Effects of glutamine supplementation, GH, and IGF-I on glutamine metabolism in critically ill patients

During critical illness glutamine deficiency may develop. Glutamine supplementation can restore plasma concentration to normal, but the effect on glutamine metabolism is unknown. The use of growth hormone (GH) and insulin-like growth factor I (IGF-I) to prevent protein catabolism in these patients may exacerbate the glutamine deficiency. We have investigated, in critically ill patients, the effects of 72 h of treatment with standard parenteral nutrition (TPN; n = 6), TPN supplemented with glutamine (TPNGLN; 0.4 g x kg(-1) x day(-1), n = 6), or TPNGLN with combined GH (0.2 IU. kg(-1). day(-1)) and IGF-I (160 microg x kg (-1) x day(-1)) (TPNGLN+GH/IGF-I; n = 5) on glutamine metabolism using [2-(15)N]glutamine. In patients receiving TPNGLN and TPNGLN+GH/IGF-I, plasma glutamine concentration was increased (338 +/- 22 vs. 461 +/- 24 micromol/l, P < 0.001, and 307 +/- 65 vs. 524 +/- 71 micromol/l, P < 0.05, respectively) and glutamine uptake was increased (5.2 +/- 0.5 vs. 7.4 +/- 0.7 micromol x kg(-1) x min(-1), P < 0.05 and 5.2 +/- 1.1 vs. 7.6 +/- 0.8 micromol x kg(-1) x min(-1), P < 0.05). Glutamine production and metabolic clearance rates were not altered by the three treatments. These results suggest that there is an increased requirement for glutamine in critically ill patients. Combined GH/IGF-I treatment with TPNGLN did not have adverse effects on glutamine metabolism.     

Transport of glutamine by rat kidney brush-border membrane vesicles.

I studied the glycosylation in vivo of a viral envelope protein, the glycoprotein of vesicular stomatitis virus (VSV), by pulse labelling of virus-infected HeLa cells with 3H-labelled monosaccharides (mannose, glucosamine). Radioactivity was incorporated into the fraction of membrane-bound polyribosomes, although metabolic conversion of [3H]-mannose into amino acids was negligible. Dissociation of bound polyribosomes revealed that the radioactively co-purified with the peptidyl-tRNA. The nascent peptides were released by alkaline hydrolysis, immunoprecipitated and analysed by polyacrylamide-gel electrophoresis. It is apparent from the size distribution of the [3H]mannose-labelled nascent chains that attachment of carbohydrate starts when approximately half of the amino acid sequence of the viral glycoprotein has been synthesized.     

The simultaneous hydrolysis of glutathione and glutamine by rat kidney gamma-glutamyltransferase

The simultaneous hydrolysis of L-glutamine and glutathione by rat kidney gamma-glutamyltransferase has been studied. At concentrations of the two substrates giving rates approximating to Vmax, it is shown that glutamine hydrolysis predominates. Under saturating conditions for both substrates, 65% of the glutamic acid produced is derived from glutamine. Using [14C]glutamine it is also shown that at physiological plasma concentrations of glutamine (in excess of 400 microM) and glutathione (20 microM) glutamine hydrolysis is the predominating reaction. We therefore suggest that both glutamine and glutathione are important in vivo donors for gamma-glutamyltransferase.     

Ammonia production and pathways of glutamine utilization in rat kidney slices

Ammonia production from glutamine was studied in slices from non-acidotic and acidotic rat kidneys. Slices from non-acidotic kidneys made 53% as much ammonia from d-glutamine as from l-glutamine during the initial 15 min of incubation. Thereafter the production rate from the l-isomers accelerated while that from the d-isomers remained constant. The accelerated rate of ammonia production from l-glutamine was dependent upon tissue swelling since prevention of swelling reduced the production rate. Swelling activates the mitochondrial glutaminase I pathway as evidenced by the rise in ammonia produced per glutamine utilized ratio as well as by the accelerated rate of CO₂ production derived from the oxidative disposal of glutamine's carbon skeleton. Cortical slice swelling activates the mitochondrial pathway in a manner not unlike that seen in vivo during chronic acidosis and may reflect increased permeability to glutamine.Acidotic rat kidneys are not swollen in vivo while cortical slices initially produce 4-fold more ammonia than do non-acidotic slices. After 15 min, this 4-fold difference in total ammonia production drops to only a 2-fold difference due to the swelling-induced activation of the mitochondrial pathway. Consequently, slice swelling obliterates the important fact that ammonia production by the mitochondrial pathway is 15-fold greater in acidotic than in non-acidotic kidneys.     

Alterations in kidney tissue following zinc supplementation to STZ-induced diabetic rats

Diabetes mellitus is a chronic disease characterized by anomalies forming in carbohydrate, lipid, protein metabolisms and the incidence of this disease varies widely throughout the world. Zinc is an important element which is essential for life and is present in nature. In this study, the animals were divided into four groups. These groups were named as untreated; zinc sulfate; streptozotocin (STZ); STZ and zinc sulfate. STZ (65 mg/kg) was dissolved in a freshly prepared 0.01 M pH 4.5 citrate buffer and given with intraperitoneal injection in a single dose. Zinc sulfate (100 mg/kg) was dissolved in distilled water and given to the animals by gavage at a daily dose for 60 days. The rats were sacrificed under ether anesthesia. This study was aimed to investigate histological and biochemical changes of zinc supplementation on the kidney tissue in STZ-induced diabetic rats. In the current study, histological and histochemical observations showed that the occurred degenerative changes decreased after giving zinc in the kidney tissue of diabetic group. Kidney glutathione (GSH) levels decreased and lipid peroxidation (LPO), nonenzymatic glycosylation (NEG), urea and creatinine levels increased in diabetic rats. GSH levels increased, while LPO, NEG, urea and creatinine levels decreased in the kidney with administration of zinc to diabetic rats. As a result, we observed curative effects of zinc given to diabetic rats. We can say that zinc may be an important antioxidant for the treatment of secondary complications of diabetes in kidney tissue.     

Lectin binding in the diabetic rat kidney

In this study metal-conjugated concanavalin A (Con A) and Bandieraea simplicifolia isolectin II (BSA II) have been applied to sections from kidneys of control rats and rats which had untreated diabetes for 70 days or for 200 days. Lectin binding was measured by atomic absorption spectrophotometric analysis of ferritin-iron or hemocyanin-copper. Con A binding increased significantly with diabetes; was totally blocked by alpha-D-mannoside; was not inhibited by fructose lysine; and was enhanced by NaHB4 preincubation. BSA II binding also increased significantly with diabetes.     

Lectin binding in the diabetic rat kidney

Summary In this study metal-conjugated concanavalin A (Con A) andBandieraea simplicifolia isolectin II (BSA II) have been applied to sections from kidneys of controls rats and rats which had untreated diabetes for 70 days or for 200 days. Lectin binding was measured by atomic absorption spectrophotometric analysis of ferritin-iron or hemocyanin-copper.Con A binding increased significantly with diabetes; was totally blocked by alpha-D-mannoside; was not inhibited by fructose lysine; and was enhanced by NaHB₄ preincubation. BSA II binding also increased significantly with diabetes.     

Effects of benfluorex-vitamin C supplementation on cutaneous capillaries of diabetic rats

The ultrastructural changes on capillaries of the dermis in diabetic and benfluorex-vitamin C treated diabetic rats have been investigated. Three groups of 21 Wistar albino rats were used in the examination: control, diabetes, and benfluorex-vitamin C treated diabetic rats. Diabetes was induced by injection of streptozotocin. The streptozotocin-induced group was treated for 21 days with vitamin C and benfluorex, of which antidiabetic and antihyperlipidemic effects were experimentally proved. Samples taken from the skin of rats' legs were examined under transmission electron microscopy. Swollen endothelial cells, narrowed capillary lumens, a thickened basement membrane, and fusion of mitochondrial cristae in the capillaries of diabetic rat dermis were seen. In the benfluorex-vitamin C treated group, contrary to the diabetic group, neither signs of degeneration in endothelial cells nor a significant difference with the control group with regard to capillary structure were observed. Amelioration in capillaries appears to be due to benfluorex and vitamin C treatment in diabetes.     

Effects of glutamine supplementation on the immune status in weaning piglets with intrauterine growth retardation

Neonates with intrauterine growth retardation (IUGR) often suffer from impaired cellular immunity, and weaning may further aggravate adverse effects of IUGR on development and function of the immune system. In this study, we investigated effects of glutamine supplementation on immune status in the intestines of weaning pigs with IUGR, focusing on molecular mechanisms underlying altered immune response. Piglets with IUGR were weaned at 21 days of age and received orally 1.22 g alanine or 1 g glutamine per kg body weight every 12?h. Weight gain and intestinal weight of weaning piglets were increased by glutamine supplementation. Levels of serum IgG in piglets supplemented with glutamine were increased compared with Control piglets. The production of IL-1 and IL-8 in the serum and jejunum was decreased by glutamine supplementation, whereas the levels of IL-4 in the serum and the concentrations of IL-4 and IL-10 in the jejunum were increased. The expression of heat shock protein 70 (Hsp70) in the jejunum was increased by glutamine supplementation, but the degradation of inhibitor?κB and the activity of nuclear factor-κB (NF-κB) were decreased. In conclusion, glutamine supplementation enhanced immune response in weaning piglets with IUGR. The effects of glutamine in IUGR are associated with increased Hsp70 expression and suppression of NF-κB activation.     

Ammonia production and pathways of glutamine utilization in rat kidney slices.

Ammonia production from glutamine was studied in slices from non-acidotic and acidotic rat kidneys. Slices from non-acidotic kidneys made 53% as much ammonia from D-glutamine as from L-glutamine during the initial 15 min of incubation. Thereafter the production rate from the L-isomer accelerated while that from the D-isomer remained constant. The accelerated rate of ammonia production from L-glutamine was dependent upon tissue swelling since prevention of swelling reduced the production rate. Swelling activates the mitochondrial glutaminase I pathway as evidenced by the rise in ammonia produced per glutamine utilized ratio as well as by the accelerated rate of CO2 production derived from the oxidative disposal of glutamin's carbon skeleton. Cortical slice swelling activates the mitochondrial pathway in a manner not unlike that seen in vivo during chronic acidosis and may reflect increased permeability to glutamine. Acidotic rat kidneys are not swollen in vivo while cortical slices initially produce 4-fold more ammonia than do non-acidotic slices. After 15 min, this 4-fold difference in total ammonia production drops to only a 2-fold difference due to the swelling-induced activation of the mitochondrial pathway. Consequently, slice swelling obliterates the important fact that ammonia production by the mitochondrial pathway is 15-fold greater in acidotic than in non-acidotic kidneys.     

Dietary glutamine supplementation enhances endothelial progenitor cell mobilization in streptozotocin-induced diabetic mice subjected to limb ischemia

Diabetes is a metabolic disorder with increased risk of vascular diseases. Tissue ischemia may occur with diabetic vascular complications. Bone marrow-derived endothelial progenitor cells (EPCs) constitute a reparative response to ischemic injury. This study investigated the effects of oral glutamine (GLN) supplementation on circulating EPC mobilization and expression of tissue EPC-releasing markers in diabetic mice subjected to limb ischemia. Diabetes was induced by a daily intraperitoneal injection of streptozotocin for 5 days. Diabetic mice were divided into 2 nonischemic groups and 6 ischemic groups. One of the nonischemic and 3 ischemic groups were fed the control diet, while the remaining 4 groups received diets with identical components except that part of the casein was replaced by GLN. The respective diets were fed to the mice for 3 weeks, and then the nonischemic mice were sacrificed. Unilateral hindlimb ischemia was created in the ischemic groups, and mice were sacrificed at 1, 7 or 21 days after ischemia. Their blood and ischemic muscle tissues were collected for further analyses. Results showed that plasma matrix metallopeptidase (MMP)-9 and the circulating EPC percentage increased after limb ischemia in a diabetic condition. Compared to groups without GLN, GLN supplementation up-regulated plasma stromal cell-derived factor (SDF)-1 and muscle MMP-9, SDF-1, hypoxia-inducible factor-1 and vascular endothelial growth factor gene expression. The CD31-immunoreactive intensities were also higher in the ischemic limb. These findings suggest that GLN supplementation enhanced circulating EPC mobilization that may promote endothelium repair at ischemic tissue in diabetic mice subjected to limb ischemia.     

Effect of glutamine supplementation on neutrophil function in male judoists

Glutamine is an important amino acid for immune function. Though high intensity and prolonged exercise decreases plasma glutamine concentration and causes immune suppression, the relationship between neutrophil functions and glutamine has not yet been found. The purpose of this study was to investigate the impacts of glutamine supplementation on neutrophil function. Twenty‐six male university judoists were recruited. Subjects were classified into glutamine and control groups. The glutamine group ingested 3000 mg of glutamine per day and the control group ingested placebo for 2 weeks. Examinations were performed at the start of preunified loading exercise (pre‐ULE), then 1 and 2 weeks after ULE (post‐ULE). Reactive oxygen species (ROS) production, phagocytic activity, serum opsonic activity and serum myogenic enzymes were measured. Differences between the levels obtained in pre‐ULE and post‐ULE for the two groups were compared. In the glutamine group, ROS production activity increased 1 week after ULE, whereas it was not observed in the control group (P < 0.001). Though myogenic enzymes increased significantly after ULE (P < 0.001), the glutamine group remained unchanged by supplementation during ULE. Glutamine supplementation has prevented excessive muscle damage and suppression of neutrophil function, especially in ROS production activity, even during an intensive training period. Copyright © 2013 John Wiley & Sons, Ltd.     

Effect of glutamine supplementation on exercise-induced changes in lymphocyte function

The purpose of this study was toinvestigate the possible role of glutamine in exercise-inducedimpairment of lymphocyte function. Ten male athletesparticipated in a randomized, placebo-controlled, double-blindcrossover study. Each athlete performed bicycle exercise for 2 hat 75% of maximum O₂ consumption on 2 separate days.Glutamine or placebo supplements were given orally during and up to2 h postexercise. The trial induced postexercise neutrocytosisthat lasted at least 2 h. The total lymphocyte count increased bythe end of exercise due to increase of bothCD3⁺TCR⁺ andCD3⁺TCR⁺ T cells as well asCD3CD16⁺CD56⁺ naturalkiller (NK) cells. Concentrations of CD8⁺ andCD4⁺ T cells lacking CD28 and CD95 on their surfaceincreased more than those of cells expressing these receptors. Withinthe CD4⁺ cells, only CD45RA memory cells, butnot CD45RA⁺ naive cells, increased in response to exercise.Most lymphocyte subpopulations decreased 2 h after exercise.Glutamine supplementation abolished the postexercise decline in plasmaglutamine concentration but had no effect on lymphocyte trafficking, NKand lymphokine-activated killer cell activities, T cell proliferation,catecholamines, growth hormone, insulin, or glucose. Neutrocytosis wasless pronounced in the glutamine-supplemented group, but it is unlikelythat this finding is of any clinical significance. This study does notsupport the idea that glutamine plays a mechanistic role inexercise-induced immune changes.

     

Effects of fucoidan on diabetic rat testicular tissue

Increased reactive oxygen species (ROS) resulting from hyperglycemia and inadequate endogenous antioxidant systems are responsible for the complications of diabetes. ROS accumulate in the cell and stimulate apoptosis, which compromises sperm quality and function. We investigated the possible effects of fucoidan, a potent antioxidant with a regulatory effect on blood glucose homeostasis, on the testicular tissues of rats with experimental diabetes. Diabetes was induced by administering 40 mg/kg streptozotocin (STZ) on five consecutive days. Twenty-four Wistar albino male rats were divided into four groups: group 1, control group (CG); group 2, diabetes group (DG); group 3, early fucoidan group (EFG) treated with 50 mg/kg fucoidan after diabetes induction; group 4, late fucoidan group (LFG) treated with the same dose of fucoidan 15 days after diabetes induction. Fucoidan was administered intraperitoneally every two days for four weeks. Basement membrane thickness and Johnsen scores were higher in the DG than in the CG; no difference was found for either the EFG or LFG compared to the CG. Seminiferous tubule diameters of EFG were significantly greater than for the DG. Apoptotic tubule and apoptotic cell indexes were significantly greater in the DG and significantly less in the EFG and LFG groups compared to the CG. Early use of fucoidan in diabetic individuals may minimize damage to testicular tissue.     

Effect of vanadate administration on polyol pathway in diabetic rat kidney.

Effect of oral administration of sodium orthovanadate for three weeks on polyol pathway in renal cortex and medulla was studied in control and alloxan diabetic rats. An enhancement in aldose reductase in cortex and medulla and sorbitol dehydrogenase in cortex was observed in alloxan diabetic rats. Despite depressed insulin secretion, vanadate treatment to diabetic rats counteracted hyperglycemia, normalized elevated enzyme activities and glucose level, prevented medullary sorbitol accumulation and markedly checked increase in kidney weight. These results show that vanadate causes marked improvement in renal hypertrophy and has an antidiabetogenic effect on polyol pathway in diabetic kidney.     

The effect of ascorbate supplementation on oxidative stress in the streptozotocin diabetic rat.

An increase in oxidative stress may contribute to the development of diabetic complications. The key aqueous-phase chain-breaking antioxidant ascorbate is known to be deficient in diabetes, and we have therefore investigated the effects of ascorbate supplementation on oxidative stress in the streptozotocin diabetic rat. Markers of lipid peroxidation (malondialdehyde [MDA] and diene conjugates) were increased in plasma and erythrocytes of untreated diabetic animals, and levels of the antioxidants ascorbate and retinol were reduced. Plasma tocopherol was unchanged. Insulin treatment normalized MDA and ascorbate levels, although ascorbate metabolism remained disturbed, as indicated by increased levels of dehydroascorbate. High-dose ascorbate supplementation in the absence of insulin treatment restored plasma ascorbate to normal and increased plasma retinol and tocopherol levels. However, MDA and diene conjugate levels remained unchanged, possibly as a result of increased iron availability. High-dose ascorbate supplementation should be approached with caution in diabetes, as ascorbate may exert both antioxidant and prooxidant effects in vivo.