Liver Glycogen Levels Research Articles

A novel layered double hydroxide-hesperidin nanoparticles exert antidiabetic, antioxidant and anti-inflammatory effects in rats with diabetes.

Incidence of diabetes has increased significantly worldwide over recent decades. Our objective was to prepare and characterize a novel nano-carrier of hesperidin to achieve a sustained release of hesperidin and to explore the potency of the novel formula as an antidiabetic agent compared to metformin in type 2 diabetic rats. Hesperidin was loaded on MgAl-layered double hydroxide (LDH). The formula was characterized using Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), transmission electron microscopy, and dynamic light scattering. The release profile of hesperidin and MgAl-LDH-Hesperidin were studied in vitro. The parameters studied in vivowere blood glucose, glycated hemoglobin (HbA1c), insulin, lipid profile, and liver glycogen levels. We also investigated the levels of interleukin (IL)-17, tumor necrosis factor-Alfa (TNF-α), malondialdehyde (MDA), catalase, and the mRNA expression of peroxisome proliferator-activated receptor-gamma (PPARγ) and nuclear factor erythroid 2-related factor-2 (NrF2). There were variations in the XRD patterns and FTIR confirming the physical adsorption of hesperidin on the surface of LDH. The results indicated that the diabetic rats treated with administration of antidiabetic formula, MgAl-LDH-Hesperidin, showed a beneficial effect on the levels of blood glucose, insulin, HbA1c%, and lipid profile, comparing to diabetic control rats. The antidiabetic agent also showed a significant decrease in the levels of TNF-α, IL-17, and MDA, and an increase in the level of catalase. Marked upregulation of the expression levels of mRNA for PPARγ andNrF2 were recorded. The novel nano-hesperidin formula MgAl-LDH-Hesperidin revealed a sustained release of hesperidin and exhibited antidiabetic, antihyperlipidemic, antioxidant, and anti-inflammatory properties, and also is a promising agent for effective delivery of drugs to treat type 2 diabetes.

Growth performance, fillet quality and immunity after challenge with streptococcus agalactiae of red tilapia ( oreochromis sp .) fed diet containing cysteamine and creatine

This study aimed to evaluate the effect of creatine and cysteamine supplementation on the growth performance, fillet quality and resistance to bacterial infection of Streptococcus agalactiae in red tilapia. Five types of experimental diet (quadruplicate) with the same energy and protein content were tested. These consist of N (negative control), A (0.5 g/kg cysteamine), B (0.4 g/kg creatine), C (0.5 g/kg cysteamine plus 0.4 g/kg creatine) and D (0.25 g/kg cysteamine plus 0.2 g/kg creatine). Male red tilapia (Oreochromis sp.), with average body weight of 49.51 ± 4.90 g, were kept in 175-L tank at 10 fish/tank density for 56 days rearing period. The challenge test was carried out through injecting fish (body weight: 152.05 ± 12.2 g) with S. agalactiae 107 CFU/ml as much as 0.10 ml/fish. The results showed that treatment C (0.5 g/kg cysteamine plus 0.4 g/kg creatine) increased (p < 0.05) levels of protein retention, activity of superoxide dismutase (SOD) of the liver (p < 0.05), but decreased levels of malondialdehyde (MDA) and liver glycogen (p < 0.05). This last condition was followed by a decrease (p < 0.05) in plasma protein levels that resulted in a decline (p < 0.05) of water content and MDA of fillet after stored for 24 h. In addition, treatment C increased fish resistance to S. agalactiae infection, demonstrated by higher survival rate and lysozyme activity (p < 0.05). In conclusion, the treatment C boosts protein retention, fillet quality and robustness to pathogenic bacterial infections.

The ameliorative activity of Chrysobalanus orbicularis in streptozotocin-induced type II diabetes mellitus rat model

Chrysobalanus orbicularis is a medicinal plant present in West Africa in the Itsekiri speaking part of Nigeria. It is used conventionally in diabetes mellitus management. This research investigates the ameliorative activity of the aqueous leaf extract of C. orbicularis in a streptozotocin-induced diabetic rat model. Freshly prepared streptozotocin (40 mg/kg body weight [BW]) was administered intraperitoneally to induce diabetes. Three diabetic groups were placed on aqueous leaf extract of C. orbicularis at 11.076, 22.134, and 44.268 mg/kg BW respectively; a group was placed on metformin (44.28 mg/kg BW), and the other two groups were the diabetic control and normal control. The experiment lasted for 28 days, thereafter, fasting blood glucose levels and body weight variations were recorded. Also, glycogen level, antioxidant enzyme, hexokinase and glucose-6-phosphatase activities, malonaldehyde (MDA) as well as glucose transporters 2 and 4 levels were analyzed using standard procedures. Diabetic rats administered aqueous extract of C. orbicularis leaf significantly (p < 0.05) decreased the fasting blood glucose and MDA levels, and glucose-6-phosphatase activity. In addition, administration of aqueous extract of C. orbicularis leaf to diabetic rats demonstrated a momentous increase in liver glycogen level, superoxide dismutase, catalase, glutathione peroxidase, reduced glutathione, glutathione transferase, and hexokinase activities as well as GLUT-2 and GLUT-4 levels. The data from this study suggest that the aqueous extract of C. orbicularis leaf may be beneficial in the management of diabetic mellitus and its secondary effects.

The Mediator complex kinase module is necessary for fructose regulation of liver glycogen levels through induction of glucose-6-phosphatase catalytic subunit (G6pc)

ObjectiveLiver glycogen levels are dynamic and highly regulated by nutrient availability as the levels decrease during fasting and are restored during the feeding cycle. However, feeding in the presence of fructose in water suppresses glycogen accumulation in the liver by upregulating the expression of the glucose-6-phosphatase catalytic subunit (G6pc) gene, although the exact mechanism is unknown. We generated liver-specific knockout MED13 mice that lacked the transcriptional Mediator complex kinase module to examine its effect on the transcriptional activation of inducible target gene expression, such as the ChREBP- and FOXO1-dependent control of the G6pc gene promoter. MethodsThe relative changes in liver expression of lipogenic and gluconeogenic genes as well as glycogen levels were examined in response to feeding standard low-fat laboratory chow supplemented with water or water containing sucrose or fructose in control (Med13fl/fl) and liver-specific MED13 knockout (MED13-LKO) mice. ResultsAlthough MED13 deficiency had no significant effect on constitutive gene expression, all the dietary inducible gene transcripts were significantly reduced despite the unchanged insulin sensitivity in the MED13-LKO mice compared to that in the control mice. G6pc gene transcription displayed the most significant difference between the Med13 fl/fl and MED13-LKO mice, particularly when fed fructose. Following fasting that depleted liver glycogen, feeding induced the restoration of glycogen levels except in the presence of fructose. MED13 deficiency rescued the glycogen accumulation defect in the presence of fructose. This resulted from the suppression of G6pc expression and thus G6PC enzymatic activity.Among two transcriptional factors that regulate G6pc gene expression, FOXO1 binding to the G6pc promoter was not affected, whereas ChREBP binding was dramatically reduced in MED13-LKO hepatocytes. In addition, there was a marked suppression of FOXO1 and ChREBP-β transcriptional activities in MED13-LKO hepatocytes. ConclusionsTaken together, our data suggest that the kinase module of the Mediator complex is necessary for the transcriptional activation of metabolic genes such as G6pc and has an important role in regulating glycogen levels in the liver through altering transcription factor binding and activity at the G6pc promoter.

Effects of different ratios of crude protein and non-fibrous carbohydrates on growth, metabolism, physiology, nutrient utilization and muscle cellularity of Lophiosilurus alexandri, a carnivorous freshwater fish

The aim of this research was to evaluate the effects of different ratios of crude protein/non-fibrous carbohydrates (CP/NFC) on growth, metabolism, physiology, nutrient utilization efficiency and muscle cellularity of juvenile Lophiosilurus alexandri. Treatments consisted of isoenergetic diets (4500 kcal/kg) with CP/NFC ratios of 0.78; 1.02; 1.26 and 1.60, delivered for 75 days. Polynomial regression analysis showed that the different CP/NFC ratios affected weight gain and specific growth rate with estimated maximum values of 1.27 and 1.25, respectively. Feed conversion rate and the hepatosomatic index had minimum values with CP/NFC ratios of 1.33 and 1.02, respectively. Muscle and liver glycogen levels were maximized with CP/NFC content of 0.87 and 1.26, respectively. The CP/NFC ratio of 1.20 maximized the deposition of lipids in the muscle while that of 1.11 maximized the concentration of total soluble proteins in the same tissue. Plasma total cholesterol, non-HDL cholesterol and triglycerides increased linearly with increasing dietary CP/NFC ratio. Ratios estimated at 0.99, 0.94, 0.93 and 0.98 minimize the concentrations of HDL, total protein, alanine aminotransferase and creatinine in fish plasma, respectively. Nucleus diameter of hepatocytes increased linearly with increasing dietary CP/NFC ratio, while percentage of liver tissue nuclei decreased linearly. Hypertrophic muscle fibers were influenced by diets with the estimated ratio of 1.21 CP/NFC maximizing the occurrence of these fibers. Fish body crude protein increased linearly with increasing CP/NFC ratios in the diet. Fish body content of mineral matter, total phosphorus and visceral fat were minimized with the ratios of 1.34, 1.38 and 1.12, respectively. There were no effects of CP/NFC ratios on the viscerosomatic index, feed intake, carcass yield, the enzymes aspartate aminotransferase and gamma-glutamyl transferase, percentages of cytoplasmic and capillary sinusoids in liver tissue, and body dry matter, crude energy, total lipids, viscera dry matter. It is concluded that diets containing CP/NFC within 1.11–1.33 improve productive performance and nutrient use efficiency of fish. The different CP/NFC ratios also interfere with some plasma and metabolic variables that, associated with performance results, indicate a better physiological state of juvenile L. alexandri with an initial average weight of 30 g.

Antidiabetic effects of fermented milk contained with Gardenia jasminoides water extracts on streptozotocin‐induced mice

Gardenia jasminoides, a kind of flowering plant, possesses numerous bio-activities. The purpose of this study was to investigate the characteristics and hypoglycemic effects of fermented milk (CFM, GFM1, GFM2,and GFM3) contained with 0, 0.1%, 0.2%, and 0.3% of G. jasminoides water extracts. We demonstrated that GFM3 has the highest total count of lactic acid bacteria, water-holding capacity, and antioxidant activities (DPPH, ABTS, and FRAP) among four kinds of fermented milk. In vivo hypoglycemic activity showed that GFM3 notably reduced blood glucose levels, insulin, weight loss rate, food intake, and organ indexes compared with model group. GFM3 could improve the diabetes by reducing the levels of serum TC, TG, LDL-C, and increasing HDL-C content (p < .01). Moreover, it can also increase the levels of hepatic T-SOD, GSH-Px, CAT, hexokinase, and glycogen and decrease MDA content in liver (p < .05 or p < .01). Furthermore, GFM3 stimulated the phosphorylations of p-adenosine 5′-monophosphate-activated protein kinase (AMPK) and Acetyl-CoA carboxylase (ACC), but an inhibitor of Fatty acid synthase (FAS), p < .01. Overall, GFM3 treatment protected the mice better than CFM, so Gardenia jasminoides water extracts is expected to become a hypoglycemic agent. Practical applications With the increasing awareness of nutrition among consumers, the food processing industry is supplementing existing foods, including natural plants or their extracts, to eventually form functional foods. Our results indicated that novel fermented milk contained with Gardenia jasminoides water extracts has better qualities and activities when compared with traditional fermented milk.

Anti-diabetic effects of the soluble dietary fiber from tartary buckwheat bran in diabetic mice and their potential mechanisms.

BackgroundTartary buckwheat has beneficial effects on glucose and lipid metabolism of patients with type 2 diabetes mellitus. However, the physiological effects of a soluble dietary fiber (SDF) from tartary buckwheat have rarely been studied, especially in vivo.ObjectiveThis study aimed to examine the hypoglycemic and hypolipidemic effects of SDF from tartary buckwheat bran on high-fat diet/streptozotocin-induced diabetic mice.DesignThe SDF of tartary buckwheat bran was collected according to the Association of Official Analytical Chemists method 991.43. Diabetic mice were treated with high-fat diets supplemented with 0.5, 1, and 2% SDF for 8 weeks. Parameters related to glucose and lipid metabolism and relevant mechanisms, including the excretion of short-chain fatty acids and the glycemic signaling pathway in the liver, were investigated. In addition, the structural characterization of a purified polysaccharide from SDF of tartary buckwheat bran was illustrated.ResultSupplementation with SDF in the diet resulted in reduced levels of fasting blood glucose, improved oral glucose tolerance, increased levels of liver glycogen and insulin, as well as improved lipid profiles in both the serum and liver, in diabetic mice. The amelioration of glucose and lipid metabolism by SDF was accompanied by an increase in the short-chain fatty acid levels in the cecum and co-regulated by hepatic adenosine-5′-monophosphate-activated protein kinase (AMPK) phosphorylation. A neutral tartary buckwheat polysaccharide with an average molecular weight of 19.6 kDa was purified from the SDF, which consisted mainly of glucose with α-glycosidic bonds.ConclusionsThe SDF of tartary buckwheat bran exhibits hypoglycemic and hypolipidemic effects in diabetic mice, contributing to the anti-diabetic mechanisms of tartary buckwheat.

Effects of glucose ingestion at different frequencies on glycogen recovery in mice during the early hours post exercise

BackgroundWhen a high-carbohydrate diet is ingested, whether as small frequent snacks or as large meals, there is no difference between the two with respect to post-exercise glycogen storage for a period of 24 h. However, the effect of carbohydrate intake frequency on glycogen recovery a few hours after exercise is not clear. Athletes need to recover glycogen quickly after physical exercise as they sometimes exercise multiple times a day. The aim of this study was to determine the effect of carbohydrate intake at different frequencies on glycogen recovery during the first few hours after exercise.MethodsAfter 120 min of fasting, 6-week-old male ICR mice were subjected to treadmill running exercise (20 m/min for 60 min) to decrease the levels of muscle and liver glycogen. Mice were then given glucose as a bolus (1.2 mg/g of body weight [BW], immediately after exercise) or as a pulse (1.2 mg/g of BW, every 15 min × 4 times). Following this, the blood, tissue, and exhaled gas samples were collected.ResultsIn the bolus group, blood glucose concentration was significantly lower and plasma insulin concentration was significantly higher than those in the pulse group (p < 0.05). The plantaris muscle glycogen concentration in the bolus group was 25.3% higher than that in the pulse group at 60 min after glucose ingestion (p < 0.05). Liver glycogen concentration in the pulse group was significantly higher than that in the bolus group at 120 min after glucose ingestion (p < 0.05).ConclusionsThe present study showed that ingesting a large amount of glucose immediately after exercise increased insulin secretion and enhanced muscle glycogen recovery, whereas frequent and small amounts of glucose intake was shown to enhance liver glycogen recovery.

Lactose-riched Mongolian mare's milk improves physical fatigue and exercise performance in mice.

Fatigue may cause the efficiency of the organ in human body to decrease, which may affect the daily life and exercise performance of the general people and athletes. Mare's milk powder (MMP) is a lactose rich supplement. The research of the study is to evaluate the whether MMP has anti-fatigue effect. Forty male ICR mice were randomly divided into four group to receive vehicle or MMP by oral gavage at 0 (Vehicle), 0.27 (MMP-1X), 0.54 (MMP-2X), 1.35 (MMP-5X) g/kg/day for 14 days. The forelimb grip of the MMP-2X, and MMP-5X group were significantly higher than the vehicle group. The swim-to-exhaustion times of the MMP-1X, MMP-2X, and MMP-5X group were significantly greater than the vehicle group. Glycogen levels in liver and muscle were significantly larger in the MMP-1X, MMP-2X, and MMP-5X groups than the vehicle group. Receive MMP supplement for 14 days can promoting exercise performance and amelioration of exercise-induced fatigue.

Influence of hypoxia on biochemical aspects and on expression of genes related to oxygen-homeostasis of the Amazonian cichlid Astronotus ocellatus (Agassiz, 1831).

Variations in dissolved oxygen levels are common in the Amazonian aquatic environments and the aquatic organisms that inhabit these environments developed a variety of adaptive responses to deal with such conditions. Some Amazonian fish species are tolerant to low oxygen levels and the cichlid Astronotus ocellatus is one of the most hypoxia-tolerant species. Herein, we aimed to unveil the biochemical and molecular responses that A. ocellatus presents when submitted to hypoxia. Hypoxia indicators were measured, such as plasma glucose, plasma lactate, hepatic glycogen and relative transcript levels of prolyl hydroxylase 2 (phd2) and hypoxia-inducible factor-1α (hif-1α) in juveniles of approximately 50 g exposed to 1, 3, and 5 hours of hypoxia (0.7 mg O2.L-1), followed by 3 hours of recovery in normoxia (6 mg O2.L-1). Fish exposed to hypoxia reduced liver glycogen levels within 3 hours of hypoxia, when comparing with 1 hour, and increased plasma glucose and lactate. Under the same condition, phd2 transcripts levels increased in gills, but decreased in liver. In contrast, hypoxia did not affect relative gene expression of hif-1α in both tissues. Based on the transcription pattern of phd2, these results showed that liver and gills of A. ocellatus have different molecular strategies to cope with environmental hypoxia.

Friedelin exhibits antidiabetic effect in diabetic rats via modulation of glucose metabolism in liver and muscle

Ethnopharmacological relevanceDemand for plant-based medications and therapeutics is increasing worldwide as of its potential effects and no toxic. Traditionally, so many medicinal plants are used to treat diabetes. Subsequently, investigation on medicinal plants was enduring to discover potential antidiabetic drugs. A. tetracantha is used traditionally to cure diabetes mellitus, cough, dropsy, chronic diarrhea, rheumatism, phthisis and smallpox. Scientifically, A. tetracantha has been reported as an antidiabetic agent. Friedelin, the isolated compound has been reported as hypolipidemic, antioxidant, scavenging of free radicals, antiulcer, anti-inflammatory, analgesic and antipyretic agent. Aim of the studyTo scrutinize the mechanism of antidiabetic activity of friedelin isolated from the leaves of A. tetracantha. Materials and methodsA. tetracantha leaves powder (5 kg) was soaked in hexane (15 L) to obtain hexane extract. Using column chromatography, the hexane extract was fractionated using a combination of solvents like hexane and ethyl acetate. 25 fractions were obtained and the fractions 13 and 14 yielded the compound, friedelin. Friedelin at the doses of 20 and 40 mg/kg was used to treated STZ -induced diabetic rats for 28 days. Later 28 days of treatment, the bodyweight changes, levels of blood glucose, insulin, SGOT, SGPT, SALP, liver glycogen and total protein were assessed. ResultsFriedelin significantly brought these altered levels to near normal. Moreover, friedelin also enhanced the translocation as well as activation of GLUT2 and GLUT4 through PI3K/p-Akt signaling cascade in skeletal muscles and liver on diabetic rats. ConclusionThis finding proved that friedelin has an anti-diabetic effect through insulin-dependent signaling cascade mechanism, thus it may lead to establishing a drug to treat type 2 diabetes mellitus.

Industrial applications of selected JFS articles

Industrial applications of selected <i>JFS</i> articles

Comparative Study on Antidiabetic Effect of Ethanolic Extract of Jute Leaf on Neonatal Streptozotocin- Induced Type-2 Diabetic Model Rat

Aim: Functional food and their bioactive compounds have been considered as a new approach for the prevention and management of type 2 diabetes and its complications. According to this approach current study was carried out as an elucidation of antidiabetic properties of Corchorus capsularis and Corchorus olitorius varieties of jute leaf (ethanolic extract) on nSTZ-induced type-2 diabetic rats.&#x0D; Methodology: The type-2 diabetic model rat was developed by a single intraperitoneal injection of freshly prepared STZ (90 mg/kg/10 ml) in sterile citrate buffer (0.1 M, pH 4.5) to rat pups (48 hour old). After three months, OGTT was performed to select diabetic (FSG &gt; 6.5mmol/L and after 90 min of glucose load &gt; 14 mmol/L) experimental rats. The rats were randomly divided into four groups [DWC, GT, Ext-1 and Ext-2 represent, diabetic water control, glybenclamide treated (20 mg/5 ml/kg body weight), C. capsularis treated and C. olitorius treated group (1.25 g/10 ml/kg body weight) respectively]. One group was kept with normal rats [normal water control, NWC]. The treatment was given once daily or 28 consecutive days. Fasting serum glucose, liver glycogen and lipid profile were estimated by using standard methods.&#x0D; Results: The results showed that Ext-1 and Ext-2 treated groups gradually decreased serum glucose level (7.15 ±0.67 to 5.94 ± 1.19 and 7.20 ± 0.93 to 5.28 ±1.03 respectively) and reducing effect by Ext-2 was significant (p=0.001). Both extract showed lower liver glycogen level compared with GT group [5.0±2.5 Vs 17.7±6.5 (Ext-1 vs GT) and 7.5±6.4 Vs 17.7±6.5 (Ext-2 vs GT)] and even Ext-1 manifested significant effect (p=0.05). Additionally, lipid profile estimation revealed no significant improvement by the consumption of both the extracts.&#x0D; Conclusion: On the basis of current investigations, it may be concluded that both variety of jute’s leaf demonstrated hypoglycemic properties in Type 2 diabetic model rats; further in-depth studies are recommended to explore the exact mechanism(s) of hypoglycemic effect.

Antifatigue effect of functional cookies fortified with mushroom powder (Tricholoma Matsutake) in mice.

Bakery products made by cereal and edible fungi powder have a unique flavor and health benefits, dramatically enhancing the nutritional value of the products. In this study, we investigated the antifatigue effect of a novel Tricholoma matsutake cookie (TMC) by the exhaustive swimming test. Male Kunming ICR mice were randomly divided into seven groups (each group, n = 10), fed with saline, ordinary cookies (4, 8, 16g/kg B.W./day), and TMC (4, 8, 16g/kg B.W./day) by gavage. After 30-day administration, the weight-loaded swimming test was carried out on the mice to evaluate the antifatigue effect of TMC. In comparison with the effect of ordinary cookies, the intake of TMC significantly prolonged the exhaustive swimming time of mice and increased the level of muscle glycogen and liver glycogen, accompanied by the reduction of lactic acid and urea nitrogen level in serum. Additionally, TMC dramatically improved the activity of superoxide dismutase and glutathione peroxidase in serum and largely decreased the level of malondialdehyde. All in all, TMC could enforce exhaustive swimming tolerance, accelerate the decomposition of sports-related metabolites such as lactic acid and urea nitrogen, and increase the activity of the antioxidant enzyme, thereby improving sports-related energy storage and relieving fatigue. Our findings broadened the application of T. matsutake in the processing of bakery products and provided the theoretical basis and technical support for the development of antifatigue products. PRACTICAL APPLICATION: In this study, we investigated the antifatigue effect of a novel Tricholoma matsutake cookie by the exhaustive swimming test. Collectively, the results of the present study suggested that the cookies fortified with T. matsutake could be considered as an antifatigue bakery product. Furthermore, our findings broadened the application of T. matsutake in the processing of bakery products and provided the theoretical basis and technical support for the development of antifatigue products.

Probing metabolic memory in the hepatic response to fasting.

Tissues may respond differently to a particular stimulus if they have been previously exposed to that same stimulus. Here, we tested the hypothesis that a strong metabolic stimulus such as fasting may influence the hepatic response to a subsequent fast and thus elicit a memory effect. Overnight fasting in mice significantly increased plasma free fatty acids, glycerol, β-hydroxybutyrate, and liver triglycerides, and decreased plasma glucose, plasma triglycerides, and liver glycogen levels. In addition, fasting dramatically changed the liver transcriptome, upregulating genes involved in gluconeogenesis and in uptake, oxidation, storage, and mobilization of fatty acids, and downregulating genes involved in fatty acid synthesis, fatty acid elongation/desaturation, and cholesterol synthesis. Fasting also markedly impacted the liver metabolome, causing a decrease in the levels of numerous amino acids, glycolytic-intermediates, TCA cycle intermediates, and nucleotides. However, these fasting-induced changes were unaffected by two previous overnight fasts. Also, no significant effect was observed of prior fasting on glucose tolerance. Finally, analysis of the effect of fasting on the transcriptome in hepatocyte humanized mouse livers indicated modest similarity in gene regulation in mouse and human liver cells. In general, genes involved in metabolic pathways were upregulated or downregulated to a lesser extent in human liver cells than in mouse liver cells. In conclusion, we found that previous exposure to fasting in mice did not influence the hepatic response to a subsequent fast, arguing against the concept of metabolic memory in the liver. Our data provide a useful resource for the study of liver metabolism during fasting.

Transcriptomic and Proteomic Analysis of Clear Cell Foci (CCF) in the Human Non-Cirrhotic Liver Identifies Several Differentially Expressed Genes and Proteins with Functions in Cancer Cell Biology and Glycogen Metabolism.

Clear cell foci (CCF) of the liver are considered to be pre-neoplastic lesions of hepatocellular adenomas and carcinomas. They are hallmarked by glycogen overload and activation of AKT (v-akt murine thymoma viral oncogene homolog)/mTOR (mammalian target of rapamycin)-signaling. Here, we report the transcriptome and proteome of CCF extracted from human liver biopsies by laser capture microdissection. We found 14 genes and 22 proteins differentially expressed in CCF and the majority of these were expressed at lower levels in CCF. Using immunohistochemistry, the reduced expressions of STBD1 (starch-binding domain-containing protein 1), USP28 (ubiquitin-specific peptidase 28), monad/WDR92 (WD repeat domain 92), CYB5B (Cytochrome b5 type B), and HSPE1 (10 kDa heat shock protein, mitochondrial) were validated in CCF in independent specimens. Knockout of Stbd1, the gene coding for Starch-binding domain-containing protein 1, in mice did not have a significant effect on liver glycogen levels, indicating that additional factors are required for glycogen overload in CCF. Usp28 knockout mice did not show changes in glycogen storage in diethylnitrosamine-induced liver carcinoma, demonstrating that CCF are distinct from this type of cancer model, despite the decreased USP28 expression. Moreover, our data indicates that decreased USP28 expression is a novel factor contributing to the pre-neoplastic character of CCF. In summary, our work identifies several novel and unexpected candidates that are differentially expressed in CCF and that have functions in glycogen metabolism and tumorigenesis.

A negative reciprocal regulatory axis between cyclin D1 and HNF4α modulates cell cycle progression and metabolism in the liver

Hepatocyte nuclear factor 4α (HNF4α) is a master regulator of liver function and a tumor suppressor in hepatocellular carcinoma (HCC). In this study, we explore the reciprocal negative regulation of HNF4α and cyclin D1, a key cell cycle protein in the liver. Transcriptomic analysis of cultured hepatocyte and HCC cells found that cyclin D1 knockdown induced the expression of a large network of HNF4α-regulated genes. Chromatin immunoprecipitation-sequencing (ChIP-seq) demonstrated that cyclin D1 inhibits the binding of HNF4α to thousands of targets in the liver, thereby diminishing the expression of associated genes that regulate diverse metabolic activities. Conversely, acute HNF4α deletion in the liver induces cyclin D1 and hepatocyte cell cycle progression; concurrent cyclin D1 ablation blocked this proliferation, suggesting that HNF4α maintains proliferative quiescence in the liver, at least, in part, via repression of cyclin D1. Acute cyclin D1 deletion in the regenerating liver markedly inhibited hepatocyte proliferation after partial hepatectomy, confirming its pivotal role in cell cycle progression in this in vivo model, and enhanced the expression of HNF4α target proteins. Hepatocyte cyclin D1 gene ablation caused markedly increased postprandial liver glycogen levels (in a HNF4α-dependent fashion), indicating that the cyclin D1-HNF4α axis regulates glucose metabolism in response to feeding. In AML12 hepatocytes, cyclin D1 depletion led to increased glucose uptake, which was negated if HNF4α was depleted simultaneously, and markedly elevated glycogen synthesis. To summarize, mutual repression by cyclin D1 and HNF4α coordinately controls the cell cycle machinery and metabolism in the liver.

Effects of different dietary carbohydrate‐to‐lipid ratios on growth, plasma biochemical indexes, digestive and immune enzymes activities of juvenile orange‐spotted grouper Epinephelus coioides

An 8-week feeding trial was conducted to investigate the effects of dietary carbohydrate-to-lipid (CHO:L) ratios on growth, liver and muscle glycogen content, haematological indices, and liver and intestinal enzyme activity of juvenile grouper (Epinephelus coioides). Five isonitrogenous (496.0 g/kg protein) and isoenergetic (21.6 KJ/g gross energy) diets with varying CHO:L ratios of 0.65 (D1), 1.31 (D2), 2.33 (D3), 4.24 (D4) and 8.51 (D5), respectively, were fed to triplicate groups of 20 fish (average 10.02 ± 0.1 g) for 8 weeks. Results showed that the weight gain rate, specific growth rate and protein efficiency ratio (PER) of juvenile grouper increased first and then decreased with the increase in CHO:L ratio, reaching a maximum value in the D4 (CHO:L = 4.24) diet. The trend for the feed conversion ratio was opposite to the PER. Along with the diet CHO:L ratios, the apparent digestibility coefficients (ADC) of crude lipid and energy for the juvenile groupers decreased gradually, while the ADC of dry matter, liver and muscle glycogen level increased gradually. Total protein, triglycerides and cholesterol in serum were all maximized in the D4 diet and glucose in the D5 (CHO:L = 8.51) diet. Digestive enzyme activity in the intestine was significantly affected by dietary CHO:L ratio. Liver hexokinase, phosphofructokinase and glucose-6-phosphate dehydrogenase activity increased significantly as CHO:L ratio increased. Liver lysozyme and acid phosphatase activity in the groupers fed the D3 (CHO:L = 2.33) diet was significantly higher than that of other diets. Liver fructose-1,6-bisphosphatase and alkaline phosphatase activity reached a maximum value in the D4 diet and was significantly higher than that in the D1 diet. Taking the above results together, it can be concluded that an optimal dietary CHO:L ratio of 2.33 is suitable for grouper culture concerning growth performance and health.

Diets containing purified nucleotides reduce oxidative stress, interfere with reproduction, and promote growth in Nile tilapia females

The objective of the present study was to assess the health, growth, and reproductive performance of Nile tilapia females fed diets containing purified nucleotides (NT). A total of 420 females (286 ± 7.15 g) and 140 males (536 ± 13.4 g) were used. Fishes were fed with extruded rations (280 g of digestible protein kg−1; 11.72 MJ of digestible energy kg−1) containing 0.0, 2.5, 5.0, 7.5, and 10.0 g NT kg−1 for 150 days. After mating occurred in hapas, the eggs were harvested and incubated. At the end of the experiment, blood and tissue samples were collected for hematological, biochemical, morphological, and centesimal composition analyses. In addition, indicators of immune system function and redox balance were analyzed. The production of eggs g−1 by spawning females was significantly the highest at 7.5 g NT kg−1. The percentage of spawning females was significantly low in the 7.5 g and 10.0 g NT kg−1 groups, reflecting on fecundity. However, these females showed significantly the highest growth rates. Diets containing 5.0, 7.5, and 10.0 g NT kg−1 significantly increased hemoglobin levels. Plasma alanine aminotransferase (ALT) significantly decreased in fish fed diets containing between 2.5 and 7.5 g NT kg−1, whereas alkaline phosphatase (ALP) and plasma triglyceride levels significantly decreased with 7.5 g NT kg−1. Levels of NT higher than 5.0 g NT kg−1 led to a significantly increase of albumin and significantly reduced plasma calcium levels. Liver glycogen levels significantly increased with 5.0 g NT kg−1. The activities of liver superoxide dismutase (SOD) and glutathione S-transferase (GST) were significantly the highest in the 5.0 g NT kg−1 and 7.5 g NT kg−1 groups, respectively. The highest glycogen content in the ovaries was significantly observed with 7.5 g NT kg−1. Lipid peroxidation (LPO) in the ovaries was significantly lower with 2.5, 7.5, and 10.0 g NT kg−1, and the highest catalase activity was significantly observed with 2.5 and 7.5 g NT kg−1. Lysozyme activity in the spleen and kidney and levels of nitrite/nitrate in the kidney were not significantly altered. Only the levels of spleen nitrite/nitrate significantly decreased at 10.0 g NT kg−1. Rations containing more than 5.0 g NT kg−1 significantly increased the percentage of secondary growth oocytes, and liver and intestinal morphology was not significantly altered. Diets containing 5.0 g NT kg−1 significantly increased crude protein content in the ovaries. Supplementation with purified nucleotides, using rations containing between 5.0 and 7.5 g NT kg−1, affected energy metabolism, redox balance, and liver and gonadal status in tilapia females, thereby improving their health and, consequently, their reproduction and growth rates.

Impacts of dose and length of exposure to boldenone and stanazolol on enzymatic antioxidant systems, myeloperoxidase and NAGase activities, and glycogen and lactate levels in rat liver

We investigated the adverse effects of the anabolic androgenic steroids (AAS) boldenone (BOL) and stanazolol (ST) on the enzymatic antioxidant systems of the rat liver. Male Wistar rats were divided in three protocols (P): PI, 5 mg/kg BOL or ST once a week for 4 weeks; PII, 2.5 mg/kg BOL or ST once a week for 8 weeks; PIII, 1.25 mg/kg BOL or ST once a week for 12 weeks. AAS were administered intramuscularly (0.2 ml, olive oil vehicle) once a week in all protocols. Activities of the enzymes glutathione peroxidase (GPx), glutathione S-transferase (GST), and glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), were investigated. We assessed the content of hydrogen peroxide (H2O2), glycogen and lactate; and enzyme markers of neutrophils (myeloperoxidase, MPO) and macrophages (NAGase). PI and PII altered the SOD and CAT activities and increased the H2O2 content. PI led to increases in the MPO and NAGase activities. In contrast, changes in GPx, GST and, GR were observed under PII and, to a greater extend, under PIII. Following PIII, GPx, GR, and GST exhibited reduced activities. All protocols altered the glycogen and lactate content. The use of high doses of AAS for a short duration first alters SOD/CAT activity. In contrast, at lower doses of AAS for long periods is associated with changes in the glutathione system. Protocols with high doses of AAS for a short duration exert the most deleterious effects on redox status, markers of cellular infiltration, and the metabolic functioning of hepatic tissues.