mRNA Expression Of Glucose Transporter Research Articles

Inhibiting GLUT-1 expression and PI3K/Akt signaling using apigenin improves the radiosensitivity of laryngeal carcinoma in vivo.

Hypoxia is an important factor in radioresistance of laryngeal carcinoma. Glucose transporter-1 (GLUT-1) is an important hypoxic marker in malignant tumors, including laryngeal carcinoma. Apigenin is a natural phytoestrogen flavonoid that has potential anticancer effects. Various studies have reported that the effects of apigenin on lowering GLUT-1 expression were involved in downregulation of the PI3K/Akt pathway. Thus, apigenin may improve the radiosensitivity of laryngeal carcinoma by suppressing the expression of GLUT-1 via the PI3K/Akt pathway. The effect of GLUT-1 and PI3K/Akt pathway-related factor expressions by apigenin or antisense oligonucleotides (AS-ODNs) on the radiosensitivity of laryngeal carcinoma in vivo was assessed. The xenograft volume, xenograft weight and apoptosis detection were performed to determine radiosensitivity. The results showed that apigenin or apigenin plus GLUT-1 AS-ODNs improved the radiosensitivity of xenografts. Apigenin or apigenin plus GLUT-1 reduced the expression of GLUT-1, Akt, and PI3K mRNA after X-ray radiation. We found similar results at the protein level. The results suggest that the effects of apigenin on inhibiting xenograft growth and enhancing xenograft radiosensitivity may be associated with suppressing the expression of GLUT-1 via the PI3K/Akt pathway. In addition, apigenin may enhance the effects of GLUT-1 AS-ODNs via the same mechanism.

Ontogenic Changes of Villus Growth, Lactase Activity, and Intestinal Glucose Transporters in Preterm and Term Born Calves with or without Prolonged Colostrum Feeding.

Oral glucose supply is important for neonatal calves to stabilize postnatal plasma glucose concentration. The objective of this study was to investigate ontogenic development of small intestinal growth, lactase activity, and glucose transporter in calves (n = 7 per group) that were born either preterm (PT; delivered by section 9 d before term) or at term (T; spontaneous vaginal delivery) or spontaneously born and fed colostrum for 4 days (TC). Tissue samples from duodenum and proximal, mid, and distal jejunum were taken to measure villus size and crypt depth, protein concentration of mucosa and brush border membrane vesicles (BBMV), total DNA and RNA concentration of mucosa, mRNA expression and activity of lactase, and mRNA expression of sodium-dependent glucose co-transporter-1 (SGLT1) and facilitative glucose transporter 2 (GLUT2) in mucosal tissue. Additionally, protein expression of SGLT1 in BBMV and GLUT2 in crude mucosal membranes and immunochemical localization of GLUT2 in the enterocytes were determined. Villus height in distal jejunum was lower in TC than in T. Crypt depth in all segments was largest and the villus height/crypt depth ratio in jejunum was smallest in TC calves. Concentration of RNA was highest in duodenal mucosa of TC calves, but neither lactase mRNA and activity nor SGLT1 and GLUT2 mRNA and protein expression differed among groups. Localization of GLUT2 in the apical membrane was greater, whereas in the basolateral membrane was lower in TC than in T and PT calves. Our study indicates maturation processes after birth for mucosal growth and trafficking of GLUT2 from the basolateral to the apical membrane. Minor differences of mucosal growth, lactase activity, and intestinal glucose transporters were seen between PT and T calves, pointing at the importance of postnatal maturation and feeding for mucosal growth and GLUT2 trafficking.

Insulin Control of Blood Glucose and GLUT4 Expression in the Skeletal Muscle of Septic Rats.

Insulin resistance is common in septic patients. The level at which the serum glucose should be maintained using insulin infusions for optimal utilization by skeletal muscles is not yet established. The objective of the present study was to compare glucose transporter 4 (GLUT4) mRNA and GLUT4 expression and glucose utilization at the recommended glucose levels of 6-8 mmol/L (110-140 mg/dL) and 8-10 mmol/L (140-180 mg/dL) in septic rats. This was a prospective randomized study using 44 Sprague-Dawley rats (260-330 g). Rats were anaesthetized with gaseous diethyl ether. Catheters were implanted into the jugular vein and artery. Following a laparotomy, rats in the experimental group (n = 36) were rendered septic by standard caecal ligation and puncture (CLP) and intraperitoneal lipopolysaccharide (LPS) infusion (O111:[B4], 1 mg/kg). Control animals (n = 8) underwent laparotomy, but no caecal ligation or puncture and no LPS injection. Four experimental groups were studied: sham-operated control, sepsis treated with fluid maintenance only, sepsis treated with fluid and insulin infusion controlling blood glucose concentration at 6-8 mmol/L and sepsis treated with fluid and insulin infusion controlling blood glucose concentration at 8-10 mmol/L. Hyperinsulinaemic-euglycaemic clamp experiment was done before fluid maintenance and insulin treatment to calculate average glucose infusion rate. All septic rats were markedly hyperglycaemic compared with sham-operated controls two hours after operation. Glucose infusion rate during hyperinsulinaemic-euglycaemic clamp experiment was slower in septic rats, suggesting that they were insulin resistant. At the 12th and 24th hour, skeletal muscle was taken to observe pathological change and analyse the GLUT4 mRNA and GLUT4 levels. There were more inflammatory cells, less GLUT4 mRNA and GLUT4 expression in the skeletal muscles of septic rats. Insulin increased the expression of GLUT4 mRNA and GLUT4 in the skeletal muscle of septic rats. Among all septic rats, the expression of GLUT4 mRNA and GLUT4 was more in the 8-10 mmol/L group. Blood glucose concentration of 8-10 mmol/L results in more glucose utilization than 6-8 mmol/L in the skeletal muscle of septic rats during insulin therapy.

Effects of different amylose to amylopectin ratios on serum indices related to glucose metabolism and glucose transporter expression in fattening lambs

The objective of this study was to determine the effects of different amylose to amylopectin ratios on serum indices related to glucose metabolism and the mRNA expression of glucose transporters in the small intestinal mucosa of fattening lambs. A total of 36 male 7-day-old lambs of similar weight were randomly assigned to four treatments of iso-starch diets containing tapioca starch, maize starch, wheat starch, and pea starch, with the determined ratio for amylose to amylopectin of 0.12, 0.23, 0.24, and 0.48, respectively. Serum glucose, creatinine, insulin, and glucagon were reduced with the advancing age of lambs. The results indicated that feeding lambs with a pea starch diet (high amylose to amylopectin ratio) had increased serum cholesterol (P=0.047), lactate dehydrogenase (LDH) (P=0.010), and growth hormone (GH) (P=0.037). Pea starch did not influence the serum glucose (P=0.160), but significantly decreased the serum insulin (P=0.021). The pea starch diet significantly up-regulated the mRNA expression of glucose transporters, which may increase intestinal glucose uptake. By contrast, the consumption of tapioca starch resulted in the opposite effects, and significantly down-regulated the mRNA expression of glucose transporters. Collectively, these data suggest that starches with higher amylose to amylopectin ratios promote higher concentrations of plasma cholesterol, LDH and GH, and up regulate the gene expression profile of glucose transporters in fattening lambs.

Effect of Solanum surattense on mitochondrial enzymes in diabetic rats and in vitro glucose uptake activity in L6 myotubes.

Background:S. surattense is widely used in Siddha medicine for various ailments.Objective:The aim was to evaluate the impact of alcoholic leaf-extract of S. surattense on mitochondrial enzymes in streptozotocin (STZ) induced diabetic rats and to study the in vitro muscle glucose uptake activity on L6 myotubes.Materials and Methods:The male albino Wistar rats were randomly divided into five groups of six animals each. Diabetes was induced by intraperitoneal injection of STZ (40 mg/kg body weight). After being confirmed the diabetic rats were treated with alcoholic leaf-extract of S. surattense (100 mg/kg body weight) for 45 days. The biochemical estimations (liver mitochondrial enzymes, antioxidants, thiobarbituric acid reactive substances [TBARS]) and histopathological studies were performed. Further, the in vitro muscle glucose uptake activity in L6 myotubes and messenger RNA (mRNA) expression of glucose transporter-4 (GLUT-4) was performed.Results:In diabetic rats, the activities of liver mitochondrial enzymes were found to be significantly lowered. The mitochondrial TBARS level increased, whereas the activities/level of enzymatic and non-enzymatic antioxidants decreased in diabetic rats. Administration of S. surattense to diabetic rats significantly reversed the above parameters toward normalcy. Furthermore in diabetic rats, the histopathological studies showed growth of adipose tissue and shrinkage of islets in the pancreas, liver showed fatty change with mild inflammation of portal triad, and kidney showed messangial capillary proliferation of glomeruli and fatty infiltration of tubules. Treatment with S. surattense brought back these changes to near normalcy. The extract was analyzed for in vitro muscle glucose uptake activity in L6 myotubes and mRNA expression of GLUT-4 by semi-quantitative reverse transcriptase-polymerase chain reaction. One nano gram per millilitre of S. surattense leaf-extract gave 115% glucose uptake on L6 myotubes. It also showed elevated levels of GLUT-4 mRNA transcripts, when compared with control cells.Conclusion:These studies strongly support the anti-diabetic nature of S. surattense.

Effect of Intermittent Hypoxia and Rimonabant on Glucose Metabolism in Rats: Involvement of Expression of GLUT4 in Skeletal Muscle.

BackgroundObstructive sleep apnea (OSA) and its main feature, chronic intermittent hypoxia (IH) during sleep, is closely associated with insulin resistance (IR) and diabetes. Rimonabant can regulate glucose metabolism and improve IR. The present study aimed to assess the effect of IH and rimonabant on glucose metabolism and insulin sensitivity, and to explore the possible mechanisms.Material/MethodsThirty-two rats were randomly assigned into 4 groups: Control group, subjected to intermittent air only; IH group, subjected to IH only; IH+NS group, subjected to IH and treated with normal saline; and IH+Rim group, subjected to IH and treated with 10 mg/kg/day of rimonabant. All rats were killed after 28 days of exposure. Then, the blood and skeletal muscle were collected. We measured fasting blood glucose levels, fasting blood insulin levels, and the expression of glucose transporter 4 (GLUT4) in both mRNA and protein levels in skeletal muscle.ResultsIH can slow weight gain, increase serum insulin level, and reduce insulin sensitivity in rats. The expressions of GLUT4 mRNA, total GLUT4, and plasma membrane protein of GLUT4 (PM GLUT4) in skeletal muscle were decreased. Rimonabant treatment was demonstrated to improve weight gain and insulin sensitivity of the rats induced by IH. Rimonabant significantly upregulated the expression of GLUT4 mRNA, PM GLUT4, and total GLUT4 in skeletal muscle.ConclusionsThe present study demonstrates that IH can cause IR and reduced expression of GLUT4 in both mRNA and protein levels in skeletal muscle of rats. Rimonabant treatment can improve IH – induced IR, and the upregulation of GLUT4 expression may be involved in this process.

Dysregulation of Glucose Homeostasis Following Chronic Exogenous Administration of Leptin in Healthy Sprague-Dawley Rats.

Impaired glucose utilization is seen in chronic hyperleptinaemia associated conditions such as obesity and type 2 diabetes mellitus. It is unclear if this impaired glucose utilization is due to the effect of persistent hyperleptinaemia on insulin secretion from the beta cells of pancreas. To examine the effects of chronic leptin administration on plasma glucose regulation in rats. Glucose challenge curves were plotted for male Sprague-Dawley rats treated with either normal saline (Control; n=8) or subcutaneous leptin injection for 42 days (60 μg/kg body weight/day; n=8). Plasma glucose and plasma insulin levels were measured at 0, 5, 10, 15, 20 and 25 minutes after glucose challenege. Skeletal muscle tissue was collected at the end of a glucose challenge for glucose transporter-4 protein content, insulin receptor and glucose transporter-4 mRNA expression. Data were analysed using repeated measures and one-way ANOVA with post-hoc analysis. Chronic leptin treatment caused significantly higher fasting insulin level. Post glucose challenge, there was a significant increase in blood glucose levels and insulin level in the leptin treated rats. There was no significant difference in the skeletal muscle glucose transporter-4 content. However, leptin treated rats showed decreased mRNA expression of Insulin Receptor and glucose transporter-4 in the skeletal muscle. Leptin administration for 42 days caused hyperinsulinaemia and decreased the expression of insulin receptors in insulin sensitive tissues leading to the development of an insulin resistance-like state in the rats.

A gold bullet to treat obesity related metabolic disorders

Objectives: Obesity is associated with chronic low grade inflammation. Adipose resident macrophages (ATMs) in obese individuals are known to produce increased levels of proinflammatory cytokines. We have previously shown that intraperitoneal (IP) injection of 20—30 nm gold nanoparticles (AuNPs) in lean mice can induce significant fat loss along with reduced adiposetissue-derived, tumour necrosis factor (TNF) mRNA expression without a concomitant change in ATM number. This study aimed to investigate whether AuNPs also reduce fat mass and metabolic disorders in mice fed a high-fat diet (HFD). Method: Male C57BL/6 mice (6 weeks) were fed a HFD with/without daily IP injection of AuNPs (LAu 0.785 g/g/d, HAu 7.85 g/g/d) for 9 weeks. Control group was fed standard chow with vehicle injection. IP-glucose tolerance test (IP-GTT) was performed a week before the endpoint. mRNA expression of pro-inflammatory cytokines and fat metabolic markers were determined in retroperitoneal fat and liver using real-time PCR. Results: At the end point, body weight was similar between AuNP-treated and non-treated HFD-fed mice, but adiposity and liver weight were reduced by AuNP treatment. Blood glucose levels during IPGTT were significantly lower in AuNP-treated mice compared to the non-treated HFD-fed group. In addition, AuNP treatment improved plasma free fatty acid, triglyceride and cholesterol levels in HFD-fed mice. In the fat tissue, mRNA expression of pro-inflammatory markers was significantly reduced by AuNP treatment, including TNF . In the liver, AuNP treatment downregulated inflammatory markers and improved lipid metabolic markers. Interestingly, liver mRNA expression of glucose transporter-4 was upregulated by AuNP treatment. Conclusion: AuNPs treatment in HFD-fed mice prevented their development of glucose intolerance with improved fat metabolism and reduced inflammation in both the fat and liver. Our results therefore support the potential development of AuNPs as a new therapeutic strategy for obesity related metabolic disorders.

Glucose Transporter 1 Expression Is Regulated by Aged Garlic Extract during Cerebral Ischemia

Aged garlic extract is obtained after prolonged hydroethanolic extraction of garlic cloves and is recognized for its high antioxidant content. Although evidence indicates that aged garlic extract delays brain ischemia-induced damage, the molecular mechanisms of this effect are unclear. Moderate concentration of reactive oxygen species induces glucose transporter 1 (GLUT1) expression that in astrocytessupports brainenergy metabolism. This has suggested that reduction of excessive reactive oxygen species formation during severe ischemia maycontribute to neuroprotection. Thus, in view of the high antioxidant properties of aged garlic extract, we investigated the effect of ischemia on GLUT1 expression and its regulation after aged garlic extract treatment. Rats were subjected to middle cerebral artery occlusion during 2 hto produce ischemia and then were sacrificed after different times of reperfusion. In fronto-parietal cortex GLUT1 mRNA expression peaked after 1and 24h of reperfusion. Nevertheless, no significant change on total protein level was observed by Western blotting, buta severe reduction on GLUT1 positive astrocytes was detected by immunofluorescence after 2 h of reperfusion. Remarkably, aged garlic extract (360 mg/kg;i.p.) administered after is chemia increased GLUT1 mRNA expressionin cortex after 2 h of reperfusion and partially prevented the middle cerebral artery occlusion-induced reduction of GLUT1 in astrocytes. These results suggest that GLUT1 over expression in astrocytes may underlie some of the beneficial effects on neuronal survival observed with aged garlic extract treatment.

Long-acting hypoglycemic effects of PEGylated FGF21 and insulin glargine in mice with type 1 diabetes

ObjectiveIn this study, we compared the long-acting hypoglycemic effect of PEGylated FGF21 (PEG-FGF21) with insulin glargine in mice with STZ-induced type 1 diabetes. MethodsPEG-FGF21 and insulin glargine were administered once daily for two months, and blood glucose was measured prior to the next administration. Real-time PCR was used to measure mRNA expression of glucokinase (GK), glucose 6-phosphatase (G6pase), phosphoenolpyruvate carboxykinase (PEPCK), glucose transporter 1 (GLUT1) and glucose transporter 4 (GLUT4). ResultsDuring long-term treatment, the blood glucose of untreated mice remained at 25.0 to 28.0mmol/L for the whole experiment, and the blood glucose of mice treated with insulin glargine remained at 16.5 to 18.0mmol/L. However, mice treated with PEG-FGF21 had lower blood glucose levels of 8.0 to 9.0mmol/L on day 10 and maintained this level until the end of the experiment. qRT-PCR showed that PEG-FGF21 up-regulated mRNA expression of GK and GLUT1, and down-regulated mRNA expression of G6Pase and PEPCK. Insulin glargine up-regulated mRNA expression of GLUT4, but had no effect on GK, G6Pase, PEPCK or GLUT1. ConclusionsPEG-FGF21 has a better long-acting efficacy than insulin glargine. PEG-FGF21 achieves glucose clearance by accelerating glycolysis by up-regulating expression of GK and GLUT1 and inhibiting gluconeogenesis via down-regulation of G6Pase and PEPCK expression.

Intrafetal glucose infusion alters glucocorticoid signaling and reduces surfactant protein mRNA expression in the lung of the late-gestation sheep fetus.

Increased circulating fetal glucose and insulin concentrations are potential inhibitors of fetal lung maturation and may contribute to the pathogenesis of respiratory distress syndrome (RDS) in infants of diabetic mothers. In this study, we examined the effect of intrafetal glucose infusion on mRNA expression of glucose transporters, insulin-like growth factor signaling, glucocorticoid regulatory genes, and surfactant proteins in the lung of the late-gestation sheep fetus. The numerical density of the cells responsible for producing surfactant was determined using immunohistochemistry. Glucose infusion for 10 days did not affect mRNA expression of glucose transporters or IGFs but did decrease IGF-1R expression. There was reduced mRNA expression of the glucocorticoid-converting enzyme HSD11B-1 and the glucocorticoid receptor, potentially reducing glucocorticoid responsiveness in the fetal lung. Furthermore, surfactant protein (SFTP) mRNA expression was reduced in the lung following glucose infusion, while the number of SFTP-B-positive cells remained unchanged. These findings suggest the presence of a glucocorticoid-mediated mechanism regulating delayed maturation of the surfactant system in the sheep fetus following glucose infusion and provide evidence for the link between abnormal glycemic control during pregnancy and the increased risk of RDS in infants of uncontrolled diabetic mothers.

Short communication: Protein kinase C regulates glucose uptake and mRNA expression of glucose transporter (GLUT) 1 and GLUT8 in lactating bovine mammary epithelial cells

The aim of this study was to determine the role of protein kinase C (PKC) in regulating glucose uptake in lactating bovine mammary epithelial cells (BMEC). The BMEC were cultured and treated with different concentrations of phorbol 12-myristate 13-acetate (PMA;0, 10, 25, 50, 100, and 200ng/mL), the classic activator of PKC, for 48h. Compared with the cells with no PMA treatment, 50 and 100ng of PMA/mL significantly stimulated the glucose uptake of the BMEC, whereas the glucose uptake by the cells treated with the lowest and the highest amounts of PMA (25 and 200ng/mL, respectively) did not show a significant difference. Consistently, the mRNA expression of glucose transporter (GLUT) 1 and 8 showed a similar pattern of increase under the treatments of PMA. Furthermore, when the cells were pretreated with GF1090203X (0, 0.25, 0.5, 1, and 2μM), an inhibitor of PKC, for 30min before exposed to PMA (50ng/mL), the PMA-induced glucose uptake and GLUT1 and GLUT8 expression were decreased by GF1090203X in a dose-dependent manner. These results demonstrate that PKC is involved in the regulation of glucose uptake by BMEC, and this function may work, at least partly, through upregulating the expression of GLUT1 and GLUT8.

Hyperoxia reverses glucotoxicity-induced inhibition of insulin secretion in rat INS-1 β cells.

Chronic hyperglycemia has deleterious effects on pancreatic β-cell function, a process known as glucotoxicity. This study examined whether chronic high glucose (CHG) induces cellular hypoxia in rat INS-1 β cells, and whether hyperoxia (35% O2) can reverse glucotoxicity-induced inhibition of insulin secretion. CHG (33.3 mm, 96 h) reduced insulin secretion, and down-regulated insulin and pancreatic duodenal homeobox factor 1 gene expression. CHG also increased intracellular pimonidazole-protein adducts, a marker for hypoxia. CHG also enhanced hypoxia-inducible factor 1α (HIF-1α) protein expression and its DNA-binding activity, which was accompanied by a decrease in mRNA expression of glucose transporter 2 (GLUT2), glucokinase and uncoupling protein-2 and an increase in mRNA expression of GLUT1 and pyruvate dehydrogenase kinase 1. Hyperoxia restored the decrease in insulin secretion and the gene expression except for GLUT2, and suppressed intracellular hypoxia and HIF-1α activation. These results suggest that glucotoxicity may cause β-cell hypoxia. Hyperoxia might prevent glucotoxicity-induced β-cell dysfunction and improve insulin secretion.

Improved glucose tolerance with restored expression of glucose transporter 4 in C57BL/6 mice after a long period of high-fat diet feeding

This study examined the effects of long-term consumption of a high-fat diet (HFD) on glucose metabolism in C57BL/6 mice. A total of 95 male 6-week-old mice were divided into normal diet (ND) and HFD groups until 16, 26, 36, 47, or 77 weeks of age. Plasma and blood glucose and insulin levels and area under the glucose curve (AUCglucose) during an intraperitoneal glucose tolerance test were measured. Quantitative real-time PCR was performed to determine the mRNA expression levels of glucose transporter 4 (GLUT4) in adipose tissue. AUCglucose levels in HFD mice at 16, 26, and 36 weeks were significantly higher than those in ND mice (p < 0.001) and significantly reduced in HFD mice at 47 and 77 weeks. Nonfasting plasma glucose concentrations in both groups were significantly decreased at 47 and 77 weeks (p ≤ 0.05). Insulin levels in HFD mice at 47 and 77 weeks were 12.5% lower than those at 36 weeks. Relative GLUT4 mRNA expression in adipose tissue of HFD mice was significantly decreased at 26 and 36 weeks and increased at 47 and 77 weeks. These data suggest that the glucose tolerance was impaired by relatively short-term high-fat feeding and significantly improved by very long periods of high-fat feeding with restored GLUT4 expression in adipose tissue in HFD mice. The differential effects of short- and long-term HFD on glucose metabolism must be considered for recharacterizing the diet-induced obesity and clarify the pathogenesis of metabolic disorders in this animal model.

Myostatin alters glucose transporter-4 (GLUT4) expression in bovine skeletal muscles and myoblasts isolated from double-muscled (DM) and normal-muscled (NM) Japanese shorthorn cattle

The purpose of this study was to determine whether myostatin alters glucose transporter-4 (GLUT4) expression in bovine skeletal muscles and myoblasts isolated from double-muscled (DM) and normal-muscled (NM) Japanese Shorthorn cattle. Plasma concentrations of glucose were lower in DM cattle than in NM cattle (P < 0.01). The expression of GLUT4 messenger RNA (mRNA) in the skeletal muscle ex vivo and in myoblasts at 72 h after differentiation in vitro was higher in DM cattle than in NM cattle (P < 0.01). In contrast, the NM and DM cattle did not differ with respect to skeletal muscle expression of GLUT1 and myocyte enhancer factor-2c (MEF2c), a transcription factor of GLUT4. In differentiated myoblasts, the expression of GLUT1, GLUT4, and MEF2c mRNAs was greater in DM cattle than in NM cattle (P < 0.01). In the presence and absence of insulin, glucose uptake in myoblasts was increased in DM cattle relative to that of NM cattle (P < 0.01). The addition of myostatin decreased the expression of GLUT4 and MEF2c mRNAs in DM myoblasts (P < 0.05). Results of the present study suggest that myostatin inhibits the expression of GLUT4 mRNA possibly via MEF2c and that the greater ability of the DM cattle to produce muscle relative to the NM cattle may be due to their greater sensitivity to insulin and greater use of glucose.

Urtica dioica extract attenuates depressive like behavior and associative memory dysfunction in dexamethasone induced diabetic mice

Evidences suggest that glucocorticoids results in depression and is a risk factor for type 2 diabetes. Further diabetes induces oxidative stress and hippocampal dysfunction resulting in cognitive decline. Traditionally Urtica dioica has been used for diabetes mellitus and cognitive dysfunction. The present study investigated the effect of the hydroalcoholic extract of Urtica dioica leaves (50 and 100 mg/kg, p.o.) in dexamethasone (1 mg/kg, i.m.) induced diabetes and its associated complications such as depressive like behavior and cognitive dysfunction. We observed that mice administered with chronic dexamethasone resulted in hypercortisolemia, oxidative stress, depressive like behavior, cognitive impairment, hyperglycemia with reduced body weight, increased water intake and decreased hippocampal glucose transporter-4 (GLUT4) mRNA expression. Urtica dioica significantly reduced hyperglycemia, plasma corticosterone, oxidative stress and depressive like behavior as well as improved associative memory and hippocampal GLUT4 mRNA expression comparable to rosiglitazone (5 mg/kg, p.o.). Further, Urtica dioica insignificantly improved spatial memory and serum insulin. In conclusion, Urtica dioica reversed dexamethasone induced hyperglycemia and its associated complications such as depressive like behavior and cognitive dysfunction.

Long-term intake of rice improves insulin sensitivity in mice fed a high-fat diet

ObjectiveThe aim of this study was to evaluate the effects of rice as a carbohydrate source and its molecular mechanisms on insulin resistance induced by a high-fat diet (HFD). MethodsC57 BL/6 J mice were divided into three groups and were fed a low-fat diet (LFD); a HFD (with 18% fat, 0.5% cholesterol, 51.5% w/w cornstarch and sucrose); or a HFD with rice (HFD-CR, with 18% fat, 0.5% cholesterol and 51.5% w/w rice powder) for 12 wk. In the HFD-CR diet, cooked rice powder was substituted for cornstarch and sucrose in the HFD as a carbohydrate source. ResultsHFD-CR–fed mice had significantly lower body weight, blood glucose, insulin and leptin levels and ameliorated glucose responses with decreased homeostasis model assessment-insulin resistance compared with HFD-fed mice. Hepatic mRNA levels of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase were down-regulated in the HFD-CR group. The hypertrophied islet size and the decreased pancreatic mRNA expression of glucose transporter 2 in the HFD group were normalized with cooked rice consumption. Rice promoted glucose uptake by activating AMP-activated protein kinase and downstream glucose transporter 4 in the skeletal muscle. ConclusionRice consumption as a carbohydrate source might potentiate improvements in glucose uptake via AMP-activated protein kinase activation and glucose transporter 4 expression in the skeletal muscles, thereby improving insulin sensitivity.

Hypoxia-inducible Factor-1α (HIF-1α) Protein Diminishes Sodium Glucose Transport 1 (SGLT1) and SGLT2 Protein Expression in Renal Epithelial Tubular Cells (LLC-PK1) under Hypoxia

In this work, we demonstrated the regulation of glucose transporters by hypoxia inducible factor-1α (HIF-1α) activation in renal epithelial cells. LLC-PK1 monolayers were incubated for 1, 3, 6, or 12 h with 0% or 5% O2 or 300 μm cobalt (CoCl2). We evaluated the effects of hypoxia on the mRNA and protein expression of HIF-1α and of the glucose transporters SGLT1, SGLT2, and GLUT1. The data showed an increase in HIF-1α mRNA and protein expression under the three evaluated conditions (p < 0.05 versus t = 0). An increase in GLUT1 mRNA (12 h) and protein expression (at 3, 6, and 12 h) was observed (p < 0.05 versus t = 0). SGLT1 and SGLT2 mRNA and protein expression decreased under the three evaluated conditions (p < 0.05 versus t = 0). In conclusion, our results suggest a clear decrease in the expression of the glucose transporters SGLT1 and SGLT2 under hypoxic conditions which implies a possible correlation with increased expression of HIF-1α.

Inhibition of Glucose Transporter 1 (GLUT1) Chemosensitized Head and Neck Cancer Cells to Cisplatin

Glucose transporter 1 (GLUT1) facilitates the cellular uptake of glucose and is overexpressed in most cancers. The altered expression of GLUT1 may influence the sensitivity of tumor cells to chemotherapy. This study investigated whether the knockdown of GLUT1 expression to sensitize head and neck cancer cells to the chemotherapy drug cisplatin in vitro. Anti-GLUT1 antibody was used to block activity of GLUT1 protein, and GLUT1-shRNA was used to knock down its mRNA expression in Cal27 cells. Immunocytochemistry, Western blot, and qRT-PCR were used to detect expression of GLUT1 mRNA and protein, respectively. Lentivirus was used to carrying GLUT1-shRNA to knockdown GLUT1 expression in Cal27 cells for MTT and flow cytometry analyses of cell viability and apoptosis, respectively. Glucose uptake assay was used to assess the changes in glucose levels in Cal27 cells. It showed that GLUT1 mRNA and protein were expressed in Cal27 cells, and GLUT1 protein was localized on the cell membrane. Both anti-GLUT1 antibody and GLUT1-shRNA sensitized Cal27 cells to cisplatin treatment under both normoxia and hypoxia conditions. Anti- GLUT1 antibody and GLUT1-shRNA inhibited tumor cell growth in vitro and induced them to undergo apoptosis. GLUT1-shRNA also suppressed tumor cell uptake of glucose into the cells. Our findings suggest that inhibition of GLUT1 activity and expression can sensitize Cal27 cells to cisplatin treatment in both normoxic and hypoxic conditions. These data could be further verified in animal xenografts before potential application as a clinical adjuvant or neoadjuvant therapy of head and neck cancer with cisplatin.

Effects of orexin A on GLUT4 expression and lipid content via MAPK signaling in 3T3-L1 adipocytes

Orexin A regulates food intake, energy metabolism and gastrointestinal function; it also increases glucose uptake and inhibits lipolysis, suggesting a role for orexin A in glucose and lipid metabolism. In this study, the effects of orexin A on glucose transporter 4 (GLUT4) mRNA level and lipid content were explored in 3T3-L1 preadipocytes and adipocytes. Orexin receptor 1 (OX1R) protein expression was determined in the adipose tissue of normal and obese rats. In addition, 3T3-L1 preadipocytes and differentiated 3T3-L1 adipocytes were incubated with different concentrations of orexin A (10−9 to 10−7M), without or with OX1R specific antagonist, then the peroxisome proliferator-activated receptor-γ2 (PPARγ2) mRNA expression was analyzed. Differentiated 3T3-L1 adipocytes were exposed to orexin A, without or with MAPK and OX1R antagonist, after which the GLUT4 and ERK1/2, JNK, and p38 MAPK activation, and triglyceride (TG) content were measured. We observed that OX1R protein expression was decreased in obese rats, and OX1R protein level was negatively correlated with body fat, Lee's index, TG, total cholesterol, and fasting insulin levels. Orexin A enhanced PPARγ2 mRNA expression in a dose-dependent manner in 3T3-L1 preadipocytes through OX1R. In differentiated 3T3-L1 adipocytes, orexin A significantly increased GLUT4 mRNA levels, which was blocked by the ERK1/2, JNK, and p38 MAPK inhibitors as well as OX1R antagonist. Furthermore, orexin A increased cellular TG content via ERK1/2, JNK, and p38 MAPK as well as OX1R. Thus, orexin A increases GLUT4 mRNA expression and lipid accumulation in differentiated 3T3-L1 adipocytes via ERK1/2, JNK, and p38 MAPK signaling. In addition, orexin A increases PPARγ2 mRNA expression in 3T3-L1 preadipocytes. Further studies are necessary to elucidate the impact of orexin A in metabolic disorders and adipocyte differentiation.