Jane Bond

EFFECT OF FATTY ACIDS ON THE PROLIFERATION OF CONCANAVALIN A-STIMULATED RAT LYMPH NODE LYMPHOCYTES

1. The effect of a range of fatty acids upon concanavalin A-stimulated [3H]thymidine incorporation into rat lymphocytes was investigated. 2. All fatty acids tested inhibited the response to mitogen but the extent of the inhibition was dependent upon the fatty acid concentration used, the time of addition of fatty acid and the duration of exposure of the cells to fatty acid. 3. All fatty acids were inhibitory at concentrations of 50 microM or above; at lower concentrations some were inhibitory and some were stimulatory. Above 50 microM the inhibitory effect was concentration dependent; the greater the fatty acid concentration, the greater the inhibition. 4. The longer the lymphocytes were exposed to the fatty acid the greater was the inhibitory effect. This was true if the fatty acids were added at the same time as the mitogenic stimulus or if they were added before or after the stimulus. Some fatty acids maintained their inhibitory effect when added 24 or 48 hr after the mitogenic stimulus. 5. Generally unsaturated fatty acids were more inhibitory than saturated fatty acids; the greatest inhibition of proliferation was caused by eicosapentaenoate and arachidonate and the least inhibition by myristate and palmitate. 6. Inhibition was greater in the absence of serum. 7. Inhibition by unsaturated fatty acids could be partially or totally relieved by addition in combination with myristate or palmitate, suggesting that the inhibitory effect of fatty acids may be due to alteration of membrane fluidity caused by an imbalance of fatty acids presented to the cells. 8. PGE2 levels were similar in the medium of cells grown in the presence of fatty acids with varying inhibitory effects, indicating that PGE2 production is not the sole mechanism of suppression of the proliferative response. 9. Although the mechanism by which fatty acids exert their effect remains to be determined, these results indicate that lymphocyte proliferation and so an immune response could be influenced by dietary lipid manipulation.

The effect of tumour bearing on skeletal muscle glutamine metabolism

1. The effects of tumour bearing on glutamine metabolism in rat skeletal muscle were examined using the Walker 256 carcinosarcoma. 2. There was a rapid and marked decrease in skeletal muscle glutamine content, which was correlated with the size of the tumour, and a decrease in plasma glutamine concentration. 3. The rate of release of glutamine from EDL muscle in vitro was increased in cachectic, tumour bearing animals, but was unaffected from the soleus muscle of the same animals. 4. It is hypothesized that the increase in the rate of muscle glutamine release during cachexia represents a response of this tissue in order to satisfy the demand for glutamine by the tumour or by cells of the immune system.

Effects of physiological and pathological levels of glucocorticoids on skeletal muscle glutamine metabolism in the rat

The effects of physiological and pathological concentrations of glucocorticoids were investigated using the glucocorticoid antagonist RU486 and the synthetic glucocorticoid dexamethasone, respectively. The effects of these treatments on the concentrations of glutamine and other amino acids in skeletal muscle and plasma and on the rates of release of glutamine and alanine from incubated preparations of skeletal muscle of the rat were investigated. Dexamethasone treatment increased the concentration of glutamine and the rate of release of this amino acid from incubated soleus muscle preparations. This treatment decreased the concentration of glutamine in both gastrocnemius and EDL muscles, but was without effect on the rate of glutamine release from EDL muscles. In contrast, administration of the glucocorticoid antagonist RU486 decreased the rate of glutamine release from muscle. It is concluded that glucocorticoids have marked effects on the metabolism of glutamine by skeletal muscle per se and that these hormones may be important in the control of the rate of glutamine release from muscle in both physiological and pathological conditions.

Propionate regulates lymphocyte proliferation and metabolism

1. The effect of propionate on lymphocyte proliferation and metabolism was investigated. Lymphocytes obtained from human blood and rat mesenteric lymph nodes were utilized. 2. Propionate at concentrations of 0.04 and 1.0 mmol/l stimulated the amount of [3H]thymidine incorporated either in cultured human T lymphocytes or rat T and B lymphocytes. 3. Concentrations of propionate between 2 and 5 mmol/l caused a marked inhibition of lymphocyte proliferation. 4. This short-chain fatty acid was metabolized by these cells and produced succinate in significant amounts; however, its oxidation was low. 5. Propionate did not alter glucose, glutamine and pyruvate utilization and oxidation in incubated rat lymphocytes but increased the formation of lactate and aspartate. 6. In contrast, propionate inhibited by 50% the synthesis of lymphocyte lipid from [1-14C]acetate at concentrations of 0.5 and 1 mmol/l and reduced by half the incorporation of 3H2O into lipids at 1 and 5 mmol/l. 7. The results suggest that inhibition of lipid synthesis is a possible mechanism leading to reduction of lymphocytes proliferation.

Studies on the effects of growth hormone administration in vivo on the rates of glucose transport and utilization in rat skeletal muscle

Abstract. The effects of growth hormone (GH) administration to rats in vivo on the sensitivity of the rate of glucose utilization to insulin were studied in soleus muscles isolated from these rats. A single injection of GH did not increase the rate of glucose transport within 1–2 h. However, 12 h after, the rate of glucose transport was increased at 10 mU insulin l‐1 and was accompanied by a similar increase in the rate of lactate formation but no change in the rate of glycogen synthesis. Prolonged treatment with GH decreased the rate of glucose transport and glycogen synthesis and increased the content of glucose 6‐phosphate at physiological levels of insulin but did not affect the rate of lactate formation. These results suggest that: (a) GH does not increase the rate of glucose transport acutely; however, after several hours, the sensitivity of glucose transport and glycolysis to insulin are increased; (b) prolonged elevations of the level of GH in plasma decrease the sensitivity of the rate of glucose transport and glycogen synthesis to insulin. However, redirection of glucose residues away from the pathway of glycogen synthesis towards that of glycolysis and a possible increase in the rate of glycogenolysis maintain a normal rate of lactate formation, although the rate of glucose transport is decreased.

Thyroid hormone analogue SKF L-94901: effects on amino acid and carbohydrate metabolism in rat skeletal muscle in vitro

In summary, hyperthyroidism increased the rate of glycolysis and decreased glycogen synthesis in isolated incubated rat soleus muscle preparations. SKF 901 also increased glycolysis, but the stimulation was 5-fold less than in T3-treated muscles. Hyperthyroidism increased the rate of glutamine release from skeletal muscle, but SKF 901 did not affect glutamine metabolism.

Propionate modifies lipid biosynthesis in rat peritoneal macrophages

1. This study examines the effect of propionate, normally produced in the gut, on lipid metabolism of resident macrophage. This cell is very abundant in the epithelial lining of the gut. 2. The activity of propionyl-CoA synthetase in macrophages was shown to be 0.39 nmol/min per mg protein, so this cell presents the ability to use propionate. Propionate at concentrations varying from 0.5 to 5 mM did not affect the activities of carnitine acetyltransferase, ATP-citrate lyase, acetoacetyl-CoA thiolase and 3-oxoacid-CoA transferase. 3. Thus this short chain fatty acid did not alter the capacity for transferring acetyl-CoA from mitochondria to cytosol and for ketone bodies formation and oxidation. However, propionate (40 mM) inhibited the incorporation of [1-14C]-palmitate into phospholipids, cholesterol, cholesterol ester and triacylglycerol and the incorporation of [3-14C]-pyruvate into phospholipids. 4. These findings suggest that fibre-rich diet by generating propionate may regulate macrophage lipid metabolism.

The effect of propionate on lipid synthesis in rat lymphocytes

1. The effect of propionate on lipid synthesis in lymphocytes cultured for 24 hr and incubated for 2 hr was investigated. 2. [1-14C]-propionate was incorporated mainly into phospholipids in both control and concanavalin A (Con A) stimulated cultured lymphocytes. 3. The content of free coenzyme A markedly decreased in 2 hr incubated lymphocytes when propionate was added to the medium at concentrations from 10 to 100 mmol/l. 4. Propionate at 40 mmol/l decreased the incorporation of [1-14C]-palmitate into phospholipids (86%), triacylglycerol (87%) and cholesterol ester (98%) and increased in cholesterol (133%) of cultured lymphocytes. 5. Addition of propionate into the culture medium at 2.5 and 5.0 mmol/l concentrations markedly increased the activity of hydrolases of various acylCoA derivatives. 6. The results suggest that propionate may reduce the content of acylCoA and so its esterification and this might be important for the regulation of lymphocytes proliferation.