OBJECTIVETo investigate the role of the key intracellular signaling molecule glycogen synthase kinase-3 beta in the mechanism of liver ischemia reperfusion (IR).

METHODSC57BL/6 mice were subjected to 90 min warm liver cephalad lobe ischemia, followed by various length of reperfusion. Experiment groups included sham control group, liver IRI model group and glycogen synthase kinase-3 beta inhibitor-treated group (SB216763 in DMSO, 25 g/kg, i.p, 2 hour prior to the onset of liver ischemia). The expression of glycogen synthase kinase-3 beta protein was analysed by Western blotting. The serum ALT levels were determined to reflect the function of liver. The affected liver lobes were harvested for histology analysis. The inflammatory gene expression was detected by Quantitative PCR.

RESULTSBy western blot analysis, we found that ischemia itself activated glycogen synthase kinase-3 beta by a significant decrease of its phosphorylation. Glycogen synthase kinase-3 beta inhibitor SB216763-pretreatment ameliorated the liver damages significantly as compared to the controls (sALT: 2046+/-513 U/L vs 5809+/-1689 U/L, P = 0.0153), and suppressed the gene expressions of IL-12, TNFa, IL-1b and IL-6.

CONCLUSIONSThis study demonstrated that the ischemia process modulated liver innate immune activation via a GSK-3-dependent mechanism which favored the development of a pro-inflammation response and lead to liver tissue damages. GSK-3b may be a new therapeutic target to ameliorate liver IRI in transplant patients.

Animals ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Inflammation ; metabolism ; Liver ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Reperfusion Injury ; metabolism ; pathology

OBJECTIVETo investigate the changes in the functional activity of glycogen synthase kinase-3 (GSK-3) in the hepatic tissue after endotoxin (lipopolysaccharide, LPS) tolerance and explore the effects of LPS-induced GSK-3 inhibition on glycogen metabolism in the liver.

METHODSMale SD rats were randomly divided into normal control, endotoxin pretreatment and GSK-3 inhibitor (lithium chloride) groups with corresponding pretreatments prior to a large dose of LPS challenge (10 mg/kg) to induce liver injury. Glycogen deposition and content in the hepatic tissue was detected using periodic acid-Schiff (PAS) staining and a glycogen quantification kit, respectively. Western blotting was performed for semi-quantitative analysis of protein level and inhibitory phosphorylation of GSK-3, and a Coomassie brilliant blue G-250-based colorimetric assay was used to detect calpain activity in the liver.

RESULTSGlycogen content in the liver decreased significantly after LPS challenge in all the 3 groups (P<0.05) but showed no significant difference among the groups (P>0.05). Both LPS and lithium chloride pretreatments caused a significant increase of liver glycogen content (P<0.05). LPS pretreatment induced inhibitory phosphorylation of GSK-3β (P<0.05) and partial cleavage of GSK-3α but did not affect the expression of GSK-3 protein (P>0.05). Large-dose LPS challenge significantly increased the activity of calpain in the liver tissue (P<0.05) to a comparable level in the 3 groups (P>0.05).

CONCLUSIONEndotoxin pretreatment induces inhibitory phosphorylation of GSK-3β and partial cleavage of GSK-3α and promotes the deposition of liver glycogen but does not affect the activity of calpain, which may contribute to an increased glycogen reserve for energy supply in the event of large-dose LPS challenge.

Animals ; Calpain ; metabolism ; Glycogen ; metabolism ; Glycogen Synthase Kinase 3 ; antagonists & inhibitors ; metabolism ; Lipopolysaccharides ; adverse effects ; Lithium Chloride ; pharmacology ; Liver ; drug effects ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

The purpose of this study was to investigate the effect of butanol (BuOH) fraction of Alisma canaliculatum (Ac) and/or selenium (Se) treatment on glycogen level, lipid metabolism and lipid peroxidation in streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley rats were assigned to one of the five groups: normal, STZ-control, and three experimental groups (Ac group, Ac-Se group, and Se group). Diabetes was experimentally induced by intravenous administration of 45 mg/kg of STZ in citrate buffer. The BuOH fraction of Ac (400 mg/kg bw) was orally administered for 3 weeks. The Se group were fed a AIN-93 recommended diet mixed with Na(2)SeO(3) (2 mg/kg diet). The liver glycogen level of Ac and Ac-Se groups were significantly higher, when compared with the STZ-control groups. The muscle glycogen level was not significantly differ among all groups. The levels of liver triglyceride were higher in Ac-Se group than the STZ-control group. Pancreas protein levels were significantly increased in Ac-Se group than STZ-control group. The concentration of liver malondialdehyde (MDA) was significantly decreased in Ac and Se groups and decreased in Ac-Se group. Administration of BuOH fraction of Alisma canaliculatum and selenium supplementation increased the liver glycogen and triglyceride levels, and reduced peroxidative liver damage in STZ induced diabetic rats. These results suggest that treatment with a BuOH fraction of Alisma canaliculatum in combination with selenium has no synergistic antioxidative effect. Selenium supplementation may lead a decrease MDA of liver in diabetic rats.
Administration, Intravenous ; Alisma* ; Animals ; Citric Acid ; Diet ; Glycogen* ; Humans ; Lipid Metabolism* ; Lipid Peroxidation* ; Liver ; Liver Glycogen ; Male ; Malondialdehyde ; Pancreas ; Rats* ; Rats, Sprague-Dawley ; Selenium* ; Streptozocin ; Triglycerides

Blood Glucose ; metabolism ; Female ; Glycogen Storage Disease ; diagnosis ; Humans ; Infant, Newborn ; Liver ; pathology

Daily administration of glucocorticoids for 10 days to dogs resulted in a significant increase in the hepatic bile secretion in response to secretory stimulants. The response of hepatic bile in testosterone-treated animals was not changed and the response was increased in DOCA--treated animals. A significant increase of liver weight was induced by the animals receiving glucocorticoids. Other organ weight was not changed; however, a slight reduction of kidney weight was seen in prednisolone, dexamethasone, and DOCA treated animals and also in animals supplemented with cortisone following adrenalectomy. The presence of large areas of ballooning and vesicular changes of liver cells was seen in glucocorticoid treated animals, particularly in cases of dexamethasone and prednisolone. Both vesicular changes of liver cell and its glycogen content were increased by the repeated administration of prednisolone and reduced by the cessation of treatment. Special stain and liver glycogen determination demonstrated the material distending the liver cell was glycogen. These findings indicate that long term administration of glucocorticoids results in an increase of liver weight and hepatic glycogen content as well as increased bile secretion.
Animal ; Bile/secretion* ; Bile Acids and Salts/metabolism ; Bilirubin/secretion ; Cholagogues and Choleretics/pharmacology ; Dogs ; Glucocorticoids/pharmacology* ; Liver/drug effects* ; Liver/pathology ; Liver Glycogen/metabolism ; Organ Weight ; Substances: ; Bile Acids and Salts ; Cholagogues and Choleretics ; Glucocorticoids ; Liver Glycogen ; Bilirubin

Glycogen storage diseases are inherited disorders of carbohydrate metabolism caused by a deficiency of enzymes that are involved in degradation of glycogen in the liver. The accumulation of glycogen occurs in the liver and other organs. Type Ia is the most common form and clinically may manifest of glycogen storage disease itself rather than growth hormone deficiency. But in this case the patient showed exceptional extreme growth retardation. Growth hormone stimulation test with clonidine and L-dopa revealed that the patient had growth hormone deficiency. Therefore, we report of a case of glycogen storage disease type Ia with the presence of GH deficiency with review of literature. A 16-year-old male was admitted for the evaluation of hepatomegaly and extreme short stature. The height was 113.5cm, less than third percentile of same age group, and compatible with fiftieth percentile of height of 6 years of age. After laboratory work up including liver biopsy, he was diagnosed with type I glycogen storage disease. The patient was presented with metabolic acidosis, hyperuricemia, and hypoglycemia. Hypoglycemia was managed with frequent feeding with high starch diet and intravenous glucose infusion. Metabolic acidosis was treated with sodium bicarbonate. Secondary hyperuricemia was treated with allopurinol. The patient is being followed at out-patient clinic with clinical improvement after of GH administration.
Acidosis ; Adolescent ; Allopurinol ; Biopsy ; Carbohydrate Metabolism ; Clonidine ; Diet ; Glucose ; Glycogen Storage Disease* ; Glycogen* ; Growth Hormone ; Hepatomegaly ; Humans ; Hyperuricemia ; Hypoglycemia ; Levodopa ; Liver ; Male ; Outpatients ; Sodium Bicarbonate ; Starch

The purpose of this study was to determine the effect of releasing of immobilization on glycogen metabolism of hindlimb muscle after 7days use of a hindlimb casting in mice. The experimental group was divided into control group and recovery groups after removal of left hindlimb casting. The recovery groups, were subivided into the 0, 3rd and 5th day after removal of left hindlimb casting. The results were as follows; 1. The degree of atrophy of hindlimb muscles by 7 days immobilization was measured by ratio of muscle to body weight in plantaris and soleus muscles. The muscle to body weight ratios of plantaris and soleus muscles were decreased by 88% and 74%, respectively on the day of cast removal. The ratios of the both muscles were increased to the level of the control values on the 3rd and 5th day removal of casting. 2. A significant reduction of the glycogen concentration in gastrocnemius muscle occurred after 7 days hindlimb immobilization. The glycogen concentration in gastrocnemius muscle was decreased by 63% on the day of cast removal. The glycogen concentration was recovered to the values of the control group on the 3rd and 5th day after removal of hindlimb casting. 3. The level of muscle glycogen concentration of 25% glucose ingested control group was almost twice that of the normal diet control group. The muscle glycogen concentration of glucose ingested group was significantly less by 81% after 7 days of immobilization compared with the respective control gmup. The concentration recovered to the values of control on the 3rd and 5th day after removal of hindlimb casting. In contrast, there was no significant difference in the liver glycogen concentration between the immobilized grop and the cast removed group in which was removed. 4. The effects of releasing of hindlimb immobilization on plasma glucose, insulin concentration and insulin
Animals ; Atrophy ; Blood Glucose ; Body Weight ; Diet ; Glucose ; Glycogen ; Hindlimb Suspension ; Hindlimb ; Hypokinesia ; Immobilization ; Insulin ; Liver Glycogen ; Metabolism ; Mice ; Muscle, Skeletal ; Muscles

OBJECTIVETo determine the mechanism underlying the therapeutic activities of glycogen synthase kinase 3b (GSK3b) against hepatic ischemia-reperfusion (H-IR) injury by investigating the inhibitive effects of GSK3b on inflammation mediated by Toll-like receptor 4 (TLR4).

METHODSC57BL/6 male mice were subjected to 90 min of warm liver cephalad lobe ischemia, followed by reperfusion for various lengths of time. The mice were divided into three groups: the H-IR untreated model (control group), and the H-IR inflammation-induced models that received an intraperitoneal injection of purified lipopolysaccharide (LPS) endotoxin alone (inflammation group) or with pretreatment of the SB216763 GSK3b-specific inhibitor (intervention group). To create a parallel isolated cell system for detailed investigations of macrophages, marrow-derived stem cells were isolated from femurs of the H-IR control group of mice and used to derive primary macrophages. The cells were then divided into the same three groups as the whole mouse system: control, LPS-induced inflammation model, and inflammation model with SB216763 intervention. Differential expressions of inflammation-related proteins and genes were detected by Western blotting and real-time quantitative PCR, respectively.

RESULTSThe phosphorylation levels of ERK, JNK and p38 MAPK were induced in liver at 1 h after reperfusion, but then steadily decreased and returned to baseline levels by 4 h after reperfusion. In addition, the phosphorylation levels of ERK and JNK were induced in macrophages at 15 min after LPS stimulation, while the phosphorylation level of p38 MAPK was induced at 1 h; SB216763 pretreatment suppressed the LPS-stimulated ERK, JNK and p38 phosphorylation in macrophages. In the mouse model, GSK3b activity was found to promote the gene expression of anti-inflammatory cytokine IL-10 (control: 0.21 ± 0.08, inflammation: 0.83 ± 0.21, intervention: 1.76 ± 0.67; F = 3.16, P = 0.027) but to significantly inhibit the gene expression of pro-inflammatory cytokines IL-12 (control: 0.11 ± 0.05, inflammation: 0.85 ± 0.11, intervention: 0.43 ± 0.10; F = 2.67, P = 0.038), TNF-a, (control: 0.052 ± 0.012, inflammation: 8.11 ± 0.98, intervention: 3.9 ± 0.82; F = 4.13, P = 0.016), IL-6 (control: 0.22 ± 0.08, inflammation: 6.37 ± 0.81, intervention: 2.11 ± 0.63; F = 3.21, P = 0.024), and IL-1b (control: 0.12 ± 0.07, inflammation: 2.51 ± 0.62, and intervention: 1.28 ± 0.33; F = 2.22, P = 0.030).

CONCLUSIONInhibition of GSK3b selectively regulates the expression of anti-inflammatory and pro-inflammatory cytokines in liver Kupffer cells (liver macrophages). This process may be one of the mechanisms underlying the ability of GSK3b to ameliorate hepatic ischemia-reperfusion injury, possibly because inhibition of pro-inflammatory cytokines may indirectly mediate liver cell apoptosis.

Animals ; Cells, Cultured ; Cytokines ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Inflammation ; metabolism ; pathology ; Lipopolysaccharides ; adverse effects ; Liver ; pathology ; Macrophages ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Reperfusion Injury ; Toll-Like Receptor 4 ; metabolism

Prolonged administration of anterior hypophyseal, adrenocortical, or thyroid hormones is known to cause degeneration, degranulation and necrosis of the beta-cells in the Langerhans islets of the pancreas. However, the effects of extirpation of these endocrine glands upon the Langerhans islets has not been reported, a1though it is known that removal of any of these glands bring about hypoglycemia, decreased tissue uptake of glucose, and increased tissue sensitivity to insulin. The present investigation is studies of the morphologic alterations of the beta-cells in the Langerhans islets following hypophysectomy, adrenalectomy, or thyroidectomy in rats. Hypophysectomy, adrenalectomy, and thyroidectomy, all induce similar morphologic alterations in the beta-cells of the islets. These consist of increased beta-cell population, the accumlnation of beta-granules, and atrophy of the individual betacell. Therefore, these changes are considered to be not specific following the withdrawal of specific hormones but a common effect of the hypoglycemia due to removal of the hypophysis, adrenals, or thyroid glands. A similar common degeneration of the beta-cells due to hyperglycemia occurs when hormones of these endocrine glands are given excessively.
Adrenal Cortex Hormones/physiology ; Adrenalectomy* ; Animal ; Atrophy/etiology ; Blood Glucose ; Diabetes Mellitus/etiology ; Glycogen/metabolism ; Hyperglycemia/etiology ; Hypoglycemia/etiology ; Hypophysectomy* ; Insulin/secretion ; Islets of Langerhans/pathology* ; Liver Glycogen/metabolism ; Muscles/metabolism ; Myocardium/metabolism ; Necrosis/etiology ; Rats ; Staining and Labeling ; Thyroidectomy* ; Thyroxine/physiology

OBJECTIVETo investigate if higher hepatocellular glycogen contents can alleviate hepatic ischemia reperfusion injury and its relationship to ICAM-1 gene expression in hepatic sinusoidal cells (HSCs).

METHODSTwenty-one rabbits fed with a standard diet were randomly divided into three groups (n=7 in each). All the animals were subjected to hepatic ischemia reperfusion injury then sacrificed. Before the injury, group A rabbits fasted for 24 hours; group C rabbits had 6 intravenous glucose solution (25%, 20 ml) injections, 4 hours between two injections. Hepatic enzymological changes, hepatic ICAM-1 mRNA expressions and leukocytic counts in the sinusoids were observed.

RESULTSThe liver glycogen contents of the three groups were significantly different. Livers of group A had higher contents of glycogen (9.85+/-0.91 mg/g. wet tissue); in group B they were 38.93+/-5.72; and in group C they were 48.31+/-6.58. Group C animals had the slightest liver function damage. There were no differences in the pre- and post-ischemic ICAM-1 mRNA contents in the three groups. However, livers with a higher content of glycogen showed less expression of ICAM-1 mRNA (group A: 1.398+/-0.365 ng/mg wet tissue; group B: 0.852+/-0.297; group C: 0.366+/-0.183) and lower leukocytic counts. The relationship analysis showed a negative relationship between hepatocellular glycogen and hepatic ICAM-1 mRNA contents (r= -0.965, P less than 0.01).

CONCLUSIONSHepatocellular glycogen is important in protecting liver ischemic reperfusion injury. Also hepatocellular glycogen decreases the expression of ICAM-1 mRNA of HSCs.

Animals ; Female ; Glycogen ; pharmacology ; Hepatocytes ; chemistry ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Liver ; chemistry ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Rabbits ; Reperfusion Injury ; genetics ; metabolism ; pathology