Liver Glycogen Deposition Research Articles

Nephelometric determination of glycogen in the rat liver glycogen deposition assay of adrenal steroids.

A method is described for rapid and precise nephelometric determination of rat liver glycogen. Instrument readings are made following addition of diluted alkaline liver digests to 57.5% alcohol. Results indicate good recovery and reproducibility, with adherence to conditions used for the std. curve. Fifteen solutions containing varying amounts of glycogen showed an average recovery of 98.19% ± 0.62%. The procedure was used in bio-assay of adrenal steriods and found to compare favorably to that recommended by NF XI.

The effects of some 21-methyl-substituted corticoids on inflammation, liver glycogen and electrolyte-regulating activity in the rat.

ABSTRACT The anti-inflammatory, liver glycogen deposition and electrolyte-regulating activities in the rat have been studied for the following new steroids: 21-Me-9α-F-Δ1-F 21α-Ac; its 21β-Ac; 21-Me-6α,9α-diF-Δ1-F 21α-Ac; and its 21β-Ac. Using the granuloma pouch assay, results show that there is a linear relationship between degree of inhibition of inflammatory exudate and logarithm of dose assayed for each steroid. The most potent of the new steroids is 21-Me-6α,9α=diF-Δ1-F 21α-Ac (ca. four times Δ1-F Ac), next is its 21β-Ac epimer (ca. twice Δ1-F Ac) and the lowest in activity is 21-Me-9α-F-Δ1-F 21β-Ac (almost equipotent to Δ1-F Ac). Results of liver glycogen deposition assay reveal an even greater potency of the new steroids in relation to Δ1-F Ac, although the rank of activity is the same as that obtained from the granuloma pouch test. The intense sodium retaining-potassium excreting activity of 9α-F-Δ1-F Ac, the parent compound of the new steroids, is modified by the 21-methyl substituent with the result that the 21-methyl steroids induce more natriuresis with the exception of 21-Me-9α-F-Δ1-F 21β-Ac, and less kaliuresis, than the parent compound. The introduction of a 21-methyl grouping creates a new center of asymmetry at C-21. Results from the present investigation indicate that for a given steroid the 21α-ol configuration has more anti-inflammatory and glucogenic activities than the corresponding 21β-ol. The 21-methyl corticoids may be of usefulness to treat inflammatory disorders in man.

CORTISOL STIMULATION OF GLYCOGEN SYNTHESIS IN FASTED RATS1

The acute effects of cortisol that result in an increase in liver glycogen deposition were studied in fasted rats, using C14O2 and glucose-6-C14 as labeled substrates. The incorporation of labeled carbon into blood glucose, liver glycogen and liver dicarboxylic acids was determined. The results of these experiments indicate that cortisol increases the incorporation of label from radioactive bicarbonate into liver glycogen without any significant increase in the activity of either blood glucose or liver dicarboxylic acids. Studies with labeled glucose showed increased deposition of glycogen from this substrate. There was little recycling of C14 from glucose via the Cori cycle. Under these conditions cortisol did not appear to increase hepatic glucose production as measured by changes in specific activity of blood glucose. Evidence is presented that in the fasting adrenalectomized animal there is some specific impairment in glycogen formation apart from the ability of the liver to form glucose. It is sugges...

Metabolism of acetate in scorbutic guinea pigs.

The effects of feeding acetate on the glucose tolerance, deposition of liver glycogen; urinary excretion of citric acid, malic acid and ketone bodies; total body lipid, phospholipid and cholesterol; and cholesterol content of different tissues were studied in normal and scorbutic guinea pigs. The effects of prolonged insulin treatment on the urinary excretion of citrate, malate and ketone bodies were also studied in normal and scorbutic guinea pigs fed acetate. It was observed that when normal guinea pigs were fed acetate, total body lipid and liver glycogen increased and there was no change in body cholesterol, tissue cholesterol and glucose tolerance. In scorbutic guinea pigs total body cholesterol and cholesterol contents of blood, small intestine and testes were increased and were further enhanced when the animals were fed acetate. Lowered glucose tolerance, diminished liver glycogen, total body lipid and phospholipid observed in scorbutic guinea pigs did not change after acetate feeding. The increased urinary excretions of ketone bodies, citric acid and malic acid by scorbutic guinea pigs came down to normal levels when they were injected with insulin. The increased cholesterogenesis in scurvy seems to be due to hypoinsulinism.

THE DURATION OF GLUCAGON EFFECT1

The duration of glucagon effect was determined by measuring the interval after an injection of glucagon into fasted rats, before glucose feeding resulted in liver glycogen deposition. Doses of 0.2–0.4 mg./kg. inhibited liver glycogen deposition for 2–2.5 hours. Larger doses (1.0 mg./kg.); administered either intravenously or subcutaneously, produced inhibition of glycogenesis for 4-7 hours. However, increasing the intraperitoneal dose resulted in less prolongation of effect. Zinc glucagon induced more prolonged effects than glucagon. Large doses (5.0 mg./kg.) of zinc glucagon inhibited liver glycogen deposition for at least S hours. The effects observed were not due to depletion of “primer glycogen.”

Relationship of vitamin B6 to gluconeogenic action of cortisol.

Recent investigations have shown that cortisol administration results in a marked increase in liver glutamic-pyruvic transaminase enzyme activity. Because of this observation it has been suggested that gluconeogenic action of adrenal steroids may depend upon elevation of this transaminase level. Since pyridoxal phosphate is the cofactor for the enzyme, effects of cortisol on liver glycogen deposition and glutamic-pyruvic transaminase activity were studied in vitamin B6 deficient and control rats. The findings demonstrate that glucocorticoid action of cortisol is diminished in B6 depleted animals as is the effect of the hormone on transaminase activity. Other parameters of cortisol action such as involution of lymphoid tissue and inhibition of ACTH release were not altered by the vitamin deficiency. It is concluded that gluconeogenic activity of adrenal corticoids depends not only on their molecular configuration but also upon nutritional status of the animal. The findings also support the concept that glu...

Adrenal Corticoid Activities of 1, 9 fluoro-hydrocortisone 16 , 17 isopropylidenedioxy.

Glucocorticoid properties of subcutaneously and orally administered triamcinolone acetonide were assayed in immature adrenalectomized rats by liver glycogen deposition and thymus involution. Subcutaneously, triamcinolone acetonide is about 3 times as potent as triamcinolone on liver glycogen assay and 7 times triamcinolone on the thymolytic assay. Oral activities of triamcinolone acetonide also were greater than those of the parent steroid. Triamcinolone acetonide was capable of inducing a diuretic, natriuretic and kaliuretic response in adrenalectomized rats. Triamcinolone acetonide was more active than either desoxycorticosterone or triamcinolone for maintaining the life of immature, adrenalectomized male rats.

Biological effects of hypoglycin A in hydrocortisone-treated adrenalectomized rats.

SummaryAdministration of hypoglycin A (cyclopropanepropionic acid, α-amino-2-methylene) to adrenalectomized male rats reduced the degree of hydrocortisone-induced liver glycogen deposition and hyperglycemia, without altering either urinary glucose or nitrogen excretion.

ADRENAL CORTICOID ACTIVITIES OF 9α-FLUOROllβ,16α7α,21-TETRAHYDROXY-l,4-PREGNADIENE-3,20-DIONE1231,2,3

Triamcinolone (9α-fluoro-l1β,16α,17α,21-tetrahydroxy-l,4-pregnadiene-3, 20-dione) was studied in rats to assess its adrenocortical activities using the following assays: electrolyte excretion, liver glycogen deposition, growth-survival and anti-inflammatory activity (asbestos pellet technique). Subcutaneously administered triamcinolone was 3 to 12 times more potent than prednisolone and 10 to 40 times more active than hydrocortisone in liver glycogen deposition tests. On oral assay, the relative potency of triamcinolone was higher. Triamcinolone proved to be 5 to 10 times more active than hydrocortisone acetate in the anti-inflammatory assay. Triamcinolone was more efficacious than hydrocortisone and at least as effective as DOC in maintaining the life of immature, male, adrenalectomized rats. Although survival time was prolonged, there was a suppression of growth when compared with the DOC-treated rats. Triamcinolone was found to be a potent diuretic and natriuretic steroid.

Adrenal, Hypophyseal and Pancreatic Hormones in the Liver Glycogen Response to Low Atmospheric Pressure

The administration of 3 cc of adrenocortical extract (Upjohn) to adrenalectomized rats fasted at sea-level fails to cause liver glycogen deposition. Such animals under low atmospheric pressure accumulate about 1% liver glycogen. Adrenalectomized-hypophysectomized rats maintained on a minimal dose of hypophyseal extract, given 3 cc of adrenocortical extract and stressed do not deposit liver glycogen. These results suggest that the hypophyseal hormones function not only to control the adrenal cortices under these conditions, but also may contribute directly to the observed carbohydrate alterations. When larger doses of hypophyseal hormones are used there is a significant difference in liver glycogen between the stressed and nonstressed groups indicating the possible implication of still another agent, perhaps insulin, since alloxan diabetic rats do not accumulate liver glycogen in response to low atmospheric pressure.

Effect of Alterations in Side Chain upon Anti-inflammatory and Liver Glycogen Activities of Hydrocortisone Esters

A number of esters of hydrocortisone and related steroids have been examined for activity in the granuloma inhibition (subcutaneous cotton pellet) test, both after local application of the steroids and after systemic administration. Liver glycogen deposition activity was examined by a one-day oral test in mice, a six-day test in rats, and for duration of action after a single subcutaneous injection. Rates of absorption and activity per mg. of steroid absorbed also were determined. Wide differences in activity were found, depending upon the configuration at C-21. Part of this difference was due to diverse rates of absorption, but there were also differences in activity per mg. of steroid absorbed. It is concluded that ease of hydrolysis of the ester is also a factor in determining potency. The tertiary butyl acetate is particularly effective when locally applied, but systemic activity of this ester is relatively low because of reduced rate of absorption and decreased activity per mg. absorbed. After subcutaneous injection, the free alcohol of hydrocortisone is more rapidly absorbed and gives a more prompt response than any of its esters examined. The 9α-fluoro derivative is more gradually absorbed and gives a more sustained response than does hydrocortisone.

Adrenal Corticoid Activities of 6-Methyl- 1-Hydrocortisone

Comparisons are presented on relative potencies of Δ1-F and a new analogue, 6-methyl-Δ1-F, on 3 different assays for corticoid activity: anti-inflammatory (granuloma pouch technic), liver glycogen deposition, and sodium retention. 6-Methyl-Δ1-F is 2 times as potent as Δ1-F on the anti-inflammatory test and about 3 times Δ1-F on the glycogen deposition assay. 6-Methyl-Δ1-F, like Δ1-F, does not induce sodium and water retention on the assay for electrolyte activity, but in contrast to Δ1-F, this steroid acts as a diuretic and natriuretic agent.

Role of the Adrenal in Response of Rats to Orinase

SummaryIn the intact rat low doses of Orinase produced a significant depression of blood sugar and deposition of liver glycogen while higher doses were ineffective. The adrenalectomized rat was found to be approximately 10 times as sensitive and the adrenal demedullated rat 3-6 times more sensitive to Orinase than the intact rat. Hydrocortisone partially and epinephrine completely counteracted the hypoglycemic effects of Orinase. Failure of high doses of this sulfonylurea to produce hypoglycemia and liver glycogen deposition in the intact rats may be due to a stimulation of epinephrine secretion by the adrenal medulla. It is also suggested that glucocorticoid secretion is also stimulated by high doses of Orinase in the intact and adrenal demedullated animals.

Hypoglycemic Action of Orinase. Effect on Output of Glucose by Liver

Conclusions1. A simple method for gauging changes in the output of glucose by the liver has been described. Throughout post-absorption, there are present in hepatic venous blood aperiodic fluctuations in glucose which are reflected throughout the major peripheral arterial tree. Under the influence of the drug, these fluctuations at both sites promptly disappear. The quantitated changing rate of output of glucose by the liver after Orinase (33% fall in 10 minutes) is drastic. A small dose of glucagon by vein immediately reverses this trend with a return in exaggerated form of the normal postabsorptive fluctuatinos. As a corollary one may infer that long use of the drug does not impair liver glycogen deposition. 2. It is suggested that the site of action of Orinase is proximal to the phosphorylase enzyme systems and takes the form of a suppression of glucagon production at source with a resulting reduction of glucose output by the liver. After 70 days of intensive exposure to the drug in the normal dog, acu...

Adrenocortical properties of delta1, 4-pregnadiene-17alpha, 21-diol-3, 11, 20-trione (meticorten) and delta1, 4-pregnadiene-11beta, 17alpha, 21-triol-3, 20-dione (meticortelone).

Summary(1) Meticorten and Meticortelone, 2 new steroids, were studied in animals to assess their adrenocortical activities. (2) Glucocorticoid properties were assayed by the eosinophil response in adrenalectomized mice, by the liver glycogen deposition in adrenalectomized rats, and by the involution of the thymus in immature rats. (3) By all 3 methods, the glucocorticoid activity of Meticorten and Meticortelone was found to be 3 to 4 times greater than that of cortisone and hydrocortisone. (4) Meticorten and Meticortelone were more active than cortisone and hydrocortisone in maintaining the life of immature, male, adrenalectomized rats. (5) Prolonged administration of Meticorten, Meticortelone, cortisone and hydrocortisone to young, male rats produced the following biological changes: a) suppression of somatic growth, b) decrease in thymus, spleen and adrenal weights on an absolute basis as well as per 100 g body weight, c) increase in the relative weights of liver, kidney, testes, thyroid and pituitary a...

Maintenance of adrenalectomized dogs with 9 alpha halo adrenal and other steroids.

The 9 alpha halogenated adrenal steroids such as cortisone, hydrocortisone, corticosterone and certain derivatives of progesterone originally prepared by Fried et al. (1953, 1954, 1955a, 1955b), constitute a remarkable group of physiologically active compounds. They are unique in that some of them display potencies considerably in excess of the naturally occurring parent steroids both as to glucocorticoid and mineralocorticoid activity. The surprising feature of these 9α-halo corticoids consists primarily of the fact that substitution of a halogen atom at the 9 alpha position should so greatly enhance their potency both as to liver glycogen deposition and sodium retention (Fried et al., 1953, 1954, 1955a, 1955b). Through the cortesty of Dr. J. Fried of the Squibb Institute for Medical Research, New Brunswick, New Jersey, the writers obtained sufficient of these modified steroids for testing their life-maintaining and mineralocorticoid activity in adrenalectomized dogs. Few investigations have appeared to ...

Glucocorticoid and Mineralocorticoid Activities of 1-Fluorohydrocortisonei

Summary(1) Comparisons have been made of relative potencies of hydrocortisone, fluorohydrocortisone, Δ1-hydrocortisone, and Δ1-fluorohydrocortisone in causing deposition of liver glycogen and reduction of sodium excretion in the adrenalectomized rat. (2) In glycogen deposition potency, the compounds have activity of the following magnitude: Δ1-fluorohydrocortisone acetate-50; fluorohy-drocortisone acetate-12.6; Δ1-hydrocorti-sone-2.9; hydrocortisone-1. (3) In sodium-retaining potency, the compounds have activity of the following magnitude: Δ1-fluorohydrocortisone acetate-4.6; fluorohydro-cortisone acetate-5.0; Δ1-hydrocortisone acetate-very slight; hydrocortisone acetate -very slight; DOCA-1.

Effect of mesoxalate upon the carbohydrate figures of liver in normal and alloxan diabetic rabbits.

1. Contents of various carbohydrate in liver and blood sugar values are measured with the purpose to find out the effect of mesoxalate on the deposition of glycogen in livers of normal and alloxan diabetic rabbits.2. Injection of mesoxalate solution accelerates the deposition of glycogen in both livers of normal and of diabetic rabbits, and in the latter case the deposition of glycogen is more significant.

The adrenal cortex and sulfur metabolism; the effect of ACTH on the sulfhydryl concentration of rat adrenal, liver and muscle.

As early as 1909 it was noted that the adrenal cortex was unexpectedly active in sulfur metabolism: both its sulfur content and its ability to store such compounds were greater than those of other tissues studied (Aufrecht and Diesing, 1909; Loeper et al 1926). In 1928 it was shown that treatment of rats and rabbits with colloidal sulfur resulted in adrenal cortical hypertrophy and sulfur storage, together with massive accumulation of liver glycogen (Lurie, 1928; Foldes, 1928). At that time, however, the changes in these two organs could not be interpreted, for the relation between liver glycogen deposition and 11-oxycorticosteroids was as yet unsuspected. In recent years, further evidence has accumulated to show a relation between sulfur metabolism and adrenal cortical function. Studies with S35 for example, have confirmed the great avidity of the cortex for cystine sulfur (Lee et al., 1951).

The adrenal cortex and sulfur metabolism; the influence of amino acid sulfur on the formation of liver glycogen.

A NUMBER of years ago it was demonstrated that oral or percutaneous administration of sulfur to rats or rabbits resulted in adrenal hyperplasia and marked deposition of liver glycogen (Burgi and Gordonoff, 1926; Foldes, 1928). The possible interrelation of these two observations remained obscure at that time, for the glycogenic activity of 11-oxy corticoids was as yet unsuspected. In retrospect, it seems likely that the administered sulfur induced an adrenal hyperplasia and an outpouring of adrenal cortical hormones which resulted in excessive liver glycogen deposition. Recent studies indicating an intimate association between adrenal cortical function and sulfur metabolism (Goldzieher and Edlin, 1953; Goldzieher, Rawls and Goldzieher, 1953) have suggested further exploration of these findings. In view of the fact that colloidal sulfur, though of a very low order of toxicity, is still a foreign substance and its administration may constitute a nonspecific stress, it was decided first to investigate the ef...