The effects of progesterone on isolated rat adipocytes were studied in vitro during various steps of glucose metabolism, transport, lipogenesis and lipolysis. Progesterone decreased the phosphorylation of glucose into glucose-6-phosphate as assessed by measuring the uptake of 2-deoxyglucose but it had no effect on transmembrane transport of glucose as determined by measuring the entry of 3-0-methylglucose into the cell. As glucose phosphorylation is a rate-limiting step of the pentose-phosphate pathway, these data could explain the inhibition of lipogenesis and the enhancement of lipolysis observed when progesterone is present in incubation medium. Progesterone might thus modulate a regulatory step of glucose metabolism and antagonize insulin action in the fat cell.
M.-TH. SUTTER-DUB, B. DAZEY, E. HAMDAN and M.-TH. VERGNAUD
I. Gause and S. Edén
The hormonal regulation of GH binding and the effects of GH on glucose metabolism were studied in hypophysectomized rats. Male rats (130–140 g) were hypophysectomized and on the day after the operation treatment with one or a combination of two hormones was started and continued for 7 days. The different hormonal treatments were (1) cortisone acetate, (2) insulin, (3) insulin plus cortisone acetate, (4) thyroxine, (5) thyroxine plus cortisone acetate and (6) GH. Glucose metabolism was studied by determining the production of CO2 from [14C]glucose in epididymal fat pads and adipocytes and the incorporation of [14C]glucose into lipids in adipocytes. Binding of GH was measured in cell aliquots using 125I-labelled human GH.
In hypophysectomized control animals, GH binding was decreased to approximately 25% of the binding observed in adipocytes of normal rats. Insulin treatment increased GH binding by approximately 100% and the response to GH was markedly increased. Similar effects were achieved by thyroxine treatment. Basal levels of glucose oxidation were markedly decreased after hypophysectomy but were increased towards normal by insulin or thyroxine treatment. Neither cortisone nor GH treatment had any effect on GH binding or glucose metabolism.
The results show that insulin and thyroxine may be important for the GH receptor and the insulin-like effect of GH in adipocytes.
J. Endocr. (1985) 105, 331–337
S Ranganathan and PA Kern
Treatment of HIV infection using protease inhibitors is frequently associated with lipodystrophy and impaired lipid and glucose metabolism. We examined the effect of saquinavir, one of the protease inhibitors, on lipid metabolism and glucose transport in cultured adipocytes. Saquinavir inhibited lipoprotein lipase (LPL) activity in 3T3-F442A and 3T3-L1 adipocytes. The inhibition of LPL was 81% at a concentration of 20 microg/ml. Another closely related drug, indinavir, had a small inhibitory effect. Saquinavir also inhibited the biosynthesis of lipids from [(14)C]-acetate. Saquinavir increased the lipolysis. Saquinavir had no significant effect on the cellular protein synthesis or protein content. Saquinavir increased the basal glucose transport threefold and decreased insulin-stimulated glucose transport by 35%. These studies suggest that some HIV protease inhibitors have direct effects on lipid and glucose metabolism. This inhibition of lipogenesis and glucose transport may explain some of the lipodystrophy, dyslipidemia and disturbed glucose metabolism with the clinical use of these drugs.
M. H. VAN WOERT, P. S. MUELLER, L. M. AMBANI and U. RATHEY
Insulin and glucose metabolism were studied in parkinsonian patients before and during treatment with l-DOPA and in a chronically ill, elderly, control group of patients. The parkinsonian patients had a low absolute glucose disappearance rate and a low serum insulin response to intravenous glucose compared with controls, which was not altered by l-DOPA therapy. The serum glucose and insulin responses to the oral glucose tolerance tests in parkinsonian patients were similar to those of the control group. Normal immediate insulin responses were observed after infusions of tolbutamide and glucagon.
l-DOPA decreased serum insulin and glucose levels during the first 90 min of the oral glucose tolerance test and produced an increase in concentration of human growth hormone in serum of some parkinsonian patients. l-DOPA therapy had no effect on intravenous tolbutamide and glucagon tolerance tests. Our results indicate a selective defect in the mechanism of intravenous glucose-induced insulin release in patients with Parkinson's disease.
R. Gross and P. Mialhe
To elucidate the hypolipacidaemic effect of insulin in ducks, its action on the uptake of free fatty acids (FFA) by duck hepatocytes was determined. At low doses (10 mu./l) insulin stimulated FFA uptake. This effect was not observed with higher doses of insulin (20, 30 and 50 mu./l). Growth hormone at physiological concentrations and corticosterone (14·4 nmol/l) decreased basal activity, probably by reducing glucose metabolism and consequently α-glycerophosphate (α-GP) supply. Insulin was able to reverse the inhibition induced by GH and corticosterone on both FFA uptake and α-GP production. These results therefore suggest that the hypolipacidaemic effect of insulin may be partly mediated by its action on hepatic FFA uptake.
J. Endocr. (1984) 102, 381–386
Daugaard JR, JL Laustsen, BS Hansen and EA Richter
The isolated effect of growth hormone on carbohydrate metabolism in rat skeletal muscle was studied in growth hormone-deficient dwarf rats (dw/dw) treated with either recombinant human growth hormone or saline for 10 days. In addition, age-matched heterozygous (DW/dw) (normal weight and plasma IGF-I) control rats were treated with saline. Growth hormone increased weight gain from 0.1+/-0.1 (s.e.m) to 3.6+/-0.1 g/day and plasma IGF-I concentration from 364+/-23 to 451+/-32 ng/ml. Glucose metabolism in skeletal muscle perfused with basal, submaximal and maximal concentrations (0, 600 and 60 000 pmol/l respectively) of insulin was not changed by growth hormone. No change could be detected in the total number of glucose transporters (GLUT1 and GLUT4) in the skeletal muscles, except from a lower amount of GLUT4 in the soleus muscle in the heterozygous control group. However, at submaximal insulin concentrations, skeletal muscle glucose uptake and transport were significantly lower in the heterozygous control group compared with the growth hormone-deficient group. This could indicate either a direct long-term effect of growth hormone or more likely a secondary effect attributable to the difference in body weight (205.2+/-3.1 vs 361. 6+/-5.9 g for dwarf rats and heterozygous controls respectively), and thereby muscle fibre size, between the groups probably resulting in lower average interstitial insulin and glucose concentrations at a given plasma concentration in the heterozygous rats. It is concluded that restoration of subnormal growth hormone concentrations for 10 days has no effect on insulin-stimulated glucose metabolism in skeletal muscle in vitro.
C. Svensson, S. Sandler and C. Hellerström
Previous studies have shown that 4 weeks after syngeneic transplantation of a suboptimal number of islets into either C57BL/6J (BL/6J) or C57BL/KsJ (BL/KsJ) diabetic mice there is an impaired insulin secretion by the perfused grafts. After normalization of the blood glucose level with a second islet graft, the BL/6J strain showed restored insulin secretion whilst that of the BL/KsJ strain remained impaired. The aim of the present work was to study the effects of glucose on the in-vitro function of islet β-cells from these two mouse strains, with different sensitivities of their β-cells to glucose in vivo. Isolated pancreatic islets from each strain were kept for 1 week in tissue culture at 5·6, 11, 28 or 56 mmol glucose/l and were subsequently analysed with regard to insulin release, (pro)-insulin and total protein biosynthesis, insulin, DNA and insulin mRNA contents and glucose metabolism. Islets from both strains cultured at 28 or 56 mmol glucose/l showed an increased accumulation of insulin in the culture medium and an enhanced glucose-stimulated insulin release compared with corresponding control islets cultured at 11 mmol glucose/l. After culture at either 5·6 or 56 mmol/l, rates of (pro)insulin biosynthesis were decreased in BL/KsJ islets in short-term incubations at 17 mmol glucose/l, whereas islets cultured at 56 mmol glucose/l showed a marked increase at 1·7 mmol glucose/l. In BL/6J islets, the (pro)insulin biosynthesis rates were similar to those of the BL/KsJ islets with one exception, namely that no decrease was observed at 56 mmol glucose/l. Islets of both strains showed a decreased insulin content after culture with 56 mmol glucose/l. Insulin mRNA content was increased in islets cultured in 28 or 56 mmol glucose/l from both mouse strains. Glucose metabolism showed no differences in the rates of glucose oxidation, however, in islets cultured in 56 mmol glucose/l the utilization of glucose was increased in both BL/6J and BL/KsJ animals. There were no differences in DNA content in islets cultured at different glucose concentrations, suggesting no enhancement of cell death.
The present study indicates that, irrespective of genetic background, murine β-cells can adapt to very high glucose concentrations in vitro without any obvious signs of so-called glucotoxicity. Previously observed signs of glucotoxicity in vivo in BL/KsJ islets appear not to be related only to glucose but rather to an additional factor in the diabetic environment.
Journal of Endocrinology (1993) 136, 289–296
D. J. Jerry, L. C. Griel Jr, J. F. Kavanaugh and R. S. Kensinger
Differential binding of homologous and heterologous prolactin was investigated in porcine mammary tissue. Specific binding of ovine prolactin to porcine mammary membranes or tissue slices was significantly greater than specific binding of the homologous porcine prolactin. Ovine prolactin was also more potent than porcine prolactin in stimulating proliferation of Nb2 cells. In contrast, stimulation of glucose metabolism in porcine mammary explants by porcine prolactin was greater than that by ovine prolactin. Differences in specific binding were probably not due to damage during iodination, as low concentrations of iodinated prolactins were similar to unlabelled prolactins in their abilities to stimulate proliferation of Nb2 cells. Furthermore, electrophoretic analysis of medium from binding reactions suggested that differences in specific binding were not due to proteolytic cleavage of the homologous prolactin into large (> 10 kDa) fragments. These studies suggest that ovine prolactin either binds to sites in addition to the authentic lactogenic receptor in porcine mammary tissue or that a significantly higher affinity of ovine prolactin for the porcine lactogenic receptor has little effect on its biological activity.
Journal of Endocrinology (1991) 130, 43–51
S Yamada, M Komatsu, T Aizawa, Y Sato, H Yajima, T Yada, S Hashiguchi, K Yamauchi and K Hashizume
When isolated rat pancreatic islets are treated with 16.7 mM glucose, a time-dependent potentiation (TDP) of insulin release occurs that can be detected by subsequent treatment with 50 mM KCl. It has been thought that TDP by glucose is a Ca2+-dependent phenomenon and only occurs when exposure to glucose is carried out in the presence of Ca2+. In contrast to this, we now demonstrate TDP under stringent Ca2+-free conditions (Ca2+-free buffer containing 1 mM EGTA). In fact, under these Ca2+-free conditions glucose caused an even stronger TDP than in the presence of Ca2+. TDP induced by glucose in the absence of extracellular Ca2+ was unaffected by inhibitors of protein kinase C (PKC). However, cerulenin or tunicamycin, two inhibitors of protein acylation, eradicated TDP without affecting glucose metabolism. The TDP by glucose was not associated with an increase in the cytosolic free Ca2+ concentration ([Ca2+]i) during subsequent treatment with high K+. Exposure of islets to forskolin under Ca(2+)-free conditions did not cause TDP despite a large increase in the cellular cAMP levels. In conclusion, glucose alone induces TDP under stringent Ca2+-free conditions when [Ca2+]i was significantly lowered. Protein acylation is implicated in the underlying mechanism of TDP.
P. E. B. REILLY and E. J. H. FORD
Continuous infusions of tracer amounts of [U-14C]glucose were used to measure the effect of a single intramuscular injection of 0·5 mg betamethasone/kg on the rates of glucose entry in six sheep eating 900 g/day chopped hay containing 49 g protein. In four of the sheep the rates of oxidation of glucose to CO2 were also measured. Hyperglycaemia occurred with a maximum plasma glucose concentration occurring 24 h after administration of the steroid. The mean glucose entry rates, expressed as mg/min/kg ± s.e.m., rose from 1·27 ± 0·11 (6) to 1·75 ± 0·16 (6). The difference between these means is significant (P < 0·001). The mean proportion of CO2 derived from glucose after administration of betamethasone (11·64 ± 1·4% (4)) was not significantly different (P > 0·5) from that derived before its administration (12·47 ± 0·34% (4)). The amounts of glucose available for synthetic purposes increased because the total production rates increased. The mean extra amount available was 11·3 ± 2·1 (4) mg C/min. These results are discussed in the light of other findings relating to the actions of the glucocorticosteroids on glucose metabolism.