RESUMO
Glycogen, a complex branched glucose polymer, is responsible for sugar storage in blood glucose homeostasis. It comprises small ß particles bound together into composite α particles. In diabetic livers, α particles are fragile, breaking apart into smaller particles in dimethyl sulfoxide, DMSO; they are however stable in glycogen from healthy animals. We postulate that the bond between ß particles in α particles involves hydrogen bonding. Liver-glycogen fragility in normal and db/db mice (an animal model for diabetes) is compared using various hydrogen-bond breakers (DMSO, guanidine and urea) at different temperatures. The results showed different degrees of α-particle disruption. Disrupted glycogen showed changes in the mid-infra-red spectrum that are related to hydrogen bonds. While glycogen α-particles are only fragile under harsh, non-physiological conditions, these results nevertheless imply that the bonding between ß particles in α particles is different in diabetic livers compared to healthy, and is probably associated with hydrogen bonding.
Assuntos
Ligação de Hidrogênio , Animais , Camundongos , Dimetil Sulfóxido/química , Glicogênio Hepático/metabolismo , Ureia/química , Guanidina/química , Guanidina/farmacologia , Fígado/metabolismo , MasculinoRESUMO
Liver glycogen is a highly branched glucose polymer found as ß 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.
Assuntos
Diabetes Mellitus Tipo 2 , Glicogênio Hepático , Humanos , Camundongos , Animais , Glicogênio Hepático/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Projetos Piloto , Proteômica , Glicogênio/metabolismo , Fígado/metabolismoRESUMO
Liver fibrosis (LF) leads to liver failure and short survival. Liver glycogen is a hyperbranched glucose polymer, comprising individual ß particles, which can bind together to form aggregated α particles. Glycogen functionality depends on its molecular structure. This study compared the molecular structure of liver glycogen from both LF and healthy rats, and explored underlying mechanisms for observed differences. Glycogen from both groups contained α and ß particles; the LF group contained a higher proportion of ß particles, with the glycogen containing fewer long chains than seen in the control group. Both glycogen branching enzyme and glycogen phosphorylase showed a significant decrease of activity in the LF group. Transcriptomics and proteomics revealed a functional deficiency of mitochondria in the LF group, which may lead to changes in glycogen structure. These results provide for the first time an understanding of how liver fibrosis affects liver glycogen metabolism and glycogen structure. HYPOTHESIS: We hypothesized that the molecular structure of liver glycogen from a rat model of liver fibrosis would be altered compared to the control group.
Assuntos
Cirrose Hepática/metabolismo , Glicogênio Hepático/metabolismo , Animais , Configuração de Carboidratos , Cirrose Hepática/patologia , Glicogênio Hepático/química , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Glycogen is a complex branched glucose polymer found in many tissues and acts as a blood-glucose buffer. In the liver, smaller ß glycogen particles can bind into larger composite α particles. In mouse models of diabetes, these liver glycogen particles are molecularly fragile, breaking up into smaller particles in the presence of solvents such as dimethyl sulfoxide (DMSO). If this occurs in vivo, such a rapid enzymatic degradation of these smaller particles into glucose could exacerbate the poor blood-glucose control that is characteristic of the disease. High-amylose resistant starch (RS) can escape digestion in the small intestine and ferment in the large intestine, which elicits positive effects on glycemic response and type 2 diabetes. Here we postulate that RS would help attenuate diabetes-related liver glycogen fragility. Normal maize starch and two types of high-amylose starch were fed to diabetic and non-diabetic mice. Molecular size distributions and chain-length distributions of liver glycogen from both groups were characterized to test glycogen fragility before and after DMSO treatment. Consistent with the hypothesis that high blood glucose is associated with glycogen fragility, a high-amylose RS diet prevented the fragility of liver-glycogen α particles. The diets had no significant effect on the glycogen chain-length distributions.
Assuntos
GlicogênioRESUMO
Liver glycogen α particles are molecularly fragile in diabetic mice, and readily form smaller ß particles, which degrade more rapidly to glucose. This effect is well associated with the loss of blood-glucose homeostasis in diabetes. The biological mechanism of such fragility is still unknown; therefore, there are perceived opportunities that could eventually lead to new means to manage type 2 diabetes. The hierarchical structures of glycogen particles are controlled by the underlying biosynthesis/degradation process that involves various enzymes, including, for example, glycogen synthase (GS) and glycogen-branching enzyme (GBE). Recent studies have shown that fragile glycogen α particles in diabetic mice have longer chains and a higher molecular density compared to wild-type mice, indicating an enhanced enzymatic activity ratio of GS to GBE in diabetes. Furthermore, it has been shown that with an improved blood glucose homeostasis, the glycogen fragility in diabetic mice can be restored by treatment with active ingredients from traditional Chinese medicine, yet the underlying mechanism is unknown. In this review, we summarize recent advances in understandings glycogen fragility from the perspectives of glycogen biosynthesis/degradation, glycogen hierarchical structures, and its relation to diabetes. Importantly, we for the first time set GS/GBE activity ratio as the therapeutic target for diabetes.
Assuntos
Diabetes Mellitus/tratamento farmacológico , Glicogênio/química , Glicogênio/metabolismo , Hipoglicemiantes/farmacologia , Fígado/metabolismo , Animais , Sistemas de Liberação de Medicamentos , Glucose/metabolismo , HumanosRESUMO
Glycogen is a complex branched glucose polymer. Liver glycogen in db/db mouse, a type-2 diabetic mouse model, has been found to be more molecularly fragile than in healthy mice. Size-exclusion chromatography was employed in this study to investigate the molecular structure of liver glycogen in two types of type 1 diabetic mouse models (NOD and C57BL/6J mice), sacrificed at various times throughout the diurnal cycle, and the fragility of liver glycogen after exposure to a hydrogen-bond disruptor were tested. Type 1 diabetic mice exhibit a similar glycogen fragility with that observed for db/db mice. This eliminates many of the potential causes for glycogen molecular fragility; the most likely explanation is that it is caused by high blood-glucose level and/or insulin deficiency, both phenotypes being common to both type 1 and type 2 diabetic mice. This result suggests ways towards new drug targets for the management of diabetes.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Glicogênio/metabolismo , Animais , Glicemia/metabolismo , Peso Corporal , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Tipo 1/sangue , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Glycogen is a complex branched glucose polymer functioning as a blood-sugar reservoir in animals. Liver glycogen ß particles can bind together to form α particles, which have a slower enzymatic degradation to glucose. The linkage between ß particles in α particles in diabetic liver breaks (is fragile) in dimethyl sulfoxide (DMSO), a H-bond disruptor, consistent with blood-sugar homeostasis loss in diabetes. We examined diurnal changes in the molecular structure of healthy and diabetic mouse-liver glycogen. Healthy mouse glycogen was fragile to DMSO during glycogen synthesis but not degradation; diabetic glycogen was always fragile. Two alternative mechanisms for this are suggested: healthy glycogen is fragile when formed and becomes stable during subsequent degradation, a process damaged in diabetes; alternatively, there are two types of glycogen: one compact but fragile and the other loose but non-fragile. This suggests potential types of diabetes drug targets through modifying the activities of glycogen synthesis enzymes.
Assuntos
Relógios Circadianos , Diabetes Mellitus Experimental/metabolismo , Glicogênio/metabolismo , Fígado/metabolismo , Animais , Diabetes Mellitus Experimental/genética , Glicogênio/química , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Correlations among barley protein, starch molecular structure and grain size were determined using 30 barley samples with variable protein contents. Starch molecular structure was characterized by fluorophore-assisted carbohydrate electrophoresis and by size-exclusion chromatography (SEC, also termed GPC). The chain-length distributions of amylopectin were fitted using a mathematical model reflecting the relative activities of starch branching enzymes and starch synthase enzymes. Increased protein content significantly and negatively correlated with higher amounts of amylose with longer chains (degree of polymerization, DP 1600-40000) while barley grain sizes positively associated with starch contents. Protein content also positively correlated with the proportion of longer chains of amylopectin (DP 34-100). These results showed that the enzyme activities of starch synthases change with protein content, leading to altered starch contents, structures and grain sizes. From this perspective, selecting for large grain size (or low protein content) does not necessarily relate to starch structure, although may suggest long chains of amylopectin. Measuring starch structure could give a good indication of process performance in human food, animal feed and brewing, as all these structural features contribute to significant functional properties.
RESUMO
Glycogen is a highly branched glucose polymer which is involved in maintaining blood-sugar homeostasis. Liver glycogen contains large composite α particles made up of linked ß particles. Previous studies have shown that the binding which links ß particles into α particles is impaired in diabetic mice. The present study reports the first molecular structural characterization of human-liver glycogen from non-diabetic patients, using transmission electron microscopy for morphology and size-exclusion chromatography for the molecular size distribution; the latter is also studied as a function of time during acid hydrolysis in vitro, which is sensitive to certain structural features, particularly glycosidic vs. proteinaceous linkages. The results are compared with those seen in mice and pigs. The molecular structural change during acid hydrolysis is similar in each case, and indicates that the linkage of ß into α particles is not glycosidic. This result, and the similar morphology in each case, together imply that human liver glycogen has similar molecular structure to those of mice and pigs. This knowledge will be useful for future diabetes drug targets.