Liver Glycogen Synthesis Research Articles

Dietary trimethylamine N-oxide exacerbates impaired glucose tolerance in mice fed a high fat diet

Trimethylamine N-oxide (TMAO) is an oxidation product of trimethylamine (TMA) and is present in many aquatic foods. Here, we investigated the effects of TMAO on glucose tolerance in high fat diet (HFD)-fed mice. Male C57BL/6 mice were randomly assigned to the control, high fat (HF), and TMAO groups. The HF group was fed a diet containing 25% fat, and the TMAO group was fed the HFD plus 0.2% TMAO for 4 weeks. After 3 weeks of feeding, oral glucose tolerance tests were performed. Dietary TMAO increased fasting insulin levels and homeostasis model assessment-estimated insulin resistance (HOMA-IR) and exacerbated the impaired glucose tolerance in HFD-fed mice. These effects were associated with the expression of genes related to the insulin signal pathway, glycogen synthesis, gluconeogenesis and glucose transport in liver. mRNA levels of the pro-inflammatory cytokine MCP-1 increased significantly and of the anti-inflammatory cytokine IL-10 greatly decreased in adipose tissue. Our results suggest that dietary TMAO exacerbates impaired glucose tolerance, obstructs the hepatic insulin signaling pathway, and causes adipose tissue inflammation in mice fed a high fat diet.

Sterol Regulatory Element-binding Protein-1 (SREBP-1) Is Required to Regulate Glycogen Synthesis and Gluconeogenic Gene Expression in Mouse Liver

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.

Activity of glycogen synthase and glycogen phosphorylase in normal and cirrhotic rat liver during glycogen synthesis from glucose or fructose

Cirrhotic patients often demonstrate glucose intolerance, one of the possible causes being a decreased glycogen-synthesizing capacity of the liver. At the same time, information about the rates of glycogen synthesis in the cirrhotic liver is scanty and contradictory. We studied the dynamics of glycogen accumulation and the activity of glycogen synthase (GS) and glycogen phosphorylase (GP) in the course of 120min after per os administration of glucose or fructose to fasted rats with CCl4-cirrhosis or fasted normal rats. Blood serum and liver pieces were sampled for examinations. In the normal rat liver administration of glucose/fructose initiated a fast accumulation of glycogen, while in the cirrhotic liver glycogen was accumulated with a 20min delay and at a lower rate. In the normal liver GS activity rose sharply and GPa activity dropped in the beginning of glycogen synthesis, but 60min later a high synthesis rate was sustained at the background of a high GS and GPa activity. Contrariwise, in the cirrhotic liver glycogen was accumulated at the background of a decreased GS activity and a low GPa activity. Refeeding with fructose resulted in a faster increase in the GS activity in both the normal and the cirrhotic liver than refeeding with glucose. To conclude, the rate of glycogen synthesis in the cirrhotic liver is lower than in the normal one, the difference being probably associated with a low GS activity.

The Glycogen Synthase 2 Gene (Gys2) Displays Parallel Evolution Between Old World and New World Fruit Bats

Frugivorous and nectarivorous bats rely largely on hepatic glycogenesis and glycogenolysis for postprandial blood glucose disposal and maintenance of glucose homeostasis during short time starvation, respectively. The glycogen synthase 2 encoded by the Gys2 gene plays a critical role in liver glycogen synthesis. To test whether the Gys2 gene has undergone adaptive evolution in bats with carbohydrate-rich diets in relation to their insect-eating sister taxa, we sequenced the coding region of the Gys2 gene in a number of bat species, including three Old World fruit bats (OWFBs) (Pteropodidae) and two New World fruit bats (NWFBs) (Phyllostomidae). Our results showed that the Gys2 coding sequences are highly conserved across all bat species we examined, and no evidence of positive selection was detected in the ancestral branches leading to OWFBs and NWFBs. Our explicit convergence test showed that posterior probabilities of convergence between several branches of OWFBs, and the NWFBs were markedly higher than that of divergence. Three parallel amino acid substitutions (Q72H, K371Q, and E666D) were detected among branches of OWFBs and NWFBs. Tests for parallel evolution showed that two parallel substitutions (Q72H and E666D) were driven by natural selection, while the K371Q was more likely to be fixed randomly. Thus, our results suggested that the Gys2 gene has undergone parallel evolution on amino acid level between OWFBs and NWFBs in relation to their carbohydrate metabolism.

Molecular mechanisms of the alternative lipogenic function of insulin

The proposed hypothesis suggests that major function of insulin is stimulation of triglyceride accumulation in adipose tissue and glycogen synthesis in liver and muscles. The impairment of insulin functioning diminishes triglyceride storage in adipose tissue, elevates the level of its metabolite in periphery and suppresses glucose intake by cells. Leptin disturbs direct insulin action on adipocytes, and prevents fat accumulation. Leptin deficiency or impairment of its functioning facilitate lipogenic effect of insulin, and induce obesity. Lipodystrophy decreases leptin secretion and enhances triglyceride production activated by insulin. Triglycerides are not accumulated in adipose tissue because of its deficiency, and overwhelm peripheral tissues. Lipid metabolites decrease glucose consumption and induce lipoatrophic diabetes. The hypothesis on the lipogenic insulin functioning is confirmed by specific knockout of Insr gene in only tissue: muscles, adipose tissue and other, and by the restoration of its expression in transgenic mice.

Is There a Specific Role for Sucrose in Sports and Exercise Performance?

The consumption of carbohydrate before, during, and after exercise is a central feature of the athlete's diet, particularly those competing in endurance sports. Sucrose is a carbohydrate present within the diets of athletes. Whether sucrose, by virtue of its component monosaccharides glucose and fructose, exerts a meaningful advantage for athletes over other carbohydrate types or blends is unclear. This narrative reviews the literature on the influence of sucrose, relative to other carbohydrate types, on exercise performance or the metabolic factors that may underpin exercise performance. Inference from the research to date suggests that sucrose appears to be as effective as other highly metabolizable carbohydrates (e.g., glucose, glucose polymers) in providing an exogenous fuel source during endurance exercise, stimulating the synthesis of liver and muscle glycogen during exercise recovery and improving endurance exercise performance. Nonetheless, gaps exist in our understanding of the metabolic and performance consequences of sucrose ingestion before, during, and after exercise relative to other carbohydrate types or blends, particularly when more aggressive carbohydrate intake strategies are adopted. While further research is recommended and discussed in this review, based on the currently available scientific literature it would seem that sucrose should continue to be regarded as one of a variety of options available to help athletes achieve their specific carbohydrate-intake goals.

Liver Glycogen Loading Dampens Glycogen Synthesis Seen in Response to Either Hyperinsulinemia or Intraportal Glucose Infusion

The purpose of this study was to determine the effect of liver glycogen loading on net hepatic glycogen synthesis during hyperinsulinemia or hepatic portal vein glucose infusion in vivo. Liver glycogen levels were supercompensated (SCGly) in two groups (using intraportal fructose infusion) but not in two others (Gly) during hyperglycemic-normoinsulinemia. Following a 2-h control period during which fructose infusion was stopped, there was a 2-h experimental period in which the response to hyperglycemia plus either 4× basal insulin (INS) or portal vein glucose infusion (PoG) was measured. Increased hepatic glycogen reduced the percent of glucose taken up by the liver that was deposited in glycogen (74 ± 3 vs. 53 ± 5% in Gly+INS and SCGly+INS, respectively, and 72 ± 3 vs. 50 ± 6% in Gly+PoG and SCGly+PoG, respectively). The reduction in liver glycogen synthesis in SCGly+INS was accompanied by a decrease in both insulin signaling and an increase in AMPK activation, whereas only the latter was observed in SCGly+PoG. These data indicate that liver glycogen loading impairs glycogen synthesis regardless of the signal used to stimulate it.

Skeletal muscle insulin resistance promotes increased hepatic de novo lipogenesis, hyperlipidemia, and hepatic steatosis in the elderly.

Aging is closely associated with muscle insulin resistance, hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 diabetes. We examined the hypothesis that muscle insulin resistance in healthy aging promotes increased hepatic de novo lipogenesis (DNL) and hyperlipidemia by altering the distribution pattern of postprandial energy storage. Healthy, normal weight, sedentary elderly subjects pair-matched to young subjects were given two high-carbohydrate meals followed by 13C/1H magnetic resonance spectroscopy measurements of postprandial changes in muscle and liver glycogen and lipid content, and assessment of DNL using 2H2O. Net muscle glycogen synthesis was reduced by 45% (P < 0.007) in the elderly subjects compared with the young, reflecting severe muscle insulin resistance. Net liver glycogen synthesis was similar between groups (elderly, 143 ± 23 mmol/L vs. young, 138 ± 13 mmol/L; P = NS). Hepatic DNL was more than twofold higher in the elderly than in the young subjects (elderly, 14.5 ± 1.4% vs. young, 6.9 ± 0.7%; P = 0.00015) and was associated with approximately threefold higher postprandial hepatic triglyceride (TG) content (P < 0.005) and increased fasting plasma TGs (elderly, 1.19 ± 0.18 mmol/L vs. young, 0.74 ± 0.11 mmol/L; P = 0.02). These results strongly support the hypothesis that muscle insulin resistance in aging promotes hyperlipidemia and NAFLD by altering the pattern of postprandial carbohydrate storage away from muscle glycogen and into hepatic DNL.

Preventive effect of phycocyanin from Spirulina platensis on alloxan-injured mice

The preventive effect of phycocyanin (obtained from Spirulina platensis) on alloxan-injured mice is investigated. Oral administration of phycocyanin was started two weeks before an alloxan injury and continued until four weeks later. Tests resulted in the following positive results of oral phycocyanin administration on alloxan-injured mice: decrease fasting blood glucose and glycosylated serum protein (GSP); maintain total antioxidative capability (T-AOC); avert malondialdehyde (MDA) formation in the liver, kidney, and pancreas; decrease total cholesterol (TC) level and triglycerides (TG) level in serum and liver; increase the levels of hepatic glycogen level; maintain glucokinase (GK) expression in the liver and decrease p53 expression in the pancreas at mRNA level. The histological observations also supported the above results. Acute toxicity study further shows that phycocyanin is relatively safe. These results led to the conclusion that phycocyanin has significant preventive effect on alloxan-injured mice. The inhibition of p53 pathway could be one of the mechanisms that led to the protection of pancreatic islets from alloxan injury. We also proposed that GK expression that functions to promote liver glycogen synthesis could be the reason for reduced blood glucose level. The encouraging results are the first step in studying the potential of phycocyanin as a clinical measure in preventing diabetes.

KNOXing on the Door of Selective Insulin Resistance

Insulin controls systemic nutrient homeostasis by promoting anabolic processes in various tissues, including the stimulation of glucose influx (into muscle and adipose), protein and glycogen synthesis (in muscle and liver), lipid synthesis and storage (in liver and adipose), and the inhibition of fatty acid oxidation, glycogenolysis, gluconeogenesis, and apoptosis and autophagy (especially in a damaged heart).1–3 Resistance to the action of insulin is associated with chronic physiological and inflammatory stress that develops during obesity and inactivity. In too many instances it progresses to type 2 diabetes mellitus when compensatory hyperinsulinemia fails to maintain glucose homeostasis. A combination of compensatory hyperinsulinemia and inconsistent levels of insulin resistance across body tissues can cause life-threatening sequela, especially nonalcoholic fatty liver disease and atherosclerosis.3,4 Thus, dissection of the insulin signaling pathways and the molecular mechanisms of tissue-specific insulin resistance might reveal novel strategies to arrest or reverse the progression of metabolic disease. See accompanying article on page 1236 Cell-based studies initiated decades ago and extended most recently with mouse genetics reveal a common insulin signaling cascade that begins by activation of the insulin receptor tyrosine kinase (IR) and propagates through the insulin receptor substrates (IRS1 and IRS2) to the phosphatidyl inositol 3 kinase→ v-akt murine thymoma viral oncogene homolog (AKT) cascade 1. AKT plays a particularly broad role as it phosphorylates many protein substrates—including the direct phosphorylation and inactivation of FoxO (forkhead box protein O1 family of transcription factors, FoxO1 and FoxO3) and the indirect phosphorylation of CRTC2 (cAMP response element binding protein–regulated transcription coactivator 2) that inactivates cAMP response element binding protein. Inactivation of these factors suppresses the expression of many hepatic genes, including phosphoenolpyruvate carboxykinase and glucose-6-phosphatase that promote gluconeogenesis (Figure).5,6 Genetic disruption of hepatic insulin signaling by the ablation of the …

Exposure of perfluorononanoic acid suppresses the hepatic insulin signal pathway and increases serum glucose in rats

Exposure to perfluorononanoic acid (PFNA), an increasingly persistent organic pollutant that has been detected in abiotic and biotic matrices, has been demonstrated to cause hepatotoxicity in animals. However, the effects of PFNA on hepatic glucose metabolism have not been fully characterized. In this study, male rats were exposed to 0, 0.2, 1 or 5mg/kg/d PFNA for 14 days to explore the specific effect of PFNA on hepatic glycometabolism and its underlying mechanisms. The results showed that administration of 5mg/kg/d PFNA significantly increased serum glucose and hepatic glycogen in rats. Quantitative real-time PCR analysis showed that PFNA exposure changed the expression levels of several genes related to hepatic glucose metabolism, such as the glucose-6-phosphatase (G6PC) gene and the glucose transporter 2 (GLUT2) gene, which were upregulated, and the glucokinase (GCK) gene and the phosphoinositide-3-kinase, catalytic, alpha polypeptide (PI3Kca) gene, which were decreased. The protein expression levels of phospho-insulin receptor 1(IRS1), phospho-PI3K, phospho-AKT and phospho-phosphoinositide-dependent kinase 1 (PDK1) were decreased in the livers of rats that received 5mg/kg/d PFNA. The expression of phospho-glycogen synthase kinase-3 beta (GSK3β, Ser 9) was increased, which explains the augment of hepatic glycogen. Significant increases in hydrogen peroxide (H2O2) and malondialdehyde (MDA) were found in the livers of 5mg/kg/d PFNA-treated rats. Thus, exposure to PFNA disordered glucose metabolism via inhibiting hepatic insulin signal pathway, accelerating the output of glucose and increasing glycogen synthesis in the rat liver. Furthermore, the oxidative stress induced by PFNA may be involved in this process.

Experimental hyperprolinemia induces mild oxidative stress, metabolic changes, and tissue adaptation in rat liver

The present study investigated the effects of chronic hyperprolinemia on oxidative and metabolic status in liver and serum of rats. Wistar rats received daily subcutaneous injections of proline from their 6th to 28th day of life. Twelve hours after the last injection the rats were sacrificed and liver and serum were collected. Results showed that hyperprolinemia induced a significant reduction in total antioxidant potential and thiobarbituric acid-reactive substances. The activities of the antioxidant enzymes catalase and superoxide dismutase were significantly increased after chronic proline administration, while glutathione (GSH) peroxidase activity, dichlorofluorescin oxidation, GSH, sulfhydryl, and carbonyl content remained unaltered. Histological analyses of the liver revealed that proline treatment induced changes of the hepatic microarchitecture and increased the number of inflammatory cells and the glycogen content. Biochemical determination also demonstrated an increase in glycogen concentration, as well as a higher synthesis of glycogen in liver of hyperprolinemic rats. Regarding to hepatic metabolism, it was observed an increase on glucose oxidation and a decrease on lipid synthesis from glucose. However, hepatic lipid content and serum glucose levels were not changed. Proline administration did not alter the aminotransferases activities and serum markers of hepatic injury. Our findings suggest that hyperprolinemia alters the liver homeostasis possibly by induction of a mild degree of oxidative stress and metabolic changes. The hepatic alterations caused by proline probably do not implicate in substantial hepatic tissue damage, but rather demonstrate a process of adaptation of this tissue to oxidative stress. However, the biological significance of these findings requires additional investigation.

Fructose and Galactose Enhance Postexercise Human Liver Glycogen Synthesis

Both liver and muscle glycogen stores play a fundamental role in exercise and fatigue, but the effect of different CHO sources on liver glycogen synthesis in humans is unclear. The aim was to compare the effect of maltodextrin (MD) drinks containing galactose, fructose, or glucose on postexercise liver glycogen synthesis. In this double-blind, triple crossover, randomized clinical trial, 10 well-trained male cyclists performed three experimental exercise sessions separated by at least 1 wk. After performing a standard exercise protocol to exhaustion, subjects ingested one of three 15% CHO solutions, namely, FRU (MD + fructose, 2:1), GAL (MD + galactose, 2:1), or GLU (MD + glucose, 2:1), each providing 69 g CHO·h(-1) during 6.5 h of recovery. Liver glycogen changes were followed using (13)C magnetic resonance spectroscopy. Liver glycogen concentration increased at faster rates with FRU (24 ± 2 mmol·L(-1)·h(-1), P < 0.001) and with GAL (28 ± 3 mmol·L(-1)·h(-1), P < 0.001) than with GLU (13 ± 2 mmol·L(-1)·h(-1)). Liver volumes increased (P < 0.001) with FRU (9% ± 2%) and with GAL (10% ± 2%) but not with GLU (2% ± 1%, NS). Net glycogen synthesis appeared linear and was faster with FRU (8.1 ± 0.6 g·h(-1), P < 0.001) and with GAL (8.6 ± 0.9 g·h(-1), P < 0.001) than with GLU (3.7 ± 0.5 g·h(-1)). When ingested at a rate designed to saturate intestinal CHO transport systems, MD drinks with added fructose or galactose were twice as effective as MD + glucose in restoring liver glycogen during short-term postexercise recovery.

Tracing metabolic routes of dietary carbohydrate and protein in rainbow trout (Oncorhynchus mykiss) using stable isotopes ([13C]starch and [15N]protein): effects of gelatinisation of starches and sustained swimming

Here we examined the use of stable isotopes, [¹³C]starch and [¹⁵N]protein, as dietary tracers to study carbohydrate assimilation and distribution and protein utilisation, respectively, by rainbow trout (Oncorhynchus mykiss). The capacity of glucose uptake and use by tissues was studied, first, by varying the digestibility of carbohydrate-rich diets (30 % carbohydrate), using raw starch and gelatinised starch (GS) and, second, by observing the effects of two regimens of activity (voluntary swimming, control; sustained swimming at 1·3 body lengths/s, exercise) on the GS diet. Isotopic ratio enrichment (¹³C and ¹⁵N) of the various tissue components (protein, lipid and glycogen) was measured in the liver, muscles, viscera and the rest of the fish at 11 and 24 h after a forced meal. A level of 30 % of digestible carbohydrates in the food exceeded the capacity of rainbow trout to use this nutrient, causing long-lasting hyperglycaemia that raises glucose uptake by tissues, and the synthesis of glycogen and lipid in liver. Total 13C recovered 24 h post-feeding in the GS group was lower than at 11 h, indicating a proportional increase in glucose oxidation, although the deposition of lipids in white muscle (WM) increased. Prolonged hyperglycaemia was prevented by exercise, since sustained swimming enhances the use of dietary carbohydrates, mainly through conversion to lipids in liver and oxidation in muscles, especially in red muscle (RM). Higher recoveries of total 15N for exercised fish at 24 h, mainly into the protein fraction of both RM and WM, provide evidence that sustained swimming improves protein deposition, resulting in an enhancement of the protein-sparing effect.

Glycerol is a major substrate for glucose, glycogen, and nonessential amino acid synthesis in late-term chicken embryos1,2,3

The objective was to determine the contributions of glucose to glycogen synthesis and glycerol to glycogen, glucose, and nonessential AA (NEAA) synthesis on embryonic day (e) 14/15 and e19/20. Chicken embryos from small (56.6 ± 0.88 g) and large eggs (71.7 ± 1.09 g) were repeatedly dosed with either [(13)C(3)]glycerol (14 mg/d for 4 d) or [(13)C(6)]glucose (15 mg/d for 3 d) into the chorio-allantoic fluid before blood and tissue collection. (13)C-Mass isotopomer enrichments in blood glucose, liver, and muscle glycogen, and blood and tissue NEAA were analyzed by mass spectrometry. Glucose metabolism did not differ between small- and large-egg embryos. Although glucose entry was 60% less for e20 compared with e15 embryos, e20 embryos conserved glucose more efficiently as a result of 2- to 3-fold greater (P < 0.001) rates of glucose carbon recycling. Importantly, the direct contribution of glucose to liver glycogen synthesis was minimal on e15, and on e20 direct incorporation of glucose into liver glycogen was only 17%. By comparison, [(13)C(3)]glycerol dosing led to the appearance of [M + 1], [M + 2], and [M + 3] isotopomers in blood glucose and in liver and muscle glycogen on e14 and e19. Here, the (13)C-isotopomer enrichments in blood glucose were ~2-fold greater (P < 0.05) in small- than in large-egg embryos on e14 and e19. Furthermore, [(13)C(3)]glycerol dosing led to substantial labeling of [M + 1], [M + 2], and [M + 3] isotopomers of alanine, aspartate, and glutamate in blood and in tissues where (13)C enrichments were greater (P < 0.05) in liver of small-egg embryos. In summary, this study provides unequivocal evidence that glycerol is a precursor for glucose and NEAA synthesis. Furthermore, glycerol, but not egg-derived glucose, is a major substrate for synthesis of liver and muscle glycogen and is an important anaplerotic substrate for the tricarboxylic acid cycle of embryos during later development.

Hibernating above the permafrost: effects of ambient temperature and season on expression of metabolic genes in liver and brown adipose tissue of arctic ground squirrels

Hibernating arctic ground squirrels (Urocitellus parryii), overwintering in frozen soils, maintain large gradients between ambient temperature (T(a)) and body temperature (T(b)) by substantially increasing metabolic rate during torpor while maintaining a subzero T(b). We used quantitative reverse-transcription PCR (qRT-PCR) to determine how the expression of 56 metabolic genes was affected by season (active in summer vs hibernating), metabolic load during torpor (imposed by differences in T(a): +2 vs -10°C) and hibernation state (torpid vs after arousal). Compared with active ground squirrels sampled in summer, liver from hibernators showed increased expression of genes associated with fatty acid catabolism (CPT1A, FABP1 and ACAT1), ketogenesis (HMGCS2) and gluconeogenesis (PCK1) and decreased expression of genes associated with fatty acid synthesis (ACACB, SCD and ELOVL6), amino acid metabolism, the urea cycle (PAH, BCKDHA and OTC), glycolysis (PDK1 and PFKM) and lipid metabolism (ACAT2). Stage of hibernation (torpid vs aroused) had a much smaller effect, with only one gene associated with glycogen synthesis (GSY1) in liver showing consistent differences in expression levels between temperature treatments. Despite the more than eightfold increase in energetic demand associated with defending T(b) during torpor at a T(a) of -10 vs +2°C, transcript levels in liver and brown adipose tissue differed little. Our results are inconsistent with a hypothesized switch to use of non-lipid fuels when ambient temperatures drop below freezing.

Berberine ameliorates hyperglycemia in alloxan-induced diabetic C57BL/6 mice through activation of Akt signaling pathway

Recently, it is implicated that the abnormality of Akt signaling pathway is involved in the diabetic pathology. Previous studies have demonstrated that berberine could decrease blood glucose by elevating liver glycogen synthesis. However, the underlying mechanism is still unclear. In the present study, we investigated the effects of berberine on fasting blood glucose, liver glycogen, Akt, Glycogen synthase kinase-3, glucokinase and insulin receptor substrate (IRS) in alloxan-induced diabetic mice, exploring its possible hypoglycemic mechanism. We found that in alloxan-induced diabetic mice, the high blood glucose was significantly lowered by berberine treatment. Liver glycogen content, the expression and activity of glucokinase and the phosphorylated Akt and IRS were all significantly reduced in diabetic mice whereas berberine blocked these changes. Berberine also depressed the increasing of phosphorylated GSK-3β in diabetic mice. Collectively, Berberine upregulates the activity of Akt possibly via insulin signaling pathway, eventually lowering high blood glucose in alloxan-induced diabetic mice.

Mechanism of anti-hyperglycemic action of Vatairea macrocarpa (Leguminosae): Investigation in peripheral tissues

Aim of the study Previous studies in our laboratory have demonstrated that the treatment of diabetic rats during 21 days with V. macrocarpa stem-bark ethanolic extract (VmE), reduced glycemia, urinary glucose and urea, increased liver glycogen content and improved other parameters diabetes related. The objective of this study was to evaluate if the anti-hyperglicemic mechanisms of VmE could be caused by improvement in the insulin signaling pathway in the peripheral tissues (liver, adipose and skeletal muscle). Material and methods Streptozotocin-diabetic rats were separated into two groups: diabetic control (DC) and diabetic treated with VmE (DT) during 21 days. The alterations on the insulin signaling in liver, retroperitoneal adipose tissue (RET) and extensor digitorum longus (EDL) muscles were investigated through determination of insulin receptor (IR), protein kinase B/AKT content and AKT phosphorylation levels using Western blotting analysis. This same methodology was used to evaluate the phosphoenolpyruvate carboxykinase (PEPCK) levels in the liver from these animals. Results The treatment with the extract increased the content of IR and the basal phosphorylation of AKT in the three tissues. In the liver from diabetic treated group, the insulin-stimulated AKT phosphorylation was higher and the PEPCK protein levels were reduced. Conclusions Data from this work suggest that the anti-hyperglycemic activity of stem-bark extract of V. macrocarpa can occur through stimulation of insulin signaling pathways in peripheral tissues from diabetic rats, mainly in liver and adipose tissue, probably promoting increase in the glucose uptake and liver glycogen synthesis. The concomitant decreasing in hepatic PEPCK levels could be associated to inhibition of gluconeogenesis, which can also contribute to glycemia reduction.

Impaired Glucose Tolerance and Predisposition to the Fasted State in Liver Glycogen Synthase Knock-out Mice

Conversion to glycogen is a major fate of ingested glucose in the body. A rate-limiting enzyme in the synthesis of glycogen is glycogen synthase encoded by two genes, GYS1, expressed in muscle and other tissues, and GYS2, primarily expressed in liver (liver glycogen synthase). Defects in GYS2 cause the inherited monogenic disease glycogen storage disease 0. We have generated mice with a liver-specific disruption of the Gys2 gene (liver glycogen synthase knock-out (LGSKO) mice), using Lox-P/Cre technology. Conditional mice carrying floxed Gys2 were crossed with mice expressing Cre recombinase under the albumin promoter. The resulting LGSKO mice are viable, develop liver glycogen synthase deficiency, and have a 95% reduction in fed liver glycogen content. They have mild hypoglycemia but dispose glucose less well in a glucose tolerance test. Fed, LGSKO mice also have a reduced capacity for exhaustive exercise compared with mice carrying floxed alleles, but the difference disappears after an overnight fast. Upon fasting, LGSKO mice reach within 4 h decreased blood glucose levels attained by control floxed mice only after 24 h of food deprivation. The LGSKO mice maintain this low blood glucose for at least 24 h. Basal gluconeogenesis is increased in LGSKO mice, and insulin suppression of endogenous glucose production is impaired as assessed by euglycemic-hyperinsulinemic clamp. This observation correlates with an increase in the liver gluconeogenic enzyme phosphoenolpyruvate carboxykinase expression and activity. This mouse model mimics the pathophysiology of glycogen storage disease 0 patients and highlights the importance of liver glycogen stores in whole body glucose homeostasis.

Insulin-Like Growth Factor 2 and the Insulin Receptor, But Not Insulin, Regulate Fetal Hepatic Glycogen Synthesis

Whether insulin or IGFs regulate glycogen synthesis in the fetal liver remains to be determined. In this study, we used several knockout mouse strains, including those lacking Pdx-1 (pancreatic duodenal homeobox-1), Insr (insulin receptor), and Igf2 (IGF-II) to determine the role of these genes in the regulation of fetal hepatic glycogen synthesis. Our data show that insulin deficiency does not alter hepatic glycogen stores, whereas Insr and Igf2 deficiency do. We found that both insulin receptor isoforms (IR-A and IR-B) are present in the fetal liver, and their expression is gestationally regulated. IR-B is highly expressed in the fetal liver; nonetheless, the percentage of hepatic IR-A isoform, which binds Igf2, was significantly higher in the fetus than the adult. In vitro experiments demonstrate that Igf2 increases phosphorylation of hepatic Insr, insulin receptor substrate-2, and Akt proteins and also the activity of glycogen synthase. Igf2 ultimately increased glycogen synthesis in fetal hepatocytes. This increase could be blocked by the phosphoinositide 3-kinase inhibitor LY294008. Taken together, we propose Igf2 as a major regulator of fetal hepatic glycogen metabolism, the insulin receptor as its target receptor, and phosphoinositide 3-kinase as the signaling pathway leading to glycogen formation in the fetal liver.