Effects of hypo- and hyper-thyroidism on liver composition, blood glucose, ketone bodies and insulin in the male rat

1. Thyroidectomized rats injected daily with 0, 0.1, 2 or 25mug of l-thyroxine/100g body wt. were compared with intact controls. In plasma, the protein-bound iodine was decreased in the rats given the 0 or 0.1mug doses and increased in those given the 25mug dose. 2. Blood glucose decreased in those given 2mug and was augmented in those given 25mug, and ketone bodies were the same in all the groups. 3. Plasma insulin was lowest in the rats given the 0 or 0.1mug doses and was highest in those given the 2 or 25mug doses of thyroxine. 4. After 48h starvation, the decrease in blood glucose and increase in ketone bodies observed in all the groups was greatest in the group not supplemented with thyroxine. 5. Plasma insulin concentrations remained at the value for fed animals in the rats given the 25mug dose of thyroxine but decreased in the other groups. 6. In fed animals, concentrations of hepatic DNA P, citrate, total fatty acids and acetyl-CoA were similar in all the groups, and glycogen was low only in the rats given the 25mug dose of thyroxine. 7. After 48h starvation, liver DNA P, total fatty acids and acetyl-CoA increased in all the groups, except in the rats given the 25mug dose, where both total fatty acids and acetyl-CoA remained at the value for fed animals. Liver citrate did not change in the groups given the 0 or 25mug doses of thyroxine, but decreased in the other groups. 8. The results are discussed in relation to the regulation of intermediary metabolism in hypo- and hyper-thyroidism.     

Effects of hypo- and hyper-thyroidism on in vivo lipogenesis in fed and fasted rats

Thyroidectomized rats (T) daily injected with either 0, 0.1, 1.8 or 25 microgram of L-thyroxine/100 g body wt. were compared with intact controls (C). The appearance of radioactivity in fatty acids 30 min after the i.p. injection of (3-14C)pyruvate was reduced in adipose tissue and enhanced in liver of T+25, being no differences between the other groups and C. (14C)-Fatty acids are reduced with 3 h of fasting only in the adipose tissue of T+1.8 and C, while 24 h produces a reduction in liver in the T+1.8, T+25 and C, and in adipose tissue in the T+1.8 and C animals. The highest percentage of radioactivity was observed in the liver glyceride glycerol fraction, being greater in T+25 than in the other groups. Fasting produces an increment in the (14C)-glyceride glycerol fraction. Being significant only in thehypothyroid animals in both liver and adipose tissue. The most sensitive parameter to fasting was the formation of (14C)-non-saponifiable lipid in both the C and T+1.8 animals, while it does not change in T+0 or T+0.1, but is enhanced within 24 h in the adipose tissue of T+25. It is proposed that most of the observed changes are due to the other endocrine disfunction s which appear in hypo- and hyperthyroidism, as the in vivo results do not comply with in vitro effects of thyroxine onlipogenesis of others.     

Effect of dietary supply of calcium on thyroid function in rats

To determine if calcium had a goitrogenic effect on the thyroid function in rats, weanling rats were fed, for three weeks, a diet containing either 0.5 microgram or 0.04 microgram iodine per gram of diet, or an adequate (0.47%) or an excessive (2%) amount of calcium. With an adequate iodine diet, the calcium load did not induce an increase in the weight of the thyroid or a decrease in serum thyroid hormone concentration. However, the rats given a calcium load had a lighter body weight and a lower iodine content in the thyroid tissue; they also had a higher thyroxine (T4) content in the liver and kidney tissues than the rats receiving an adequate calcium diet. With a low iodine diet, the calcium load brought out a decrease in growth and a lower serum triiodothyronine (T3) concentration and liver and kidney T3 contents. These changes suggest that the calcium load might have acted on the thyroid function through an inhibition of T4-T3 conversion in the serum as well as in liver and kidney tissues.     

Immunocytochemical localization of hepatic fatty acid binding protein in the liver of fed and fasted rats

Summary The immunocytochemical localization of fatty acid binding protein (FABP) of liver type was studied at light and electron microscopic levels by the peroxidase-antiperoxidase (PAP) method using a specific polyclonal antibody against FABP in the liver of fed and fasted rats. In the liver of rats fed ad libitum, the intense immunoreactivity was confined to portions of the liver cell cytoplasm adjacent to the glycogen area. After 2-days' fasting, such a focal intracellular localization of the immunoreactivity was abolished, in association with the disappearance of the glycogen area, and was replaced by a diffuse distribution of the immunoreactivity throughout the cytoplasm, with higher intensity at the periphery of the cells. In liver cells exhibiting an overall hypertrophy of smooth endoplasmic reticulum (SER) induced by the treatment of fasted rats with phenobarbital, the peripheral localization of FABP immunoreactivity ramained unchanged compared with that obtained in the case of fasting alone, and the immunoreactivity did not occur in association with the proliferated SER in the central cytoplasm. These results suggest that FABP, although cytosolic in nature, changes its localization within the liver cells in response to the general metabolic alterations caused by the starvation, inferring that FABP is intimately involved in the intracellular transport and metabolism of free fatty acids.     

Immunocytochemical localization of hepatic fatty acid binding protein in the liver of fed and fasted rats

The immunocytochemical localization of fatty acid binding protein (FABP) of liver type was studied at light and electron microscopic levels by the peroxidase-antiperoxidase (PAP) method using a specific polyclonal antibody against FABP in the liver of fed and fasted rats. In the liver of rats fed ad libitum, the intense immunoreactivity was confined to portions of the liver cell cytoplasm adjacent to the glycogen area. After 2-days' fasting, such a focal intracellular localization of the immunoreactivity was abolished, in association with the disappearance of the glycogen area, and was replaced by a diffuse distribution of the immunoreactivity throughout the cytoplasm, with higher intensity at the periphery of the cells. In liver cells exhibiting an overall hypertrophy of smooth endoplasmic reticulum (SER) induced by the treatment of fasted rats with phenobarbital, the peripheral localization of FABP immunoreactivity remained unchanged compared with that obtained in the case of fasting alone, and the immunoreactivity did not occur in association with the proliferated SER in the central cytoplasm. These results suggest that FABP, although cytosolic in nature, changes its localization within the liver cells in response to the general metabolic alterations caused by the starvation, inferring that FABP is intimately involved in the intracellular transport and metabolism of free fatty acids.     

Effect of L-alanine infusion on gluconeogenesis and ketogenesis in the rat in vivo.

1. In 48 h-starved 6-week-old rats the 14C incorporation in vivo into blood glucose from a constant-specific-radioactivity pool of circulating [14c]actateconfirmed that lactate is the preferred gluconeogenic substrate. 2. Increasing the blood [alanine] to that occurrring in the fed state increased 14C incorporation into blood glucose 2.3-fold from [14c]alanine and 1.7-fold from [14c]lactate. 3. When the blood [alanine] was increased to that in the fed state, the 14C incorporation into liver glycogen from circulating [14c]alanine or [14c]lactate increased 13.5- and 1.7-fold respectively. 4. The incorporation of 14C into blood acetoacetate and 3-hydroxybutyrate from a constant-specific-radioactivity pool of circulating [14c]oleate was virtually abolished by increasing the blood [alanine] to that existing in the fed state. However, the [acetoacetate] remained unchanged, whereas [3-hydroxybutyrate] decreased, although less rapidly than did its radiochemical concentration. 5. It is concluded that during starvation in 6-week-old rats, the blood [alanine] appears to influence ketogenesis for circulating unesterfied fatty acids and inversely affects gluconeogenesis from either lactate or alanine. A different pattern of gluconeogenesis may exist for alanine and lactate as evidenced by comparative 14C incorporation into liver glycogen and blood glucose.     

The influence of diet on the lipogenic response to thyroxine in rat liver.

ATP citrate lyase, acetyl-CoA synthetase, malic enzyme and hexose monophosphate dehydrogenase activities and rates of $\text{denovo}$ synthesis of long chain fatty acids from labeled acetate and citrate were measured in cell-free fractions of liver from rats fed various diets, with and without D- or L- thyroxine. Diets containign sucrose (vs. isocaloric glucose) or lard (vs. isocaloric corn oil) stimulated hepatic lipogenesis both in control and in thyroxine-treated rats. The lipogenic response to thyroxine was greatly modified by diet, except for an invariable rise in malic enzyme activity. With diets providing less than 6% of calories as linoleic acid, thyroxine increased fatty acid synthesis, depleted liver glycogen and retarded growth; when linoleic acid was increased to 16% of calories, thyroxine had no effect on fatty acid synthesis or growth and liver glycogen depletion was significantly attenuated. This response to dietary linoleic acid suggests that these phenomena may be largely secondary to the increased requirement for essential fatty acid in thyrotoxicosis. Further study should reveal the extent to which observed effects of excess thyroid hormone are amenable to control by dietary polyunsaturated fat.     

Time-course of changes in blood glucose and ketone bodies, plasma lipids and liver fatty acid composition in streptozotocin-diabetic male rats.

Male rats were given streptozotocin (100 mg/kg) by intraperitoneal injection. Groups of control and streptozotocin-treated animals were sacrificed at daily intervals for 4 days after injection. Over this period, treated rats lost weight continuously while control animals progressively gained weight. Within 24 h of treatment blood glucose and plasma free fatty acids were raised to levels which were sustained for the remainder of the experiment. After 48 h blood ketone bodies, plasma cholesterol and triglycerides were maximally raised and liver glycogen and blood lactate similarly lowered. The percentage composition of major fatty acids in liver lipids was unchanged until 4 days after treatment when there were significant increases in the proportion of oleate and linoleate and reductions in stearate and arachidonate. The data confirm that streptozotocin induces a rapid and sustained diabetes. It is suggested that metabolic experiments, in streptozotocin-diabetic rats, may be performed 48 h after treatment.     

Influence of fasting on glycogen depletion in rats during exercise

We recently observed that a 24-h fasted group of rats could run longer than an ad libitum fed control group before becoming exhausted. Because of the demonstrated importance of glycogen levels and free fatty acid availability during endurance exercise, we have investigated several parameters of carbohydrate and lipid metabolism in exercised and nonexercised rats that were either fed ad libitum or fasted for 24 h. A 24-h fast depleted liver glycogen, lowered plasma glucose concentration, decreased muscle glycogen levels, and increased free fatty acid and beta-hydroxybutyrate concentrations in plasma. During exercise the fasted group had lower plasma glucose concentration, higher plasma concentration of free fatty acids and beta-hydroxybutyrate, and a lower muscle glycogen depletion rate than did the ad libitum fed group. Since fasted rats were able to continue running even when plasma glucose had dropped to levels lower than those of fed-exhausted rats, it seems unlikely that blood glucose level, per se, is a factor in causing exhaustion. These results suggest that fasting increases fatty acid utilization during exercise and the resulting "glycogen sparing" effect may result in increased endurance.     

Effect of maternal exercise on fetal liver glycogen late in gestation in the rat

To determine the effect of maternal exercise on fetal liver glycogen content, fed and fasted rats that were pregnant for 20.5 or 21.5 days were run on a rodent treadmill for 60 min at 12 m/min with a 0% grade or 16 m/min up a 10% grade. The rats were anesthetized by intravenous injection of pentobarbital sodium, and fetal and maternal liver and plasma samples were collected and frozen. Fetal liver glycogenolysis did not occur as a result of maternal exercise. Fetal blood levels of lactate increased 22-60%, but glucose, plasma glucagon, and insulin were unchanged during maternal exercise. Maternal liver glycogen decreased as a result of exercise in all groups of rats except the fasted 20.5-day-pregnant group. Plasma free fatty acids increased in all groups and blood lactate increased in fed (20.5 days) and fasted (21.5 days) pregnant rats. Maternal glucose, glucagon, and insulin values remained constant during exercise. The fetus appears to be well-protected from metabolic stress during moderate-intensity maternal exercise.     

Pyruvate dehydrogenase activities during the fed-to-starved transition and on re-feeding after acute or prolonged starvation.

We investigated the temporal relationship between hepatic glycogen depletion and cardiac and hepatic PDH (pyruvate dehydrogenase complex) activities during the acute phase of starvation. There was a striking correlation between the decline in hepatic glycogen and PDH inactivation during the first 10 h of starvation. Re-feeding after 6 h starvation was associated with complete re-activation of PDH in liver and re-activation to approx. 75% of the fed value in heart, whereas in rats previously starved for 24-48 h re-activation was delayed in liver and diminished in heart. The results are discussed with reference to the fate of dietary carbohydrate after re-feeding.     

Metabolic effects of ethanol in the rat liver.

The NADPH is one of the cofactors in ethanol metabolism. The aim of the study was to investigate the effect of ethanol on a NADPH generating enzyme (G6P-DH) and on some metabolic parameters of the liver. After a 2-day starvation period rats were fed a lipid free diet for three days. During this refeeding period the animals were divided into three groups; they received a single daily dose of 4 g per kg b.w. ethanol, isocaloric aqueous glucose solution or water by gastric tube. In response to ethanol the activity of hepatic G6P-DH decreased. The amount of triglyceride remained unchanged, certain changes occurred in the fatty acid composition of total lipid. The liver glycogen content was elevated. In female rats treated with ethanol the activity of glucose-6-phosphatase increased.     

Changes of glycogen content in liver,skeletal muscle,and heart from fasted rats

Glycogen content of white and red skeletal muscles, cardiac muscle, and liver was investigated in conditions where changes in plasma levels of non‐esterified fatty acids (NEFA) occur. The experiments were performed in fed and 12 and 48 h‐fasted rats. The animals were also submitted to swimming for 10 and 30 min. Glycogen content was also investigated in both pharmacologically induced low plasma NEFA levels fasted rats and pharmacologically induced high plasma NEFA levels fed rats. The participation of Akt and glycogen synthase kinase‐3 (GSK‐3) in the changes observed was investigated. Plasma levels of NEFA, glucose, and insulin were determined in all conditions. Fasting increased plasma NEFA levels and reduced glycogen content in the liver and skeletal muscles. However, an increase of glycogen content was observed in the heart under this condition. Akt and GSK‐3 phosphorylation was reduced during fasting in the liver and skeletal muscles but it remained unchanged in the heart. Our results suggest that in conditions of increased plasma NEFA levels, changes in insulin‐stimulated phosphorylation of Akt and GSK‐3 and glycogen content vary differently in liver, skeletal muscles, and heart. Akt and GSK‐3 phosphorylation and glycogen content are decreased in liver and skeletal muscles, but in the heart it remain unchanged (Akt and GSK‐3 phosphorylation) or increased (glycogen content) due to consistent increase of plasma NEFA levels. Copyright © 2009 John Wiley & Sons, Ltd.     

The effect of starvation and refeeding on lipogenic enzymes in mammary glands and livers of lactating rats.

Lactating rats were starved for 48 h and refed a high-carbohydrate diet for a further 48 h. Starvation stops milk secretion, which resumes shortly after refeeding. Three lipogenic enzymes, fatty acid synthase, glucose 6-phosphate dehydrogenase (EC 1.1.1.49) and 'malic' enzyme (EC 1.1.1.40) all decrease in the mammary gland during starvation and are restored to the pre-starvation levels 48 h after refeeding. The same enzymes in liver also decrease during starvation, but increase to values significantly higher than those for the normal fed rats after refeeding the high-carbohydrate diet. For the fatty acid synthase these values were four times the pre-starvation values. Serum insulin and prolactin concentrations also increased upon refeeding the high-carbohydrate diet.     

Glycogen-storage disease in rats, a genetically determined deficiency of liver phosphorylase kinase.

Rats from an inbred strain (NZR/Mh) were found to have high concentrations of glycogen in their livers, even after 24 h of starvation. Despite this, blood glucose concentrations were well maintained on starvation for up to 72 h. The primary defect is a deficiency of liver phosphorylase kinase, causing a lack of active glycogen phosphorylase, although total phosphorylase is normal. The intravenous injection of glucagon caused a rapid activation of cyclic AMP-dependent protein kinase in the liver, but no increase in either phosphorylase kinase or phosphorylase a activity. Although total glycogen synthase activity in the livers of affected rats was higher than normal, glycogen synthase in the active form was very low, presumably as a result of the high liver glycogen content. The condition is transmitted as autosomal recessive and, apart from hepatomegaly, the affected rats appear healthy.     

Metabolic utilization of muscular L-proline in 24-hr starved rats.

1. The aim of this paper was to study the in vivo skeletal muscle L-proline related to its destination to other key tissues such as liver and intestine as well as to give some insight into the role of blood cells in proline handling. 2. L-U-[14C]Proline was injected intramuscularly and following by sampling of blood, liver, intestine and contralateral muscle at 20 and 30 min after injection. 3. The distribution of radioactivity between blood cells and plasma and in total and individual amino acids, protein and glycogen fractions was determined in the above tissues. 4. The pattern of well fed rats was compared with those submitted to 24-hr complete starvation. 5. During starvation a minor degree of proline oxidation occurs. 6. The main destruction of proline in the liver seem to be the synthesis of proteins. 7. The radioactivity recovered in the blood proline fraction of starved rats is twice that of the fed rats and that it could be attributed mainly to plasma protein. 8. We have obtained in vivo evidence for the role of erythrocyte in the interorgan proline transport.     

Effects of Feeding a Histidine-excess Diet and Subsequent Starvation on Liver and Muscle Glycogen,and on Serum Glucose

The effects of feeding with a histidine-excess diet and subsequent starvation on liver and muscle glycogen, and on serum glucose were investigated in young and adult rats.

Feeding with a histidine-excess diet resulted in the accumulation of liver glycogen in both young and adult rats. The hepatic glycogen continued to decrease during starvation, and the liver became almost totally depleted of glycogen after starvation for 48 hr. Glycogen in the liver of young rats starved for 24 hr after previous feeding with a histidine-excess diet was significantly higher than that of young rats starved for 24 hr after previous feeding with a basal diet.

Muscle glycogen after feeding and subsequent starvation was not affected by the types of diets fed previously, muscle glycogen during starvation showing a slight decrease in young rats and a slight increase in adult rats.

Feeding with a histidine-excess diet caused a significant decrease of serum glucose in young rats, but not in adult rats. Serum glucose in young rats was markedly reduced by starvation after previous feeding with a basal diet, but not after previous feeding with a histidine-excess diet. In adult rats, there were no changes in serum glucose between rats starved after feeding with either a basal diet or a histidine-excess diet, and serum glucose was decreased slightly by starvation after feeding with the test diets.

The overall results indicate that the maintenance of serum glucose in young rate even during starvation after previous feeding with a histidine-excess diet might be partially concerned with the export of glucose from the accumulated glycogen in the liver due to the diet.     

The Effect of Lipid Peroxide on the Lipid and Carbohydrate Metabolism in Rat Liver

Feeding tests were carried out on rats to clarify the mechanisms of fatty liver formation induced by autoxidized methyl linoleate. Lipid peroxides prepared by autoxidation of highly purified methyl linoleate were given orally to rats. Triglyceride and glycogen contents in liver were determined and enzyme activities including triglyceride synthetase and α-glycerophosphate dehydrogenase were also examined. The following results were obtained. 1. Triglyceride accumulation in rat liver fed autoxidized methyl linoleate was observed. 2. Increase in triglyceride content in rat liver was soon followed by the decrease of hepatic glycogen. 3. When rats were starved prior to introduction of autoxidized methyl linoleate, hepatic triglyceride accumulation did not occur. 4. The activities of α-glycerophosphate dehydrogenase and triglyceride synthetase in liver, and those of glutamic oxalacetic transaminase and leucine aminopeptidase in plasma were practically similar among the rats of test groups fed fresh or autoxidized methyl linoleate and the control fed diet without methyl linoleate. 5. The addition of l-carnitine which is a stimulator of fatty acid oxidation retarded the accumulation of the hepatic triglyceride mentioned above.     

Endocrine-metabolic relations in rats with genetic arterial hypertension

Levels of triiodothyronine, thyroxine, insulin, glucose and free fatty acids in the blood; contents of adrenaline, noradrenaline in adrenals and glycogen in the liver; activity of phenylethanolamine-N-methyltransferase in adrenals, hexokinase and glucose-6-phosphatase in the liver were studied in male Wistar rats and rats with inherited stress-induced arterial hypertension /ISIAH/. It was found that genetically caused rise of hypophyseal-thyroid systems activity in ISIAH-rats leads to a decrease of insulin blood level, activation of lipolysis and breach of glucose tolerance.     

Induction of fatty liver by fasting in suncus

We found that a fatty liver was easily induced in a novel experimental animal, Suncus murinus (suncus), by withholding food. Hepatic triglyceride content increased linearly for up to 24 h after fasting in these animals. Serum levels of neutral lipids are very low in the fed state compared with those in rats, and decreased significantly after 24 h fasting. On the other hand, serum free fatty acids, which are at the same level in fed animals as in rats, increased threefold in the fasting suncus. In order to learn whether the fatty liver induced by fasting is an unusual physiological state or a pathological on-going state in suncus, they were refed after 24 h fasting. Refeeding resulted in a decrease in hepatic triglyceride content to the level of fed animals. Serum lipid levels, which decreased with fasting, returned to those of fed animals. This evidence indicates that hepatic lipid secretion is impaired even in a physiological state to some extent and that starvation causes increasing influx of free fatty acid to the liver, which might be followed by esterification and result in triglyceride accumulation in the liver. In conclusion, hepatic lipid and lipoprotein metabolism is unique to the suncus, which is a useful animal model for the study of intra-hepatic lipid transport.