Epinephrine-stimulated Lipolysis Research Articles

Abdominal Fat Cell Lipolysis, Body Fat Distribution, and Metabolic Variables in Premenopausal Women*

It is well established that abdominal obesity is related to numerous metabolic abnormalities and that this correlation represents a significant risk factor for coronary heart disease and related mortality. In the present study the relationships among the regional distribution of body fat, selected metabolic variables, and abdominal adipose cell lipolysis were investigated in 30 premenopausal women, 34 +/- 8 yr (mean +/- SD) of age, with body mass indices ranging from 17-45 kg/m2. Basal as well as epinephrine- and isoproterenol-stimulated lipolyses were positively correlated with fasting plasma insulin and triglyceride levels (0.48 less than r less than 0.64; 0.05 greater than P less than 0.0005 and 0.46 less than r less than 0.60; 0.05 greater than P less than 0.005, respectively) and with the insulin area measured during an oral glucose tolerance test (0.49 less than r less than 0.67; 0.005 greater than P less than 0.0005). With the exception of epinephrine-stimulated lipolysis, these correlations remained significant when lipolysis was corrected for cell surface area. Basal and maximal epinephrine- and isoproterenol-induced lipolyses were also negatively related to plasma high density lipoprotein cholesterol (-0.52 less than r less than -0.36; 0.05 greater than P less than 0.005). However, these relationships were no longer significant after control for fat cell surface. The associations between abdominal lipolysis and fat distribution did not remain significant when data were adjusted for total adiposity. Taken together, these results support the notion that variations in abdominal adipocyte lipolysis 1) depend more on total body fatness than on fat distribution, and 2) may be involved in the metabolic complications associated with abdominal obesity, particularly those pertaining to plasma insulin and triglyceride metabolism.

Importance of blood glucose concentration in regulating lipolysis during fasting in humans

The importance of the decline in blood glucose concentration on lipolysis and the lipolytic effect of epinephrine was evaluated during short-term fasting. Lipolytic rates were determined by infusing [2H5]glycerol and [1-13C]palmitic acid. Five volunteers were studied after 12 h of fasting before and during epinephrine infusion and after 84 h of fasting, before and during glucose infusion when plasma glucose was restored to postabsorptive values, and during glucose plus epinephrine infusion. In another protocol, five volunteers were given glucose intravenously throughout fasting to maintain plasma glucose at postabsorptive levels and isotopic studies were performed after 12 and 84 h of fasting before and during epinephrine infusion. Glucose infusion after 84 h of fasting restored glucose and insulin concentrations and lipolytic rates toward 12-h fasting values. When euglycemia was maintained throughout fasting, plasma insulin still declined (P less than 0.05) and lipolytic rates still increased (P less than 0.05). Despite similar glucose concentrations, the lipolytic response to epinephrine infusion was greater after 84 h than after 12 h of fasting in both protocols (P less than 0.05). These studies demonstrate that the decline in plasma glucose contributes to, but is not required for, the increase in lipolysis during fasting. The increase in epinephrine-stimulated lipolysis that occurs during fasting is not dependent on a decrease in plasma glucose concentration.

Determination of the Affinity of Ractopamine and Clenbuterol for the Beta-Adrenoceptor of the Porcine Adipocyte2

Dissociation constants (Kd) of ractopamine and clenbuterol for the swine adipocyte beta-adrenergic receptor were estimated from competition studies with epinephrine for the stimulation of lipolysis. Both compounds competitively inhibited epinephrine-stimulated lipolysis in the absence of adenosine deaminase. Three methods for estimating Kd values were used and similar estimates were obtained with each method. Ractopamine and clenbuterol showed greater affinity for the beta-receptor than did epinephrine and had similar Kd values of 1 to 2 x 10(-7) M. The low capacity of ractopamine and clenbuterol to stimulate lipolysis in vitro does not result from poor coupling to the beta-receptor. Ractopamine and clenbuterol may be considered partial agonists, possessing high affinity for the beta-adrenoceptor but exhibiting a relative ineffectiveness for adenylate cyclase activation.

Pervanadate [peroxide(s) of vanadate] mimics insulin action in rat adipocytes via activation of the insulin receptor tyrosine kinase

Both vanadate and hydrogen peroxide (H2O2) are known to have insulin-mimetic effects. We previously reported that the mixture of vanadate plus H2O2 results in the generation of a peroxide(s) of vanadate, which strongly enhances IGF-II binding to rat adipocytes (Kadota et al., 1987b). We now report that pervanadate mimics insulin in isolated rat adipocytes to (1) stimulate lipogenesis, (2) inhibit epinephrine-stimulated lipolysis, and (3) stimulate protein synthesis. The efficacy of pervanadate is comparable to that of insulin. However, it is 10(2)-10(3) times more potent than vanadate alone. Exposure of intact rat adipocytes to pervanadate was found to activate the WGA-purified insulin receptor tyrosine kinase assayed with the exogenous substrate poly(Glu80/Tyr20) in a dose-dependent manner to a maximum of 1464% of control at 10(-3) M compared with a maximum insulin effect of 1046% at 10(-6) M. In contrast, in vitro assayed autophosphorylation of the WGA-purified extract was increased 3-fold after exposure of intact cells to insulin but not significantly increased after pervanadate. Furthermore, high concentrations of pervanadate (10(-5) M) inhibited subsequent in vitro added insulin-stimulated autophosphorylation. In vitro addition of pervanadate to WGA-purified receptors could not stimulate autophosphorylation or exogenous tyrosine kinase activity and did not inhibit insulin-stimulated autophosphorylation. Labeling of intact adipocytes with [32P]orthophosphate followed by exposure to 10(-4) M pervanadate increased insulin receptor beta-subunit phosphorylation (7.9 +/- 3.0)-fold, while 10(-7) M insulin and 10(-4) vanadate increased labeling (5.3 +/- 1.8)- and (1.1 +/- 0.2)-fold, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of a High Fat Diet on Rat Adipocyte Lipolysis: Responses to Epinephrine, Forskolin, Methylisobutylxanthine, Dibutyryl Cyclic AMP, Insulin and Nicotinic Acid

An earlier report from this laboratory showed that feeding rats a high fat diet decreased epinephrine-stimulated lipolysis in their adipose tissue. Experiments were designed to explore further the effects of such diets on adipocyte response to epinephrine and to several other lipolytic and antilipolytic agents. Rats were fed diets with 67% of energy consisting of glucose or lard for 5 to 7 d. Adipocytes were prepared from epididymal fat pads and lipolysis measured by the release of glycerol into the medium during 1-h incubations. The cells from the rats fed the high fat diet showed lower lipolytic responses to stimulation by epinephrine, forskolin and dibutyryl cyclic AMP than those from rats fed the high glucose diet. The lard diet effect on the lipolytic response to isobutylmethylxanthine varied among experiments, but it also decreased it in some of them. However, the high fat diet did not induce decreased sensitivity or responsiveness to the antilipolytic effect of insulin, although previous reports have demonstrated resistance to other actions of insulin in rats fed a high fat diet. The antilipolytic effect of nicotinic acid was also similar in cells from rats fed a high fat diet to that found for cells from rats fed the high glucose diet.

Requirement for bivalent cations in the actions of insulin and sodium nitroprusside on metabolism in rat adipocytes

The requirement for Ca 2+ and Mg 2+ in the actions of insulin and sodium nitroprusside on rat adipocyte metabolism was investigated: (i) sodium nitroprusside, but not insulin, increased cGMP levels in cells incubated in the absence of Ca 2+ and/or Mg 2+; (ii) sodium nitroprusside and insulin are unable to increase the incorporation of [ 14C]glucose into triglycerides and [ 14C]leucine into proteins in the absence of Ca 2+ and Mg 2+; (iii) sodium nitroprusside and insulin showed antilipolytic actions in Ca 2+- and Mg 2+-free medium. We conclude that in the absence of Ca 2+ and Mg 2+, sodium nitroprusside and insulin have very similar regulatory properties on triglyceride, protein synthesis and adrenaline-stimulated lipolysis, but not on cGMP levels in rat adipocytes. This could provide evidence that omission of bivalent cations was inhibitory at more than one site, or that sodium nitroprusside mimics insulin's actions by another mechanism that does not involve cGMP.

Evaluation of Prostaglandin E2 as a Regulator of Lipolysis in Bovine Adipose Tissue1

Effects of exogenous prostaglandin E2 (PGE2) on rates of lipolysis in sections of subcutaneous adipose tissue biopsied from fed and fasted Holstein steers were determined. The interaction of PGE2 with several exogenous effectors of lipolysis and of the adenylate cyclase-cAMP system also was measured. Epinephrine increased basal (nonstimulated) lipolysis approximately one-fold. Prostaglandin E2 had no effect on either basal or epinephrine-stimulated lipolysis. Dibutyryl cAMP increased rate of lipolysis .4-fold, whereas theophylline increased lipolysis more than one-fold. Theophylline had an additive effect on epinephrine-stimulated lipolysis. Dibutyryl cAMP increased theophylline-stimulated lipolysis but not epinephrine-stimulated lipolysis. Prostaglandin E2 had no effect on epinephrine-, dibutyryl cAMP- or theophylline-stimulated lipolysis. Fasting decreased basal lipolysis by 40%. Furthermore, lipolysis in tissue incubated with PGE2, epinephrine or PGE2 plus epinephrine decreased from 30 to 50% upon fasting. As also shown with tissue from fed steers, PGE2 did not alter basal or epinephrine-stimulated lipolysis in tissue from fasted steers. Influences of exogenous effectors on lipolysis in adipose tissue from fed and fasted steers indicate that PGE2 does not control the adenylate cyclase-cAMP system that regulates lipolysis in bovine adipose tissue.

Regulation of glucose utilization in adipose cells and muscle after long-term experimental hyperinsulinemia in rats.

The effects of chronic insulin administration on the metabolism of isolated adipose cells and muscle were studied. Adipose cells from 2 and 6 wk insulin-treated and control rats, fed either chow or chow plus sucrose, were prepared, and insulin binding, 3-O-methylglucose transport, glucose metabolism, and lipolysis were measured at various insulin concentrations. After 2 wk of treatment, adipose cell size and basal glucose transport and metabolism were unaltered, but insulin-stimulated transport and glucose metabolism were increased two- to threefold when cells were incubated in either 0.1 mM glucose (transport rate limiting) or 10 mM glucose (maximum glucose metabolism). Insulin binding was increased by 30%, but no shift in the insulin dose-response curve for transport or metabolism occurred. After 6 wk of treatment, the effects of hyperinsulinemia on insulin binding and glucose metabolism persisted and were superimposed on the changes in cell function that occurred with increasing cell size in aging rats. Hyperinsulinemia for 2 or 6 wk did not alter basal or epinephrine-stimulated lipolysis in adipose cells or the antilipolytic effect of insulin. In incubated soleus muscle strips, insulin-stimulated glucose metabolism was significantly increased after 2 wk of hyperinsulinemia, but these increases were not observed after 6 wk of treatment. We conclude that 2 wk of continuous hyperinsulinemia results in increased insulin-stimulated glucose metabolism in both adipose cells and soleus muscle. Despite increased insulin binding to adipose cells, no changes in insulin sensitivity were observed in adipose cells or muscle. In adipose cells, the increased glucose utilization resulted from both increased transport (2 wk only) and intracellular glucose metabolism (2 and 6 wk). In muscle, after 2 wk of treatment, both glycogen synthesis and total glucose metabolism were increased. These effects of hyperinsulinemia were lost in muscle after 6 wk of treatment, when compared with sucrose-supplemented controls.

Ventromedial and dorsomedial hypothalamic syndromes in the weanling rat: Is the “center” concept really outmoded?

This report juxtaposes findings from weanling rats with precise lesions in the Ventromedial (VMNL rats) to data of weanling rats with lesions in the dorsomedial (DMNL) hypothalamic nuclei. Despite the proximity of the two nuclei their destruction produces opposite effects in most cases but similar responses in other parameters. Absolute and relative food intake are normal in VMNL rats yet they become obese in the face of normal body weight gains. DMNL rats show both reduced absolute food intake and body weight but normal relative food intake and body composition. Both VMNL and DMNL cause reduced linear growth and running wheel activity. DMNL rats defend their lower body weight set point against various challenges and maintain normal body composition. Organ growth in both absolute and relative terms is reduced in VMNL rats. In DMNL rats relative organ growth is normal. Pancreatic growth, protein/pancreas and content and concentrations of several pancreatic enzymes are normal in DMNL but reduced in VMNL rats. Mean 24-hour plasma growth hormone (GH) and corticosterone (B) levels are reduced and insulin levels are greatly elevated in VMNL rats; prolactin (PRL) levels are normal. In DMNL rats. GH, B, insulin and somatomedin activity are normal but PRL is elevated. Circadian rhythms of GH, insulin and triiodothyronine are normal in DMNL rats but B levels are disrupted, as they are in VMNL rats. Glucose incorporation and oxidation in adipose tissue of VMNL rats are enhanced in VMNL rats but normal in DMNL rats. Gluconeogenesis in VMNL rats is enhanced as early as 4 hours post-operatively; in DMNL rats it is normal at this time and several weeks thereafter. Basal lipolysis in epididymal fat pads is elevated in both VMNL and DMNL rats but epinephrine-stimulated lipolysis is elevated in VMNL and decreased in DMNL rats. Both VMNL and DMNL rats show normal basal and epinephrine-stimulated lipolysis in interscapular brown adipose tissue. Several hepatic enzymes are normal in DMNL and depressed in VMNL rats. The above data suggest that the DMN and its circuitry are part of an “organismic” set point system with a “true” body weight and not fat set point, as seems to be the case in the VMNL rat.

Lack of relationship between changes in adiposity and plasma lipids following endurance training

In order to study the effect of alterations in fat morphology and metabolism induced by exercise-training on plasma lipids, 13 healthy young men were subjected to a 20-week aerobic training program on bicycle. Training significantly increased maximal aerobic power (V̇O 2 max) ( P < 0.001) and decreased per cent body fat ( P < 0.001). A significant reduction of mean adipocyte diameter and an increase in isolated fat cell epinephrine-stimulated lipolysis were also observed following training. However, with the exception of total cholesterol, no changes were noted in plasma lipids. Neither before nor after training were triglycerides and high density lipoprotein cholesterol (HDL-C) correlated with V̇O 2 max, fat cell diameter and adipocyte-stimulated lipolysis. The present study demonstrates that an important fat loss (mean loss of fat = 3 kg) and a significant gain in V̇O 2 max induced by endurance training do not necessarily produce an increase in HDL-C levels in normal male subjects. Moreover, changes in V̇O 2 max, body fatness and in fat cell epinephrine-stimulated lipolysis produced by endurance training are not related to modifications in plasma lipids in healthy men.

In vitro effect of thyroid hormones on lipolysis of rat fat cells during aging

The in vitro effect of triiodothyronine (T 3) 10 −5 M upon lipolysis was studied on white fat cells isolated from 1.5–6- and 30-month-old rats. We couldn't see any consistent effect of this hormone upon the basal lipolysis. We observed a T 3 effect on epinephrine-stimulated lipolysis on the three groups of animals. After 1 h of incubation the increase of glycerol release varies with the dose of epinephrine; after 3 h the T 3 effect persisted only in the 6-month and 30-month groups.

Level of physical fitness and adipocyte lipolysis in humans.

The present experiment was conducted to study the influence of exercise training on adipose tissue lipolytic activity and to identify the amount of training required to induce maximal adaptation in humans. Fifty-one male subjects were divided into three groups according to their training regimen: 1) sedentary subjects (SS) (n = 21); 2) trained subjects (TS) (n = 15) who had exercised during a period of 20 wk, 5 days/wk, 45 min/session; and 3) experienced marathon runners (MR) (n = 15) who ran an average of 120 km/wk for many years. Biopsies of fat were performed in the suprailiac region after an overnight fast. Adipocyte diameter (AD) and epinephrine-stimulated lipolysis ( ESL ) were assessed on collagenase-isolated fat cells. A lower AD was noted in the MR group compared with the two other groups. Basal lipolysis (BL) and ESL were significantly higher in TS and MR than in controls. Moreover, BL values were comparable in the two trained groups, whereas ESL in the TS group was higher than in the MR group. These results indicate that training increases suprailiac fat cell lipolysis, which seems to adapt maximally within about 4 mo.

Correlation of lipolytic and lipogenic actions of synthetic peptides with structure.

The effects of various synthetic peptides on basal and ACTH-stimulated lipolysis in fat cells were examined. Glu-Arg-Gly-Phe-Phe-Phe possessed lipolytic activity and increased ACTH-stimulated lipolysis at concentrations higher than 0.5 mumol/ml. Glu-Arg-Gly-Phe-Phe-Tyr did not cause any release of FFA from fat cells. Glu-Arg-Gly-Leu-Leu-Leu had no lipolytic activity but inhibited ACTH-stimulated lipolysis at concentrations higher than 0.25 mumol/ml. Glu-Arg-Gly-Leu-Leu-Leu also inhibited epinephrine-stimulated lipolysis. The effects of the peptides on basal and insulin-stimulated lipogenesis in fat cells were examined. Glu-Arg-Gly-Phe-Phe-Tyr increased both basal and insulin-stimulated lipogenesis. A tripeptide, Glu-Arg-Gly, inhibited both basal and insulin-stimulated lipogenesis. Glu-Arg-Gly-Leu-Leu-Leu had no effect on either basal or insulin-stimulated lipogenesis. Glu-Arg-Gly-Phe-Phe-Phe and ACTH, which elicit FFA release from fat cells, also stimulated formation of [14C]triglyceride from [14C]glucose.

Biological activity of bacterial derived human growth hormone in adipose tissue of hypophysectomized rats.

Segments of epididymal fat from hypophysectomized rats were incubated in Krebs-Ringer bicarbonate buffer for 1-4 h. Responses to three bacterially synthesized human GH (hGH) preparations were compared with responses to two highly purified pituitary hGH preparations. Hormones of both bacterial and pituitary origin increased the rate of oxidation of [U14C] glucose to 14CO2 and antagonized epinephrine-stimulated lipolysis. These insulin-like effects were produced over a range of 30-300 ng/ml hGH, and their concentration dependence was quite similar regardless of the source of hormone. Exposure of tissues to GH in the first hour of incubation makes them refractory to the insulin-like actions of a test dose of GH added in the fourth hour of incubation. hGH of either pituitary or bacterial origin also appeared to be equipotent in producing such refractoriness. The minimal effective concentration for this response fell between 10 and 30 ng/ml. Hormones from both sources were also capable of displacing 125I-labeled hGH from specific binding sites on isolated adipocytes, and the displacement curves were superimposable. Fifty percent displacement was seen at concentrations in the range of 10-20 ng/ml. Cell preparations produced a delayed increase in lipolysis which was evident in the presence of 0.3 mg/ml of theophylline in the fourth hour of incubation. In this regard, two of the bacterial hGH preparations were more potent than pituitary hGH and produced statistically significant increases in lipolysis at concentrations 10-30 times lower than pituitary hGH studied simultaneously in tissues of the same rats. Despite the presence of less than 1% contamination with bacterial protein, the greater potency of the bacterial hGH may be due to material of bacterial origin, since one bacterial hGH preparation, which was further purified to remove 99% of the remaining bacterial contamination was equipotent with pituitary hGH. These data indicate that a protein hormone synthesized in bacteria, and therefore likely to be free of peptides which might contaminate hGH purified from pituitary extracts, has all of the diverse physiological effects in adipose tissue that have been observed with preparations of GH derived from pituitary glands. These findings lend strong support to the conclusion that both insulin-like and lipolytic actions of GH are intrinsic properties of the molecule and cannot be attributed to contaminants present in the pituitary extract.

Insulin-like activity of photochemically obtained monovalent monomeric concanavalin A in the presence of anti-concanavalin A antibodies: dependence on multivalency for stimulation of glucose oxidation of rat fat cells.

Insulin-like action of monovalent monomeric concanavalin A (m-Con A) was examined in rat adipocytes in the presence of anti-m-Con A antiserum. The antisera from rabbits injected with m-Con A reacted with not only monovalent monomeric but also tetravalent tetrameric concanavalin A (alpha-Con A) in Ouchterlony double diffusion analysis. m-Con A alone did not show any appreciable effect on glucose oxidation of adipocytes while it slightly inhibited glycerol release stimulated by epinephrine. In contrast, exposure of adipocytes to m-Con A in the presence of antibodies to m-Con A resulted in stimulation of glucose oxidation and inhibition of epinephrine-stimulated lipolysis. The stimulation and the inhibition with m-Con A in the presence of the antibodies were of the same degree as those with alpha-Con A. Both alpha- and m-Con A were slightly active in inhibiting 125I-labeled insulin binding. These results demonstrate that the ability of anti-m-Con A antiserum to aggregate m-Con A bound to receptors on the isolated-adipocyte plasma membrane allowed m-Con A to mimic the biological activity of insulin and that the aggregation of receptors for ligands other than insulin can induce insulin-like action in rat fat cells.

Oxytocin action: lipid metabolism in adipocytes from homozygous diabetes insipidus rats (Brattleboro strain).

Oxytocin, like insulin, stimulates glucose oxidation in normal rat adipocytes. Fat cells from homozygous Brattleboro rats that exhibit diabetes insipidus (HoDI animals) and that have a normal number of oxytocin receptors, however, are unable to respond to oxytocin in terms of glucose oxidation. We now report that in adipocytes from HoDI animals that are responsive to insulin, oxytocin was also unable to stimulate lipogenesis. In contrast, oxytocin like insulin was able to inhibit epinephrine-stimulated lipolysis in adipocytes from HoDI animals. Thus, in HoDI adipocytes, the results indicate that the receptor-effector system is only partially defective.

The reversible inhibition by carbonyl cyanide m-chlorophenyl hydrazone of epinephrine-stimulated lipolysis in perifused isolated fat cells

Lipolysis stimulated in perifused isolated fat cells by 0.5 μm epinephrine is an ATP-dependent process which can be monitored by measuring the release of glycerol. The stimulated lipolysis is inhibited by 10 μM carbonyl cyanide m-chlorophenyl hydrazone (CCCP), an uncoupler of oxidative phosphorylation. If 20 mM glucose is continuously present in the perifusion medium during and after treatment with epinephrine and CCCP, the inhibition of the stimulated lipolysis is reversible when the CCCP is discontinued; otherwise it is not readily reversible. Since 20 mM 2-deoxyglucose will not substitute for glucose, metabolism of glucose beyond phosphorylation by hexokinase is concluded to be necessary in order to maintain the reversibility of the inhibition by CCCP. Substitution of 10 mM succinate for glucose also did not preserve the reversibility of the CCCP inhibition, and there was no significant difference in the amount of decrease of ATP in fat cells incubated with CCCP and epinephrine in the presence of glucose as compared to the decrease observed in the presence of succinate. The mechanism by which glucose maintains reversibility of the inhibition of stimulated lipolysis by CCCP is therefore not clear.

Inhibition of the lipolytic action of beta-adrenergic agonists in human adipocytes by alpha-adrenergic agonists.

The aim of this study was to define the role of the alpha-adrenergic receptor in the regulation of lipolysis by human adipocytes. Glycerol production by isolated human adipocytes was stimulated by the pure beta-adrenergic agonist isoproterenol in a dose-dependent fashion. This stimulation of lipolysis was inhibited by the alpha-adrenergic agonists methoxamine, phenylephrine, and clonidine. Epinephrine-stimulated lipolysis was potentiated by the alpha-adrenergic antagonists, dihydroergocryptine, phentolamine, phenoxybenzamine, and yohimbine. Whereas the attenuation of beta-adrenergic agonist-stimulated lipolysis by alpha-adrenergic agonists was reversed completely by the alpha 2-adrenergic antagonist yohimbine, the alpha 1-antagonist prazosin did not reverse such attenuation. It is concluded that alpha-adrenergic agonists act as antilipolytic agents in human adipocytes and that this action may result from the interaction of these compounds with a population of alpha 2-adrenergic receptors.

Weight reduction in young obese children. I. Effects on adipose tissue cellularity and metabolism.

A 10-year longitudinal study was conducted on 16 prepubescent youngsters who had undergone successful weight reduction. Their ages ranged from 2 to 10 years when the study began. In all subjects, weight reduction proceeded only by a decrease in adipose cell size (from 0.62 +/- 0.02 to 0.46 +/- 0.02 micrograms lipid per cell) and resulted in a corresponding 33% decrease (from 177 +/- 6 to 144 +/- 5%) in percent ideal body weights. Cell numbers did not change appreciably during the period of weight loss (29.4 +/- 2.6 versus 28.7 +/- 2.3 x 10(9) total adipocytes). Three years after the start of the study, 14 of 20 youngsters had maintained their reduced percent ideal body weights, including eight who remained below 130% ideal body weight. Ten years later, only four remained below 130% ideal body weight. All four children had total adipose cell numbers below 20 x 10(9) total adipocytes at the start of the weight reduction program, a value below the lower limit for adult normal weight subjects. Thirteen other children have maintained or decreased their initial percent ideal weights. The remaining nine youngsters have further increased their percent ideal body weights. In vitro metabolic studies of the patient's adipocytes revealed a greater than 50% depression of epinephrine-stimulated lipolysis pre- and immediately postweight reduction; this decrease persisted for the entire period of study, irrespective of the maintenance of a normal percent ideal body weight. At the same time, normal 150% increases in the in vitro production of 14CO2 from [1-14C]glucose in the presence of insulin occurred.

Mechanism of the age-related decrease of epinephrine-stimulated lipolysis in isolated rat adipocytes: beta-adrenergic receptor binding, adenylate cyclase activity, and cyclic AMP accumulation.

beta-adrenergic binding ([3H]dihydroalprenolol), adenylate cyclase activity, and cAMP accumulation were measured in adipocytes to investigate whether the mechanism of decreased hormone-sensitive lipolytic response with age was mediated through membrane-associated events. The dose of epinephrine required for half maximal stimulation of glycerol release (ED50) was significantly lower in 2-month-old rats (0.8 +/- 0.2 microM)than in matur (6- and 12-month-old) rats (5.2 +/- 1.5 and 6.2 +/- 1.5 microM, respectively). In 24-month-old rats the ED50 (0.7 +/- 0.2 microM) was less than in mature rats. maximum rates of hormone-stimulated glycerol release (per 10(6) cells) was highest in the two mature groups and decreased by 50% in the old rats (P less than 0.01). Lipolytic changes were independent of cell size. beta-adreanergic raeceptor number (50-90 thousand sites/call) and affinity (KD 4-5 nM) were the same in each age group. ED50 and maximum level of hormone-stimulated adenylatae cyclase activity did not change with age. The ED50 of cAMP accumulation of young rats was 3 +/- 5 microM compared with 24 +/- 4 and 25 +/- 5 microM in 6- and 12-month-old rats, respectively. In old rats, the ED50 of cAMP accumulation was 2 +/- 1 microM (P less than 0.001 compared with mature rats). Maximally stimulated cAMP levels were the same in old and mature animals. Phosphodiesterase activity in the presence and absence of 10(-5) M isoproterenol did not change with age. The results suggest that age-related decrease of epinephrine-sensitive lipolysis in old rats may be due to alterations of the lipolytic pathway distal to the receptor-adenylate cyclase complex and the generation of cyclic AMP.