling protein 2 and 3. Interestingly, in response to activation of LXRs, myotubes from patients with type 2 diabetes showed an elevated uptake and incorporation of palmitate into complex lipids but an absence of palmitate oxidation to CO(2). These results provide evidence for a functional role of LXRs in both lipid and glucose metabolism and energy uncoupling in human myotubes. Furthermore, these data suggest that increased intramyocellular lipid content in type 2 diabetic patients may involve an altered response to activation of components in the LXR pathway.

Topics: Anticholesteremic Agents; Cells, Cultured; Diabetes Mellitus, Type 2; DNA-Binding Proteins; Gene Expression; Glucose; Glycogen; Humans; Hydrocarbons, Fluorinated; Lipid Metabolism; Liver X Receptors; Middle Aged; Muscle, Skeletal; Obesity; Orphan Nuclear Receptors; Receptors, Cytoplasmic and Nuclear; Sulfonamides

Variation in the calpain 10 gene has been reported to increase susceptibility to type 2 diabetes. Part of this susceptibility appears to be mediated by a decrease in whole body insulin sensitivity. As skeletal muscle is the primary tissue site of the peripheral insulin resistance in type 2 diabetes, the aim of this study was to use a human skeletal muscle cell culture system to explore the effects of calpain inhibition on insulin action. Calpain 10 mRNA and protein expression was examined in cultured myoblasts, myotubes, and whole skeletal muscle from non-diabetic subjects using RT-PCR and Western blotting. Changes in insulin-stimulated glucose uptake and glycogen synthesis in response to the calpain inhibitors ALLN and ALLM were measured. Calpain 10 expression was confirmed in cultured human myoblasts, myotubes, and native skeletal muscle. Insulin-stimulated glucose uptake was significantly decreased following preincubation with ALLN [404+/-40 vs 505+/-55 (mean+/-SEM)pmol/mg/min; with vs without ALLN: p = 0.04] and ALLM [455+/-38 vs 550+/-50 pmol/mg/min; with vs without ALLM: p = 0.025] in day 7 fused myotubes, but not in myoblasts. Neither ALLN nor ALLM affected insulin-stimulated glycogen synthesis in myoblasts or myotubes. These studies confirm calpain 10 expression in cultured human muscle cells and support a role for calpains in insulin-stimulated glucose uptake in human skeletal muscle cells that may be relevant to the pathogenesis of the peripheral insulin resistance in type 2 diabetes.

Topics: Base Sequence; Biological Transport; Calpain; Cell Culture Techniques; Cells, Cultured; Diabetes Mellitus, Type 2; DNA Primers; Gene Expression Regulation, Enzymologic; Genetic Predisposition to Disease; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Muscle, Skeletal; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription, Genetic

To determine whether the hepatic insulin resistance of obesity and type 2 diabetes is due to impaired insulin-induced suppression of glycogenolysis as well as gluconeogenesis, 10 lean nondiabetic, 10 obese nondiabetic, and 11 obese type 2 diabetic subjects were studied after an overnight fast and during a hyperinsulinemic-euglycemic clamp. Gluconeogenesis and glycogenolysis were measured using the deuterated water method. Before the clamp, when glucose and insulin concentrations differed among the three groups, gluconeogenesis was higher in the diabetic than in the obese nondiabetic subjects (P < 0.05) and glycogenolysis was higher in the diabetic than in the lean nondiabetic subjects (P < 0.05). During the clamp, when glucose and insulin concentrations were matched and glucagon concentrations were suppressed, both glycogenolysis and gluconeogenesis were higher (P < 0.01) in the diabetic versus the obese and lean nondiabetic subjects. Furthermore, glycogenolysis and gluconeogenesis were higher (P < 0.01) in the obese than in the lean nondiabetic subjects. Plasma free fatty acid concentrations correlated (P < 0.001) with glucose production and gluconeogenesis both before and during the clamp and with glycogenolysis during the clamp (P < 0.01). We concluded that defects in the regulation of glycogenolysis as well as gluconeogenesis cause hepatic insulin resistance in obese nondiabetic and type 2 diabetic humans.

Topics: Blood Glucose; Body Mass Index; Body Weight; Diabetes Mellitus, Type 2; Female; Gluconeogenesis; Glucose Clamp Technique; Glycogen; Humans; Hyperinsulinism; Insulin; Male; Middle Aged; Obesity

Protein tyrosine phosphatase 1B (PTP1B) acts as a physiological negative regulator of insulin signaling by dephosphorylating the activated insulin receptor (IR). Here we examine the role of PTP1B in the insulin-sensitizing action of rosiglitazone (RSG) in skeletal muscle and liver. Fat-fed, streptozotocin-treated rats (10-week-old), an animal model of type II diabetes, and age-matched, nondiabetic controls were treated with RSG (10 micromol kg(-1) day(-1)) for 2 weeks. After RSG treatment, the diabetic rats showed a significant decrease in blood glucose and improved insulin sensitivity. Diabetic rats showed significantly increased levels and activities of PTP1B in the skeletal muscle (1.6- and 2-fold, respectively) and liver (1.7- and 1.8-fold, respectively), thus diminishing insulin signaling in the target tissues. We found that the decreases in insulin-stimulated glucose uptake (55%), tyrosine phosphorylation of IRbeta-subunits (48%), and IR substrate-1 (IRS-1) (39%) in muscles of diabetic rats were normalized after RSG treatment. These effects were associated with 34 and 30% decreases in increased PTP1B levels and activities, respectively, in skeletal muscles of diabetic rats. In contrast, RSG did not affect the increased PTP1B levels and activities or the already reduced insulin-stimulated glycogen synthesis and tyrosine phosphorylation of IRbeta-subunits and IRS-2 in livers of diabetic rats. RSG treatment in normal rats did not significantly change PTP1B activities and levels or protein levels of IRbeta, IRS-1, and -2 in diabetic rats. These data suggest that RSG enhances insulin activity in skeletal muscle of diabetic rats possibly by ameliorating abnormal levels and activities of PTP1B.

Topics: Animals; Blotting, Western; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dietary Fats; Glucose; Glucose Tolerance Test; Glycogen; Hepatocytes; Hypoglycemic Agents; Insulin; Insulin Resistance; Liver; Liver Glycogen; Male; Muscle, Skeletal; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Protein Tyrosine Phosphatases; Rats; Rats, Sprague-Dawley; Rosiglitazone; Signal Transduction; Thiazolidinediones

Changes in the activity of glycogen synthase a and related kinases (phosphatidylinositol-3-kinase, protein kinase B, p44/42 MAP kinases and p70s6 kinase) evoked by GLP-1 in human myocytes from normal subjects were recently implied in the effect of this hormone upon D-glucose transport and glycogen synthesis in the same cells. The major aims of the present study were i) to investigate the possible extension of this knowledge to myocytes obtained from type 2 diabetic patients, ii) to compare in these patients the response to GLP-1, insulin or the structurally related GLP-1 peptides, exendin (1-39)amide and exendin(9-39)amide, and iii) to explore possible differences in the responsiveness to these agents between normal and diabetic subjects. Apart from the much higher basal PI3K activity and impaired response to insulin of p44/42 MAP kinases in the diabetic patients, the changes in enzyme activity caused by either hormone or peptide, although not identical, were essentially comparable. Nevertheless, significant differences in glucose transport and metabolism parameters were observed in the diabetic patients vs. normal subjects: in the diabetic patients, basal 2-deoxy-glucose uptake and glycogen synthase a activity were lower, accompanied by a similar increasing effect of GLP-1 or insulin; yet, the basal value for glycogen synthesis was higher, coinciding with a lesser relative increment in response to GLP-1 or insulin.

Topics: Aged; Aged, 80 and over; Cells, Cultured; Deoxyglucose; Diabetes Mellitus, Type 2; Exenatide; Female; Glucose; Glycogen; Glycogen Synthase; Humans; Immunoblotting; Insulin; Male; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Muscle Cells; Peptide Fragments; Peptides; Phosphatidylinositol 3-Kinases; Ribosomal Protein S6 Kinases, 70-kDa; Signal Transduction; Venoms

In obesity, skeletal muscle accumulates triglyceride. Recent work from Hulver and colleagues (2005) in the October issue of Cell Metabolism implicates stearoyl-CoA desaturase as part of the underlying molecular mechanism.

Topics: Carnitine O-Palmitoyltransferase; Diabetes Mellitus, Type 2; Fatty Acids; Glycogen; Humans; Insulin Resistance; Malonyl Coenzyme A; Muscle Fibers, Skeletal; Muscle, Skeletal; Obesity; Oxidation-Reduction; Stearoyl-CoA Desaturase; Triglycerides

Hepatic glucose fluxes and intracellular movement of glucokinase (GK) in response to increased plasma glucose and insulin were examined in 10-wk-old, 6-h-fasted, conscious Zucker diabetic fatty (ZDF) rats and lean littermates. Under basal conditions, plasma glucose (mmol/l) and glucose turnover rate (GTR; micromol.kg(-1).min(-1)) were slightly higher in ZDF (8.4 +/- 0.3 and 53 +/- 7, respectively) than in lean rats (6.2 +/- 0.2 and 45 +/- 4, respectively), whereas plasma insulin (pmol/l) was higher in ZDF (1,800 +/- 350) than in lean rats (150 +/- 14). The ratio of hepatic uridine 5'-diphosphate-glucose 3H specific activity to plasma glucose 3H specific activity ([3H]UDP-G/[3H]G; %), total hepatic glucose output (micromol.kg(-1).min(-1)), and hepatic glucose cycling (micromol.kg(-1).min(-1)) were higher in ZDF (35 +/- 5, 87 +/- 16, and 33 +/- 10, respectively) compared with lean rats (18 +/- 3, 56 +/- 6, and 11 +/- 2, respectively). [3H]glucose incorporation into glycogen (micromol glucose/g liver) was similar in lean (1.0 +/- 0.7) and ZDF (1.6 +/- 0.8) rats. GK was predominantly located in the nucleus in both rats. With elevated plasma glucose and insulin, GTR (micromol.kg(-1).min(-1)), [3H]UDP-G/[3H]G (%), and [3H]glucose incorporation into glycogen (micromol glucose/g liver) were markedly higher in lean (191 +/- 22, 62 +/- 3, and 5.0 +/- 1.4, respectively) but similar in ZDF rats (100 +/- 6, 37 +/- 3, and 1.4 +/- 0.4, respectively) compared with basal conditions. GK translocation from the nucleus to the cytoplasm occurred in lean but not in ZDF rats. The unresponsiveness of hepatic glucose flux to the rise in plasma glucose and insulin seen in prediabetic ZDF rats was associated with impaired GK translocation.

Topics: Animals; Biological Transport; Blood Glucose; Carrier Proteins; Diabetes Mellitus; Diabetes Mellitus, Type 2; Glucagon; Glucokinase; Glucose; Glucose-6-Phosphate; Glycogen; Insulin; Intracellular Membranes; Intracellular Signaling Peptides and Proteins; Liver; Male; Muscle, Skeletal; Obesity; Rats; Rats, Zucker; Thinness; Tissue Distribution

We previously reported that the Goto-Kakizaki (GK) rat, an animal model of type 2 diabetes, has hepatic insulin resistance using a perfused rat liver model. Pioglitazone, eicosapentaenoic acid (EPA), and fenofibrate are antihyperlipidemic agents and improve glucose tolerance. There have been few studies showing that these agents directly improve hepatic insulin sensitivity in type 2 diabetes mellitus. The aim of this study was to explore the effects of these agents on hepatic insulin sensitivity directly using a perfused GK rat liver model.. GK rats were treated with oral pioglitazone (6 or 10 mg/kg body weight), EPA (1 or 2 g/kg body weight), or fenofibrate (30 mg/kg body weight) for 2 weeks. Livers were perfused in situ with glucagon or with glucagon and insulin, and hepatic glucose outputs were measured.. In the pioglitazone-treated GK rats, blood glucose levels were significantly decreased. In the pioglitazone- and EPA-treated GK rats, insulin infusion significantly attenuated hepatic glucose output stimulated by glucagon. In the fenofibrate-treated GK rats, fat deposits in the hepatocytes were decreased, and glucose output elicited by glucagon was significantly decreased compared with that in the untreated GK rats, whereas insulin infusion did not affect glucose output by glucagon.. These findings suggest that pioglitazone and EPA may improve glucose tolerance by directly increasing hepatic insulin sensitivity, while fenofibrate may improve glucose tolerance by improving hepatic glycogen metabolism in the GK rats. We previously reported that the Goto-Kakizaki (GK) rat, an animal model of type 2 diabetes, has hepatic insulin resistance using a perfused rat liver model. Pioglitazone, eicosapentaenoic acid (EPA), and fenofibrate are antihyperlipidemic agents and improve glucose tolerance. There have been few studies showing that these agents directly improve hepatic insulin sensitivity in type 2 diabetes mellitus. The aim of this study was to explore the effects of these agents on hepatic insulin sensitivity directly using a perfused GK rat liver model.

Topics: Animals; Diabetes Mellitus, Type 2; Eicosapentaenoic Acid; Fenofibrate; Glycogen; Hypoglycemic Agents; Hypolipidemic Agents; Insulin Resistance; Liver; Male; Models, Animal; Perfusion; Pioglitazone; Rats; Thiazolidinediones

Insulin resistance is one of the primary characteristics of type 2 diabetes. Mice overexpressing a dominant-negative IGF-I receptor specifically in muscle (MKR mice) demonstrate severe insulin resistance with high levels of serum and tissue lipids and eventually develop type 2 diabetes at 5-6 wk of age. To determine whether lipotoxicity plays a role in the progression of the disease, we crossed MKR mice with mice overexpressing a fatty acid translocase, CD36, in skeletal muscle. The double-transgenic MKR/CD36 mice showed normalization of the hyperglycemia and the hyperinsulinemia as well as a marked improvement in liver insulin sensitivity. The MKR/CD36 mice also exhibited normal rates of fatty acid oxidation in skeletal muscle when compared with the decreased rate of fatty acid oxidation in MKR. With the reduction in insulin resistance, beta-cell function returned to normal. These and other results suggest that the insulin resistance in the MKR mice is associated with increased muscle triglycerides levels and that whole-body insulin resistance can be, at least partially, reversed in association with a reduction in muscle triglycerides levels, although the mechanisms are yet to be determined.

Topics: Animals; CD36 Antigens; Diabetes Mellitus, Type 2; Fatty Acids; Glucose; Glucose Clamp Technique; Glycogen; Hyperglycemia; Hyperinsulinism; In Vitro Techniques; Insulin; Insulin Resistance; Insulin Secretion; Islets of Langerhans; Liver; Male; Mice; Mice, Transgenic; Muscle, Skeletal; Oxidation-Reduction; Triglycerides

Impaired insulin action is a characteristic feature of type 2 diabetes. The study aims were to investigate whether after prolonged culture skeletal muscle cultures from insulin-resistant, type 2 diabetic patients (taking >100 U insulin/d) displayed impaired insulin signaling effects compared with cultures from nondiabetic controls and to determine whether retained abnormalities were limited to insulin action by studying an alternative pathway of stimulated glucose uptake. Studies were performed on myotubes differentiated for 7 d between passages 4 and 6. Insulin-stimulated glucose uptake (100 nm; P < 0.05) and insulin-stimulated glycogen synthesis (1 nm; P < 0.01) were significantly impaired in the diabetic vs. control cultures. Protein kinase B (PKB) expression and phosphorylated PKB levels in response to insulin stimulation (20 nm) were comparable in the diabetic and control cultures. 5-Amino-4-imidazolecarboxamide riboside (AICAR) mimics the effect of exercise on glucose uptake by activating AMP-activated protein kinase. There was no difference in AICAR (2 mm)-stimulated glucose uptake between diabetic vs. control myotube cultures (P = not significant). In conclusion, diabetic muscle cultures retain signaling defects after prolonged culture that appear specific to the insulin signaling pathway, but not involving PKB. This supports an intrinsic abnormality of the diabetic muscle cells that is most likely to have a genetic basis.

Topics: Aged; Aminocaproic Acid; Cells, Cultured; Diabetes Mellitus, Type 2; Female; Glucose; Glycogen; Glycogen Synthase; Humans; Insulin; Insulin Resistance; Male; Middle Aged; Muscle Fibers, Skeletal; Phosphorylation; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Time Factors

The contribution of increased gluconeogenesis (GNG) to the excessive rate of endogenous glucose production (EGP) in type 2 diabetes (T2DM) is well established. However, the separate effects of obesity (total body fat), visceral adiposity, and T2DM have not been investigated. We measured GNG (by the (2)H(2)O technique) and EGP (with 3-(3)H-glucose) after an overnight fast in 44 type 2 diabetic and 29 gender/ethnic-matched controls. Subjects were classified as obese (body mass index 30 kg/m(2) or greater) or nonobese (body mass index < 30 kg/m(2)); diabetic subjects were further subdivided according to the severity of fasting hyperglycemia [fasting plasma glucose (FPG) < 9 mm or >or= 9 mm]. EGP was similar in nondiabetic controls and T2DM with FPG less than 9 mm but was increased in T2DM with FPG >or= 9 mm (P < 0.001). Within the diabetic groups, obesity had an independent effect to further increase basal EGP (P < 0.01). In both nonobese diabetic groups, both the percent GNG and gluconeogenic flux were increased, compared with nonobese nondiabetic controls. In both diabetic groups, obesity further increased both percent GNG and gluconeogenic flux. In obese and nonobese T2DM, the increase in gluconeogenic flux was not accompanied by a reciprocal decrease in glycogenolysis, indicating a loss of hepatic autoregulation. By multivariate analysis, gluconeogenic flux was positively correlated with percent body fat, visceral fat, and the fasting plasma free fatty acid and glucose concentrations (all P

Topics: Abdomen; Adipose Tissue; Adult; Anthropometry; Case-Control Studies; Diabetes Mellitus; Diabetes Mellitus, Type 2; Female; Gluconeogenesis; Glucose; Glucose Clamp Technique; Glycogen; Humans; Magnetic Resonance Imaging; Male; Middle Aged; Multivariate Analysis; Obesity

To assess the effect of taurine supplementation on respiratory gas exchange, which might reflect the improved metabolism of glucose and/or lipid in the type 2 diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats.. Male OLETF rats (16 weeks of age) were randomly divided into two groups: unsupplemented group and taurine-supplemented (3% in drinking water) group. After 9 weeks of treatment, indirect calorimetry and insulin tolerance tests were conducted. The amounts of visceral fat pads, tissue glycogen, the blood concentrations of glucose, triacylglycerol, taurine, and electrolytes, and the level of hematocrit were compared between groups. A nondiabetic rat strain (Long-Evans Tokushima Otsuka) was used as the age-matched normal control.. The indirect calorimetry showed that the treatment of OLETF rats with taurine could reduce a part of postprandial glucose oxidation possibly responsible for the increase of triacylglycerol synthesis in the body. Taurine supplementation also improved hyperglycemia and insulin resistance and increased muscle glycogen content in the OLETF rats. Supplementation with taurine increased the blood concentration of taurine and electrolyte and fluid volume, all of which were considered to be related to the improvement of metabolic disturbance in OLETF rats.. Taurine supplementation may be an effective treatment for glucose intolerance and fat/lipid accumulation observed in type 2 diabetes associated with obesity. These metabolic changes might be ascribed, in part, to the alteration of circulating blood profiles, where the improved hyperglycemia and/or the blood accumulation of taurine itself would play roles.

Topics: Animal Nutritional Physiological Phenomena; Animals; Blood; Blood Glucose; Blood Pressure; Body Weight; Calorimetry, Indirect; Diabetes Mellitus, Type 2; Dietary Supplements; Drinking; Eating; Electrolytes; Food; Glycogen; Hematocrit; Insulin; Insulin Resistance; Male; Osmolar Concentration; Pulmonary Gas Exchange; Rats; Rats, Inbred OLETF; Taurine

Several defects in response to insulin have been described in vivo and in vitro in type 2 diabetes: a decreased glucose transport, defective glucose oxidation and altered glycogen synthesis. At present, it is unknown whether glucose oxidation is primarily affected or secondarily affected by, e.g. increased free fatty acids (FFA). The aim of this study was to evaluate whether myotubes established from type 2 diabetic subjects express a primarily or a FFA-induced reduced insulin-mediated glucose oxidation. We have therefore investigated glucose oxidation under basal, physiological conditions and during acute insulin stimulation with/without FFA. We found that myotubes established from type 2 diabetic subjects express a reduced insulin-stimulated increase in glucose oxidation. Moreover, an acute exposure to FFA reduces insulin-mediated glucose oxidation without alterations in glucose uptake and glycogen synthesis. Thus, we conclude that the diminished increase in insulin-stimulated glucose oxidation seen in type 2 diabetic subjects in vivo may be of genetic origin. Moreover, the glucose-fatty acid cycle seems not to be crucial for the pathophysiology of insulin resistance.

Topics: Cells, Cultured; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Glucose; Glycogen; Humans; Insulin; Muscle Fibers, Skeletal; Muscle, Skeletal; Oxidation-Reduction; Palmitic Acid

The aim of this study was to examine the mechanisms by which dietary carbohydrate and fat modulate fasting glycemia. We compared the effects of an eucaloric high-carbohydrate (89% carbohydrate) and high-fat (89% fat) diet on fasting glucose metabolism and insulin sensitivity in seven obese patients with type 2 diabetes using stable isotopes and euglycemic hyperinsulinemic clamps. At basal insulin levels glucose concentrations were 148 +/- 11 and 123 +/- 11 mg/dl (8.2 +/- 0.6 and 6.8 +/- 0.6 mmol/liter) on the high-carbohydrate and high-fat diet, respectively (P < 0.001), with insulin concentrations of 12 +/- 2 and 10 +/- 1 microIU/ml (82 +/- 11 and 66 +/- 10 pmol/liter) (P = 0.08). Glucose production was higher on the high-carbohydrate diet (1.88 +/- 0.06 vs. 1.55 +/- 0.05 mg/kg.min (10.44 +/- 0.33 vs. 8.61 +/- 0.28 micromol/kg.min) (P < 0.001) because of higher glycogenolysis. Gluconeogenic rates were not different between the diets. During the use of hyperinsulinemic euglycemic clamps, insulin-mediated suppression of glucose production and stimulation of glucose disposal were not different between the diets. Free fatty concentrations were suppressed by 89 and 62% (P < 0.0001) on the high-carbohydrate and high-fat diet, respectively. We conclude that short-term variations in dietary carbohydrate to fat ratios affect basal glucose metabolism in people with type 2 diabetes merely through modulation of the rate of glycogenolysis, without affecting insulin sensitivity of glucose metabolism.

Topics: Diabetes Mellitus, Type 2; Dietary Carbohydrates; Dietary Fats; Dose-Response Relationship, Drug; Female; Glucose; Glucose Clamp Technique; Glycogen; Humans; Insulin; Male; Middle Aged; Osmolar Concentration; Postprandial Period

To understand the day-to-day pathophysiology of impaired muscle glycogen storage in type 2 diabetes, glycogen concentrations were measured before and after the consumption of sequential mixed meals (breakfast: 190.5 g carbohydrate, 41.0 g fat, 28.8 g protein, 1253 kcal; lunch: 203.3 g carbohydrate, 48.1 g fat, 44.0 g protein, 1497.5 kcal) by use of natural abundance (13)C magnetic resonance spectroscopy. Subjects with diet-controlled type 2 diabetes (n = 9) and age- and body mass index-matched nondiabetic controls (n = 9) were studied. Mean fasting gastrocnemius glycogen concentration was significantly lower in the diabetic group (57.1 +/- 3.6 vs. 68.9 +/- 4.1 mmol/l; P < 0.05). After the first meal, mean glycogen concentration in the control group rose significantly from basal (97.1 +/- 7.0 mmol/l at 240 min; P = 0.005). After the second meal, the high level of muscle glycogen concentration in the control group was maintained, with a further rise to 108.0 +/- 11.6 mmol/l by 480 min. In the diabetic group, the postprandial rise was markedly lower than that of the control group (65.9 +/- 5.2 mmol/l at 240 min, P < 0.005, and 70.8 +/- 6.7 mmol/l at 480 min, P = 0.01) despite considerably greater serum insulin levels (752.0 +/- 109.0 vs. 372.3 +/- 78.2 pmol/l at 300 min, P = 0.013). This was associated with a significantly greater postprandial hyperglycemia (10.8 +/- 1.3 vs. 5.3 +/- 0.2 mmol/l at 240 min, P < 0.005). Basal muscle glycogen concentration correlated inversely with fasting blood glucose (r = -0.55, P < 0.02) and fasting serum insulin (r = -0.57, P < 0.02). The increment in muscle glycogen correlated with initial increment in serum insulin only in the control group (r = 0.87, P < 0.002). This study quantitates for the first time the subnormal basal muscle glycogen concentration and the inadequate glycogen storage after meals in type 2 diabetes.

Topics: Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Female; Glycogen; Humans; Insulin; Insulin Resistance; Lactic Acid; Magnetic Resonance Spectroscopy; Male; Middle Aged; Muscle, Skeletal; Postprandial Period; Triglycerides

Improvement of insulin sensitivity and lipid and glucose metabolism by coactivation of both nuclear peroxisome proliferator-activated receptor (PPAR)gamma and PPARalpha potentially provides beneficial effects over existing PPARgamma and alpha preferential drugs, respectively, in treatment of type 2 diabetes. We examined the effects of the dual PPARalpha/gamma agonist ragaglitazar on hyperglycemia and whole body insulin sensitivity in early and late diabetes stages in Zucker diabetic fatty (ZDF) rats and compared them with treatment with the PPARgamma preferential agonist rosiglitazone. Despite normalization of hyperglycemia and Hb A(1c) and reduction of plasma triglycerides by both compounds in both prevention and early intervention studies, ragaglitazar treatment resulted in overall reduced circulating insulin and improved insulin sensitivity to a greater extent than after treatment with rosiglitazone. In late-intervention therapy, ragaglitazar reduced Hb A(1c) by 2.3% compared with 1.1% by rosiglitazone. Improvement of insulin sensitivity caused by the dual PPARalpha/gamma agonist ragaglitazar seemed to have beneficial impact over that of the PPARgamma-preferential activator rosiglitazone on glycemic control in frankly diabetic ZDF rats.

Topics: Animals; Body Composition; Body Weight; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Eating; Fatty Acids, Nonesterified; Glucose Clamp Technique; Glycated Hemoglobin; Glycogen; Hypoglycemic Agents; Insulin Resistance; Islets of Langerhans; Liver; Male; Oxazines; Phenylpropionates; Rats; Rats, Zucker; Receptors, Cytoplasmic and Nuclear; Rosiglitazone; Thiazoles; Thiazolidinediones; Transcription Factors

Caesalpinia bonducella, widely distributed throughout the coastal region of India and used ethnically by the tribal people of India for controlling blood sugar was earlier reported by us to possess hypoglycemic activity in animal model. This prompted us to undertake a detail study with the aqueous and ethanolic extracts of the seeds of this plant in both type 1 and 2 diabetes mellitus in Long Evans rats. Significant blood sugar lowering effect (P < 0.05) of C. bonducella was observed in type 2 diabetic model. Special emphasis was given on the mechanistic study by gut absorption of glucose and liver glycogen.

Topics: Animals; Blood Glucose; Body Weight; Caesalpinia; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Female; Glycogen; Hypoglycemic Agents; Intestinal Absorption; Lipids; Liver; Male; Plant Extracts; Rats; Rats, Long-Evans

Intrauterine growth retardation (IUGR) has been linked to the development of type 2 diabetes in later life. We have developed a model of uteroplacental insufficiency, a common cause of intrauterine growth retardation, in the rat. Early in life, the animals are insulin resistant and by 6 mo of age they develop diabetes. Glycogen content and insulin-stimulated 2-deoxyglucose uptake were significantly decreased in muscle from IUGR rats. IUGR muscle mitochondria exhibited significantly decreased rates of state 3 oxygen consumption with pyruvate, glutamate, alpha-ketoglutarate, and succinate. Decreased pyruvate oxidation in IUGR mitochondria was associated with decreased ATP production, decreased pyruvate dehydrogenase activity, and increased expression of pyruvate dehydrogenase kinase 4. Such a defect in IUGR mitochondria leads to a chronic reduction in the supply of ATP available from oxidative phosphorylation. Impaired ATP synthesis in muscle compromises energy-dependent GLUT4 recruitment to the cell surface, glucose transport, and glycogen synthesis, which contribute to insulin resistance and hyperglycemia of type 2 diabetes.

Topics: Aconitate Hydratase; Adenosine Triphosphate; Animals; Blood Glucose; Blotting, Western; Citrate (si)-Synthase; Cytochromes; Diabetes Mellitus, Type 2; Electron Transport; Fatty Acids, Nonesterified; Female; Fetal Growth Retardation; Glucose; Glycogen; Homeostasis; Lactic Acid; Microscopy, Electron; Mitochondria, Muscle; Muscle Proteins; Muscle, Skeletal; Oxidative Phosphorylation; Oxygen Consumption; Pregnancy; Pyruvate Dehydrogenase Complex; Rats; Rats, Sprague-Dawley

Lipid accumulation in nonadipose tissues is closely related to the development of type 2 diabetes in obese subjects. We examined the potential preventive effect of peroxisome proliferator-activated receptor (PPAR)-alpha and PPAR-gamma stimulation on the development of diabetes in obese diabetes-prone OLETF rats. Chronic administration of a PPAR-alpha agonist (0.5% [wt/wt] fenofibrate) or a PPAR-gamma agonist (3 mg x kg(-1) x day(-1) rosiglitazone) completely prevented the development of glycosuria. Pancreatic islets from untreated OLETF rats underwent sequential hypertrophy and atrophy, which was completely prevented by chronic fenofibrate treatment. In contrast, rosiglitazone treatment did not affect islet hypertrophy at earlier stages but prevented beta-cell atrophy at later stages. Fenofibrate treatment decreased body weight and visceral fat, whereas rosiglitazone treatment increased body weight. Despite the opposite effects on adiposity, both drugs were equally effective in improving insulin actions in skeletal muscle. Furthermore, both drugs significantly decreased the triglyceride content in the soleus muscle and pancreatic islets. The present study demonstrates that the PPAR-alpha agonist fenofibrate prevents the development of diabetes in OLETF rats by reducing adiposity, improving peripheral insulin action, and exerting beneficial effects on pancreatic beta-cells.

Topics: Adipose Tissue; Animals; Basal Metabolism; Blood Glucose; Diabetes Mellitus; Diabetes Mellitus, Type 2; Fatty Acids; Fenofibrate; Glycogen; Hypoglycemic Agents; Hypolipidemic Agents; Insulin; Islets of Langerhans; Male; Muscle, Skeletal; Obesity; Rats; Rats, Inbred OLETF; Receptors, Cytoplasmic and Nuclear; Rosiglitazone; Thiazoles; Thiazolidinediones; Transcription Factors; Triglycerides; Viscera

Defects in liver and muscle glycogen synthesis are major factors contributing to postprandrial hyperglycemia in patients with type 2 diabetes. Therefore, activation of glycogen synthase through inhibition of glycogen synthase kinase (GSK)-3 represents a potential new therapeutic target. To examine this possibility, we performed oral glucose tolerance tests (OGTTs) and euglycemic-insulinemic clamp studies in Zucker diabetic fatty (fa/fa) rats before and after treatment with novel GSK-3 inhibitors. GSK-3 inhibition caused a 41 +/- 2% (P < 0.001) and 26 +/- 4% (P < 0.05) reduction in the area under the glucose and insulin concentration curves, respectively, during the OGTT. This improvement in glucose disposal could mostly be attributed to an approximate twofold increase in liver glycogen synthesis. In contrast, there was no significant increase in muscle glycogen synthesis despite an approximate threefold activation of muscle glycogen synthase activity. GSK-3 inhibitor treatment increased liver glycogen synthesis about threefold independent of insulin concentration during the clamp studies. In contrast, muscle glucose uptake and muscle glycogen synthesis were independent of drug treatment. GSK-3 inhibitor treatment lowered fasting hyperglycemia in diabetic rats by 6.0 +/- 1.3 mmol/l but had no significant effect on glucose disposal during the clamp. In conclusion, GSK-3 inhibition significantly improved oral glucose disposal, mostly by increasing liver glycogen synthesis. These studies suggest that GSK-3 inhibition may represent an important new therapeutic target for treatment of patients with type 2 diabetes.

Topics: Animals; Diabetes Mellitus, Type 2; Enzyme Inhibitors; Glucose; Glucose Clamp Technique; Glucose Tolerance Test; Glycogen; Glycogen Synthase Kinase 3; Hypoglycemic Agents; Imidazoles; Insulin; Liver; Male; Muscle, Skeletal; Pyridines; Pyrimidines; Rats; Rats, Zucker

alpha-Lipoic acid (LA) is currently being investigated as a glucose-lowering agent for diabetes control; it is also considered a powerful dietary antioxidant. The objective of this study was to investigate the fate of glucose in isolated rat muscles incubated with LA and determine its effects on intramuscular redox status. Rat soleus muscles were incubated for up to 60 min with 2.4 mmol/L LA in the presence or absence of insulin. Intramuscular concentrations of LA were evaluated (uptake and reduction), and glycogen synthesis, glucose oxidation, intramuscular reactive oxygen species (ROS) production and mitochondrial membrane potential investigated. Insulin enhanced glycogen synthesis, whereas LA decreased rates by >50%. LA elevated ROS production and in combination with t-butylhydroperoxide, an oxidant, additively inhibited glycogen synthesis rates by 80%. Insulin acted as an antioxidant and attenuated ROS production by 30%. LA uncoupled the mitochondria and accelerated glucose oxidation 1.5-fold relative to the control. The glycogen synthesis pathway was found to be dependent on mitochondrial function because treatment with mitochondrial inhibitors eliminated the majority of glycogen synthesis. These data show that in this model, LA acts as a mild prooxidant, causing mitochondrial uncoupling and inhibition of glycogen synthesis. It appears that LA regulates glucose metabolism in the muscle differently than insulin.

Topics: Animals; Antioxidants; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; Culture Techniques; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Glucose; Glycogen; Glycolysis; Hypoglycemic Agents; Insulin; Male; Membrane Potentials; Mitochondria, Muscle; Muscle, Skeletal; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Rotenone; Thioctic Acid; Uncoupling Agents

Recently, we observed that impairments exist in skeletal muscle free fatty acid (FFA) utilization during exercise in obese subjects with Type II diabetes. The main objective of the present study was to investigate whether plasma FFA oxidation is impaired during exercise in non-obese Type II diabetic patients. Stable isotope tracers of palmitate and glucose were infused for 2 h at rest and 1h of bicycle exercise at 40% peak oxygen consumption ( V*O(2)max) in volunteers with Type II diabetes and a healthy control group. At rest, plasma FFA oxidation was not significantly different between subjects with Type II diabetes and control subjects (2.13+/-0.51 versus 1.93+/-0.54 micromol.kg(-1).min(-1) respectively). During exercise, Type II diabetic patients and control subjects had similar rates of total fat [Type II diabetes, 9.62+/-1.84 micromol.kg(-1).min(-1); control, 12.08+/-4.59 micromol.kg(-1).min(-1); not significant (NS)] and glucose oxidation (Type II diabetes, 44.24+/-10.36 micromol.kg(-1).min(-1); control, 57.37+/-14.54 micromol.kg(-1).min(-1); NS). No aberrations were present in plasma FFA uptake [rate of disappearance ( Rd ); Type II diabetes, 11.78+/-4.82; control, 10.84+/-3.39; NS] and oxidation rates (Type II diabetes 8.10+/-1.44; control 8.00+/-3.12, NS) in Type II diabetic patients; triacylglycerol-derived fatty acid oxidation was 2.6-fold lower in Type II diabetic patients than in control subjects, but this difference was not statistically significant. Muscle glycogen oxidation was lower in diabetes patients than in control subjects (Type II diabetes, 25.16+/-13.82 micromol.kg(-1).min(-1); control, 42.04+/-10.58 micromol.kg(-1).min(-1); P <0.05) and plasma glucose contributed more to energy expenditure in Type II diabetes (26+/-3% in diabetic versus 15+/-2% in control, P <0.05). We conclude that plasma FFA oxidation is not impaired during exercise in non-obese Type II diabetic patients. The data confirm that Type II diabetes is a heterogeneous disease, and that the adaptation at the substrate level differs between obese and non-obese patients and may contribute to differences in the final appearance of the various phenotypes.

Topics: Adult; Blood Glucose; Case-Control Studies; Diabetes Mellitus, Type 2; Energy Metabolism; Exercise; Fatty Acids, Nonesterified; Glycogen; Humans; Insulin; Lipids; Male; Middle Aged; Muscle, Skeletal; Oxidation-Reduction; Oxygen Consumption

To examine the effect of glimepiride on insulin-stimulated glycogen synthesis in cultured human skeletal muscle cells in comparison with glibenclamide.. Myotubes derived from glucose-tolerant subjects were incubated with glimepiride or glibenclamide (0-100 micro mol/l) for 4 h and with or without insulin (100 nmol/l) for 2 h, and subsequently glycogen synthesis was determined.. Glimepiride had no significant effect on basal glycogen synthesis; in contrast, glimepiride caused a dose-dependent increase of insulin-stimulated glycogen synthesis, with a maximal effect of 39.97 +/- 8.4% (mean +/- SEM, n = 4, P < 0,02). The time course of this glimepiride effect on insulin-stimulated glycogen synthesis showed a peak after 12 h incubation with a half maximal effect after 4 h. Preincubation of the myotubes with wortmannin (100 nmol/l), an inhibitor of phosphatidylinositol (PI)- 3 kinase, caused an inhibition of this glimepiride effect on insulin-stimulated glycogen synthesis. In contrast to glimepiride, incubation of myotubes with glibenclamide (0-100nmol/l), a second generation sulfonylurea, had no significant effect on basal or insulin-stimulated glycogen synthesis.. Incubation of cultured human skeletal muscle cells derived from glucose-tolerant subjects with glimepiride caused a dose-dependent increase of insulin-stimulated glycogen synthesis using therapeutic glimepiride concentrations. This glimepiride effect seems to be mediated via the PI3 kinase pathway. In contrast to glimepiride, glibenclamide had no significant effect on basal or insulin-stimulated glycogen synthesis. These results suggest that glimepiride, beside its well-known effect to stimulate insulin secretion, possess an insulin-sensitizing action in cultured human skeletal muscle cells in support of the concept of an extrapancreatic action of glimepiride.

Topics: Androstadienes; Cell Culture Techniques; Cells, Cultured; Diabetes Mellitus, Type 2; Female; Glyburide; Glycogen; Humans; Hypoglycemic Agents; Insulin; Kinetics; Male; Muscle, Skeletal; Sulfonylurea Compounds; Wortmannin

It has been shown that tungstate is an effective hypoglycemic agent in several animal models of diabetes. In this study, we examined the effectiveness of oral tungstate treatment in a new experimental diabetic syndrome, induced by streptozotocin (STZ) and nicotinamide in adult rats, that shares several features with human type 2 diabetes. Sodium tungstate was administered in the drinking water (2 mg/mL) of control and diabetic rats for 15, 30, 60, and 90 d. Glucose metabolism was explored in vivo by intravenous glucose tolerance test. Insulin secretion and action were assessed in vitro in the isolated perfused pancreas and isolated adipocytes, respectively. Two weeks of tungstate treatment did not modify the moderate hyperglycemia of diabetic rats but reduced their intolerance to glucose, owing to an enhancement of postloading insulin secretion. However, this effect was transient, since it declined after 30 d and vanished after 60 and 90 d of tungstate administration, whereas a trend toward a reduction in basal hyperglycemia was observed on prolonged treatment. Oral tungstate was unable to modify glucose-stimulated insulin secretion in the isolated perfused pancreas, as well as muscle glycogen levels, hepatic glucose metabolism, and insulin-stimulated lipogenesis in isolated adipocytes. Nevertheless, the decreased insulin content of pancreatic islets of diabetic rats was partially restored on prolonged tungstate treatment. In conclusion, in the STZ-nicotinamide model of diabetes, tungstate was unable to permanently correct the alterations in glucose metabolism, despite some indirect evidence of a trophic effect on beta-cells. The ineffectiveness of tungstate could be related to the absence, in this diabetic syndrome, of relevant metabolic alterations in the liver, which thus appear to constitute the major target of tungstate action.

Topics: Adipocytes; Administration, Oral; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Drug Administration Schedule; Glucose; Glucose Tolerance Test; Glycogen; In Vitro Techniques; Insulin; Insulin Secretion; Islets of Langerhans; Liver; Male; Muscle, Skeletal; Pancreas; Rats; Rats, Wistar; Time Factors; Tungsten Compounds

Hepatic glucose production is increased as a metabolic consequence of insulin resistance in type 2 diabetes. Because fructose 2,6-bisphosphate is an important regulator of hepatic glucose production, we used adenovirus-mediated enzyme overexpression to increase hepatic fructose 2,6-bisphosphate to determine if the hyperglycemia in KK mice, polygenic models of type 2 diabetes, could be ameliorated by reduction of hepatic glucose production. Seven days after treatment with virus encoding a mutant 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase designed to increase fructose 2,6-bisphosphate levels, plasma glucose, lipids, and insulin were significantly reduced in KK/H1J and KK.Cg-A(y)/J mice. Moreover, high fructose 2,6-bisphosphate levels downregulated glucose-6-phosphatase and upregulated glucokinase gene expression, thereby reversing the insulin-resistant pattern of hepatic gene expression of these two key glucose-metabolic enzymes. The increased hepatic fructose 2,6-bisphosphate also reduced adiposity in both KK mice. These results clearly indicate that increasing hepatic fructose 2,6-bisphosphate overcomes the impairment of insulin in suppressing hepatic glucose production, and it provides a potential therapy for type 2 diabetes.

Topics: Adipose Tissue; Animals; Blood; Body Weight; Diabetes Mellitus, Type 2; Epididymis; Female; Fructosediphosphates; Glucokinase; Glucose-6-Phosphatase; Glycogen; Insulin; Insulin Resistance; Liver; Male; Mice; Mice, Inbred Strains; Phosphofructokinase-2

Considerable evidence suggests that skeletal muscle insulin resistance is an inherent feature of type 2 diabetes and contributes to the pathogenesis of the disease. In patients with poorly controlled diabetes, hyperglycemia is thought to produce additional insulin resistance in muscle. The magnitude and nature of hyperglycemia-induced insulin resistance is not known. The purpose of the present study was to determine the biochemical mechanisms responsible for increased insulin-stimulated glucose disposal after the achievement of tight glycemic control with a mixed-split regimen. We performed hyperinsulinemic-euglycemic clamps with indirect calorimetry and vastus lateralis muscle biopsies in eight type 2 diabetic patients who had poor glycemic control (HbA(1c) 10.1%) and again after 3 months of intensive insulin therapy designed to produce near-normoglycemia (HbA(1c) 6.6%). Improved glycemic control increased insulin-stimulated glucose disposal (5.16 +/- 0.32 vs. 3.69 +/- 0.33 mg x kg(-1) x min(-1); P < 0.01); nonoxidative glucose disposal, which primarily reflects glycogen synthesis (2.11 +/- 0.26 vs. 0.90 +/- 0.16 mg x kg(-1) x min(-1); P < 0.01); and glycogen synthase fractional velocity (0.094 +/- 0.017 vs. 0.045 +/- 0.007; P < 0.05). There was no improvement in insulin-stimulated glucose oxidation (3.05 +/- 0.25 vs. 2.79 +/- 0.20 mg x kg(-1) x min(-1)), hexokinase II mRNA expression (increase over basal values), or hexokinase II enzymatic activity (0.51 +/- 0.16 vs. 0.42 +/- 0.18 pmol x min(-1) x microg(-1) protein). All of the increase in insulin-stimulated glucose disposal could be accounted for by increased glycogen synthesis, which is likely attributable to increased activation of glycogen synthase by insulin.

Topics: Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Female; Glycogen; Glycogen Synthase; Hexokinase; Humans; Insulin; Male; Middle Aged; Osmolar Concentration; Reference Values; RNA, Messenger

The 5'AMP-activated protein kinase is an important mediator of muscle contraction-induced glucose transport and a target for pharmacological treatment of Type II (non-insulin-dependent) diabetes mellitus. The 5'AMP-activated protein kinase can be activated by 5-aminoimidazole-4-carboxamide ribonucleoside. We hypothesised that 5-aminoimidazole-4-carboxamide ribonucleoside treatment could restore glucose homeostasis in ob/ob mice.. Lean and ob/ob mice were given 5-aminoimidazole-4-carboxamide ribonucleoside (1 mg.g body wt(-1).day(-1) s.c) or 0.9 % NaCl (vehicle) for 1-7 days.. Short-term 5-aminoimidazole-4-carboxamide ribonucleoside treatment normalised glucose concentrations in ob/ob mice within 1 h, with effects persisting over 4 h. After 1 week of daily injections, 5-aminoimidazole-4-carboxamide ribonucleoside treatment corrected hyperglycaemia, improved glucose tolerance, and increased GLUT4 and hexokinase II protein expression in skeletal muscle, but had deleterious effects on plasma non-esterified fatty acids and triglycerides. Treatment with 5-aminoimidazole-4-carboxamide ribonucleoside increased liver glycogen in fasted and fed ob/ob mice and muscle glycogen in fasted, but not fed ob/ob and lean mice. Defects in insulin-stimulated phosphatidylinositol 3-kinase and glucose transport in skeletal muscle from ob/ob mice were not corrected by 5-aminoimidazole-4-carboxamide ribonucleoside treatment. While ex vivo insulin-stimulated glucose transport was reduced in isolated muscle from ob/ob mice, the 5-aminoimidazole-4-carboxamide ribonucleoside stimulated response was normal.. The 5-aminoimidazole-4-carboxamide ribonucleoside mediated improvements in glucose homeostasis in ob/ob mice can be explained by effects in skeletal muscle and liver. Due to the apparently deleterious effects of 5-aminoimidazole-4-carboxamide ribonucleoside on the blood lipid profile, strategies to develop tissue-specific and pathway-specific activators of 5'AMP-activated protein kinase should be considered in order to improve glucose homeostasis.

Topics: Aminoimidazole Carboxamide; Animals; Biological Transport; Blood Glucose; Diabetes Mellitus; Diabetes Mellitus, Type 2; Glucose; Glucose Tolerance Test; Glycogen; Homeostasis; Hypoglycemic Agents; Injections, Subcutaneous; Insulin; Insulin Resistance; Liver; Liver Glycogen; Mice; Mice, Inbred C57BL; Mice, Obese; Muscle, Skeletal; Obesity; Ribonucleotides

Topics: Acetyl-CoA Carboxylase; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin Resistance; Multienzyme Complexes; Muscle, Skeletal; Peroxisome Proliferators; Transcription Factors

The effect of inhibition of glycogen phosphorylase by 1,4-dideoxy-1,4-imino-d-arabinitol on rates of gluconeogenesis, gluconeogenic deposition into glycogen, and glycogen recycling was investigated in primary cultured hepatocytes, in perfused rat liver, and in fed or fasted rats in vivo clamped at high physiological levels of plasma lactate. 1,4-Dideoxy-1,4-imino-d-arabinitol did not alter the synthesis of glycerol-derived glucose in hepatocytes or lactate-derived glucose in perfused liver or fed or fasted rats in vivo. Thus, 1,4-dideoxy-1,4-imino-d-arabinitol inhibited hepatic glucose output in the perfused rat liver (0.77 +/- 0.19 versus 0.33 +/- 0.09, p < 0.05), whereas the rate of lactate-derived gluconeogenesis was unaltered (0.22 +/- 0.09 versus 0.18 +/- 0.08, p = not significant) (1,4-dideoxy-1,4-imino-d-arabinitol versus vehicle, micromol/min * g). Overall, the data suggest that 1,4-dideoxy-1,4-imino-d-arabinitol inhibited glycogen breakdown with no direct or indirect effects on the rates of gluconeogenesis. Total end point glycogen content (micromol of glycosyl units/g of wet liver) were similar in fed (235 +/- 19 versus 217 +/- 22, p = not significant) or fasted rats (10 +/- 2 versus 7 +/- 2, p = not significant) with or without 1,4-dideoxy-1,4-imino-d-arabinitol, respectively. The data demonstrate no glycogen cycling under the investigated conditions and no effect of 1,4-dideoxy-1,4-imino-d-arabinitol on gluconeogenic deposition into glycogen. Taken together, these data also suggest that inhibition of glycogen phosphorylase may prove beneficial in the treatment of type 2 diabetes.

Topics: Animals; Arabinose; Blood Glucose; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Female; Glycogen; Glycogen Phosphorylase; Hepatocytes; Imino Furanoses; Kinetics; Lactic Acid; Liver; Magnetic Resonance Spectroscopy; Male; Perfusion; Rats; Rats, Sprague-Dawley; Sugar Alcohols; Time Factors

To determine the immediate effect of thiazolidinediones on human skeletal muscle, differentiated human myotubes were acutely (1 day) and myoblasts chronically (during the differentiation process) treated with troglitazone (TGZ). Chronic TGZ treatment resulted in loss of the typical multinucleated phenotype. The increase of muscle markers typically observed during differentiation was suppressed, while adipocyte markers increased markedly. Chronic TGZ treatment increased insulin-stimulated phosphatidylinositol (PI) 3-kinase activity and membranous protein kinase B/Akt (PKB/Akt) Ser-473 phosphorylation more than 4-fold. Phosphorylation of p42/44 mitogen-activated protein kinase (42/44 MAPK/ERK) was unaltered. Basal glucose uptake as well as both basal and insulin-stimulated glycogen synthesis increased approximately 1.6- and approximately 2.5-fold after chronic TGZ treatment, respectively. A 2-fold stimulation of PI 3-kinase but no other significant TGZ effect was found after acute TGZ treatment. In conclusion, chronic TGZ treatment inhibited myogenic differentiation of that human muscle while inducing adipocyte-specific gene expression. The effects of chronic TGZ treatment on basal glucose transport may in part be secondary to this transdifferentiation. The enhancing effect on PI 3-kinase and PKB/Akt involved in both differentiation and glycogen synthesis appears to be pivotal in the cellular action of TGZ.

Topics: Adipocytes; Base Sequence; Biomarkers; Cell Differentiation; Cells, Cultured; Chromans; Diabetes Mellitus, Type 2; DNA Primers; Gene Expression; Glucose; Glucose Transporter Type 1; Glucose Transporter Type 4; Glycogen; Humans; Hypoglycemic Agents; Insulin; Insulin Resistance; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Monosaccharide Transport Proteins; Muscle Proteins; Muscle, Skeletal; Phosphatidylinositol 3-Kinases; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Receptors, Cytoplasmic and Nuclear; RNA, Messenger; Signal Transduction; Thiazoles; Thiazolidinediones; Transcription Factors; Troglitazone

In situations of sustained hyperglycemia, much larger amounts of glycogen accumulate in islet B-cells than in other pancreatic cells. The labelling of such a glycogen pool could thus conceivably provide a mean for assessing the relative contribution of insulin-producing cells to the total pancreatic mass. In such a perspective, the present study aims at investigating pancreatic glycogen accumulation in hereditarily diabetic Goto-Kakizaki (GK) rats. When cultured at 30 mM D-glucose in the presence of D-[U-14C]glucose, pancreatic islets from GK rats accumulated 14C-labelled glycogen in a manner comparable to that previously documented in islets from normal rats. Likewise, the glycogen content of the pancreatic gland, relative to the plasma D-glucose concentration, was not different in GK and normal rats. The GK rats thus apparently represent a suitable model for further studies on the in vivo labelling of B-cell glycogen in the perspective of the non-invasive imaging and quantification of the endocrine pancreas.

Topics: Animals; Carbon Radioisotopes; Cell Culture Techniques; Diabetes Mellitus, Type 2; Female; Glucose; Glycogen; Hyperglycemia; Islets of Langerhans; Liver; Male; Models, Animal; Organ Size; Pancreas; Rats

Chronic calorie restriction in primates has been shown to have profound and unexpected effects on basal and on in vivo insulin action on skeletal muscle glycogen synthase (GS) activity. The decreased ability of insulin to activate skeletal muscle GS is a hallmark of insulin resistance and type 2 diabetes. The mechanism and role of in vivo insulin regulation of skeletal muscle GS are not fully understood. Two pathways for the activation of GS by insulin have been described by Larner and others: 1) insulin activates glucose transport that results in an increase in glucose-6-phosphate (G6P), thereby activating protein phosphatase-1, which in turn dephosphorylates and activates GS, therefore, pushing substrate into glycogen; and 2) insulin activates GS (perhaps by forming low-molecular-weight mediators which may activate protein phosphatase-1 and 2C) and activated GS subsequently pulls intermediates (e.g., G6P and uridine 5'-diphosphoglucose) into glycogen. To determine whether in vivo insulin regulates glycogen synthesis primarily via a push or pull mechanism and how this mechanism might be affected by long-term calorie restriction, skeletal muscle samples were obtained before and during a euglycemic hyperinsulinemic clamp from 41 rhesus monkeys. The monkeys varied widely in their degree of insulin sensitivity and age and included chronically calorie-restricted (CR) monkeys and ad libitum-fed monkeys. The ad libitum-fed monkeys included spontaneously type 2 diabetic, prediabetic and clinically normal animals. The apparent affinity of GS for the allosteric activator G6P (G6P Ka of GS) was measured and compared with G6P content in the muscle samples. Basal G6P Ka of GS was lower in the CR monkeys compared with the 3 ad libitum-fed groups (P: < or = 0.05). Only the normal ad libitum-fed monkeys had a decrease in the G6P Ka of GS with insulin (P: < 0.005). The insulin effect (insulin-stimulated minus basal) on the G6P Ka of GS was strongly positively related to the insulin effect on G6P content (r = 0.80, P: < 0.0001) across the entire group of monkeys. This finding supports the hypothesis that activation/dephosphorylation of GS by insulin is related to a decrease in G6P content and that paradoxical inactivation/phosphorylation of GS by insulin is related to an increase in G6P content (as demonstrated in 4 of 6 CR monkeys). Therefore, during a euglycemic hyperinsulinemic clamp, insulin regulates skeletal muscle glycogen synthesis primarily via a pull mechanism i

Topics: Animals; Basal Metabolism; Blood Glucose; Diabetes Mellitus, Type 2; Food Deprivation; Glucose Clamp Technique; Glucose-6-Phosphate; Glycogen; Glycogen Synthase; Insulin; Insulin Resistance; Kinetics; Macaca mulatta; Muscle, Skeletal; Phosphoprotein Phosphatases; Phosphorylation; Protein Phosphatase 1

Effects of endogenously derived free fatty acids (FFAs) on rates of gluconeogenesis (GNG) (determined with 2H2O), glycogenolysis (GL), and endogenous glucose production (EGP) were studied in 18 type 2 diabetic patients and in 7 nondiabetic control subjects under three experimental conditions: 1) during an 8-h fast (from 16-24 h after the last meal), when plasma FFA levels increased slowly; 2) during 4 h (from 16-20 h) of nicotinic acid (NA) administration (fasting plus NA), when plasma FFAs decreased acutely; and 3) during 4 h (from 20-24 h) after discontinuation of NA (FFA rebound), when plasma FFAs increased acutely. During fasting, FFAs increased from 636 to 711 micromol/l in type 2 diabetic patients and from 462 to 573 micromol/l in control subjects (P < 0.04), but GNG did not change in diabetic patients (6.9 vs. 6.5 micromol x kg(-1) x min(-1), P > 0.05) or in control subjects (5.1 vs. 5.4 micromol x kg(-1) x min(-1), P > 0.05). During fasting plus NA, FFAs decreased in diabetic patients and control subjects (from 593 to 193 and from 460 to 162 micromol/l, respectively); GNG decreased (from 6.1 to 4.2 and from 4.7 to 3.5 micromol x kg(-1) x min(-1)), whereas GL decreased in diabetic patients (from 5.3 to 4.4 micromol x kg(-1) x min(-1)) but increased in control subjects (from 5.4 to 7.2 micromol x kg(-1) min(-1)). During the FFA rebound, FFAs increased in diabetic patients and control subjects (from 193 to 1,239 and from 162 to 1,491 micromol/l, respectively); GNG increased (from 4.2 to 5.4 and from 3.4 to 5.3 micromol x kg(-1) x min(-1) respectively), and GL decreased (from 4.4 to 3.4 and from 7.3 to 4.3 micromol x kg(-1) x min(-1), respectively). In summary, during an extended overnight fast, increasing plasma FFA levels stimulated GNG, whereas decreasing FFA levels inhibited GNG in both diabetic and control subjects; 20 h after the last meal, approximately one-third of GNG in both diabetic and control subjects was dependent on FFAs; and autoregulation of EGP by GL in response to decreasing GNG was impaired in diabetic patients.

Topics: Diabetes Mellitus, Type 2; Fasting; Fatty Acids, Nonesterified; Female; Gluconeogenesis; Glucose; Glycerol; Glycogen; Homeostasis; Hormones; Human Growth Hormone; Humans; Insulin; Male; Middle Aged

Insulin-stimulated GLUT4 translocation is impaired in people with type 2 diabetes. In contrast, exercise results in a normal increase in GLUT4 translocation and glucose uptake in these patients. Several groups have recently hypothesized that exercise increases glucose uptake via an insulin-independent mechanism mediated by the activation of AMP-activated protein kinase (AMPK). If this hypothesis is correct, people with type 2 diabetes should have normal AMPK activation in response to exercise. Seven subjects with type 2 diabetes and eight matched control subjects exercised on a cycle ergometer for 45 min at 70% of maximum workload. Biopsies of vastus lateralis muscle were taken before exercise, after 20 and 45 min of exercise, and at 30 min postexercise. Blood glucose concentrations decreased from 7.6 to 4.77 mmol/l with 45 min of exercise in the diabetic group and did not change in the control group. Exercise significantly increased AMPK alpha2 activity 2.7-fold over basal at 20 min in both groups and remained elevated throughout the protocol, but there was no effect of exercise on AMPK alpha1 activity. Subjects with type 2 diabetes had similar protein expression of AMPK alpha1, alpha2, and beta1 in muscle compared with control subjects. AMPK alpha2 was shown to represent approximately two-thirds of the total alpha mRNA in the muscle from both groups. In conclusion, people with type 2 diabetes have normal exercise-induced AMPK alpha2 activity and normal expression of the alpha1, alpha2 and beta1 isoforms. Pharmacological activation of AMPK may be an attractive target for the treatment of type 2 diabetes.

Topics: AMP-Activated Protein Kinases; Blood Glucose; Diabetes Mellitus, Type 2; Enzyme Activation; Exercise; Gene Expression Regulation, Enzymologic; Glucose Transporter Type 4; Glycated Hemoglobin; Glycogen; Humans; Kinetics; Male; Middle Aged; Monosaccharide Transport Proteins; Multienzyme Complexes; Muscle Proteins; Muscle, Skeletal; Physical Exertion; Protein Serine-Threonine Kinases; Reference Values; Rest; RNA, Messenger; Transcription, Genetic

The aim of this study was to ascertain whether the presence of hypertension conveys a more severe degree of insulin resistance in type 2 diabetes mellitus and, if so, which biochemical pathways are involved. We quantitated the rates of total glucose disposal, glycogen synthesis (GS), glycolysis, glucose oxidation, endogenous glucose production, and LOX in the basal state and during a 4-h euglycemic ( approximately 5 mM) hyperinsulinemic ( approximately 300 pM) clamp carried out in combination with a dual-tracer infusion ([(3)H]-3- and [(14)C]-U-D-glucose) and indirect calorimetry in 42 nonobese noninsulin-treated type 2 diabetic subjects (22 hypertensive and 20 normotensive) and 23 nonobese nondiabetic subjects (9 without and 14 with essential hypertension). Compared with normotensive controls, both groups of diabetic subjects were markedly insulin resistant. In the basal state, all glucose fluxes were similar in diabetic subjects with or without hypertension. During insulin infusion, total glucose disposal was significantly reduced in hypertensive diabetic subjects, compared with their normotensive counterparts (18.7 +/- 1.0 vs. 28.6 +/- 3.0 micromol/min.kg lean body mass; P < 0.01). This difference was almost entirely explained by a marked reduction in GS (4.5 +/- 2.0 vs. 12.5 +/- 3.3 micromol/min.kg lean body mass; P < 0.01). Endogenous glucose production was not different in the two diabetic groups during insulin infusion and was significantly higher than in normotensive controls. Lipid oxidation was less suppressed by hyperinsulinemia in hypertensive than in normotensive diabetic subjects (1.46 +/- 0.1 vs. 0.91 +/- 0.1 micromol/min.kg lean body mass; P < 0.01). Glucose fluxes were not significantly different in nondiabetic subjects with essential hypertension and in normotensive diabetic individuals. These results indicate that hypertension markedly aggravates insulin resistance featuring type 2 diabetes mellitus. The molecular defects underlying this phenomenon involve primarily GS.

Topics: Aged; Blood Glucose; Diabetes Mellitus, Type 2; Female; Glycogen; Humans; Hypertension; Insulin Resistance; Lipid Metabolism; Male; Middle Aged; Oxidation-Reduction

A subgroup of patients with type 2 diabetes shows a clustering of abnormalities such as peripheral insulin resistance, hypertension, and microalbuminuria. To evaluate whether these traits reflect intrinsic disorders of cell function rather than in vivo environmental effects, we studied a group of 7 nondiabetic hypertensive subjects with an altered albumin excretion rate (AER) (HyMA+) and 3 groups of patients with type 2 diabetes: 7 with normal blood pressure and normal AER (DH-MA-), 7 with high blood pressure and normal AER (DH+MA-), and 7 with both high blood pressure and altered AER (DH+MA+). Glucose disposal was measured during an hyperinsulinemic clamp (40 mU. m(2)(-1). min(-1)) with primed deuterated [6.6 (2)H(2)] glucose infusion. In the same subjects, a skin biopsy was performed and the following parameters were investigated: glucose transport (as determined by [(3)H]2-deoxyglucose uptake); glycogen synthase activity (as determined by [(14)C] glucose incorporation from UDP-[U-(14)C] glucose into glycogen); glycogen phosphorylase activity (as measured by the incorporation of [U-(14)C]glucose 1-phosphate into glycogen); and total glycogen content. In vivo glucose disposal was significantly reduced in DH+MA- and DH+MA+, with respect to DH-MA-, HyMA+, and controls. Insulin-stimulated glucose transport was similar in the 3 groups of patients with diabetes. A significant reduction of intracellular glycogen content was observed in DH+MA- and DH+MA+ compared with DH-MA- in both basal and insulin-stimulated conditions, probably because of a major impairment of glycogen synthase activity. Glycogen phosphorylase activity did not show differences between the groups. These results suggest that (1) the combination of type 2 diabetes with hypertension and altered AER is associated with impaired insulin sensitivity, and (2) intrinsic, possibly genetic, factors may account for increased peripheral insulin resistance in hypertensive microalbuminuric patients with type 2 diabetes, pointing to the reduction of glycogen synthase activity as a shared common defect.

Topics: Albuminuria; Cells, Cultured; Deoxyglucose; Diabetes Mellitus, Type 2; Female; Fibroblasts; Glucose; Glucose Clamp Technique; Glycogen; Glycogen Synthase; Humans; Hypertension; Insulin; Insulin Resistance; Liver; Male; Middle Aged

Gluconeogenesis (GNG) is enhanced in type 2 diabetes. In experimental animals, insulin at high doses decreases the incorporation of labeled GNG precursors into plasma glucose. Whether physiological hyperinsulinemia has any effect on total GNG in humans has not been determined. We combined the insulin clamp with the (2)H(2)O technique to measure total GNG in 33 subjects with type 2 diabetes (BMI 29.0 +/- 0.6 kg/m(2), fasting plasma glucose 8.1 +/- 0.3 mmol/l) and in 9 nondiabetic BMI-matched subjects after 16 h of fasting and after euglycemic hyperinsulinemia. A primed-constant infusion of 6,6-(2)H-glucose was used to monitor endogenous glucose output (EGO); insulin (40 mU. min(-1). m(-2)) was then infused while clamping plasma glucose for 2 h (at 5.8 +/- 0.1 and 4.9 +/- 0.2 mmol/l for diabetic and control subjects, respectively). In the fasting state, EGO averaged 15.2 +/- 0.4 micromol. min(-1). kg(-1)(ffm) (62% from GNG) in diabetic subjects and 12.2 +/- 0.7 micromol. min(-1). kg(-1)(ffm) (55% from GNG) in control subjects (P < 0.05 or less for both fluxes). Glycogenolysis (EGO - GNG) was similar in the two groups (P = NS). During the last 40 min of the clamp, both EGO and GNG were significantly (P < 0.01 or less, compared with fasting) inhibited (EGO 7.1 +/- 0.9 and 3.6 +/- 0.5 and GNG 7.9 +/- 0.5 and 4.5 +/- 1.0 respectively) but remained significantly (P < 0.05) higher in diabetic subjects, whereas glycogenolysis was suppressed completely and equally in both groups. During hyperinsulinemia, GNG micromol. min(-1). kg(-1)(ffm) in diabetic and control subjects, was reciprocally related to plasma glucose clearance. In conclusion, physiological hyperinsulinemia suppresses GNG by approximately 20%, while completely blocking glycogenolysis. Resistance of GNG (to insulin suppression) and resistance of glucose uptake (to insulin stimulation) are coupled phenomena. In type 2 diabetes, the excess GNG of the fasting state is carried over to the insulinized state, thereby contributing to glucose overproduction under both conditions.

Topics: Adult; Blood Glucose; Blood Pressure; Body Constitution; Body Mass Index; Deuterium Oxide; Diabetes Mellitus, Type 2; Fasting; Female; Gluconeogenesis; Glucose Clamp Technique; Glycated Hemoglobin; Glycogen; Humans; Hyperinsulinism; Insulin; Kinetics; Male; Middle Aged; Reference Values; Regression Analysis; Time Factors

The effects of glucagon and insulin on glucose production were explored directly using the isolated perfused liver of the Goto-Kakizaki (GK) rat, an animal model of type-2 diabetes. In the perfused liver of control rats, infusion of glucagon (0.06-1.0 nM) into the portal vein dose-dependently increased glucose output. In the GK rat liver, in which the intracellular distribution of glycogen was heterogeneous, basal glucose output during perfusion was significantly higher than in control, whereas the effect of glucagon on the maximum glucose output was not different. Infusion of insulin inhibited the glucagon-induced hepatic glucose output by 30-40% in control livers, but had no effect on that from the GK rat liver. The increase in hepatic cAMP content after glucagon infusion was antagonized by insulin in control livers, but not in the livers of GK rats. These results indicate that the antagonistic effect of insulin on glucagon-induced hepatic glucose production was attenuated in the isolated liver of the GK rat and suggest that this insulin resistance appeared in the signal transduction process of glucagon upstream from cAMP production.

Topics: Animals; Cyclic AMP; Diabetes Mellitus, Type 2; Gastrointestinal Agents; Glucagon; Glucose; Glycogen; Hypoglycemic Agents; Insulin; Liver; Male; Microscopy, Electron; Organ Size; Perfusion; Rats; Rats, Mutant Strains

Type 2 diabetes is characterized by peripheral tissue insulin resistance. The present study was carried out to determine the insulin sensitizing action of vanadium using dexamethasone-treated 3T3 adipocytes as an in-vitro model of insulin resistance.. Fully differentiated 3T3 adipocytes were incubated with or without 100 nM dexamethasone in the presence or absence of 200 nM insulin for 6 days. Sodium orthovanadate (0-1000 microM) was added on day 2 and was present during the last 4 days. At the end, insulin (100 nM) stimulated glycogen synthesis was determined.. Vanadate treatment for 4 days, caused 2-3-fold increase in glycogen synthesis in dexamethasone treated adipocytes. At 100 microM, vanadate completely reversed dexamethasone-induced insulin resistance (by increasing the levels from 9.65 +/- 0.80 to 28.4 +/- 4.9 nmol/h). In cells treated with dexamethasone and insulin, vanadium was partially active and it caused only 30% increase in glycogen synthesis. Exposure of dexamethasone treated cells for 24 h with vanadium did not affect glycogen synthesis. Under identical condition, vanadium had no significant effect in the normal insulin sensitive adipocytes. Vanadium at 100 microM had no effect on 125I-insulin binding to insulin-resistant adipocytes. Glycogen synthesis in the normal and insulin-resistant adipocytes was stimulated by lithium, an inhibitor of glycogen synthase kinase 3 beta, suggesting the involvement of phosphorylation events in dexamethasone-induced insulin resistance.. Since vanadium was active only in the insulin-resistant adipocytes it is likely that vanadium acts by relieving dexamethasone actions rather than having independent effects. These results provide evidence for the novel insulin sensitizing action of vanadium which might be of future clinical relevance.

Topics: 3T3 Cells; Adipocytes; Animals; Carbon Radioisotopes; Cell Differentiation; Dexamethasone; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Kinetics; Lithium; Mice; Vanadates

Physical exercise is frequently recommended for the treatment of type 2 diabetes, whether as primary therapy with diet modification or as an adjunct to drug therapy. We hypothesized that mild exercise would enhance the glucose-lowering effects of 2 oral antihyperglycemic drugs, metformin and acarbose, in an animal model of type 2 diabetes. Eight-week-old male C57BL/Ks (db/db) mice were sorted into control and exercise groups and dosed daily for 4 weeks with vehicle, metformin (150 mg/kg/d), or acarbose (40 mg/kg/d). Exercise consisted of swimming (initially 5 min/d and ultimately 1 h/d for the last 2 weeks). Exercise, metformin, and acarbose independently reduced serum glucose concentrations 15% to 25% compared with the respective controls (P <.0001), but the effect on glucose concentration of combining drug therapy with exercise was no greater than the sum of the individual effects. Exercise training independently increased muscle glycogen (30%; P <.05) and liver glycogen (250%; P <.05) levels and slightly reduced serum high-density lipoprotein cholesterol (-8%; P <.05), whereas drug treatment had no effect on these variables. In addition, exercise but not drug treatment prevented the approximately 30% decline in serum insulin concentrations that occurred in the control animals (P <.05). Twenty-four hours after the last drug or exercise treatment, oral glucose tolerance and hemoglobin A1c were not significantly different between groups. Treatment also did not greatly affect triglyceride, glycerol, or total cholesterol concentrations. In conclusion, exercise and drug therapy independently decreased serum glucose in db/db mice, and these effects did not appear to be synergistic. In addition, exercise training maintained serum insulin concentrations and increased tissue glycogen storage. These results suggest that exercise has the potential to add to the efficacy of oral antihyperglycemic drugs.

Topics: Acarbose; Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Disease Models, Animal; Eating; Glucose Tolerance Test; Glycated Hemoglobin; Glycogen; Hypoglycemic Agents; Insulin; Lipids; Liver; Male; Metformin; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Muscle, Skeletal; Physical Exertion; Swimming; Treatment Outcome

Selenium in serum and selenium and glycogen in erythrocytes were determined in diabetic patients divided into noninsulin-dependent (n = 50) and insulin-dependent (n = 31) groups according to the etiopathogenesis of their diabetes. Selenium was determined by the method of atomic absorption spectrometry. Serum level of selenium was statistically significantly different in patients with either noninsulin-dependent (59.23 +/- 12.2 microg/L) or insulin-dependent (58.23 +/- 16.7 microg/L) diabetes mellitus as compared with the control group of 62 subjects (64.2 +/- 11.5 microg/L; p < 0.05). There was no statistically significant difference in the serum levels of selenium between the groups of patients with noninsulin-dependent and insulin-dependent diabetes mellitus. The levels of erythrocyte glycogen were 2.0580 +/- 1.326, 2.0380 +/- 1.735, and 2.0036 +/- 1.3537 microg/g Hb in the control group, noninsulin-dependent group, and insulin-dependent group, respectively, with no statistically significant between-group difference. The decreased levels of selenium in serum and erythrocytes of diabetic patients suggest the possible role of glutathione peroxidase activity.

Topics: Adult; Case-Control Studies; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Erythrocytes; Female; Glycogen; Humans; Male; Selenium

To examine the metabolic pathways by which troglitazone improves insulin responsiveness in patients with type 2 diabetes, the rate of muscle glycogen synthesis was measured by 13C-nuclear magnetic resonance (NMR) spectroscopy. The rate-controlling steps of insulin-stimulated muscle glucose metabolism were assessed using 31P-NMR spectroscopic measurement of intramuscular glucose-6-phosphate (G-6-P) combined with a novel 13C-NMR method to assess intracellular glucose concentrations. Seven healthy nonsmoking subjects with type 2 diabetes were studied before and after completion of 3 months of troglitazone (400 mg/day) therapy. After troglitazone treatment, rates of insulin-stimulated whole-body glucose uptake increased by 58+/-11%, from 629+/-82 to 987+/-156 micromol x m(-2) x min(-1) (P = 0.008), which was associated with an approximately 3-fold increase in rates of insulin-stimulated glucose oxidation (from 119+/-41 to 424+/-70 micromol x m(-2) x min(-1); P = 0.018) and muscle glycogen synthesis (26+/-17 vs. 83+/-35 micromol x l(-1) muscle x min(-1); P = 0.025). After treatment, muscle G-6-P concentrations increased by 0.083+/-0.019 mmol/l (P = 0.008 vs. pretreatment) during the hyperglycemic-hyperinsulinemic clamp, compared with no significant changes in intramuscular G-6-P concentrations in the pretreatment study, reflecting an improvement in glucose transport and/or hexokinase activity. The concentrations of intracellular free glucose did not differ between the pre- and posttreatment studies and remained >50-fold lower in concentration (<0.1 mmol/l) than what would be expected if hexokinase activity was rate-controlling. These results indicate that troglitazone improves insulin responsiveness in skeletal muscle of patients with type 2 diabetes by facilitating glucose transport activity, which thereby leads to increased rates of muscle glycogen synthesis and glucose oxidation.

Topics: Body Composition; Chromans; Diabetes Mellitus, Type 2; Female; Glucose; Glucose-6-Phosphate; Glycogen; Hormones; Humans; Hypoglycemic Agents; Intracellular Membranes; Male; Middle Aged; Muscle, Skeletal; Thiazoles; Thiazolidinediones; Troglitazone

To investigate the contribution of inherited biochemical defects to the peripheral insulin resistance of type 2 diabetes, we studied cultured skeletal muscle from 10 insulin-resistant nondiabetic first-degree relatives of type 2 diabetic families and 6 control subjects. Insulin stimulation of glucose uptake and glycogen synthesis was maximal in myoblasts. Insulin-stimulated glucose uptake (fold-stimulation over basal uptake) was decreased in relative compared with control myoblasts at 0.001 micromol/l (0.93 +/- 0.05 [mean +/- SE] vs. 1.15 +/- 0.06, P < 0.05) and 0.1 micromol/l (1.38 +/- 0.10 vs. 1.69 +/- 0.08, P = 0.025) insulin. Insulin responsiveness was markedly impaired in 5 of the relative myoblast cultures, and in 4 of these, there was an associated increase in basal glucose uptake (76.7 +/- 7.0 vs. 47.4 +/- 5.5 pmol x min(-1) x mg(-1) protein, relative vs. control; P < 0.02). Expression of insulin receptor substrate 1, phosphatidylinositol 3-kinase, protein kinase B, and glycogen synthase was normal in the relative cultures with impaired insulin responsiveness. Glycogen synthesis was also normal in the relative cultures. We conclude that the persistence of impaired insulin responsiveness in some of the relative cultures supports the role of inherited factors in the insulin resistance of type 2 diabetes and that the association with increased basal glucose uptake suggests that the 2 abnormalities may be linked.

Topics: Adult; Biological Transport; Blood Glucose; Cells, Cultured; Diabetes Mellitus, Type 2; Europe; Female; Glucose; Glucose Transporter Type 1; Glucose Transporter Type 4; Glycogen; Humans; Insulin; Insulin Resistance; Male; Middle Aged; Monosaccharide Transport Proteins; Muscle Proteins; Muscle, Skeletal; Reference Values; Tritium

The aim of the present study was to estimate whether a single bout of exhaustive exercise influences the glycogen and triglyceride (TG) content in red and white gastrocnemius muscle and in the liver of rats with experimental type 2 diabetes. Experiments were carried out on male Wistar rats fed from 8 to 11 weeks of age with isocaloric standard or high-fat diet (HFD) with a previous injection of low-dose of streptozotocin (STZ) or vehicle at 2 days of age (I, control group; II, HFD; III, STZ; IV, STZ + HFD). Group IV (STZ + HFD) represents a model of type 2 diabetes. Basal liver glycogen was markedly lower in all the studied groups compared to controls. Glycogen concentration after exercise fell significantly in the examined tissues in all groups in comparison to basal conditions. A significant TG accumulation in examined tissues was observed in all the studied groups in comparison to controls. Exercise decreased tissue TG content in all the groups, but it remained significantly higher in the experimental groups vs. control. We conclude that in this model of type 2 diabetes, a single bout of exercise reveals defective utilization of tissue carbohydrates and lipids.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Disease Models, Animal; Glycogen; Insulin; Liver; Male; Muscle Fibers, Fast-Twitch; Muscle, Skeletal; Physical Exertion; Rats; Rats, Wistar; Triglycerides

Although diabetic retinopathy has been thoroughly studied, little attention has been given to the corneal changes of diabetic patients. Pathophysiologic and clinical findings may be related to the ultrastructural changes found in these corneas.. To investigate the ultrastructural corneal changes of diabetic patients.. Transmission electron microscopic ultrathin sections were prepared from corneas of 16 noninsulin-dependent diabetic patients (mean age, 65 years; range, 40-82 years) who suffered from the disease for a mean period of 22 years (range, 10-30 years). We used 16 corneas from healthy age-matched donors as normal controls.. In addition to the epithelial changes that include accumulation of glycogen granules, occasional focal epithelial cell degeneration, and irregular thickening and multilamination of the epithelial basement membrane, unusual 120-nm wide-spaced collagen fibril bundles were observed scattered among both Descemet's membrane and stromal matrix.. The aggregates of wide-spaced collagen fibrils, which have not been described in other basement membranes of diabetic patients, may reflect an excessive glycosylation rate.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Basement Membrane; Cadaver; Collagen; Cornea; Corneal Diseases; Diabetes Mellitus, Type 2; Glycogen; Humans; Microscopy, Electron; Middle Aged

The effects of 1,4-dideoxy-1,4-imino-d-arabinitol (DAB) were investigated on preparations of glycogen phosphorylase (GP) and in C57BL6J (ob/ob) mice by (13)C NMR in vivo. Independent of the phosphorylation state or the mammalian species or tissue from which GP was derived, DAB inhibited GP with K(i)-values of approximately 400 nM. The mode of inhibition was uncompetitive or noncompetitive, with respect to glycogen and P(i), respectively. The effects of glucose and caffeine on the inhibitory effect of DAB were investigated. Taken together, these data suggest that DAB defines a novel mechanism of action. Intraperitoneal treatment with DAB (a total of 105 mg/kg in seven doses) for 210 min inhibited glucagon-stimulated glycogenolysis in obese and lean mice. Thus, liver glycogen levels were 361 +/- 19 and 228 +/- 19 micromol glucosyl units/g with DAB plus glucagon in lean and obese mice, respectively, compared to 115 +/- 24 and 37 +/- 8 micromol glucosyl units/g liver with glucagon only. Moreover, with glucagon only end-point blood glucose levels were at 29 +/- 2 and 17.5 +/- 2 mM in obese and lean mice, respectively, compared to 17.5 +/- 1 and 12 +/- 1 mM with glucagon plus DAB. In conclusion, DAB is a novel and potent inhibitor of GP with an apparently distinct mechanism of action. Further, DAB inhibited the hepatic glycogen breakdown in vivo and displayed an accompanying anti-hyperglycemic effect, which was most pronounced in obese mice. The data suggest that inhibition of GP may offer a therapeutic principle in Type 2 diabetes.

Topics: Animals; Arabinose; Blood Glucose; Diabetes Mellitus, Type 2; Enzyme Inhibitors; Female; Glucagon; Glycogen; Hypoglycemic Agents; Imino Furanoses; In Vitro Techniques; Kinetics; Lactic Acid; Liver; Magnetic Resonance Spectroscopy; Mice; Mice, Inbred C57BL; Mice, Obese; Muscles; Phosphorylases; Rabbits; Rats; Sugar Alcohols; Swine

Glycogen synthase kinase-3 (GSK-3) is a serine/threonine protein kinase, the activity of which is inhibited by a variety of extracellular stimuli including insulin, growth factors, cell specification factors and cell adhesion. Consequently, inhibition of GSK-3 activity has been proposed to play a role in the regulation of numerous signalling pathways that elicit pleiotropic cellular responses. This report describes the identification and characterisation of potent and selective small molecule inhibitors of GSK-3.. SB-216763 and SB-415286 are structurally distinct maleimides that inhibit GSK-3alpha in vitro, with K(i)s of 9 nM and 31 nM respectively, in an ATP competitive manner. These compounds inhibited GSK-3beta with similar potency. However, neither compound significantly inhibited any member of a panel of 24 other protein kinases. Furthermore, treatment of cells with either compound stimulated responses characteristic of extracellular stimuli that are known to inhibit GSK-3 activity. Thus, SB-216763 and SB-415286 stimulated glycogen synthesis in human liver cells and induced expression of a beta-catenin-LEF/TCF regulated reporter gene in HEK293 cells. In both cases, compound treatment was demonstrated to inhibit cellular GSK-3 activity as assessed by activation of glycogen synthase, which is a direct target of this kinase.. SB-216763 and SB-415286 are novel, potent and selective cell permeable inhibitors of GSK-3. Therefore, these compounds represent valuable pharmacological tools with which the role of GSK-3 in cellular signalling can be further elucidated. Furthermore, development of similar compounds may be of use therapeutically in disease states associated with elevated GSK-3 activity such as non-insulin dependent diabetes mellitus and neurodegenerative disease.

Topics: Adenosine Triphosphate; Aminophenols; beta Catenin; Binding, Competitive; Calcium-Calmodulin-Dependent Protein Kinases; Cell Line; Cytoskeletal Proteins; Diabetes Mellitus, Type 2; Enzyme Activation; Gene Expression Regulation; Genes, Reporter; Glycogen; Glycogen Synthase; Glycogen Synthase Kinase 3; Glycogen Synthase Kinases; Humans; Indoles; Kinetics; Liver; Maleimides; Molecular Structure; Neurodegenerative Diseases; Protein Kinases; Recombinant Proteins; Signal Transduction; Trans-Activators; Transcription, Genetic

We tested the hypothesis that a lack of suppression of glucagon causes postprandial hyperglycemia in subjects with type 2 diabetes. Nine diabetic subjects ingested 50 g glucose on two occasions. On both occasions, somatostatin was infused at a rate of 4.3 nmol/kg x min, and insulin was infused in a diabetic insulin profile. On one occasion, glucagon was also infused at a rate of 1.25 ng/kg x min to maintain portal glucagon concentrations constant (nonsuppressed study day). On the other occasion, glucagon infusion was delayed by 2 h to create a transient decrease in glucagon (suppressed study day). Glucagon concentrations on the suppressed study day fell to about 70 ng/L during the first 2 h, rising thereafter to approximately 120 ng/L. In contrast, glucagon concentrations on the nonsuppressed study day remained constant at about 120 ng/L throughout. The decrease in glucagon resulted in substantially lower (P < 0.001) glucose concentrations on the suppressed compared with the nonsuppressed study days (9.2+/-0.7 vs. 10.9+/-0.8 mmol/L) and a lower (P < 0.001) rate of release of [14C]glucose from glycogen (labeled by infusing [1-14C]galactose). On the other hand, flux through the hepatic UDP-glucose pool (and, by implication, glycogen synthesis), measured using the acetaminophen glucuronide method, did not differ on the two occasions. We conclude that lack of suppression of glucagon contributes to postprandial hyperglycemia in subjects with type 2 diabetes at least in part by accelerating glycogenolysis. These data suggest that agents that antagonize glucagon action or secretion are likely to be of value in the treatment of patients with type 2 diabetes.

Topics: Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Female; Galactose; Glucagon; Glycogen; Human Growth Hormone; Humans; Hyperglycemia; Infusions, Intravenous; Insulin; Insulin Secretion; Male; Middle Aged; Postprandial Period

To examine the mechanism by which metformin lowers endogenous glucose production in type 2 diabetic patients, we studied seven type 2 diabetic subjects, with fasting hyperglycemia (15.5 +/- 1.3 mmol/l), before and after 3 months of metformin treatment. Seven healthy subjects, matched for sex, age, and BMI, served as control subjects. Rates of net hepatic glycogenolysis, estimated by 13C nuclear magnetic resonance spectroscopy, were combined with estimates of contributions to glucose production of gluconeogenesis and glycogenolysis, measured by labeling of blood glucose by 2H from ingested 2H2O. Glucose production was measured using [6,6-2H2]glucose. The rate of glucose production was twice as high in the diabetic subjects as in control subjects (0.70 +/- 0.05 vs. 0.36 +/- 0.03 mmol x m(-2) min(-1), P < 0.0001). Metformin reduced that rate by 24% (to 0.53 +/- 0.03 mmol x m(-2) x min(-1), P = 0.0009) and fasting plasma glucose concentration by 30% (to 10.8 +/- 0.9 mmol/l, P = 0.0002). The rate of gluconeogenesis was three times higher in the diabetic subjects than in the control subjects (0.59 +/- 0.03 vs. 0.18 +/- 0.03 mmol x m(-2) min(-1) and metformin reduced that rate by 36% (to 0.38 +/- 0.03 mmol x m(-2) x min(-1), P = 0.01). By the 2H2O method, there was a twofold increase in rates of gluconeogenesis in diabetic subjects (0.42 +/- 0.04 mmol m(-2) x min(-1), which decreased by 33% after metformin treatment (0.28 +/- 0.03 mmol x m(-2) x min(-1), P = 0.0002). There was no glycogen cycling in the control subjects, but in the diabetic subjects, glycogen cycling contributed to 25% of glucose production and explains the differences between the two methods used. In conclusion, patients with poorly controlled type 2 diabetes have increased rates of endogenous glucose production, which can be attributed to increased rates of gluconeogenesis. Metformin lowered the rate of glucose production in these patients through a reduction in gluconeogenesis.

Topics: Calorimetry, Indirect; Diabetes Mellitus, Type 2; Female; Gluconeogenesis; Glucose; Glycogen; Humans; Hypoglycemic Agents; Liver; Magnetic Resonance Spectroscopy; Male; Metformin; Middle Aged

Type 2 diabetes is a polygenic disease characterized by defects in both insulin secretion and insulin action. We have previously reported that isolated insulin resistance in muscle by a tissue-specific insulin receptor knockout (MIRKO mouse) is not sufficient to alter glucose homeostasis, whereas beta-cell-specific insulin receptor knockout (betaIRKO) mice manifest severe progressive glucose intolerance due to loss of glucose-stimulated acute-phase insulin release. To explore the interaction between insulin resistance in muscle and altered insulin secretion, we created a double tissue-specific insulin receptor knockout in these tissues. Surprisingly, betaIRKO-MIRKO mice show an improvement rather than a deterioration of glucose tolerance when compared to betaIRKO mice. This is due to improved glucose-stimulated acute insulin release and redistribution of substrates with increased glucose uptake in adipose tissue and liver in vivo, without a significant decrease in muscle glucose uptake. Thus, insulin resistance in muscle leads to improved glucose-stimulated first-phase insulin secretion from beta-cells and shunting of substrates to nonmuscle tissues, collectively leading to improved glucose tolerance. These data suggest that muscle, either via changes in substrate availability or by acting as an endocrine tissue, communicates with and regulates insulin sensitivity in other tissues.

Topics: Acute-Phase Reaction; Animals; Blood Glucose; Deoxyglucose; Diabetes Mellitus, Type 2; Fasting; Glucose; Glucose Tolerance Test; Glycogen; Injections, Intraperitoneal; Insulin; Insulin Resistance; Insulin Secretion; Islets of Langerhans; Lipid Metabolism; Mice; Mice, Knockout; Muscle, Skeletal; Receptor, Insulin; Reference Values

Present animal models used to emulate type 2 diabetes may not accurately reflect the metabolic changes that occur in humans.. The purpose of this research was to evaluate diets reported to induce insulin resistance and impaired glucose metabolism in rats as a potentially useful model for studying type 2 diabetes.. Three groups of male Sprague Dawley rats (n=7) were fed either a control diet, based on AIN recommendations (53% cornstarch, 10% sucrose and 7% soybean oil), a high fat diet (25% soybean oil, 35% cornstarch) or a high fructose diet (53% fructose, 10% sucrose) for a 3 month period. Glucose tolerance tests were carried out in week 3 and week 9 of the experiment. At the termination of the experiment, serum insulin, glucose, cholesterol and triacylglycerols were measured. Glucose incorporation into glycogen and glycogen synthase activity were measured in soleus muscles.. Similar weight gain was observed for all three groups of rats. Glucose tolerance curves and fasting glucose levels were not significantly different at any time point in the experiment. Insulin levels were unchanged for the controls (171+/-21 pM), high fructose (164+/-16 pM) and high fat (181+/-30 pM) diets. Fasting serum triacylglycerols and cholesterol levels were not significantly elevated by dietary treatment. In soleus muscles, rats on all three diets had a significant increase in glycogen synthesis in response to insulin, but synthesis was similar in all three groups. Glycogen synthase activity was also not significantly affected by long-term dietary intervention.. In this study, healthy Sprague Dawley rats fed high fat or high fructose diets for 3 months adapted to the nutritional intervention without developing classical signs of insulin resistance and impaired glucose tolerance.

Topics: Adaptation, Biological; Animals; Cholesterol; Diabetes Mellitus, Type 2; Dietary Carbohydrates; Dietary Fats; Disease Models, Animal; Fructose; Glucose; Glucose Tolerance Test; Glycogen; Glycogen Synthase; Insulin; Insulin Resistance; Male; Muscles; Rats; Rats, Sprague-Dawley; Time Factors; Triglycerides

The finding of a reduced insulin-stimulated glucose uptake and glycogen synthesis in the skeletal muscle of glucose-tolerant first-degree relatives of patients with NIDDM, as well as in cultured fibroblasts and skeletal muscle cells isolated from NIDDM patients, has been interpreted as evidence for a genetic involvement in the disease. The mode of inheritance of the common forms of NIDDM is as yet unclear, but the prevailing hypothesis supports a polygenic model. In the present study, we tested the hypothesis that the putative inheritable defects of insulin-stimulated muscle glycogen synthesis might be caused by genetic variability in the genes encoding proteins shown by biochemical evidence to be involved in insulin-stimulated glycogen synthesis in skeletal muscle. In 70 insulin-resistant Danish NIDDM patients, mutational analysis by reverse transcription-polymerase chain reaction-single strand conformation polymorphism-heteroduplex analysis was performed on genomic DNA or skeletal muscle-derived cDNAs encoding glycogenin, protein phosphatase inhibitor-1, phophatase targeting to glycogen, protein kinase B-alpha and -beta, and the phosphoinositide-dependent protein kinase-1. Although a number of silent variants were identified in some of the examined genes, we found no evidence for the hypothesis that the defective insulin-stimulated glycogen synthesis in skeletal muscle in NIDDM is caused by structural changes in the genes encoding the known components of the insulin-sensitive glycogen synthesis pathway of skeletal muscle.

Topics: 3-Phosphoinositide-Dependent Protein Kinases; Carrier Proteins; Diabetes Mellitus, Type 2; DNA Mutational Analysis; Endoribonucleases; Female; Genetic Variation; Glucosyltransferases; Glycogen; Glycoproteins; Humans; Insulin; Intracellular Signaling Peptides and Proteins; Isomerism; Male; Middle Aged; Muscle, Skeletal; Phenotype; Phosphoprotein Phosphatases; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; RNA-Binding Proteins

The effect of vanadate (V) alone, magnesium (Mg) alone, and the combination of Mg plus V (MgV) on insulin-mediated glucose disposal and glucose tolerance was investigated in normal and streptozotocin-induced diabetic rats. MgV, magnesium sulfate (MgSO4) and sodium metavanadate (NaV) were added to the drinking water of normal or diabetic rats (approximately 300 g) for 3 weeks. After 3 weeks of V treatment (both MgV and NaV), diabetic rats demonstrated a normal meal tolerance test without any increase in the plasma insulin response. Rats also received a euglycemic insulin clamp (12 mU/kg x min for 120 minutes) with 3-3H-glucose infusion to quantify total body glucose disposal, glycolysis (3H2O production), and glycogen synthesis (total body glucose disposal minus glycolysis). Total glucose disposal was decreased in diabetic versus control rats (29 +/- 2 v 35 +/- 2 mg/kg x min, P < .01) and returned to levels greater than the nondiabetic control values after MgV (41 +/- 2, P < .01). Supersensitivity to insulin was not observed in diabetic rats treated with NaV (34 +/- 1). Glycogen synthesis was increased by both MgV and NaV treatment (23 +/- 21, P < .01 and 18 +/- 1, P < .05 v 14 +/- 2 mg/kg x min) in diabetic rats. A small increase in glycolysis was observed in MgSO4 and MgV rats (18 +/- 1 and 18 +/- 1 v 16 +/- 1, P < .05). NaV alone had no effect on glycolysis. Thus, Mg has a synergistic effect with V to increase muscle glycogen synthesis in diabetic rats. In normal rats, neither MgSO4 nor NaV had any effect on glucose utilization. However, MgV increased glucose disposal to rates that were significantly higher than the rate in untreated control rats (P < .05). Based on these results, MgV is superior to either V alone or Mg alone in improving insulin sensitivity and glycogen synthesis in diabetic rats.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Drug Synergism; Glucose Clamp Technique; Glycogen; Glycolysis; Hypoglycemic Agents; Magnesium Compounds; Male; Rats; Rats, Sprague-Dawley; Vanadates

The nutritional value of glycerol-1,2,3-tris(methylsuccinate), a novel ester of succinic acid with high insulinotropic efficiency both in vitro and in vivo, was assessed in both fed and starved rats. The infusion of the ester, given in a daily amount (1.2 micromol. g body wt-1) well in excess of what could result from its repeated intravenous administration as an insulinotropic agent in non-insulin-dependent diabetes (0.07 micromol. g body wt-1 for each administration), failed to prevent the fall in body weight, liver and muscle glycogen contents, and plasma d-glucose or insulin concentration, as well as the increase in plasma free fatty acid and beta-hydroxybutyrate concentrations caused by starvation. The sole indications that the ester may serve, to a limited extent, as an alternative nutrient in starved rats consisted in a somewhat higher weight of both liver and paraovarian adipose tissue and somewhat higher activity of liver glucokinase in rats receiving the ester than in animals infused with saline. The low nutritional value of this ester thus answers the objection of its possible role as an extrapancreatic nutrient or gluconeogenic precursor in the perspective of its use as an insulinotropic tool in type 2 diabetes.

Topics: 3-Hydroxybutyric Acid; Animals; Blood Glucose; Body Weight; Chloride Channels; Diabetes Mellitus, Type 2; Eating; Fatty Acids, Nonesterified; Female; Food Deprivation; Glucokinase; Glycerol; Glycogen; Insulin; Insulin Secretion; Liver; Muscle, Skeletal; Nutritive Value; Proteins; Rats; Rats, Wistar; Succinates

Insulin resistance, a major factor in the pathogenesis of type 2 diabetes mellitus, is due mostly to decreased stimulation of glycogen synthesis in muscle by insulin. The primary rate-controlling step responsible for the decrease in muscle glycogen synthesis is not known, although hexokinase activity and glucose transport have been implicated.. We used a novel nuclear magnetic resonance approach with carbon-13 and phosphorus-31 to measure intramuscular glucose, glucose-6-phosphate, and glycogen concentrations under hyperglycemic conditions (plasma glucose concentration, approximately 180 mg per deciliter [10 mmol per liter]) and hyperinsulinemic conditions in six patients with type 2 diabetes and seven normal subjects. In vivo microdialysis of muscle tissue was used to determine the gradient between plasma and interstitial-fluid glucose concentrations, and open-flow microperfusion was used to determine the concentrations of insulin in interstitial fluid.. The time course and concentration of insulin in interstitial fluid were similar in the patients with diabetes and the normal subjects. The rates of whole-body glucose metabolism and muscle glycogen synthesis and the glucose-6-phosphate concentrations in muscle were approximately 80 percent lower in the patients with diabetes than in the normal subjects under conditions of matched plasma insulin concentrations. The mean (+/-SD) intracellular glucose concentration was 2.0+/-8.2 mg per deciliter (0.11+/-0.46 mmol per liter) in the normal subjects. In the patients with diabetes, the intracellular glucose concentration was 4.3+/-4.9 mg per deciliter (0.24+/-0.27 mmol per liter), a value that was 1/25 of what it would be if hexokinase were the rate-controlling enzyme in glucose metabolism.. Impaired insulin-stimulated glucose transport is responsible for the reduced rate of insulin-stimulated muscle glycogen synthesis in patients with type 2 diabetes mellitus.

Topics: Adult; Aged; Biological Transport; Blood Glucose; Diabetes Mellitus, Type 2; Extracellular Space; Female; Glucose; Glucose-6-Phosphate; Glycogen; Hexokinase; Humans; Hyperglycemia; Hyperinsulinism; Insulin; Magnetic Resonance Spectroscopy; Male; Middle Aged; Models, Biological; Muscle, Skeletal

Young first-degree relatives of type 2 diabetic patients are insulin-resistant, with the insulin resistance mainly located in skeletal muscle due to decreased insulin-induced nonoxidative glucose metabolism and muscle glycogen synthase activation. We investigated whether the mechanism differs for dexamethasone (dex)-induced insulin resistance in first-degree relatives of type 2 diabetics versus healthy control subjects by quantifying intracellular glucose processing in muscle biopsies taken before and after 5 days of dex treatment (4 mg/d) in 20 normal glucose-tolerant relatives of type 2 diabetic patients and 20 matched controls (age, 29.4 +/- 1.7 v 29.4 +/- 1.6 years; body mass index, 25.1 +/- 1.0 v 25.1 +/- 0.9 kg/m2). In addition, an intravenous glucose tolerance test (IVGTT) combined with continuous indirect calorimetry was performed. Following 5 days of dex treatment, glucose tolerance deteriorated in both the relatives and the control subjects. Fasting dry-weight muscle glucose and fasting intracellular muscle glucose concentrations increased in response to dex only in the relatives (2.43 +/- 0.21 v 2.97 +/- 0.26 mmol/kg dry weight, P < .05; 0.28 +/- 0.07 v 0.45 +/- 0.08 mmol/L intracellular water, P < .05); no increases were observed in the control subjects. Fasting dry-weight muscle lactate also increased post-dex only in the relatives (7.37 +/- 0.40 v 10.77 +/- 1.22 mmol/kg dry weight, P < .001). Both basal muscle glucose and lactate concentrations from the IVGTT study correlated with the 2-hour post-dex glucose value obtained during the OGTT study in the relatives (R = .76 and R = .74, respectively, both P < .0001) but not in the control subjects. Basal intramuscular glycogen synthase activity decreased approximately 25% in both the relatives and control subjects post-dex; the decrement was significant (P < .01) only in control subjects. Indirect calorimetry during the post-dex IVGTT demonstrated increased glucose oxidation (P < .03) and reduced lipid oxidation (P < .03) in the relatives only. We postulate that the insulin resistance induced by dex in first-degree relatives of type 2 diabetic patients is associated with a preferential channeling of glucose into the glycolytic pathway (increased glucose oxidation and lactate production), probably associated with a preexisting downregulation of the glycosen synthase pathway.

Topics: Adult; Body Mass Index; Case-Control Studies; Dexamethasone; Diabetes Mellitus, Type 2; Female; Glucose; Glucose Tolerance Test; Glucose Transporter Type 4; Glucose-6-Phosphate; Glycogen; Glycogen Synthase; Humans; Insulin Resistance; Lactic Acid; Lipids; Male; Monosaccharide Transport Proteins; Muscle Proteins; Muscle, Skeletal; Pedigree

The influence of postprandial high intensity exercise on glycaemia was studied in patients with Type II diabetes mellitus.. Patients who were treated by diet only (n = 8) ate a standardised breakfast and 4 h later a standardised lunch. They were studied in the resting state (control day) and on another day (exercise day) when they did intermittent exercised at high intensity after breakfast) (4 bouts including 3 min at 56.5 +/- 3.9 % V.(O2) (max) (means +/- SEM), 4 min at 98.3 +/- 5.1 % V.(O2) (max) and 6 min of rest). Responses were calculated as areas under the plasma concentration curve (AUC) during 4 h after either breakfast or lunch.. Breakfast-AUCs for glucose, insulin and C peptide were lower (p < 0.05) on the exercise day compared with the control day (glucose: 538 +/- 94 vs 733 +/- 64 mmol. l(-1). 240 min; insulin: 16 +/- 4 vs 22 +/- 3 pmol. ml(-1). 240 min; C peptide: 143 +/- 22 vs 203 +/- 29 pmol. ml(-1). 240 min). After breakfast glucose appearance was unaffected by exercise, whereas disappearance and clearance increased (p < 0.05). Muscle glycogen was diminished by exercise (p < 0.05). After lunch no differences were observed between experiments. Exercise-induced reductions in glucose, insulin and C peptide responses were similar (p > 0.05) in this study of intermittent, high intensity exercise and in a previous study of isocaloric but prolonged moderate (45 min at 53 +/- 2 % V.(O2) (max)) postprandial exercise.. Postprandial high intensity exercise does not deteriorate glucose homeostasis but reduces both glucose concentrations and insulin secretion. The effect of exercise is related to energy expenditure rather than to peak exercise intensity. Finally, postprandial exercise does not influence glucose homeostasis during a subsequent main meal. [Diabetologia (1999) 42: 1282-1292]

Topics: Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Eating; Epinephrine; Exercise; Fatty Acids, Nonesterified; Glycerol; Glycogen; Homeostasis; Humans; Insulin; Kinetics; Lactates; Male; Middle Aged; Muscles; Norepinephrine; Oxygen Consumption

We investigated the effect of nutrient intake on glucose metabolism in normal Mexican-Americans (n = 6) and European-Americans (n = 6). Subjects were studied after an 18-h fast and after 5-6 h of ingestion of hourly meals that supplied 6.35 or 12.75 micromol glucose. kg(-1). min(-1). Endogenous glucose production (EGP), gluconeogenesis (GNG), and glycogenolysis (GLY) were estimated by mass isotopomer analysis with [U-(13)C]glucose infusions. Fasting EGP, GNG, and GLY did not differ between the groups. Food ingestion lowered the molar rate of GNG by only 31%. However, while consuming the lower quantity of nutrients, Mexican-Americans had higher plasma glucose (P < 0.05), a 39% higher rate of EGP (P < 0.05), and a 68% (P < 0.025) higher rate of GLY than the European-Americans. At the higher intake, EGP and GLY were suppressed completely in both groups. There was a linear relationship between insulin concentrations, EGP, and GLY in both groups, but the slope of the line was significantly (P < 0.05) greater in the European-Americans. We conclude that the sensitivity of GLY to nutrient intake differs between ethnic groups and that this may play a role in the increased predisposition of Mexican-Americans to type II diabetes.

Topics: Adult; Asian People; Blood Glucose; Carbon Isotopes; Diabetes Mellitus, Type 2; Energy Metabolism; Genetic Predisposition to Disease; Gluconeogenesis; Glucose; Glucose Tolerance Test; Glycogen; Humans; Hyperglycemia; Insulin; Kinetics; Lactic Acid; Male; Mexican Americans; Middle Aged; Postprandial Period; White People

The antidiabetic effects of 3-¿4-[2-(5-methyl-2-phenyl-oxazol-4- yl)ethoxy]phenyl¿-2S-propylamino-propionic acid (CAS 185679-16-7, HQL-975), a novel oral agent, on a genetically obese non-insulin-dependent diabetes mellitus (NIDDM) model (db/db mice) were examined. HQL-975 administration (3.7-34.1 mg/kg/d for 7 days) decreased the levels of plasma glucose, triglyceride, total cholesterol, non-esterified fatty acid and insulin in the mice. In an intraperitoneal glucose tolerance test (IPGTT), HQL-975 administration decreased the fasting plasma glucose level and improved the glucose tolerance in the mice. The HQL-975 administration also significantly increased the glycogenesis and lipogenesis from 14C-glucose in liver, but did not alter the glycogenesis in the diaphragm or the lipogenesis in adipose tissues at 2 h after the glucose loading. In the HQL-975-treated db/db mice, the radioactivity of 14C-glucose incorporated into hepatic glycogen was higher than that incorporated into hepatic total lipids. After the administration of HQL-975 (34.1 mg/kg/d for 7 days) to db/db mice, the hepatic hexokinase and fatty acid synthetase activities were significantly increased, the glycogen synthase I activity was increased but not significantly, and the glucose-6-phosphatase and the phosphoenolpyruvate carboxykinase activities were decreased. These results suggest that HQL-975 increases the hepatic glucose utilization and decreases the hepatic glucose production. Since hepatic glycogenesis is regulated by glucose itself but not by insulin in normoglycemic ICR mice, HQL-975 is thought to enhance the effect of glucose on the stimulation of hepatic glycogenesis. It is concluded that the enhancement of the hepatic glucose utilization played an important role in the hypoglycemic action of HQL-975.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Glycogen; Hypoglycemic Agents; Insulin; Lactic Acid; Lipids; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Inbred ICR

To clarify the contribution of the Asp905Tyr polymorphism of the muscle-specific glycogen-targeting subunit of protein phosphatase 1 (PP1G) to insulin resistance and related diseases.. We investigated the Asp905Tyr polymorphism of the PPP1R3 gene, which encodes the muscle-specific glycogen-targeting subunit of PP1G, in 259 Japanese patients with NIDDM and 194 healthy control subjects.. No significant difference was found in the genotype distribution between NIDDM patients (N = 259; Asp/Asp = 0.10, Asp/Tyr = 0.44, Tyr/Try = 0.46) and healthy control subjects (n = 194; Asp/Asp = 0.13, Asp/Tyr = 0.37, Tyr/Tyr = 0.50) or between patient groups subdivided by the mode of treatment: NIDDM patients with insulin therapy (Asp/Asp = 0.14, Asp/Tyr = 0.46, Tyr/Tyr = 0.40) and those without insulin therapy (Asp/Asp = 0.07, Asp/Tyr = 0.43, Tyr/Tyr = 0.50). However, NIDDM patients with the Tyr allele, which was previously reported to be associated with insulin resistance, tended to have lower BMIs than those without this allele (Asp/Asp: 24.5 +/- 1.1 kg/m2, Asp/Tyr: 22.6 +/- 0.4 kg/m2, Tyr/Tyr: 22.8 + 0.3 kg/m2, P = 0.06 by analysis of variance).. These data suggest that the Asp905Tyr polymorphism of the PPP1R3 gene is not associated with NIDDM or high BMI, both of which are known to be insulin-resistant states, in the Japanese population.

Topics: Adult; Aged; Alleles; Amino Acid Substitution; Aspartic Acid; Body Mass Index; Data Interpretation, Statistical; Diabetes Mellitus, Type 2; Female; Gene Frequency; Genes; Genotype; Glycogen; Humans; Hypoglycemic Agents; Insulin; Male; Middle Aged; Muscle, Skeletal; Phosphoprotein Phosphatases; Polymorphism, Genetic; Protein Phosphatase 1; Tyrosine

Topics: Administration, Oral; Animals; Blood Glucose; Cell Line; Diabetes Mellitus, Type 2; Enzyme Inhibitors; Glycogen; Humans; Hypoglycemic Agents; Indoles; Liver; Mice; Mice, Obese; Phosphorylases; Recombinant Proteins; Stereoisomerism; Structure-Activity Relationship

We examined the ability of an equivalent increase in circulating glucose concentrations to inhibit endogenous glucose production (EGP) and to stimulate glucose metabolism in patients with Type 2 diabetes mellitus (DM2). Somatostatin was infused in the presence of basal replacements of glucoregulatory hormones and plasma glucose was maintained either at 90 or 180 mg/dl. Overnight low-dose insulin was used to normalize the plasma glucose levels in DM2 before initiation of the study protocol. In the presence of identical and constant plasma insulin, glucagon, and growth hormone concentrations, a doubling of the plasma glucose levels inhibited EGP by 42% and stimulated peripheral glucose uptake by 69% in nondiabetic subjects. However, the same increment in the plasma glucose concentrations failed to lower EGP, and stimulated glucose uptake by only 49% in patients with DM2. The rate of glucose infusion required to maintain the same hyperglycemic plateau was 58% lower in DM2 than in nondiabetic individuals. Despite diminished rates of total glucose uptake during hyperglycemia, the ability of glucose per se (at basal insulin) to stimulate whole body glycogen synthesis (glucose uptake minus glycolysis) was comparable in DM2 and in nondiabetic subjects. To examine the mechanisms responsible for the lack of inhibition of EGP by hyperglycemia in DM2 we also assessed the rates of total glucose output (TGO), i.e., flux through glucose-6-phosphatase, and the rate of glucose cycling in a subgroup of the study subjects. In the nondiabetic group, hyperglycemia inhibited TGO by 35%, while glucose cycling did not change significantly. In DM2, neither TGO or glucose cycling was affected by hyperglycemia. The lack of increase in glucose cycling in the face of a doubling in circulating glucose concentrations suggested that hyperglycemia at basal insulin inhibits glucose-6-phosphatase activity in vivo. Conversely, the lack of increase in glucose cycling in the presence of hyperglycemia and unchanged TGO suggest that the increase in the plasma glucose concentration failed to enhance the flux through glucokinase in DM2. In summary, both lack of inhibition of EGP and diminished stimulation of glucose uptake contribute to impaired glucose effectiveness in DM2. The abilities of glucose at basal insulin to both increase the flux through glucokinase and to inhibit the flux through glucose-6-phosphatase are impaired in DM2. Conversely, glycogen synthesis is exquisitely sensitive to cha

Topics: Adult; Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Glucagon; Glucose Clamp Technique; Glycogen; Glycolysis; Human Growth Hormone; Humans; Hyperglycemia; Hypoglycemic Agents; Insulin; Middle Aged; Somatostatin; Sulfonylurea Compounds

Epidemiological studies suggest that alcohol consumption is an independent risk factor for the development of non-insulin-dependent diabetes mellitus (NIDDM). Alcoholism is known to be associated with increased plasma levels of two novel diols, 2,3-butanediol and 1,2-propanediol, metabolites known to impair insulin action in isolated adipocytes. This study examines whether 2,3-butanediol and 1,2-propanediol have the capacity to impair insulin action acutely in vivo in the rat. Using the euglycemic-hyperinsulinemic clamp, it is shown that the two diols reduce whole-body glucose utilization (by approximately 30%), with the onset of insulin resistance in vivo occurring at plasma concentrations of 2,3-butanediol (33 micromol/L) at least one order of magnitude (P < .001) lower than 1,2-propanediol (432 micromol/L). Tracer methodologies using [U-14C]glucose and 2-deoxy[1-(3)H]glucose indicate that the reduction in whole-body glucose utilization is accompanied by a reduction in glucose uptake and glycogen synthesis in the skeletal muscle and heart. The association between elevated plasma diol levels and insulin resistance demonstrated in this report raises the question of whether there is a link between the high plasma diol levels in alcohol abusers and their increased susceptibility to NIDDM.

Topics: Alcoholism; Animals; Blood Glucose; Butylene Glycols; Diabetes Mellitus, Type 2; Glycogen; Insulin; Insulin Resistance; Male; Propylene Glycol; Rats; Rats, Wistar

The spontaneously hypertensive/NIH-corpulent (SHR/N-cp) rat is a genetic model that exhibits both non-insulin-dependent diabetes mellitus (NIDDM) and hypertension. To determine the impact of long-term treatment with the long-acting angiotensin-converting enzyme (ACE) inhibitor perindopril (PE) on the glucose metabolism, lipid levels, and heart in this model, studies were performed in three groups of SHR/N-cp rats maintained on a diet containing 54% carbohydrate with 18% sucrose and 36% starch. One group of obese rats received PE (0.5 to 1.0 mg/kg body weight/d) for 3 to 4 months, a second group of obese rats received no treatment, and a third group of lean rats were used as controls. The mean systolic blood pressure (SBP) increased gradually in both untreated obese and lean rats, with lean animals showing slightly higher levels compared with untreated obese rats. By contrast, SBP was reduced to normal levels in PE-treated obese rats throughout the treatment period. Compared with lean rats, obese rats showed significantly higher body weight and fasting serum levels of glucose, insulin, total cholesterol (TC), and triglyceride (TG). However, no significant differences were observed in these metabolic parameters between PE-treated and untreated obese rats. Plasma renin activity measured at the end of the treatment period was significantly higher in PE-treated rats compared with untreated obese and untreated lean rats. The mean heart weight and left ventricular weight, expressed in absolute terms or indexed to body weight, were significantly lower in PE-treated versus untreated obese and untreated lean rats. To further determine whether glucose metabolism is directly affected by PE treatment, in vitro glycogen synthesis was evaluated in isolated soleus muscles obtained from three additional groups of animals. The basal rate of muscle glycogen synthesis was significantly lower in obese compared with lean rats (P < .05), but did not differ between PE-treated and untreated obese rats. Maximal insulin-stimulated glycogen synthesis increased threefold in PE-treated obese rats, but this increase did not differ from the increases observed in untreated obese and lean rats. In conclusion, the present study shows that long-term PE treatment in obese SHR/N-cp rats with NIDDM and hypertension effectively controlled systemic arterial pressure and resulted in a significant reduction in left ventricular weight. However, these favorable effects of PE were not associated with

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Antihypertensive Agents; Diabetes Mellitus, Type 2; Glycogen; Heart; Hypertension; Indoles; Male; Perindopril; Rats; Rats, Inbred SHR

The effects of tumor necrosis factor-alpha (TNF alpha) on glucose uptake and glycogen synthase (GS) activity were studied in human skeletal muscle cell cultures from nondiabetic and type 2 diabetic subjects. In nondiabetic muscle cells, acute (90-min) exposure to TNF alpha (5 ng/ml) stimulated glucose uptake (73 +/- 14% increase) to a greater extent than insulin (37 +/- 4%; P < 0.02). The acute uptake response to TNF alpha in diabetic cells (51 +/- 6% increase) was also greater than that to insulin (31 +/- 3%; P < 0.05). Prolonged (24-h) exposure of nondiabetic muscle cells to TNF alpha resulted in a further stimulation of uptake (152 +/- 31%; P < 0.05), whereas the increase in cells from type 2 diabetics was not significant compared with that in cells receiving acute treatment. After TNF alpha treatment, the level of glucose transporter-1 protein was elevated in nondiabetic (4.6-fold increase) and type 2 (1.7-fold) cells. Acute TNF alpha treatment had no effect on the fractional velocity of GS in either nondiabetic or type 2 cells. Prolonged exposure reduced the GS fractional velocity in both nondiabetic and diabetic cells. In summary, both acute and prolonged treatment with TNF alpha up-regulate glucose uptake activity in cultured human muscle cells, but reduce GS activity. Increased skeletal muscle glucose uptake in conditions of TNF alpha excess may serve as a compensatory mechanism in the insulin resistance of type 2 diabetes.

Topics: Adult; Cells, Cultured; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Synthase; Humans; Middle Aged; Monosaccharide Transport Proteins; Muscle, Skeletal; Reference Values; Tumor Necrosis Factor-alpha

To examine whether the hexosamine biosynthetic pathway might play a role in fat-induced insulin resistance, we monitored the effects of prolonged elevations in FFA availability both on skeletal muscle levels of UDP-N-acetyl-hexosamines and on peripheral glucose disposal during 7-h euglycemic-hyperinsulinemic (approximately 500 microU/ml) clamp studies. When the insulin-induced decrease in the plasma FFA levels (to approximately 0.3 mM) was prevented by infusion of a lipid emulsion in 15 conscious rats (plasma FFA approximately 1.4 mM), glucose uptake (5-7 h = 32.5+/-1.7 vs 0-2 h = 45.2+/-2.8 mg/kg per min; P < 0.01) and glycogen synthesis (P < 0.01) were markedly decreased. During lipid infusion, muscle UDP-N-acetyl-glucosamine (UDP-GlcNAc) increased by twofold (to 53.4+/-1.1 at 3 h and to 55.5+/-1.1 nmol/gram at 7 h vs 20.4+/-1.7 at 0 h, P < 0.01) while glucose-6-phosphate (Glc-6-P) levels were increased at 3 h (475+/-49 nmol/gram) and decreased at 7 h (133+/-7 vs 337+/-28 nmol/gram at 0 h, P < 0.01). To discern whether such an increase in the skeletal muscle UDP-GlcNAc concentration could account for the development of insulin resistance, we generated similar increases in muscle UDP-GlcNAc using three alternate experimental approaches. Euglycemic clamps were performed after prolonged hyperglycemia (18 mM, n = 10), or increased availability of either glucosamine (3 micromol/kg per min; n = 10) or uridine (30 micromol/kg per min; n = 4). These conditions all resulted in very similar increases in the skeletal muscle UDP-GlcNAc (to approximately 55 nmol/gram) and markedly impaired glucose uptake and glycogen synthesis. Thus, fat-induced insulin resistance is associated with: (a) decreased skeletal muscle Glc-6-P levels indicating defective transport/phosphorylation of glucose; (b) marked accumulation of the endproducts of the hexosamine biosynthetic pathway preceding the onset of insulin resistance. Most important, the same degree of insulin resistance can be reproduced in the absence of increased FFA availability by a similar increase in skeletal muscle UDP-N-acetyl-hexosamines. In conclusion, our results support the hypothesis that increased FFA availability induces skeletal muscle insulin resistance by increasing the flux of fructose-6-phosphate into the hexosamine pathway.

Topics: Animals; Diabetes Mellitus, Type 2; Fatty Acids; Fructosephosphates; Glucosamine; Glucose; Glucose-6-Phosphate; Glycogen; Glycolysis; Insulin; Insulin Resistance; Male; Muscle, Skeletal; Phosphorylation; Rats; Rats, Sprague-Dawley; Uridine

Human skeletal muscle cultures (HSMCs) from type II diabetic subjects were used to determine whether metabolic abnormalities such as hyperglycemia or hyperinsulinemia contribute to the defective muscle glycogen synthase (GS) activity present in this disorder. Following approximately 6 weeks of growth, diabetic cultures were fused for 4 days in normal, hyperglycemia, or hyperinsulinemia medium. Fusion of diabetic HSMCs in hyperglycemia medium (20 mmol/l vs. 5.5 mmol/l) had no effect on GS fractional velocity (FV) or mRNA levels, but impaired acute insulin-stimulation of glycogen synthesis and GS activity at 0.1 mmol/l glucose-6-phosphate, and reduced GS protein content by approximately 15% (P < 0.05). Fusion of diabetic muscle cultures in hyperinsulinemia medium (30 micromol/l vs. 22 pmol/l) improved basal GS activity, increasing the reduced GS FV by approximately 50% (P < 0.05), and decreasing the elevated Km(0.1) (half-maximal substrate concentration) by approximately 47% (P < 0.05). Hyperinsulinemia also significantly increased (P < 0.05) the reduced GS mRNA and protein levels of diabetic muscle to levels similar to that in nondiabetic subjects. In contrast to the improvements in the basal state, hyperinsulinemia completely abolished acute insulin responsiveness of GS activity and glycogen synthesis in muscle of type II diabetic subjects. The combination of hyperinsulinemia and hyperglycemia produced effects on both basal and insulin-responsive GS FV and mRNA similar to hyperinsulinemia alone, but hyperinsulinemia prevented hyperglycemia's effect of lowering GS protein and glycogen synthesis. We concluded that, in diabetic muscle, hyperinsulinemia may serve to partially compensate for the impaired basal GS activity and for the adverse effects of hyperglycemia on GS protein content, activity, and glycogen formation by both pre- and posttranslational mechanisms. Despite these beneficial effects, hyperinsulinemia also induces severe impairment of insulin-stimulated GS activity and glycogen formation, which may contribute to acquired muscle insulin resistance of type II diabetes.

Topics: Adult; Biopsy; Blotting, Northern; Cells, Cultured; Diabetes Mellitus, Type 2; Glycogen; Glycogen Synthase; Humans; Hyperglycemia; Hyperinsulinism; Immunoblotting; Insulin; Kinetics; Middle Aged; Muscle, Skeletal; Receptor, Insulin; RNA, Messenger

The relationship between clear-cell syringoma and diabetes mellitus is well established. We present a case of clear-cell porocarcinoma in a diabetic patient. The lesion consisted of a 5-cm nodule on the lateral aspect of the left leg. Histopathologically, the neoplasm was composed of irregular aggregations of neoplastic cells with striking clear-cell appearance, showing features of ductal differentiation. The clear-cell appearance of neoplastic cells was due to glycogen accumulation within their cytoplasm. Immunohistochemistry and ultrastructural studies also supported the diagnosis of a neoplasm with sweat ductal differentiation. Enzyme histochemical reactions for phosphorylase immunoreactivity on fresh, unfixed sections of the neoplasm demonstrated that this immunoreactivity was remarkably decreased. Some adnexal neoplasms of the skin mostly composed of clear cells may be cutaneous markers of diabetes mellitus. Phosphorylase activity deficiency in diabetic patients may be responsible for glycogen accumulation in neoplastic cells resulting in clear-cell appearance of these neoplasms.

Topics: Acrospiroma; Aged; Biomarkers, Tumor; Cell Differentiation; Cell Nucleus; Cytoplasm; Cytoplasmic Granules; Diabetes Mellitus, Type 2; Eccrine Glands; Glycogen; Humans; Immunohistochemistry; Male; Microvilli; Mucin-1; Phosphorylases; Skin Neoplasms

We have investigated the antidiabetic action of troglitazone in aP2/DTA mice, whose white and brown fat was virtually eliminated by fat-specific expression of diphtheria toxin A chain. aP2/DTA mice had markedly suppressed serum leptin levels and were hyperphagic, but did not gain excess weight. aP2/DTA mice fed a control diet were hyperlipidemic, hyperglycemic, and had hyperinsulinemia indicative of insulin-resistant diabetes. Treatment with troglitazone alleviated the hyperglycemia, normalized the tolerance to intraperitoneally injected glucose, and significantly decreased elevated insulin levels. Troglitazone also markedly decreased the serum levels of cholesterol, triglycerides, and free fatty acids both in wild-type and aP2/DTA mice. The decrease in serum triglycerides in aP2/DTA mice was due to a marked reduction in VLDL- and LDL-associated triglyceride. In skeletal muscle, triglyceride levels were decreased in aP2/DTA mice compared with controls, but glycogen levels were increased. Troglitazone treatment decreased skeletal muscle, but not hepatic triglyceride and increased hepatic and muscle glycogen content in wild-type mice. Troglitazone decreased muscle glycogen content in aP2/DTA mice without affecting muscle triglyceride levels. The levels of peroxisomal proliferator-activated receptor gamma mRNA in liver increased slightly in aP2/DTA mice and were not changed by troglitazone treatment. The results demonstrate that insulin resistance and diabetes can occur in animals without significant adipose deposits. Furthermore, troglitazone can alter glucose and lipid metabolism independent of its effects on adipose tissue.

Topics: Adipose Tissue; Animals; Blood Glucose; Body Weight; Cholesterol; Chromans; Diabetes Mellitus, Type 2; Eating; Fatty Acids; Glycogen; Hypoglycemic Agents; Insulin; Insulin Resistance; Insulin Secretion; Leptin; Lipoproteins, LDL; Lipoproteins, VLDL; Mice; Mice, Transgenic; Organ Size; Proteins; Receptors, Cytoplasmic and Nuclear; Thiazoles; Thiazolidinediones; Transcription Factors; Triglycerides; Troglitazone

Glycogen synthesis is impaired in first degree relatives of subjects with non-insulin-dependent diabetes mellitus and genes relevant to this metabolic pathway are considered reasonable candidates in the pathogenesis of the disease. In skeletal muscle the de novo synthesis of glycogen in primed by an enzyme named glycogenin. We have cloned the glycogenin cDNA from human skeletal muscle mRNA: human glucogenin is a 333 amino acid protein exhibiting 93% identity with rabbit glycogenin. A single transcript of about 2.4 kb, prominent in skeletal muscle, was detected by Northern blot analysis. In situ hybridization unequivocally located the human glycogenin gene to chromosome 3q25.1. Furthermore, we mapped two intronless glycogenin-related sequences to human chromosomes 12 and 13.

Topics: Amino Acid Sequence; Animals; Base Sequence; Chromosome Mapping; Chromosomes, Human, Pair 12; Chromosomes, Human, Pair 13; Chromosomes, Human, Pair 3; Cloning, Molecular; Diabetes Mellitus, Type 2; DNA Primers; DNA, Complementary; Glucosyltransferases; Glycogen; Glycoproteins; Humans; In Situ Hybridization, Fluorescence; Insulin Resistance; Molecular Sequence Data; Muscle Proteins; Muscle, Skeletal; Open Reading Frames; Rabbits; Tissue Distribution

To examine the impact of insulin resistance on the insulin-dependent and insulin-independent portions of muscle glycogen synthesis during recovery from exercise, we studied eight young, lean, normoglycemic insulin-resistant (IR) offspring of individuals with non-insulin-dependent diabetes mellitus and eight age-weight matched control (CON) subjects after plantar flexion exercise that lowered muscle glycogen to approximately 25% of resting concentration. After approximately 20 min of exercise, intramuscular glucose 6-phosphate and glycogen were simultaneously monitored with 31P and 13C NMR spectroscopies. The postexercise rate of glycogen resynthesis was nonlinear. Glycogen synthesis rates during the initial insulin independent portion (0-1 hr of recovery) were similar in the two groups (IR, 15.5 +/- 1.3 mM/hr and CON, 15.8 +/- 1.7 mM/hr); however, over the next 4 hr, insulin-dependent glycogen synthesis was significantly reduced in the IR group [IR, 0.1 +/- 0.5 mM/hr and CON, 2.9 +/- 0.2 mM/hr; (P < or = 0.001)]. After exercise there was an initial rise in glucose 6-phosphate concentrations that returned to baseline after the first hour of recovery in both groups. In summary, we found that following muscle glycogen-depleting exercise, IR offspring of parents with non-insulin-dependent diabetes mellitus had (i) normal rates of muscle glycogen synthesis during the insulin-independent phase of recovery from exercise and (ii) severely diminished rates of muscle glycogen synthesis during the subsequent recovery period (2-5 hr), which has previously been shown to be insulin-dependent in normal CON subjects. These data provide evidence that exercise and insulin stimulate muscle glycogen synthesis in humans by different mechanisms and that in the IR subjects the early response to stimulation by exercise is normal.

Topics: Adult; Analysis of Variance; Blood Glucose; Diabetes Mellitus, Type 2; Epinephrine; Female; Glucagon; Glucose; Glucose-6-Phosphate; Glucosephosphates; Glycogen; Humans; Hydrogen-Ion Concentration; Infusions, Intravenous; Insulin; Insulin Resistance; Lactates; Magnetic Resonance Spectroscopy; Male; Muscle Contraction; Muscle, Skeletal; Nuclear Family; Phosphates; Phosphocreatine; Physical Exertion; Reference Values

The effect of insulin to increase the activity of glycogen synthase (GS) in muscle has been well documented, however, the effect of in vivo insulin to inactivate glycogen phosphorylase (GP) has not been previously shown. To determine the effects of insulin on glycogenolysis in rhesus monkeys, GP and glycogen were determined in muscle samples obtained under basal fasting and insulin-stimulated conditions during a euglycemic hyperinsulinemic clamp in a group of 27 monkeys ranging from normal to overtly diabetic (NIDDM) and compared to GS activity previously examined. The diabetic monkeys had lower basal and insulin-stimulated glycogen concentrations compared to the normal and hyperinsulinemic monkeys (p < 0.05). The response of GP activity ratio (AR) to insulin (delta) was inversely correlated to delta GS fractional velocity (fv) (r = -0.57, p < 0.002) in all of the monkeys. The AR of GP was inversely correlated to the fv of GS measured under insulin-stimulated conditions (r = -0.60, p < 0.05) in the 11 normal monkeys. In the normal group, the range in response of GS to insulin (delta GSfv) was previously shown to be 3-22%, with n = 6 < 11% ('low normals') and n = 5 > 11% ('high normals'). In the present study, the low normals were shown to have (1) higher delta GP independent activity and delta GP total activity compared to the high normals and hyperinsulinemic monkeys (p less than or equal to 0.05), (2) higher insulin-stimulated GP independent activity and GP total activity compared to the other three groups (p < 0.05), (3) higher insulin-stimulated GP activity ratio compared to the high normals and hyperinsulinemic monkeys (p < 0.05), (4) and lower whole-body insulin-mediated glucose disposal rates compared to the high normals (p < 0.05). We conclude that NIDDM is accompanied by low glycogen content in the muscle, and that some clinically normal monkeys have an alteration in insulin action on muscle GS, GP, and whole-body glucose disposal rates that may precede the development of hyperinsulinemia.

Topics: Animals; Diabetes Mellitus, Type 2; Glucose Clamp Technique; Glycogen; Glycogen Synthase; Insulin; Insulin Resistance; Macaca mulatta; Muscle, Skeletal; Phosphorylases

The primary purpose of this study was to evaluate the acute effect of exercise of differing intensity on plasma glucose and insulin responses to an oral glucose challenge.. Six obese men and six obese men with NIDDM of similar age, weight, percentage body fat, and VO2peak participated in the study. Each subject underwent two 7-day exercise programs in a counterbalanced order at 2-week intervals. During each 7-day exercise period, the subjects cycled every day at a power output corresponding to 50% VO2peak for 70 min or 70% VO2peak for 50 min. Muscle glycogen utilization was estimated during exercise on day 7 using a [3H]glucose infusion technique in conjunction with indirect calorimetry. During the day before and after each 7-day exercise period, a 3-h oral glucose tolerance test (OGTT) was administered after a 12-h overnight fast.. The average caloric expenditure did not differ between exercise at 50 and 70% VO2peak in both obese and obese NIDDM subjects. However, the carbohydrate oxidation was higher (P < 0.05) during exercise at 70 than 50% VO2peak in obese subjects (77 +/- 5 vs. 68 +/- 6 g) and obese NIDDM subjects (70 +/- 4 vs. 58 +/- 6 g). Muscle glycogen utilization was also higher (P < 0.05) during exercise at 70 than 50% VO2peak in obese subjects (59 +/- 9 vs. 30 +/- 7 g) and in obese NIDDM subjects (48 +/- 5 vs. 24 +/- 5 g). In obese subjects, plasma glucose response area during the OGTT did not change after 7 days of exercise at either 50 or 70% VO2peak. Plasma insulin response area during the OGTT also did not change after 7 days of exercise at 50% VO2peak. However, plasma insulin response area was reduced (P < 0.05) after 7 days of exercise at 70% VO2peak (9,644 +/- 1,783 vs 7,538 +/- 1,522 microU.ml-1.180 min-1). In obese NIDDM subjects, both plasma glucose and insulin response areas during the OGTT did not decrease after 7 days of exercise at either 50 or 70% VO2peak.. It is concluded that the exercise-induced improvement in insulin sensitivity is influenced by exercise intensity in obese individuals. The improved insulin sensitivity after 7 days of exercise at 70% VO2peak in obese individuals may be related to greater muscle glycogen utilization during exercise. The lack of improvement in glucose tolerance and insulin sensitivity after 7 days of exercise at either 50 or 70% VO2peak in obese NIDDM patients may be due to the fact that the NIDDM patients selected in the present study were relatively hypoinsulinemic.

Topics: Adipose Tissue; Adult; Blood Glucose; Calorimetry, Indirect; Cholesterol; Diabetes Mellitus; Diabetes Mellitus, Type 2; Exercise; Glucose Tolerance Test; Glycogen; Humans; Insulin; Male; Muscle, Skeletal; Obesity; Oxygen Consumption; Physical Exertion; Triglycerides

Increased endogenous glucose production (EGP) and gluconeogenesis contribute to the pathogenesis of hyperglycaemia in non-insulin-dependent diabetes mellitus (NIDDM). In healthy subjects, however, EGP remains constant during administration of gluconeogenic precursors. This study was performed in order to determine whether administration of fructose increases EGP in obese NIDDM patients and obese non-diabetic subjects. Eight young healthy lean subjects, eight middle-aged obese NIDDM patients and seven middle-aged obese non-diabetic subjects were studied during hourly ingestion of 13C fructose (0.3 g.kg fat free mass-1.h-1) for 3 h. Fructose failed to increase EGP (measured with 6,6 2H glucose) in NIDDM (17.7 +/- 1.9 mumol.kg fat free mass-1.min-1 basal vs 15.9 +/- 0.9 after fructose), in obese non-diabetic subjects (12.1 +/- 0.5 basal vs 13.1 +/- 0.5 after fructose) and in lean healthy subjects (13.3 +/- 0.5 basal vs 13.8 +/- 0.6 after fructose) although 13C glucose synthesis contributed 73.2% of EGP in lean subjects, 62.6% in obese non-diabetic subjects, and 52.8% in obese NIDDM patients. Since glucagon may play an important role in the development of hyperglycaemia in NIDDM, healthy subjects were also studied during 13C fructose ingestion + hyperglucagonaemia (232 +/- 9 ng/l) and during hyperglucagonaemia alone. EGP increased by 19.8% with ingestion of fructose + glucagon (p < 0.05) but remained unchanged during administration of fructose or glucagon alone. The plasma 13C glucose enrichment was identical after fructose ingestion both with and without glucagon, indicating that the contribution of fructose gluconeogenesis to the glucose 6-phosphate pool was identical in these two conditions. We concluded that during fructose administration: 1) gluconeogenesis is increased, but EGP remains constant in NIDDM, obese non-diabetic, and lean individuals; 2) in lean individuals, both an increased glucagonaemia and an enhanced supply of gluconeogenic precursors are required to increase EGP; this increase in EGP occurs without changes in the relative proportion of glucose 6-phosphate production from fructose and from other sources (i.e. glycogenolysis + gluconeogenesis from non-fructose precursors).

Topics: Administration, Oral; Adult; Analysis of Variance; Diabetes Mellitus; Diabetes Mellitus, Type 2; Dietary Carbohydrates; Female; Fructose; Glucagon; Gluconeogenesis; Glucose; Glycogen; Humans; Infusions, Intravenous; Kinetics; Male; Middle Aged; Models, Theoretical; Obesity; Reference Values

Eicosanoid production, glucose (Glu), glycogen (Gly) and triglyceride (TG) metabolism, spontaneous contractile activity, PGF2 alpha and oxytocin-induced contractions have been studied in uterine tissue obtained from control (C) and non-insulin-dependent diabetic (D) rats prior to parturition. Parturition occurs on day 22 of gestation in control animals, whereas a 24 hr delay was observed in diabetic rats. Production of PGE2, PGE1, 6-keto-PGF1 alpha, PGF2 alpha, TXB2 and LTB4 was similar in uterine tissue obtained from control and diabetic rats on day 21 of pregnancy. Uterine metabolism, on day 21 of pregnancy, based on the production of 14CO2 from U14C-glucose was lower in tissues obtained from diabetic rats than in controls. Levels of TG were similar at 0 hr and after 60 min incubation in Glu or Glu-free medium in both experimental groups. Initially Gly levels in diabetic and control uteri were similar. After 60 minutes of incubation, levels of Gly in control tissue decreased only in the absence of Glu in the incubation medium. In contrast, in diabetic uterine strips, levels of Gly decreased after 60 minutes of incubation either in Glu or Glu-free medium. "In vitro" isometric-developed tension (IDT) evaluated on day 21 (C and D) and 22 (D) of pregnancy was similar at 0 hr in control and diabetic uterine preparations, but IDT in both diabetic groups was decreased after a 40 minute incubation when compared to controls. Alterations in PGF2 alpha-induced uterine responses were not seen in 21 or 22 days pregnant diabetic uterine tissue when compared to controls. In contrast, impaired oxytocin responses were observed in diabetic uteri on day 21 of gestation, but they were similar to control responses of uterine tissue from day 22 diabetic rats. We conclude that in the non-insulin-dependent late pregnant rat, there are no alterations in uterine tissue eicosanoid production, but metabolic and contractile abnormalities are present. Involvement of these alterations in the delayed initiation of parturition is discussed.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Dinoprost; Dose-Response Relationship, Drug; Eicosanoids; Female; Glucose; Glycogen; In Vitro Techniques; Isometric Contraction; Oxytocin; Pregnancy; Rats; Rats, Wistar; Streptozocin; Time Factors; Triglycerides; Uterine Contraction; Uterus

We examined the possibility that protein kinase C (PKC) is chronically activated and may contribute to impaired glycogen synthesis and insulin resistance in soleus muscles of hyperinsulinemic type II diabetic Goto-Kakizaki (GK) rats. Relative to nondiabetic controls, PKC enzyme activity and levels of immunoreactive PKC-alpha, beta, epsilon, and delta were increased in membrane fractions and decreased cytosolic fractions of GK soleus muscles. In addition, PKC-theta levels were decreased in both membrane and cytosol fractios, whereas PKC-zeta levels were not changed in either fraction in GK soleus muscles. These increases in membrane PKC (alpha, beta, epsilon, and delta) could not be accounted for by alterations in PKC mRNA or total PKC levels but were associated with increases in membrane diacylglycerol (DAG) and therefore appeared to reflect translocative activation of PKC. In evaluation of potential causes for persistent PKC activation, membrane PKC levels were decreased in soleus muscles of hyperglycemic streptozotocin (STZ)-induced diabetic rats; thus, a role for simple hyperglycemia as a cause of PKC activation in GK rats was not evident in the STZ model. In support of the possibility that hyperinsulinemia contributed to PKC activation in GK soleus muscles, we found that DAG levels were increased, and PKC was translocated, in soleus muscles of both (1) normoglycemic hyperinsulinemic obese/aged rats and (2) mildly hyperglycemic hyperinsulinemic obese/Zucker rats. In keeping with the possibility that PKC activation may contribute to impaired glycogen synthase activation in GK muscles, phorbol esters inhibited, and a PKC inhibitor, RO 31-8220, increased insulin effects on glycogen synthesis in soleus muscles incubated in vitro. Our findings suggested that: (1) hyperinsulinemia, as observed in type II diabetic GK rats and certain genetic and nongenetic forms of obesity in rats, is associated with persistent translocation and activation of PKC in soleus muscles, and (2) this persistent PKC activation may contribute to impaired glycogen synthesis and insulin resistance.

Topics: Adipocytes; Aging; Animals; Cell Membrane; Cytosol; Diabetes Mellitus, Type 2; Enzyme Activation; Glycogen; Insulin Resistance; Isoenzymes; Liver; Male; Muscle, Skeletal; Obesity; Protein Kinase C; Protein Kinase C beta; Protein Kinase C-alpha; Protein Kinase C-delta; Protein Kinase C-epsilon; Rats; Rats, Mutant Strains; Rats, Wistar; Rats, Zucker; Reference Values; RNA, Messenger; Transcription, Genetic

Insulin resistance in the offspring of parents with non-insulin-dependent diabetes mellitus (NIDDM) is the best predictor of development of the disease and probably plays an important part in its pathogenesis. We studied the mechanism and degree to which exercise training improves insulin sensitivity in these subjects.. Ten adult children of parents with NIDDM and eight normal subjects were studied before starting an aerobic exercise-training program, after one session of exercise, and after six weeks of exercise. Insulin sensitivity was measured by the hyperglycemic-hyperinsulinemic clamp technique combined with indirect calorimetry, and the rate of glycogen synthesis in muscle and the intramuscular glucose-6-phosphate concentration were measured by carbon-13 and phosphorus-31 nuclear magnetic resonance spectroscopy, respectively.. During the base-line study, the mean (+/-SE) rate of muscle glycogen synthesis was 63 +/- 9 percent lower in the offspring of diabetic parents than in the normal subjects (P < 0.001). The mean value increased 69 +/- 10 percent (P = 0.04) and 62 +/- 11 percent (P = 0.04) after the first exercise session and 102 +/- 11 percent (P = 0.02) and 97 +/- 9 percent (P = 0.008) after six weeks of exercise training in the offspring and the normal subjects, respectively. The increment in glucose-6-phosphate during hyperglycemic-hyperinsulinemic clamping was lower in the offspring than in the normal subjects (0.039 +/- 0.013 vs. 0.089 +/- 0.009 mmol per liter, P = 0.005), reflecting reduced glucose transport-phosphorylation, but this increment was normal in the offspring after the first exercise session and after exercise training. Basal and stimulated insulin secretion was higher in the offspring than the normal subjects and was not altered by the exercise training program.. Exercise increases insulin sensitivity in both normal subjects and the insulin-resistant offspring of diabetic parents because of a twofold increase in insulin-stimulated glycogen synthesis in muscle, due to an increase in insulin-stimulated glucose transport-phosphorylation.

Topics: Adult; Biological Transport, Active; Diabetes Mellitus, Type 2; Exercise; Female; Glucose; Glucose Clamp Technique; Glucose-6-Phosphate; Glycogen; Humans; Insulin; Insulin Resistance; Insulin Secretion; Male; Muscle, Skeletal; Phosphorylation; Physical Fitness

Insulin resistance is a predictor of the development of noninsulin-dependent diabetes mellitus (NIDDM) in humans. It is unclear whether insulin resistance is a primary defect leading to NIDDM or the result of hyperinsulinemia and hyperglycemia. To determine if insulin resistance is the result of extrinsic factors such as hyperinsulinemia primary skeletal muscle cell cultures were established from muscle biopsies from Pima Indians with differing in vivo insulin sensitivities. These cell cultures expressed a variety of muscle-specific phenotypes including the proteins alpha-actinin and myosin, muscle-specific creatine kinase activity, and RNA encoding GLUT4, MYF5, MYOD1, and MYOGENIN. Labeled glucose was used to measure the insulin-stimulated conversion of glucose to glycogen in these cultures. The in vivo rates of insulin-stimulated glycogen production (insulin resistance) were correlated with in vitro measures of glycogen production (P = 0.007, r = 0.58). This defect in insulin action is stable in a uniform culture environment and is retained over time. The retention of insulin resistance in myoblast derived cell cultures is consistent with the expression of an underlying biochemical defect in insulin resistant skeletal muscle.

Topics: Actinin; Adult; Blotting, Northern; Creatine Kinase; Diabetes Mellitus, Type 2; DNA-Binding Proteins; Dose-Response Relationship, Drug; Fluorescent Antibody Technique, Direct; Glucose; Glucose Transporter Type 4; Glycogen; Humans; Hypoglycemic Agents; Indians, North American; Insulin; Insulin Resistance; Monosaccharide Transport Proteins; Muscle Proteins; Muscle, Skeletal; MyoD Protein; Myogenic Regulatory Factor 5; Myogenin; Myosins; RNA, Messenger; Trans-Activators

The regulatory G-subunit of the glycogen-associated form of protein phosphatase 1 (PP1) plays a crucial part in muscle tissue glycogen synthesis and breakdown. As impaired insulin stimulated glycogen synthesis in peripheral tissues is considered to be a pathogenic factor in subsets of non-insulin-dependent diabetes mellitus (NIDDM) and obesity, the G-subunit of PP1 should be viewed as a candidate gene for inherited insulin resistance. When applying heteroduplex formation analysis and nucleotide sequencing of PP1G-subunit cDNA from 30 insulin resistant white NIDDM patients two cases were identified as heterozygous carriers of an Asp905 --> Tyr substitution. The carrier prevalence of the PP1G-subunit variant was 18% in 150 healthy subjects and 13% in 313 NIDDM subjects (chi 2 = 1.94, p = 0.16). Twenty-seven healthy subjects volunteered for a 4 h euglycaemic, hyperinsulinaemic clamp in combination with indirect calorimetry in order to elucidate the potential impact of the Tyr905 substitution on the whole body glucose metabolism. Interestingly, the Tyr905 variant was associated with altered routing of glucose: a decreased insulin stimulated non-oxidative glucose metabolism of peripheral tissues (glycogen synthesis) (p < 0.04) and an increased basal glucose oxidation rate (p < 0.04) when compared with wild type carriers. A population-based sample of 380 unrelated young healthy Caucasians was examined during a combined intravenous glucose and tolbutamide test to address whether the Asp905/Tyr905 polymorphism was associated with alterations in insulin secretion which might be secondary to the insulin resistance of skeletal muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Base Sequence; Case-Control Studies; Diabetes Mellitus, Type 2; DNA; DNA Primers; Female; Gene Frequency; Genetic Variation; Glucose; Glycogen; Humans; Insulin; Insulin Resistance; Insulin Secretion; Male; Middle Aged; Molecular Sequence Data; Nucleic Acid Heteroduplexes; Phosphoprotein Phosphatases; Polymorphism, Genetic; Protein Phosphatase 1

Myoblasts from human skeletal muscle were isolated from needle biopsy samples of vastus lateralis and fused to differentiated multinucleated myotubes. Specific high-affinity insulin and insulin-like growth factor I (IGF-I) binding, glucose transporter proteins GLUT1 and GLUT4, glycogen synthase and pyruvate dehydrogenase proteins, and their specific mRNAs were identified in fused myotubes. Insulin and IGF-I stimulated 2-deoxyglucose uptake twofold with half-maximal stimulation by insulin at 0.98 +/- 0.12 nmol/l and maximal stimulation at 17.5 nmol/l. Acute insulin treatment (33 nmol/l) doubled glycogen synthase activity and glucose incorporation into glycogen while increasing pyruvate dehydrogenase approximately 30%. In cells cultured from NIDDM subjects, both basal (6.9 +/- 1.0 vs. 13.0 +/- 1.7 pmol.mg protein-1.min-1) and acute insulin-stimulated transport (13.5 +/- 2.0 vs. 22.4 +/- 1.3 pmol.mg protein-1.min-1) were significantly reduced compared with nondiabetic control subjects (both P < or = 0.005). GLUT1 protein content of total membranes from NIDDM subjects was decreased compared with control subjects, while GLUT4 levels were similar between groups. A significant correlation (r = 0.65, P < or = 0.05) was present when maximal rates of insulin-stimulated glucose transport in cell culture from subjects were compared with their corresponding in vivo glucose disposal determined by hyperinsulinemic glucose clamp. In summary, differentiated human skeletal muscle cultures exhibit biochemical and molecular features of insulin-stimulated glucose transport and intracellular enzyme activity comparable with the in vivo situation. Defective insulin-stimulated glucose transport persists in muscle cultures from NIDDM subjects and resembles the reduced insulin-mediated glucose uptake present in vivo. We conclude that this technique provides a relevant cellular model to study insulin action and glucose metabolism in normal subjects and determine the mechanisms of insulin resistance in NIDDM.

Topics: Adult; Biological Transport; Biopsy, Needle; Cell Fusion; Cells, Cultured; Creatine Kinase; Deoxyglucose; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Synthase; Glycolysis; Humans; Insulin; Insulin-Like Growth Factor I; Middle Aged; Muscle, Skeletal; Pyruvate Dehydrogenase Complex; Receptor, IGF Type 1; Receptor, Insulin; Reference Values

Topics: Animals; Apoproteins; Blotting, Western; Cells, Cultured; Diabetes Mellitus, Type 2; Glucosyltransferases; Glycogen; Glycoproteins; Humans; Lymphocytes; Molecular Weight; Muscle Proteins; Rabbits; Sheep

Recent studies have demonstrated that reduced insulin-stimulated muscle glycogen synthesis is the major cause of insulin resistance in patients with non-insulin-dependent diabetes mellitus (NIDDM). This reduced rate has been assigned to a defect in either glucose transport or hexokinase activity. However it is unknown whether this is a primary or acquired defect in the pathogenesis of NIDDM. To examine this question, we measured the rate of muscle glycogen synthesis and the muscle glucose 6-phosphate (G6P) concentration using 13C and 31P NMR spectroscopy as well as oxidative and nonoxidative glucose metabolism in six lean, normoglycemic offspring of parents with NIDDM and seven age/weight-matched control subjects under hyperglycemic (approximately 11 mM)-hyperinsulinemic (approximately 480 pM) clamp conditions. The offspring of parents with NIDDM had a 50% reduction in total glucose metabolism, primarily due to a decrease in the nonoxidative component. The rate of muscle glycogen synthesis was reduced by 70% (P < 0.005) and muscle G6P concentration was reduced by 40% (P < 0.003), which suggests impaired muscle glucose transport/hexokinase activity. These changes were similar to those previously observed in subjects with fully developed NIDDM. When the control subjects were studied at similar insulin levels (approximately 440 pM) but euglycemic plasma glucose concentration (approximately 5 mM), both the rate of glycogen synthesis and the G6P concentration were reduced to values similar to the offspring of parents with NIDDM. We conclude that insulin-resistant offspring of parents with NIDDM have reduced nonoxidative glucose metabolism and muscle glycogen synthesis secondary to a defect in muscle glucose transport/hexokinase activity prior to the onset of overt hyperglycemia. The presence of this defect in these subjects suggests that it may be the primary factor in the pathogenesis of NIDDM.

Topics: Adenosine Diphosphate; Adult; Biological Transport; Diabetes Mellitus, Type 2; Female; Glucose; Glucose-6-Phosphate; Glucosephosphates; Glycogen; Humans; Hydrogen-Ion Concentration; Magnetic Resonance Spectroscopy; Male; Muscles; Organophosphates; Patch-Clamp Techniques; Phosphocreatine; Phosphorylation

The frequent development of Type 2 diabetes in the obese suggests a relationship between obesity and diabetes. This study presents evidence for a continuum form obesity to diabetes via glucose intolerance and hyperinsulinemic diabetes. The defect which seems to be at the origin of this development resides in the increase in lipid oxidation already present in the early stages of obesity. It reflects the increased utilisation of fatty acids for energy purpose in the obese, at the expenses of glucose. In non-diabetic obese subjects, insulin resistance can be demonstrated by the inhibition of glucose storage during a euglycemic, hyperinsulinemic, clamp. This defect in glucose storage is not observed during a oral glucose tolerance test (OGTT), as it is compensated by hyperinsulinemia and hyperglycemia during glucose tolerance. Glucose tolerance appears with the inhibition of glucose oxidation by the augmented lipid oxidation. This decreased glucose utilization causes a slowdown of the utilization of glycogen stores which leads, as a consequence, to the inhibition of glycogen synthase by its product, glycogen. Diabetes appears when the increase in glycemia and insulinemia does not compensate any more for the inhibition of glucose storage. The rise in basal glycemia simultaneously with the fall in glucose storage corresponds to the transition to diabetes. The decreased glucose mobilization together with the inhibition of glycogen phosphorylase are such in the diabetic patient that glycogen stores tend to remain full and glycogen synthase is inhibited by negative feedback. The retrograde inhibition of glycogen stores on glycogen synthase activity brings up incapacity to store glucose and leads to a rise in glycemia. Finally, the evolution of obesity to diabetes leads to a decrease in insulin secretion with increase in hepatic glucose production through gluconeogenesis and decreased capacity to store glucose. Therapeutic implications are discussed in this review.

Topics: Adult; Aged; Diabetes Mellitus; Diabetes Mellitus, Type 2; Fatty Acids; Glucose; Glycogen; Humans; Insulin; Liver; Middle Aged; Muscles; Obesity

Chronic caloric restriction (CR) prevents the development of obesity and maintains health, slows aging processes, and prevents or substantially delays the development of non-insulin-dependent diabetes. Because changes in energy metabolism could be involved in all of these positive effects of CR, we examined glycogen synthase (GS) and glycogen phosphorylase (GP) activities and glucose 6-phosphate (G6P) and glycogen concentrations in skeletal muscle samples before and during a euglycemic hyperinsulinemic clamp in 6 older aged monkeys in which CR had been continued for 10.4 +/- 2.1 years. Basal GS activity (fractional velocity and independent) was significantly higher in the CR monkeys than has been previously shown in normal, hyperinsulinemic and diabetic monkeys. The normal effect of insulin to activate GS was absent in the CR group due to the paradoxical finding in some of these monkeys of a reduction in GS activity by insulin. Insulin also had the unexpected effect of increasing the independent activity of GP above basal activity (p<0.05). There was an inverse relationship between the change (insulin-stimulated minus basal) in GS fractional velocity and GP activity ratio (r=-0.91, p<0.005). The basal independent activities of GS and GP were also inversely correlated (r=-0.79, p<0.05). The insulin-stimulated concentration of G6P tended to be higher than the basal concentration (p<0.06) and was significantly higher than that previously shown in normal monkeys (p<0.05). We suggest that long-term calorie restriction (1) results in alterations in glycogen metabolism that may be important to the anti-diabetogenic and antiaging effects of CR and (2) unmasks early defects which may indicate the likelihood of ultimately developing obesity and diabetes.

Topics: Animals; Diabetes Mellitus, Type 2; Diet, Reducing; Energy Intake; Glucose Clamp Technique; Glucose-6-Phosphate; Glycogen; Glycogen Phosphorylase; Glycogen Synthase; Macaca mulatta; Male; Muscle, Skeletal; Obesity

Impaired insulin-stimulated glycogen synthesis of peripheral tissues is a characteristic feature of many patients with non-insulin-dependent diabetes mellitus (NIDDM) and their first-degree relatives with normal glucose tolerance, suggesting putative inherited defects in this metabolic pathway. In previous studies, we have failed to reveal mutations in the coding regions of the muscle-specific glycogen synthase gene and the three genes that encode the catalytic subunits of protein phosphatase 1 (PP1) as frequent causes of insulin resistance. Because the glycogen-associated regulatory subunit of protein phosphatase 1 (PP1 G-subunit) plays a key role in the insulin stimulation of glycogen synthesis and the activity of PP1 is decreased in insulin-resistant subjects, we have now cloned the human G-subunit cDNA to search for abnormalities in the corresponding gene (designated PPP1R3 in the human genome nomenclature) in patients with NIDDM. The human cDNA was isolated from a skeletal muscle cDNA library and was found to encode a 126-kDa protein, which shows 73% amino acid identity to the rabbit PP1 G-subunit. The human G-subunit cDNA from 30 insulin-resistant NIDDM patients was analyzed for genetic variations in the G-subunit by using single-stranded conformation polymorphism (SSCP) scanning of reversely transcribed mRNA. One variant SSCP profile was detected in the region encoding the COOH-terminal part of the PP1 G-subunit in only one NIDDM patient, and subsequent nucleotide sequencing showed a C to A transversion on one allele at base position 2792. This change predicts an amino acid substitution from alanine to glutamic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Amino Acid Sequence; Animals; Base Sequence; Diabetes Mellitus, Type 2; DNA Primers; Female; Glucose; Glycogen; Humans; Macromolecular Substances; Male; Middle Aged; Molecular Sequence Data; Muscles; Phosphoprotein Phosphatases; Polymerase Chain Reaction; Polymorphism, Genetic; Protein Phosphatase 1; Rabbits; Reference Values; RNA, Messenger; Sequence Homology, Amino Acid

Skeletal muscle contributes significantly to reduced insulin-stimulated glucose disposal in patients with obesity and non-insulin-dependent (type II) diabetes mellitus (NIDDM). The biochemical basis for insulin resistance is not known but may involve reduced glucose transport and/or a defect in intracellular pathways for glucose disposal. To address this question, we measured basal and insulin-stimulated glucose oxidation, glycogen formation, and nonoxidative glycolysis (lactate and amino acid release) in an incubated muscle preparation from nonobese and morbidly obese patients with and without NIDDM. Pathways of glucose disposal were also determined in muscle of obese NIDDM patients incubated under hyperglycemic (20 mmol/L) conditions, which increases glucose uptake by mass action. Under basal conditions (no insulin present) there were no significant differences in glycogen formation or glucose oxidation between nonobese control, obese nondiabetic, or obese diabetics. Lactate release was significantly higher in obese controls compared to nonobese controls in the basal state at 5 mmol/L glucose (10.2 +/- 2.8 v 24.7 +/- 3.5 nmol/min/g, P < .05). Under maximal insulin-stimulated conditions, rates of glycogen formation, glucose oxidation, and nonoxidized glycolysis increased 1.9-, 2.3-, and 2.2-fold over basal (P < .05) in nonobese controls. By contrast, insulin was ineffective at stimulating significant increases in any metabolic pathway of glucose disposal in muscle of obese or obese NIDDM patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Analysis of Variance; Diabetes Mellitus; Diabetes Mellitus, Type 2; Female; Glucose; Glycogen; Glycolysis; Humans; In Vitro Techniques; Insulin; Male; Middle Aged; Muscles; Obesity; Obesity, Morbid; Oxidation-Reduction

1. Red blood cells can store glucose and may thus participate in blood glucose homeostasis. We investigated if a defect in this process exists in non-insulin dependent diabetes (NIDD). 2. Blood was obtained in fasting conditions from 10 normal and 10 newly diagnosed NIDD patients (before and after 4 weeks Metformin therapy). Washed erythrocytes were resuspended in media containing various glucose concentrations (4.4, 6.6, 8.8 and 13.2 mmol/L). Total glucose uptake was calculated as the sum of the measurements of lactate as well as free glucose, the latter being determined before and after addition of amyloglucosidase to the pellet. 3. Cells from diabetics showed a pronounced reduction in glucose uptake, particularly in their capacity to store glucose as glycogen (reactive to amyloglucosidase). Metformin treatment almost normalized glycogen levels, whereas lactate declined concomitantly in the pellet. 4. Our data demonstrate that a defect in glucose uptake exists in erythrocytes from NIDD patients, affecting both free and stored glucose, and that this defect is reversed by Metformin treatment, indicating that this drug can increase glycogen levels even in insulin-insensitive cells. 5. Thus, in view of their total mass, erythrocytes may be important in the impaired glucose homeostasis in NIDD, in particular in marked hyperglycaemia such as after a meal.

Topics: Adult; Diabetes Mellitus, Type 2; Erythrocytes; Glucose; Glycogen; Humans; Hyperglycemia; Lactates; Lactic Acid; Metformin; Middle Aged

Topics: Animals; Diabetes Mellitus, Type 2; Glucosyltransferases; Glycogen; Glycoproteins; Muscles; Quail

To evaluate the long-held concept that acidic guanidines lack glycemic effects, guanidinoalkanoic acids and the biguanide metformin (positive control) were administered to KKAy mice, a model of noninsulin-dependent diabetes. Two acidic guanidines, 3-guanidinopropionic acid (3-GPA) and guanidinoacetic acid, decreased the plasma glucose level; other compounds were ineffective. 3-GPA was more potent than even metformin. Insulin suppression tests in KKAy mice indicated that improved insulin sensitivity was the mode of action for 3-GPA. Glycemic effects in KKAy mice resulted from increased glucose disposal whereas gluconeogenesis, hepatic glycogen content and intestinal glucose absorption were unchanged. 3-GPA's glycemic effect was corroborated in two other models of noninsulin-dependent diabetes. In ob/ob mice, the compound reduced hyperglycemia, polyuria, glycosuria and hyperinsulinemia. In insulin-resistant rhesus monkeys, it increased the disappearance of i.v. glucose. The glycemic action of 3-GPA required the presence of some circulating insulin as well as hyperglycemia because the compound was ineffective in normoglycemic mice, insulinopenic Chinese hamsters and streptozotocin-diabetic rats. These data indicate that acidic guanidine derivatives can ameliorate hyperglycemia in animal models of noninsulin-dependent diabetes. Because acidic derivatives uniquely lack the propensity of guanidine compounds for inducing lactic acidosis, our finding suggests a new approach for developing improved antidiabetes compounds from this chemical class.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Gluconeogenesis; Glucose; Glycogen; Guanidines; Hyperglycemia; Insulin; Insulin Resistance; Intestinal Absorption; Liver Glycogen; Macaca mulatta; Male; Metformin; Mice; Mice, Inbred C57BL; Mice, Obese; Muscles; Propionates; Structure-Activity Relationship

To determine whether the tendency for NIDDM to run in families could relate to genetically determined defects in insulin stimulation of glycogen synthesis, skin fibroblasts from subjects with a strong family history of NIDDM were studied. Fibroblasts from nondiabetic subjects without any family history of NIDDM were studied as control subjects. The cells were studied after 7-16 passages in culture. Rates of glycogen synthesis were lower in fibroblasts from NIDDM subjects both basally and with maximal insulin stimulation (0.77 +/- 0.11 vs. 0.46 +/- 0.04 pmol.well-1 x h-1 [P < 0.02] and 1.49 +/- 0.26 vs. 0.69 +/- 0.05 pmol.well-1 x h-1 +adP < 0.01]). Rates of glycogen synthesis were stimulated 1.9 +/- 0.2-fold above basal in the control cells and 1.5 +/- 0.1-fold above basal in the NIDDM cells (P < 0.02). Rates of [3H]thymidine uptake were similar in control and NIDDM fibroblasts (basal, 28.3 +/- 2.8 vs. 39.2 +/- 8.0; maximum, 50.9 +/- 7.2 vs. 69.3 +/- 16.9 dpm x 10(-3), respectively). Rates of uptake increased similarly in control and NIDDM cells by 1.8 +/- 0.1- and 1.7 +/- 0.1-fold above basal. Maximum specific fibroblast insulin binding was similar for control and NIDDM subjects (194.0 +/- 29.2 vs. 176.1 +/- 24.9 fmol 125I-labeled insulin bound/mg protein respectively). The tyrosine kinase activity of insulin receptors isolated from the control and NIDDM fibroblasts was similar (basal, 135 +/- 30 vs. 149 +/- 33; submaximal, 153 +/- 28 vs. 155 +/- 30; and maximal insulin, 191 +/- 45 vs. 213 +/- 48 dpm.mg protein-1 x min-1).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Cells, Cultured; Diabetes Mellitus, Type 2; Female; Fibroblasts; Glucose; Glycogen; Humans; Insulin; Kinetics; Male; Middle Aged; Protein-Tyrosine Kinases; Receptor, Insulin; Reference Values; Skin; Thymidine

1. The relationship between hypertension, obesity, non-insulin-dependent diabetes mellitus and various parameters of glucose metabolism was studied. Lean and obese rats of the SHR/N-cp and LA/N-cp congenic strains were studied at four months of age. 2. Tritium and 14C-labeled glucoses were infused in one set of rats while tritiated water and 14C-labeled alanine were infused in a second group. 3. Glucose oxidation, turnover, conversion to glycogen, fatty acid synthesis, and alanine conversion to glucose were determined, as were blood pressure, pulse pressure and heart rate. 4. The presence of obesity influenced body weight, body fat, de novo fatty acid synthesis, organ weights, glucose mass, glucose oxidation, glucose synthesis, glucose carbon turnover and pulse pressure. 5. It had no effect on glycogen synthesis, tissue glycogen levels, blood glucose, glucose space, or blood pressure. 6. Strain differences were observed in final body weight, organ weights, blood pressure, pulse pressure, hepatic fatty acid synthesis, glucose mass, glucose space, glucose synthesis, liver glycogen levels and glucose conversion to muscle glycogen. 7. Strain-phenotype interaction effects were observed on glucose incorporation into hepatic glycogen, Cori cycle activity, hepatic de novo fatty acid synthesis, final body weight, fat pad weight, heart weight, and mean arterial pressure. 8. These results suggest that although obesity and hypertension are genetic traits in these rats, these traits are independent in their influence on the metabolism of glucose and the development of non-insulin-dependent diabetes mellitus.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Disease Models, Animal; Fatty Acids; Glucose; Glycogen; Hemodynamics; Hypertension; Liver; Male; Muscles; Obesity; Organ Size; Rats; Rats, Inbred SHR

Seven non-insulin-dependent diabetes mellitus (NIDDM) patients participated in three clamp studies performed with [3-3H]- and [U-14C]glucose and indirect calorimetry: study I, euglycemic (5.2 +/- 0.1 mM) insulin (269 +/- 39 pM) clamp; study II, hyperglycemic (14.9 +/- 1.2 mM) insulin (259 +/- 19 pM) clamp; study III, euglycemic (5.5 +/- 0.3 mM) hyperinsulinemic (1650 +/- 529 pM) clamp. Seven control subjects received a euglycemic (5.1 +/- 0.2 mM) insulin (258 +/- 24 pM) clamp. Glycolysis and glucose oxidation were quantitated from the rate of appearance of 3H2O and 14CO2; glycogen synthesis was calculated as the difference between body glucose disposal and glycolysis. In study I, glucose uptake was decreased by 54% in NIDDM vs. controls. Glycolysis, glycogen synthesis, and glucose oxidation were reduced in NIDDM patients (P < 0.05-0.001). Nonoxidative glycolysis and lipid oxidation were higher. In studies II and III, glucose uptake in NIDDM was equal to controls (40.7 +/- 2.1 and 40.7 +/- 1.7 mumol/min.kg fat-free mass, respectively). In study II, glycolysis, but not glucose oxidation, was normal (P < 0.01 vs. controls). Nonoxidative glycolysis remained higher (P < 0.05). Glycogen deposition increased (P < 0.05 vs. study I), and lipid oxidation remained higher (P < 0.01). In study III, hyperinsulinemia normalized glycogen formation, glycolysis, and lipid oxidation but did not normalize the elevated nonoxidative glycolysis or the decreased glucose oxidation. Lipid oxidation and glycolysis (r = -0.65; P < 0.01), and glucose oxidation (r = -0.75; P < 0.01) were inversely correlated. In conclusion, in NIDDM: (a) insulin resistance involves glycolysis, glycogen synthesis, and glucose oxidation; (b) hyperglycemia and hyperinsulinemia can normalize total body glucose uptake; (c) marked hyperinsulinemia normalizes glycogen synthesis and total flux through glycolysis, but does not restore a normal distribution between oxidation and nonoxidative glycolysis; (d) hyperglycemia cannot overcome the defects in glucose oxidation and nonoxidative glycolysis; (e) lipid oxidation is elevated and is suppressed only with hyperinsulinemia.

Topics: Adult; Diabetes Mellitus, Type 2; Fatty Acids, Nonesterified; Female; Glucose; Glycogen; Glycolysis; Humans; Insulin; Lactates; Lactic Acid; Lipid Metabolism; Liver; Male; Middle Aged; Oxidation-Reduction

The effect of a supraphysiological concentration of insulin on gluconeogenesis from L-[U14C] lactate was studied in hepatocytes isolated from control mice and mice made obese by a single injection of gold-thioglucose (GTG). At the time of experimentation (10-12 weeks post GTG injection) the obese mice weighted significantly more than controls (41.7 +/- 0.5 vs. 29.6 +/- 0.8 g respectively; P < 0.001), and exhibited fasting hyperinsulinaemia (35.9 +/- 4.6 vs. 21.3 +/- 4.2 microU/ml; P < 0.05) and hyperglycaemia (16.4 +/- 1.2 vs. 9.2 +/- 1.1 mmol/l; P < 0.001). The amount of lactate converted to glucose by hepatocytes isolated from GTG-obese mice was significantly greater than from lean controls (322 +/- 44 vs. 209 +/- 20 nmol/30 min/10(6) cells; P < 0.05). The addition of 10(-6)M insulin to the incubations significantly reduced lactate conversion to glucose by hepatocytes isolated from control mice (209 +/- 20 vs. 123 +/- 22 nmol/30 min/10(6) cells; P < 0.02), but there was no effect of insulin on glucose production from lactate by hepatocytes isolated from GTG-obese mice (322 +/- 44 vs. 294 +/- 47 nmol/30 min/10(6) cells). Glycogen production and triacylglycerol glycerol production from L-[U14C] lactate were also significantly increased in hepatocytes from GTG-obese mice compared with controls. There was no effect of 10(-6)M insulin on glycogen or triacylglycerol glycerol production from lactate by hepatocytes from GTG-obese mice but the addition of 10(-6)M insulin to the incubations of control hepatocytes significantly reduced the amount of lactate converted to glycogen and triacylglycerol glycerol.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Aurothioglucose; Cells, Cultured; Diabetes Mellitus, Type 2; Disease Models, Animal; Fatty Acids; Gluconeogenesis; Glucose; Glycogen; Insulin; Lactates; Liver; Male; Mice; Mice, Inbred CBA; Obesity; Triglycerides

Insulin resistance and a defective insulin activation of the enzyme glycogen synthase in skeletal muscle during euglycaemia may have important pathophysiological implications in Type 2 (non-insulin-dependent) diabetes mellitus. Hyperglycaemia may serve to compensate for these defects in Type 2 diabetes by increasing glucose disposal through a mass action effect. In the present study, rates of whole-body glucose oxidation and glucose storage were measured during fasting hyperglycaemia and isoglycaemic insulin infusion (40 mU.m-2.min-1, 3 h) in 12 patients with Type 2 diabetes. Eleven control subjects were studied during euglycaemia. Biopsies were taken from the vastus lateralis muscle. Fasting and insulin-stimulated glucose oxidation, glucose storage and muscle glycogen synthase activation were all fully compensated (normalized) during hyperglycaemia in the diabetic patients. The insulin-stimulated increase in muscle glycogen content was the same in the diabetic patients and in the control subjects. Besides hyperglycaemia, the diabetic patients had elevated muscle free glucose and glucose 6-phosphate concentrations. A positive correlation was demonstrated between intracellular free glucose concentration and muscle glycogen synthase fractional velocity insulin activation (0.1 mmol/l glucose 6-phosphate: r = 0.65, p less than 0.02 and 0.0 mmol/l glucose 6-phosphate: r = 0.91, p less than 0.0001). In conclusion, this study indicates an important role for hyperglycaemia and elevated muscle free glucose and glucose 6-phosphate concentrations in compensating (normalizing) intracellular glucose metabolism and skeletal muscle glycogen synthase activation in Type 2 diabetes.

Topics: Blood Glucose; C-Peptide; Calorimetry; Diabetes Mellitus, Type 2; Enzyme Activation; Fasting; Female; Glucose; Glucose Clamp Technique; Glycated Hemoglobin; Glycogen; Glycogen Synthase; Humans; Hyperglycemia; Insulin; Male; Middle Aged; Muscles; Reference Values

Topics: Biological Transport, Active; Diabetes Mellitus, Type 2; Glucose; Glucose-6-Phosphate; Glucosephosphates; Glycogen; Humans; Kinetics; Magnetic Resonance Spectroscopy; Male; Muscles

Peripheral insulin resistance in type II diabetes mellitus has been attributed to alterations in skeletal muscle glucose metabolism. However the direct dose-response relationship between insulin and glucose transport has not yet been studied in human skeletal muscle. We investigated 3-0-methylglucose transport in in vitro incubated skeletal muscle strips from eight healthy controls (age 61 +/- 6 yrs) and six lean type II diabetic patients treated with oral antidiabetic medication (age 73 +/- 3 yrs). Rectus abdominis muscle samples (approximately 1 g), obtained during elective abdominal surgery, were clamped at their resting length in vivo, whereupon strips (20-50 mg) were prepared for in vitro incubation. Measurements of high-energy phosphates and glycogen levels revealed that the muscle strips maintained energy levels during the incubation period. Glucose transport responded to insulin in a dose-response manner in the control group, with a 2-fold increase following maximal stimulation. Muscle strips from the diabetic group demonstrated a marked decrease in the insulin dose-response curve (P less than 0.01), when compared to healthy muscle strips. At a maximal insulin concentration (10,000 microU x ml-1), the response of the diabetic muscle tissue was 50% less than that of the healthy control tissue (P less than 0.05). This report demonstrates a dose-response curve for insulin stimulated 3-0-methylglucose transport in in vitro incubated human skeletal muscle strips. Furthermore, in type II diabetic muscle, our results provide evidence for one or several defects at a postreceptor level.

Topics: 3-O-Methylglucose; Aged; Biological Transport; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Female; Glycogen; Humans; In Vitro Techniques; Insulin; Male; Methylglucosides; Middle Aged; Muscles; Phosphates

To define the mechanisms of impaired muscle glycogen synthase and reduced glycogen formation in non-insulin dependent diabetes mellitus (NIDDM), glycogen synthase activity was kinetically analyzed during the basal state and three glucose clamp studies (insulin approximately equal to 300, 700, and 33,400 pmol/liter) in eight matched nonobese NIDDM and eight control subjects. Muscle glycogen content was measured in the basal state and following clamps at insulin levels of 33,400 pmol/liter. NIDDM subjects had glucose uptake matched to controls in each clamp by raising serum glucose to 15-20 mmol/liter. The insulin concentration required to half-maximally activate glycogen synthase (ED50) was approximately fourfold greater for NIDDM than control subjects (1,004 +/- 264 vs. 257 +/- 110 pmol/liter, P less than 0.02) but the maximal insulin effect was similar. Total glycogen synthase activity was reduced approximately 38% and glycogen content was approximately 30% lower in NIDDM. A positive correlation was present between glycogen content and glycogen synthase activity (r = 0.51, P less than 0.01). In summary, defects in muscle glycogen synthase activity and reduced glycogen content are present in NIDDM. NIDDM subjects also have less total glycogen synthase activity consistent with reduced functional mass of the enzyme. These findings and the correlation between glycogen synthase activity and glycogen content support the theory that multiple defects in glycogen synthase activity combine to cause reduced glycogen formation in NIDDM.

Topics: Diabetes Mellitus, Type 2; Enzyme Activation; Glucose; Glucose-6-Phosphate; Glucosephosphates; Glycogen; Glycogen Synthase; Humans; Insulin; Kinetics; Male; Muscles

Topics: Animals; Diabetes Mellitus; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Insulin; Islets of Langerhans

Ten newly presenting, untreated, Europid Type 2 diabetic patients were studied before and after 8 weeks treatment with intensive diet alone. Nine normal control subjects were also studied. The degree of activation of skeletal muscle glycogen synthase (GS) was used as an intracellular marker of insulin action, prior to and during a 240-min insulin infusion (100 mU kg-1 h-1). Fasting blood glucose decreased from 12.1 +/- 0.9 (+/- SE) to 9.2 +/- 0.8 mmol l-1 (p less than 0.01), but there was no change in fasting insulin concentrations, 9.9 +/- 2.3 vs 9.3 +/- 2.1 mU l-1. Fractional GS activity did not increase in the Type 2 diabetic patients during the insulin infusion either at presentation (change -1.5 +/- 1.9%) or after treatment (change +0.9 +/- 1.8%), and was markedly decreased compared with the control subjects (change +14.5 +/- 2.8%, both p less than 0.001). Glucose requirement during the clamp was decreased in the Type 2 diabetic patients at presentation (2.2 +/- 0.7 vs 7.3 +/- 0.6 mg kg-1 min-1, p less than 0.001), and despite improvement following dietary treatment to 3.3 +/- 0.6 mg kg-1 min-1 (p less than 0.01) remained lower than in the control subjects (p less than 0.001). Fasting plasma non-esterified fatty acid (NEFA) concentrations were elevated at presentation (p less than 0.05), and failed to suppress normally during the insulin infusion. After treatment fasting NEFA concentrations decreased (p less than 0.05) and suppressed normally (p less than 0.05). Insulin secretion was assessed following an intravenous bolus of glucose (0.5 g kg-1) at euglycaemia before and after treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Diet, Diabetic; Enzyme Activation; Fasting; Female; Glucose Tolerance Test; Glycated Hemoglobin; Glycogen; Glycogen Synthase; Humans; Insulin; Insulin Secretion; Male; Middle Aged; Muscles; Reference Values

Because skeletal muscle plays a major role in glucose disposal, it may be the primary site of insulin resistance in non-insulin-dependent diabetes mellitus (NIDDM). Rates of glycogen synthesis (GS), glucose utilization via glycolysis, glycolytic utilization (GU), and glucose transport (GT) were studied in epitrochlearis muscles (EMs) obtained from 10-wk-old nonfasted Sprague-Dawley rats in which NIDDM was neonatally induced with streptozocin. Plasma glucose in NIDDM rats was elevated (P less than 0.001), whereas plasma insulin was similar in NIDDM and control rats. No differences in muscle weight, protein, glycogen, ATP, phosphocreatine, lactate, lactate-pyruvate ratios, or glucose-6-phosphate were noted in EMs of control and NIDDM rats. EMs were incubated in medium containing 5.6 or 11.2 mM glucose with tracer D-[5-3H]glucose and insulin from 0 to 7.18 x 10(-7) M for 1 or 2 h, and GS, GT, and GU were evaluated. Similar rates of basal (non-insulin-mediated) and insulin-stimulated GS, GU, and GT were observed in EMs of NIDDM and control rats incubated in 5.6 mM glucose for 2 h. Insulin dose-response curves revealed similar sensitivities and responsiveness. Increasing glucose concentration (from 5.6 to 11.2 mM) induced significant increases in basal rates of GS, GU, and GT in EMs of control but not NIDDM rats. Insulin dose-response curves for GS and GT revealed decreased sensitivity and no change in responsiveness in EMs of control and NIDDM rats, even though GU of EMs of NIDDM rats was significantly lower at basal and all other insulin concentrations. These data revealed that both insulin resistance and glucose resistance contribute to the impaired glucose metabolism in EMs of the NIDDM rat.

Topics: Adenosine Triphosphate; Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glucose; Glucosephosphates; Glycogen; Humerus; Insulin; Insulin Resistance; Lactates; Lactic Acid; Muscles; Organ Size; Phosphocreatine; Proteins; Pyruvates; Pyruvic Acid; Rats; Rats, Inbred Strains

Topics: Blood Glucose; Diabetes Mellitus, Type 2; Glycogen; Humans; Muscles

Topics: Diabetes Mellitus, Type 2; Glycogen; Humans; Muscles

Increased hepatic glucose output is the main cause of fasting hyperglycemia in non-insulin dependent diabetes mellitus. Due to difficulties in obtaining a quantitative estimate of gluconeogenesis in vivo, the relative contribution of gluconeogenesis and glycogenolysis to this increased hepatic glucose output was unknown. The application in vivo of a new isotopic approach based on a mathematical model of the Krebs cycle enabled us to obtain a quantitative estimate of gluconeogenesis in vivo. Using this approach, gluconeogenesis was found to account for approximately 28% and approximately 97% of overall hepatic glucose output in healthy volunteers in the postabsorptive and in the fasted state respectively. When this technique was used to compare gluconeogenesis rates in non-insulin dependent diabetes mellitus and nondiabetic patients, gluconeogenesis was found to be increased threefold in the patients with non-insulin dependent diabetes mellitus (12.7 +/- 1.6 mu vs 3.6 +/- 0.6 mumol/Kg/min) and to be significantly correlated with fasting plasma glucose. Furthermore, the increase in gluconeogenesis could explain more than 80% of the increase in overall hepatic glucose output in patients with non-insulin dependent diabetes mellitus. In conclusion, in non-insulin dependent diabetes mellitus, gluconeogenesis, as measured by a new isotopic technique, is increased and this increase represents the main cause for increased overall hepatic glucose output and fasting hyperglycemia.

Topics: Carbon Radioisotopes; Diabetes Mellitus, Type 2; Gluconeogenesis; Glucose; Glycogen; Humans; Liver; Male

To examine the extent to which the defect in insulin action in subjects with non-insulin-dependent diabetes mellitus (NIDDM) can be accounted for by impairment of muscle glycogen synthesis, we performed combined hyperglycemic-hyperinsulinemic clamp studies with [13C]glucose in five subjects with NIDDM and in six age- and weight-matched healthy subjects. The rate of incorporation of intravenously infused [1-13C]glucose into muscle glycogen was measured directly in the gastrocnemius muscle by means of a nuclear magnetic resonance (NMR) spectrometer with a 15.5-minute time resolution and a 13C surface coil. The steady-state plasma concentrations of insulin (approximately 400 pmol per liter) and glucose (approximately 10 mmol per liter) were similar in both study groups. The mean (+/- SE) rate of glycogen synthesis, as determined by 13C NMR, was 78 +/- 28 and 183 +/- 39 mumol-glucosyl units per kilogram of muscle tissue (wet weight) per minute in the diabetic and normal subjects, respectively (P less than 0.05). The mean glucose uptake was markedly reduced in the diabetic (30 +/- 4 mumol per kilogram per minute) as compared with the normal subjects (51 +/- 3 mumol per kilogram per minute; P less than 0.005). The mean rate of nonoxidative glucose metabolism was 22 +/- 4 mumol per kilogram per minute in the diabetic subjects and 42 +/- 4 mumol per kilogram per minute in the normal subjects (P less than 0.005). When these rates are extrapolated to apply to the whole body, the synthesis of muscle glycogen would account for most of the total-body glucose uptake and all of the nonoxidative glucose metabolism in both normal and diabetic subjects. We conclude that muscle glycogen synthesis is the principal pathway of glucose disposal in both normal and diabetic subjects and that defects in muscle glycogen synthesis have a dominant role in the insulin resistance that occurs in persons with NIDDM.

Topics: Blood Glucose; Carbon Radioisotopes; Diabetes Mellitus, Type 2; Glycogen; Humans; Insulin; Magnetic Resonance Spectroscopy; Male; Middle Aged; Muscles

Vanadate has insulin-like activity in vitro and in vivo. To characterize the in vivo mechanism of action of vanadate, we examined meal tolerance, insulin-mediated glucose disposal, in vivo liver and muscle glycogen synthesis, and in vitro glycogen synthase activity in 90% partially pancreatectomized rats. Four groups were studied: group I, sham-operated controls; group II, diabetic rats; group III, diabetic rats treated with vanadate; and group IV, diabetic rats treated with phlorizin. Insulin sensitivity, assessed with the euglycemic hyperinsulinemic clamp technique in awake, unstressed rats, was reduced by approximately 28% in diabetic rats. Both vanadate and phlorizin treatment completely normalized meal tolerance and insulin-mediated glucose disposal. Muscle glycogen synthesis was reduced by approximately 80% in diabetic rats (P less than 0.01) and was completely restored to normal by vanadate, but not by phlorizin treatment. Glycogen synthase activity was reduced in skeletal muscle of diabetic rats (P less than 0.05) compared with controls and was increased to supranormal levels by vanadate treatment (P less than 0.01). Phlorizin therapy did not reverse the defect in muscle glycogen synthase. These results suggest that (a) the defect in muscle glycogen synthesis is the major determinant of insulin resistance in diabetic rats; (b) both vanadate and phlorizin treatment normalize meal tolerance and insulin sensitivity in diabetic rats; (c) vanadate treatment specifically reverses the defect in muscle glycogen synthesis in diabetic rats. This effect cannot be attributed to the correction of hyperglycemia because phlorizin therapy had no direct influence on the glycogenic pathway.

Topics: Animals; Chronic Disease; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Glucose Clamp Technique; Glycogen; Glycogen Synthase; Hyperglycemia; Insulin; Liver; Male; Muscles; Phlorhizin; Rats; Rats, Inbred Strains; Vanadates

Glucose turnover determined with tritiated isotopes of glucose is subject to potential error due to glucose/glucose-6-phosphate cycling and/or cycling through glycogen. To determine the extent to which these processes alter the apparent pattern of postprandial glucose metabolism, we measured glucose turnover simultaneously with [2(3)H] glucose (an isotope that minimally cycles through glycogen but is extensively detritiated during glucose/glucose-6-phosphate cycling) and [3(3)H] glucose (an isotope that is not detritiated during glucose/glucose-6-phosphate cycling but can cycle through glycogen). Glucose turnover was measured in patients with non-insulin-dependent diabetes mellitus (NIDDM) and nondiabetic subjects both before and after ingestion of a carbohydrate meal isotopically with labeled [6(14)C] glucose. In the postabsorptive state hepatic glucose appearance was higher (P less than .05) when determined with [2(3)H] glucose than with [3(3)H] glucose in the diabetic patients, but not in the nondiabetic subjects. After glucose ingestion the integrated responses of glucose appearance, systemic entry of ingested glucose, and hepatic glucose release all were higher (P less than .05) when determined with [2(3)H] glucose compared to [3(3)H] glucose in both the diabetic and nondiabetic subjects. However, the absolute difference between glucose turnover measured with [2(3)H] and [3(3)H] glucose were similar in the diabetic and nondiabetic subjects. Both isotopes provided a similar assessment of postprandial carbohydrate metabolism, indicating that either isotope can be used with equal efficacy to compare postprandial carbohydrate metabolism in patients with NIDDM and nondiabetic subjects.

Topics: Adult; Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Eating; Female; Glucagon; Glucose; Glycogen; Humans; Insulin; Liver; Male; Middle Aged; Random Allocation; Time Factors

Adult cats determined by clinical laboratory evaluations to be normal, impaired glucose tolerant, or overtly diabetic were used to explore prospectively the relationships among pancreatic beta cell islet amyloid polypeptide (IAPP) immunoreactivity, islet amyloid (IA) deposition, and diabetogenesis. IAPP-derived IA was found in 11 of 14 (79%) diabetic cats, in four of nine (44%) impaired glucose tolerant cats, and in two of eight (25%) normal adult cats. The presence of IA even in very small amounts, therefore, predicts a very high probability (88%) that an animal has either impaired glucose tolerance or overt DM. Although all overtly diabetic cats had a marked decrease or absence of beta cell IAPP immunoreactivity, six of six cats with impaired glucose tolerance retained IAPP immunoreactivity with 1:15,000 dilutions of antisynthetic IAPP 7-17, whereas only one of seven normal cats had IAPP immunoreactivity beyond 1:10,000 dilutions. These findings suggest that increased IAPP production preceding the development of overt DM is linked to the progressive formation of insoluble IA deposits that are apparent in most overtly diabetic individuals. Of most importance, in that IAPP has been reported to inhibit both basal and insulin-stimulated rates of glycogen synthesis, is the possibility that increased production and release of IAPP by pancreatic beta cells plays a key role in the development of the insulin resistance and impaired glucose tolerance, both of which occur in Type 2 DM.

Topics: Amyloid; Animals; Cats; Diabetes Mellitus, Type 2; Glucose; Glucose Tolerance Test; Glycogen; Immunohistochemistry; Insulin Resistance; Islets of Langerhans; Peptides

Topics: Adolescent; Adult; Aged; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glycogen; Humans; Hypoglycemic Agents; Middle Aged; Neutrophils; Peroxidase

Diabetes-associated peptide has recently been isolated and characterized from the amyloid of the islets of Langerhans in type 2 (non-insulin-dependent) diabetics, and immunoreactivity with antibodies to the peptide has been demonstrated in islet B cells of both normal and type 2 diabetic subjects. In view of the evidence presented in this paper that this 37-amino acid peptide may be a hormone present in normal individuals, we now propose the name "amylin" to replace "diabetes-associated peptide." Because increased amylin, deposited as amyloid within the islets of Langerhans, is characteristic of type 2 diabetes, the study below was performed to examine the possible effects of amylin on peripheral glucose metabolism. Whole amylin was synthesized by using solid-phase techniques, with formation of the disulfide linkage by oxidation in dilute aqueous solution and recovery of the peptide by lyophilization. The effects of amylin on glucose metabolism were studied in two preparations in vitro, isolated rat soleus muscle strips and isolated rat adipocytes. In skeletal muscle exposed to 120 nM amylin for 1 hr, there was a marked decrease in both basal and submaximally insulin-stimulated rates of glycogen synthesis, which resulted in significant reduction in the rates of insulin-stimulated glucose uptake. In muscles treated with amylin there was no response at the concentration of insulin required to stimulate glucose uptake half-maximally in untreated (control) muscles. In marked contrast, amylin had no effect on either basal or insulin-stimulated rates of glucose incorporation into either CO2 or triacylglycerol in isolated adipocytes. Therefore, amylin may be a factor in the etiology of the insulin resistance in type 2 diabetes mellitus, as both deposition of the peptide in islet amyloid and decreased rates of glucose uptake and glycogen synthesis in skeletal muscle are characteristic of this condition.

Topics: Amyloid; Animals; Diabetes Mellitus, Type 2; Glucose; Glycogen; Humans; Insulin; Islet Amyloid Polypeptide; Lactates; Male; Muscles; Rats

Mild diabetes was induced in adult rats with streptozotocin (45 mg/kg body weight), and insulin sensitivity, glycogen deposition and glycogen synthase activity assessed in liver and muscle 5 weeks later. Diabetic rats had significantly elevated fasting blood glucose concentrations (5.6 +/- 0.1 versus 3.6 +/- 0.1 mmol/l, p less than 0.001), and blood glucose concentrations 2 h after a 1 g/kg glucose load (12.0 +/- 0.6 versus 3.7 +/- 0.2 mmol/l, p less than 0.001). After a 20-h fast hepatic glucose output was significantly elevated (58 +/- 3 versus 47 +/- 3 mumol.min-1.kg-1, p less than 0.05), and failed to suppress at high insulin concentrations during a euglycaemic clamp (hepatic glucose output 21 +/- 4 versus 2 +/- 4 mumol.min-1.kg-1, p less than 0.02). Liver glycogen was lower in the diabetic rats by the end of the clamp (16 +/- 3 versus 38 +/- 6 mumol/g wet wt, p less than 0.05). At the end of the clamp total glucose turnover was lower in the diabetic rats (107 +/- 4 versus 161 +/- 17 mumol.min-1.kg-1, p less than 0.01), as was skeletal muscle glycogen synthase activity (0.46 +/- 0.04 versus 0.67 +/- 0.05 U/g wet wt, p less than 0.01) and glycogen concentration (22 +/- 2 versus 33 +/- 3 mumol/g wet wt, p less than 0.05). Blood lactate and pyruvate responses suggested that glycolytic pathways were similarly affected. Thus, insulin insensitivity develops in both liver and skeletal muscle after 5 weeks of mild streptozotocin-induced diabetes.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Fasting; Glycogen; Glycogen Synthase; Insulin; Insulin Secretion; Kinetics; Liver; Liver Glycogen; Male; Muscles; Rats; Rats, Inbred Strains; Reference Values

We studied glucose metabolism in non-insulin-dependent diabetic (NIDDM) men with and without glycogen-depleting cycle exercise 12 h beforehand and have compared the results to our previous data in lean and obese subjects. Rates of total glucose utilization, glucose oxidation, nonoxidative glucose disposal (NOGD), glucose metabolic clearance rate (MCR), and endogenous glucose production (EGP) were determined with a "two-level" insulin-clamp technique (100-min infusions at 40 and 400 mU X m-2 X min-1) combined with indirect calorimetry and D-3-[3H]glucose infusion. Muscle biopsy specimens from vastus lateralis were analyzed for glycogen content and glycogen synthase activity before and after insulin infusions. After exercise, NIDDM subjects had muscle glycogen concentrations comparable with those of lean and obese subjects. The activation of glycogen synthase both by prior exercise and insulin infusion was similar to lean controls. After exercise, total glucose disposal was significantly increased during the 40-mU X m-2 X min-1 infusion (P less than .05), but the increase observed during the 400-mU X m-2 X min-1 infusion was not significant. These increases after exercise were the result of significantly higher NOGD during both levels of insulin infusion. The MCR of glucose during both insulin infusions was reduced in NIDDM compared with lean subjects but was very similar to that in obese nondiabetics. Basal EGP was significantly reduced on the morning after exercise (4.03 +/- 0.27 vs. 3.21 +/- 0.21 mg x kg-1 fat-free mass x min-1) (P less than .05) and associated with significant reductions of fasting plasma glucose (197 +/- 12 vs. 164 +/- 9 mg/dl).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Blood Glucose; C-Peptide; Diabetes Mellitus, Type 2; Glucose; Glycogen; Glycogen Synthase; Humans; Insulin; Insulin Resistance; Male; Muscles; Physical Exertion

We studied the measurement of hepatic glucose output (HGO) with prolonged [3-3H]glucose infusion in 14 patients with non-insulin-dependent diabetes mellitus (NIDDM). Over the course of 10.5 h, plasma glucose concentration fell with fasting by one-third, from 234 +/- 21 to 152 +/- 12 mg/dl, and HGO fell from 2.35 +/- 0.18 to 1.36 +/- 0.07 mg . kg-1 . min-1 (P less than .001). In the basal state, HGO and glucose were significantly correlated (r = 0.68, P = .03), and in individual patients, HGO and glucose were closely correlated as both fell with fasting (mean r = 0.79, P less than .01). Plasma [3-3H]glucose radioactivity approached a steady state only 5-6 h after initiation of the primed continuous infusion, and a 20% overestimate of HGO was demonstrated by not allowing sufficient time for tracer labeling of the glucose pool. Assumption of steady-state instead of non-steady-state kinetics in using Steele's equations to calculate glucose turnover resulted in a 9-24% overestimate of HGO. Stimulation of glycogenolysis by glucagon injection demonstrated no incorporation of [3-3H]glucose in hepatic glycogen during the prolonged tracer infusion. In a separate study, plasma glucose was maintained at fasting levels (207 +/- 17 mg/dl) for 8 h with the glucose-clamp technique. Total glucose turnover rates remained constant during this prolonged tracer infusion. However, HGO fell to 30% of the basal value simply by maintaining fasting hyperglycemia in the presence of basal insulin levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Blood Glucose; Diabetes Mellitus, Type 2; Fasting; Female; Glucagon; Glucose; Glycogen; Humans; Insulin; Kinetics; Liver; Male; Middle Aged; Tritium

Electron microscopic studies of 13 cases of liver fibrosis associated with longstanding diabetes mellitus are presented. Comparing the centrilobular and periportal areas of the lobules there appeared to be a difference in the amounts of glycogen, fat, collagen and the character of mesenchymal cells. Centrilobularly, collagenization of sinusoids and pericellular accumulation of fibres appeared with the destruction of hepatic cells. In the periportal regions there was an accumulation of Kupffer's cells with active phagocytosis and centrally of Ito's cells and fibroblasts. The presented alterations were similar to the central pericellular fibrosis described in connection with morbid obesity. Despite the large number of available data on liver fibrosis there is still no satisfactory explanation for the pathomechanism of centrilobularly developing fibrosis.

Topics: Biopsy; Collagen; Diabetes Mellitus, Type 2; Glycogen; Humans; Liver; Liver Cirrhosis; Microscopy, Electron; Tissue Distribution

The relationship between metabolic control and glycogen lymphocyte content in diabetes mellitus, was studied. 30 insulin-treated and 30 type II diabetic subjects were evaluated with 40 age and sex matched normal controls. Glycaemic control was evaluated by a fasting and 2 h post-prandial plasma glucose and by glycosylated hemoglobin (GHb). Glycogen lymphocyte content was determined by calculation of the PAS-positive Index of the lymphocytes (PIL) according to Skrabalo. While fasting and post-prandial plasma glucose values were significantly higher in insulin-treated than in type II diabetes (p less than 0.001), no differences in GHb values were observed between the two groups (10.31 +/- 0.23% vs 9.80 +/- 0.36%). The mean PIL values were not different in these two groups (0.11 +/- 0.01 vs 0.12 +/- 0.02), but they were significantly higher when compared with control values (0.03 +/- 0.004, p less than 0.001), PIL was positively correlated with GHb in both insulin-treated (r = 0.76, p less than 0.001) and type II diabetes (r = 0.55, p less than 0.001). A correlation between PIL and plasma glucose values was observed only in the insulin-treated group and was weaker (p less than 0.005). No correlation was observed between glycogen lymphocyte content and glucose tolerance in the control group. These data confirm that diabetes mellitus is characterized by a significant increase of PAS-positive lymphocyte content and that it correlates well with glycaemic control.

Topics: Adolescent; Adult; Aged; Blood Glucose; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Female; Glycated Hemoglobin; Glycogen; Humans; Lymphocytes; Male; Middle Aged; Periodic Acid-Schiff Reaction