Preferential Stimulation of Abdominal Subcutaneous Lipolysis after Prednisolone Exposure in Humans

Preferential Stimulation of AbdominalSubcutaneous Lipolysis after PrednisoloneExposure in HumansClaus Højbjerg Gravholt, Rolf Dall, Jens Sandahl Christiansen, Niels Møller, and Ole Schmitz

AbstractGRAVHOLT, CLAUS HØJBJERG, ROLF DALL, JENSSANDAHL CHRISTIANSEN, NIELS MØLLER, ANDOLE SCHMITZ. Preferential stimulation of abdominalsubcutaneous lipolysis after prednisolone exposure inhumans. Obes Res. 2002;10:774 –781.Objective: The role of cortisol in the regulation of lipolysisis not clear. This study was undertaken to explore whethera standard dose of prednisolone for 1 week would influencelipolysis in abdominal and femoral tissue.Research Methods and Procedures: We used the microdi-alysis technique, the forearm technique, and indirect calo-rimetry, in the fasting state, after 1 week of treatment withprednisolone (30 mg daily) or placebo. Eight healthy youngmen (age: 25 � 3 years; height: 181 � 1 cm; body massindex [BMI]: 23.3 � 0.7 kg/m2) were studied.Results: Treatment with prednisolone induced insulin resis-tance (Homeostasis Model Assessment index: placebo vs.prednisolone: 7.15 � 1.63 vs. 17.00 � 14.26, p � 0.03),hyperinsulinemia (p � 0.01), and hyperglucagonemia (p �0.001), whereas growth hormone concentrations were un-affected. Abdominal adipose tissue interstitial glycerol wasincreased during treatment with prednisolone in the face ofsignificant hyperinsulinemia, although it barely reached sta-tistical significance (p � 0.06). At the femoral adiposetissue depot, no difference in lipolysis was found. Arterialand venous free fatty acids (FFA) were comparable in thetwo situations, whereas the arteriovenous difference acrossthe forearm was significantly decreased during treatmentwith prednisolone, indicating increased uptake, or decreased

release of FFA. Energy expenditure (p � 0.3), repiratoryquotient (p � 0.9), glucose oxidation (p � 0.9), lipidoxidation (p � 1.0), and protein oxidation (p � 0.1) wereunaltered on the 2 study days.Discussion: Short-term treatment with a standard dose ofcorticosteroids induces increased abdominal adipose tissuelipolysis, as well as hyperinsulinemia, hyperglucagonemia,and insulin resistance.

Key words: glucocorticoids, lipolysis, microdialysis, re-gional adipose tissue

IntroductionFree fatty acids (FFA) are postabsorptively a principal

oxidative substrate for the organism. The release of FFA istightly regulated by a number of hormones. Insulin is apotent inhibitor of lipolysis (1) and stimulates lipoproteinlipase (LPL), thus, favoring deposition of triglycerides inadipose tissue, whereas cathecolamines, thyronines, andgrowth hormone (GH) stimulate the release of FFA fromadipose tissue. Cathecolamines are very potent physiologi-cal stimulators of lipolysis in adipose tissue through stim-ulatory �1-, �2-, and �3-adrenoreceptors and inhibitory �2-adrenoreceptors (2,3), and subsequent modulation of thehormone-sensitive lipase. The lipolytic action of GH isprobably mediated through stimulation of gene expression,after binding of the GH receptor with JAK2 tyrosine kinaseand subsequent activation of the complex (4). Hereafter, theintracellular signaling process leads to the activation ofhormone-sensitive lipase. However, catecholamine and GHaction is linked, because the effect of GH on cyclic adeno-sine monophosphate (cAMP) production in adipocytes isincreased by catecholamines (5). Conversely, GH sensitizesadipocytes to the action of catecholamines (6), suggesting asynergistic effect of GH and catecholamines on lipolysis. Incontrast to insulin, cathecolamines, and GH, the role ofcortisol in this regulation is still not clear. Glucocorticoidreceptors are present ubiquitously in adipose tissue, albeit

Received for review December 17, 2001.Accepted for publication in final form April 15, 2002.Medical Department M (Endocrinology and Diabetes) and Medical Research Laboratories,Århus University Hospital, Århus C, Denmark.Address correspondence to Dr. Claus Højbjerg Gravholt, Medical Department M (Endocri-nology and Diabetes), Århus Kommunehospital, Århus University Hospital, DK-8000 ÅrhusC, Denmark.E-mail: [email protected] © 2002 NAASO

774 OBESITY RESEARCH Vol. 10 No. 8 August 2002

with increased density in visceral adipose tissue in humans(7). In vitro animal experimental studies have shown anenhancing effect of cortisol on lipolysis (8–11). In humansdexamethasone alone, and in combination with insulin, in-creases LPL activity, as well as LPL mRNA levels (12).Whereas data from in vivo studies are more ambiguous(13–18), it was demonstrated that pharmacological doses ofglucocorticoids elicit a stimulation of lipolysis in vivo(19,20). One study implicated hypercortisolemia at physio-logical levels in the increased lipolysis seen under stress(21). Furthermore, Cushing’s syndrome is associated withsignificant changes in regional fat deposits with substantialand chronic elevated levels of cortisol.

This study was undertaken to explore whether a standarddose of prednisolone for 1 week, similar to what is pre-scribed to many patients in clinical practice, would influ-ence lipolysis in abdominal and femoral tissue. To this end,we used subcutaneous microdialysis, forearm metabolic as-sessment and indirect calorimetry.

Research Methods and ProceduresDesign

The study was performed as a double-blind, placebo-controlled, cross-over trial. The two treatment periods andthe washout period lasted 1 week and 6 weeks, respectively.Test medicine was either 30-mg prednisolone tablets (Phar-macy of Aarhus University Hospital, Aarhus, Denmark; 15mg in the morning and 15 mg in the evening) or placebotablets. The medication was taken at 9:00 AM and 9:00 PM.The test medicine was packed according to clinical practicerecommendations by the Pharmacy of Aarhus UniversityHospital.

The study was approved by the local ethical committee inaccordance with the Helsinki II declaration. All subjectsgave written and oral consent before the study began.

Subjects and Experimental ProcedureEight healthy young men (age: 25 � 3 years; height:

181 � 1 cm; body mass index [BMI]: 23.3 � 0.7 kg/m2)were recruited through public advertisement.

Subjects were admitted to the Clinical Research Center inthe morning after an overnight fast (10 to 12 hours) withoutany caffeine consumption or cigarette smoking; only inges-tion of tap water was allowed. Participants were asked notto perform major physical exercise and to consume aweight-maintaining carbohydrate-rich diet for the last 3days before examination and to refrain from alcohol intakeon the day before the investigation. Participants were placedin the supine position in a bed in light clothes at roomtemperature (�22 to 24 °C) for the study. One intravenouscatheter (Viggo AB, Helsingborg, Sweden) was placed in anantecubital vein for infusion, another in the vein of a handthat was heated in a box with an air temperature of �65 °C

to provide arterialized blood for draining, and one catheterretrogradely in a deep antecubital vein for measurements offorearm arteriovenous (AV) substrate balances. Indirect cal-orimetry (Deltatrac Metabolic Monitor; Datex, Helsinki,Finland) with a ventilated hood at 40 L/min was performed;energy expenditure (EE), respiratory exchange ratio (RER),and 24-hour excretion of urea in urine were measured; andglucose, protein and lipid oxidation were calculated (22).Blood samples were drawn every 30 minutes throughout thestudy starting at t � �90. Interstitial levels of metaboliteswere sampled every 30 minutes from the abdominal andfemoral subcutaneous adipose tissue.

MicrodialysisA microdialysis catheter (CMA 60; CMA, Stockholm,

Sweden) was placed in the abdominal and the femoralsubcutaneous adipose tissue after anaesthetization of theskin with 0.05 mL lidocaine at the site of perforation of theskin. The microdialysis catheter used has a molecular cutoffof 20 kDa. Immediately after placement, perfusion of thecatheters with physiological perfusion fluid [Na�, 147 mM;K�, 4 mM; Ca2� 2.3 mM; Cl�, 156 mM; pH 6; osmolality,290 mosm/kg; Perfusion fluid T1; CMA] at a flow rate of0.3 �L/min with the use of a portable pump (CMA 106;CMA). At this flow rate, the rate of recovery with themicrodialysis catheter is �90% (23). The microdialysiscatheter was placed at t � �90 minutes. After 90 minutesof perfusion of the microdialysis catheter, allowing localedema and hemorrhage to subside, sampling started at t �0 minute and continued until t � 240 minutes every 30minutes. The first sample was, thus, withdrawn at t � 0minute. This sample reflects the integrated level of intersti-tial glucose during the preceding 30 minutes, and the samplewas assigned the time t � �15 minutes. This principle wasused for all samples. The observed changes in interstitialglycerol concentration can be seen as an index of lipolysis(24–26). The baseline values for glycerol obtained in thisstudy correspond closely with the ones available in theliterature (27–29).

Blood Flow MeasurementsThe subcutaneous adipose tissue blood flow (ATBF) in

the abdominal region, in which microdialysis was per-formed, was measured by the local 133Xe washout method(30). In short, 3.7 Mbq (0.1 mL) 133Xe was injected into thesubcutaneous area of interest, equivalent to a whole-bodyradiation dose of 0.5 mSv. Disappearance of 133Xe wasmonitored with a 2 � 2-inch NaI detector (model 905)connected to a photomultiplier base (model 276; EG&GOrtec, Wokingham, Berks, UK) covered by a cylindricalcopper collimator and coupled with a multichannel Ace-Mate (model 925) amplifier (EG&G Ortec). The system wasconnected to a computer for simultaneous sampling. Theregistration of the washout rate began at least 30 minutesafter the injection. ATBF was calculated as:

Effects of Prednisolone in Adipose Tissue, Gravholt et al.

OBESITY RESEARCH Vol. 10 No. 8 August 2002 775

ATBF � k � � � 100 (mL/100g � minute),

where k is the rate constant of the washout curve, and � isthe tissue to blood partition coefficient for 133Xe at equilib-rium; counts were collected every minute and a straight linewas fitted through the experimental points in a semiloga-rithmic diagram as a function of time. Experimental valuesof k were determined as the slope of the regression lineduring a specified T, where T is the time frame (in minutes).The time-interval was at least 15 minutes. � was calculatedas:

� � 0.22 � SFT � 2.99,

where SFT is the skinfold thickness of the abdominal adi-pose tissue (30,31).

Insulin SensitivityInsulin sensitivity was calculated using the Homeostasis

Model Assessment (HOMA) index (32). The HOMA index,which is based on simultaneous sampled fasting values ofinsulin and glucose, was previously shown to correlate wellwith the euglycemic hyperinsulinemic clamp in the assess-ment of insulin sensitivity in both normal and diabeticsubjects (32). The HOMA index (R) was calculated asfollows:

R � fasting insulin � fasting glucose/22.5.

AssaysPlasma glucose was measured immediately after sam-

pling in duplicate on an autoanalyzer (Beckman Instru-ments, Palo Alto, CA) by the glucose oxidase method. Theautoanalyzer was calibrated frequently with known humanplasma standards, as well as standards supplied by thecompany with the equipment and the intra-assay coefficientof variation (CV) was below 0.5%. GH was measured witha double monoclonal immunofluorometric assay (WallacOy, Turku, Finland). The interassay CV in samples variedbetween 1.7 and 2.4%, the intra-assay CV varied between1.9 and 3.0% for GH concentrations of 12.08 and 0.27 �g/L,and the detection limit was 0.01 �g/L. Serum insulin wasmeasured by ELISA using a two-site immunoassay (33).The intra-assay CV was 2.0% (n � 75) at a serum level of200 pM. Serum FFA was determined by a colorimetricmethod using a commercial kit (Wako Chemicals, Neuss,Germany). Blood samples were deproteinized with perchlo-ric acid for determination of glycerol, 3-hydroxybutyrate(3-OHB), and lactate was assayed by an automated fluoro-metric method (34). Plasma glucagon levels were measuredby a radioimmunoassay (35). Glycerol, glucose, and lactatein the dialysate were measured by an automated spectro-photometric kinetic enzymatic analyzer (CMA 600; CMA).

Statistical AnalysesAll statistical calculations were done with SPSS for Win-

dows, Version 8.0 (SPSS, Chicago, IL). Repeated measuresANOVA (generalized linear model) was used to test fordifferences between the prednisolone and the placebo (dis-regarding the interactions on the separate study days oftime, individuals, and treatment, because visual inspectionof the results showed that all measured variables were stableduring the study). Results are expressed as means � SEM.Significance levels under 5% were considered significant.

ResultsCirculating Hormones and Insulin Sensitivity

As expected, circulating levels of insulin, C-peptide, andglucagon were significantly higher after prednisolone exposurecompared with placebo (Figure 1, A–C), whereas GH concen-trations were unaffected (Figure 1D). The HOMA index valuesincreased significantly in response to prednisolone exposure(placebo vs. prednisolone: 7.15 � 1.63 vs. 17.00 � 14.26, p �0.03), indicating diminished insulin sensitivity.

Plasma and Interstitial GlucoseBaseline levels of arterial (t test, p � 0.5) and venous (t

test, p � 0.1) plasma glucose were similar to the corre-sponding values in the two situations. Surprisingly, arterialand venous plasma glucose were slightly lower after pred-nisolone exposure compared with corresponding values af-ter placebo (Figure 2, A and B), whereas the AV differencewas comparable in the two situations (Figure 2C). Intersti-tial glucose was comparable in both the abdominal(ANOVA, p � 0.6) and femoral (ANOVA, p � 0.2) regionsduring both placebo and prednisolone administration.

Circulating Levels of Interstitial Lipid Intermediates,and Abdominal Tissue Blood Flow

Abdominal interstitial glycerol was increased duringtreatment with prednisolone, although it barely reachedstatistical significance (p � 0.06; Figure 3A). At the fem-oral depot, there were no differences between the two treat-ments (Figure 3B). Arterial and venous FFAs were compa-rable between the two (Figure 4, A and B), whereas the AVdifference across the forearm was significantly loweredduring treatment with prednisolone (Figure 4C), indicatingincreased uptake or decreased release of FFA. Circulatinglevels of plasma glycerol, lactate, and 3-OHB were compa-rable during the 2 study days, and there were no AVdifferences (data not shown). Abdominal tissue blood flowwas similar in both situations (average blood flow: [placebovs. prednisolone: 2.25 � 0.34 vs. 2.10 � 0.33, p � 0.7]).

EEEE (placebo vs. prednisolone, 1728 � 45 vs. 1790 �

47 kcal/24 h, p � 0.3), RER (0.87 � 0.02 vs. 0.86 �



0.02, p � 0.9), glucose oxidation (753 � 101 vs. 716 �108 kcal/24 h, p � 0.9), lipid oxidation (578 � 110 vs.574 � 111 kcal/24 h, p � 1.0), and protein oxidation(397 � 61 vs. 500 � 48 kcal/24 h, p � 0.1) wereunaltered on the two study days. BMI was stable through-out the study.

DiscussionUnder circumstances similar to a clinical situation, results

from this study show that prednisolone administration leadsto hyperinsulinemia, insulin resistance, and hyperglu-cagonemia. We opted to study lipolysis under these condi-tions, allowing insulin to reach higher levels during pred-

nisolone treatment compared with placebo, and, thus,avoiding the use of a somatostatin clamp by substitutingwith insulin, glucagon, and GH. The latter approach wouldno doubt have yielded a larger difference in the indices oflipolysis studied. One asset of this approach is that it closelymimics the metabolic conditions (including hyperinsulin-emia), ultimately leading to altered fat distribution in pa-

Figure 1: (A) Insulin vs. time during the study, (B) C-peptide vs.time during the study, (C) glucagon vs. time during the study, and(D) growth hormone (GH) vs. time during the study. Filled circles(F) indicate placebo treatment, and open circles (E) indicatetreatment with prednisolone. Error bars indicate SEM. The resultof ANOVA with significance level is given in the figure.

Figure 2: (A) Arterial plasma glucose vs. time during the study.The filled circles (F) indicate placebo treatment, and open circles(E) indicate treatment with prednisolone. Error bars indicate SEM.The result of ANOVA with significance level is given in thefigure. (B) Venous plasma glucose vs. time during the study. Thefilled circles (F) indicate placebo treatment, and open circles (E)indicate treatment with prednisolone. Error bars indicate SEM.The result of ANOVA with significance level is given in thefigure. (C) Plamsa glucose AV difference vs. time during thestudy. The filled circles (F) indicate placebo treatment, and opencircles (E) indicate treatment with prednisolone. Error bars indi-cate SEM. The result of ANOVA with significance level is givenin the figure.



tients with high circulating levels of glucocorticoids. Underthe applied conditions, lipolysis was more pronounced inthe abdominal adipose depot, although barely reaching sta-tistical significance. The SD in microdialysis samples waslarger than in corresponding serum samples, and despite aseemingly clear difference in abdominal adipose glycerolvalues on the two study days at visual inspection, thedifference did not reach statistical significance (p � 0.06).Reasons for larger intra- and inter-individual CVs in micro-dialysis samples are numerous: it is impossible to positionthe catheter in exactly the same position on different studydays, both in terms of depth and point of insertion; thedialysate flow may vary slightly; the size of the subcutane-ous layer of fat influences blood flow and, thus, appearanceof substances; the effect of local trauma during insertionmay affect output, etc. Microdialysis allows continuousmonitoring of changes of fluxes of a variety of compoundsfrom interstitial fluid to the dialysate and it has been utilized

in a large number of tissues in the human body since it wasfirst introduced (36). True equilibrium can be accomplishedacross the membrane when a very low flow rate is used (29).Thus, to the extent that tissue disposal rates are constant, thechanges in interstitial glycerol concentration can be seen asan index of lipolysis because glycerol is only produced andnot taken up by adipose tissue (37), although some datacontest the assumption that glycerol is not taken up byadipose tissue, at least during noradrenaline infusion (38).The observed baseline values for glycerol obtained in thisstudy correspond closely with the ones available in theliterature (26–29). The present increase in lipolysis was

Figure 3: (A) Abdominal interstitial glycerol vs. time during thestudy, and (B) femoral interstitial glycerol vs. time during thestudy. Filled circles (F) indicate placebo treatment, and opencircles (E) indicate treatment with prednisolone. Error bars indi-cate SEM. The result of ANOVA with significance level is givenin the figure. Figure 4: (A) Arterial free fatty acid (FFA) vs. time during the

study, (B) venous FFA vs. time during the study, and (C) arterio-venous (AV) FFA difference vs. time during the study. Through-out, filled circles (F) indicate placebo treatment, and open circles(E) indicate treatment with prednisolone. Error bars indicate SEM.The result of ANOVA with significance level is given in thefigure. *The p value for the interaction between time and treatmentis given.



seen under the influence of significant hyperinsulinemia,hyperglucagonemia, and insulin resistance, and, interest-ingly, a small, but significant decrease in the level of plasmaglucose. Thus, an increase in lipolytic activity was seen, andit is conceivable that LPL activity also increased (12),indicating that treatment with prednisolone increases ab-dominal lipid turnover. Unfortunately, we could not mea-sure blood flow in more than one position (abdominaladipose tissue) and, therefore, could not calculate and com-pare the rates of appearance of glycerol from the twostudied regions. In the abdominal region, blood flow wassimilar in the two situations, although it was marginallyreduced on prednisolone, indicating that the present in-crease in abdominal lipolysis must be viewed cautiously.Interestingly, patients with Cushing’s syndrome have in-creased activity of LPL in abdominal, but not in femoral,subcutaneous tissue (39). Similarly, we were unable to seeany enhanced lipolysis in the femoral region. Although theresult during prednisolone treatment has to be seen in thecontext of hyperinsulinemia and insulin resistance—and thepresence of relatively increased lipolysis cannot be ig-nored—the usual expectation would normally be a reduc-tion in lipolysis in response to high levels of insulin, be-cause of the exquisite sensitivity of lipolysis to even minutechanges in insulinemia (40,41). The level of insulin wasparticularly elevated in the beginning of the study on theday of prednisolone treatment, attributable to the immediateeffect of the ingested prednisolone on the morning of thestudy and the evening before the investigations. Cushing’ssyndrome is characterized by redistribution of fat fromperipheral stores to the central parts of the body, primarilythe trunk, without any general increase in total fat mass orreduction in total muscle mass (42). The present finding ofincreased lipolysis in the abdominal, but not in the femoraldepot, coupled with the above-mentioned results, may ex-plain the results found in clinical practice. The results,however, are at apparent odds with previous findings bySamra et al. (18), who found that acute hypercortisolemiaincreased circulating levels of FFA and glycerol and in-creased the appearance of systemic glycerol, findings thatwould directly indicate increased whole-body lipolysis.However, Samra et al. also reported reduced AV differencein FFA when comparing arterial levels with levels in thetummy vein draining subcutaneous abdominal adipose tis-sue; this was suggestive of inhibited intracellular lipolysis.They explained their results as being due to site-specificalterations in lipolytic enzyme activities and intravascularLPL-induced triacylglycerol hydrolysis. Partly in contrast,in a short-term (hours) study, Divertie et al. (21) also foundthat acute hypercortisolemia increased palmitate appearanceand, thus, systemic lipolysis. We did not find evidence ofincreased systemic lipolysis but of increased abdominal andunchanged femoral lipolysis. The discordant results may notbe incompatible, considering the different time-courses of

cortisol/prednisolone exposure. In the studies by Samra etal. (18) and Divertie et al. (21) subjects were exposed tocortisol for hours, whereas in this study, subjects wereexposed to prednisolone for 7 days. Also, the differences inthe applied methodology may have influenced the results.For example, microdialysis vs. the tummy vein methodol-ogy has previously been shown to give only partly compa-rable results, with similar levels of adipose tissue glycerol,but with different rates of appearance of glycerol theseresults are probably attributable due to the assumptions thathave to be made when calculating the rate of appearance ofglycerol from adipose tissue, which takes into accountblood flow, skinfold thickness, and the blood partition co-efficient (43). Likewise, differences in methodology maywell explain some of the differences between the studies bySamra et al. (18) and Divertie et al. (21). Exogenous glu-cocorticoids have been shown to increase the level of serumglucagon (44), as have the physiological nocturnal rise ofcortisol (45). However, it is not likely that the elevated levelof glucagon contributed to the increased lipolysis, becausewe (46) and others (47) have shown that glucagon does notinfluence lipolysis. Likewise, the observed difference inglycemia between the placebo and the prednisolone studydays is not believed to have any influence on lipolysis(48,49).

It is not completely clear how glucocorticoids induceinsulin resistance, but insulin-mediated peripheral glu-cose disposal is impaired by an intracellular mechanismdownstream from the insulin receptor, and hepatic insulinresistance is induced (50), oxidative and nonoxidativeglucose disposal is reduced (51), as is muscle glycogensynthase activity (52). Glucocorticoids also increase glu-coneogenesis by increasing hepatic glucose output, keto-genesis, and proteolysis (45).

In summary, short-term treatment with a standard doseof corticosteroids induces increased abdominal lipolysis,and perhaps relatively increased femoral lipolysis, aswell as hyperinsulinemia, hyperglucagonemia, and insu-lin resistance.

AcknowledgmentsThe study was supported by Grant 9600822 from the

Danish Health Research Council, (Århus University,Novo Nordisk Center for Research in Growth and Re-generation), and from the Institute of Experimental Clin-ical Research, Århus University. The microdialysis cath-eters and other utensils were generously supplied byRoche Diagnostics. We thank Anette Mengel for experttechnical help.

References1. Vikman HL, Ranta S, Kiviluoto T, Ohisalo JJ. Different

metabolic regulation by adenosine in omental and subcutane-ous adipose tissue. Acta Physiol Scand. 1991;142:405–10.



2. Fain JN, Garcia-Sainz JA. Adrenergic regulation of adipo-cyte metabolism. J Lipid Res. 1983;24:945–66.

3. Enoksson S, Shimizu M, Lonnqvist F, Nordenstrom J,Arner P. Demonstration of an in vivo functional �3-adreno-ceptor in man. J Clin Invest. 1995;95:2239–45.

4. Argetsinger LS, Campbell GS, Yang X, et al. Identificationof JAK2 as a growth hormone receptor-associated tyrosinekinase. Cell. 1993;74:237–44.

5. Argetsinger LS, Carter-Su C. Mechanism of signaling bygrowth hormone receptor. Physiol Rev. 1996;76:1089–107.

6. Watt PW, Finley E, Cork S, Clegg RA, Vernon RG.Chronic control of the �- and �2-adrenergic systems of sheepadipose tissue by growth hormone and insulin. Biochem J.1991;273:39–42.

7. Rebuffe-Scrive M, Lundholm K, Bjorntorp P. Glucocorti-coid hormone binding to human adipose tissue. Eur J ClinInvest. 1985;15:267–71.

8. Goodman HM. Permissive effects of hormones on lypolysis.Endocrinology. 1970;86:1064–74.

9. Behrens CM, Ramachandran J. The effect of glucocorti-coids on adipocyte corticotropin receptors and adipocyte re-sponses. Biochim Biophys Acta. 1981;672:268–79.

10. Kawai A, Kuzuya N. Effects of glucocorticoids on hormone-stimulated lipolysis and calcium uptake in the adipose cells.Horm Metab Res. 1981;13:224–8.

11. De Mazancourt P, Giot J, Giudicelli Y. Correction by dexa-methasone treatment of the altered lipolytic cascade inducedby adrenalectomy in rat adipocytes. Horm Metab Res. 1990;22:22–4.

12. Fried SK, Russell CD, Grauso NL, Brolin RE. Lipoproteinlipase regulation by insulin and glucocorticoid in subcutane-ous and omental adipose tissues of obese women and men.J Clin Invest. 1993;92:2191–8.

13. Owen OE, Cahill GF Jr. Metabolic effects of exogenous glu-cocorticoids in fasted man. J Clin Invest. 1973;52:2596–605.

14. Saunders J, Hall SE, Sonksen PH. Glucose and free fattyacid turnover in Cushing’s syndrome. J Endocrinol Invest.1980;3:309–11.

15. Miyoshi H, Shulman GI, Peters EJ, Wolfe MH, Elahi D,Wolfe RR. Hormonal control of substrate cycling in humans.J Clin Invest. 1988;81:1545–55.

16. Birkenhager JC, Timmermans HA, Lamberts SW. De-pressed plasma FFA turnover rate in Cushing’s syndrome.J Clin Endocrinol Metab. 1976;42:28–32.

17. Horber FF, Marsh HM, Haymond MW. Differential effectsof prednisone and growth hormone on fuel metabolism andinsulin antagonism in humans. Diabetes. 1991;40:141–9.

18. Samra JS, Clark ML, Humphreys SM, MacDonald IA,Bannister PA, Frayn KN. Effects of physiological hypercor-tisolemia on the regulation of lipolysis in subcutaneous adi-pose tissue. J Clin Endocrinol Metab. 1998;83:626–31.

19. Issekutz B Jr, Borkow I. Effect of catecholamines and dibu-tyryl-cyclic-AMP on glucose turnover, plasma free fatty acids,and insulin in dogs treated with methylprednisolone. CanJ Physiol Pharmacol. 1972;50:999–1006.

20. Tanaka H, Ichikawa Y, Akama H, Homma M. In vivoresponsiveness to glucocorticoid correlated with glucocorti-coid receptor content in peripheral blood leukocytes in normalhumans. Acta Endocrinol Copenh. 1989;121:470–6.

21. Divertie GD, Jensen MD, Miles JM. Stimulation of lipolysisin humans by physiological hypercortisolemia. Diabetes.1991;40:1228–32.

22. Frayn KN. Calculation of substrate oxidation rates in vivofrom gaseous exchange. J Appl Physiol. 1983;55:628–34.

23. Moberg E, Hagstrom Toft E, Arner P, Bolinder J. Pro-tracted glucose fall in subcutaneous adipose tissue and skeletalmuscle compared with blood during insulin-induced hypogly-caemia. Diabetologia. 1997;40:1320–6.

24. Jansson PA, Smith U, Lonnroth P. Interstitial glycerol con-centration measured by microdialysis in two subcutaneousregions in humans. Am J Physiol. 1990;258:E918–E22.

25. Arner P, Bolinder J. Microdialysis of adipose tissue. J InternMed. 1991;230:381–6.

26. Hagstrom Toft E, Enoksson S, Moberg E, Bolinder J,Arner P. Absolute concentrations of glycerol and lactate inhuman skeletal muscle, adipose tissue, and blood. Am JPhysiol. 1997;273:E584–E92.

27. Samra JS, Ravell CL, Giles SL, Arner P, Frayn KN.Interstitial glycerol concentration in human skeletal muscleand adipose tissue is close to the concentration in blood. ClinSci Colch. 1996;90:453–6.

28. Hagstrom Toft E, Bolinder J, Ungerstedt U, Arner P. Acircadian rhythm in lipid mobilization which is altered inIDDM. Diabetologia. 1997;40:1070–8.

29. Rosdahl H, Hamrin K, Ungerstedt U, Henriksson J. Me-tabolite levels in human skeletal muscle and adipose tissuestudied with microdialysis at low perfusion flow. Am JPhysiol. 1998;274:E936–E45.

30. Larsen OA, Lassen NA, Quaade F. Blood flow throughhuman adipose tissue determined with radioactive xenon. ActaPhysiol Scand. 1966;66:337–45.

31. Bulow J, Jelnes R, Astrup A, Madsen J, Vilmann P. Tissue/blood partition coefficients for xenon in various adipose tissuedepots in man. Scand J Clin Lab Invest. 1987;47:1–3.

32. Matthews DR, Hosker JP, Rudenski AS, Naylor BA,Treacher DF, Turner RC. Homeostasis model assessment:insulin resistance and �-cell function from fasting plasmaglucose and insulin concentrations in man. Diabetologia.1985;28:412–9.

33. Andersen L, Dinesen B, Jorgensen PN, Poulsen F, RoderME. Enzyme immunoassay for intact human insulin in serumor plasma. Clin Chem. 1993;39:578–82.

34. Lloyd B, Burrin J, Smythe P, Alberti KG. Enzymic fluoro-metric continuous-flow assays for blood glucose, lactate,pyruvate, alanine, glycerol, and 3-hydroxybutyrate. ClinChem. 1978;24:1724–9.

35. Orskov H, Thomsen HG, Yde H. Wick chromatography forrapid and reliable immunoassay of insulin, glucagon andgrowth hormone. Nature. 1968;219:193–5.

36. Lonnroth P, Jansson PA, Smith U. A microdialysis methodallowing characterization of intercellular water space in hu-mans. Am J Physiol. 1987;253:E228–E31.

37. Coppack SW, Persson M, Judd RL, Miles JM. Glyceroland nonesterified fatty acid metabolism in human muscleand adipose tissue in vivo. Am J Physiol. 1999;276:E233–E40.



38. Kurpad A, Khan K, Calder AG, et al. Effect of noradren-aline on glycerol turnover and lipolysis in the whole body andsubcutaneous adipose tissue in humans in vivo. Clin Sci(Lond). 1994;86:177–84.

39. Rebuffe-Scrive M, Krotkiewski M, Elfverson J, Bjorntorp P.Muscle and adipose tissue morphology and metabolism in Cush-ing’s syndrome. J Clin Endocrinol Metab. 1988;67:1122–8.

40. Jensen MD, Caruso M, Heiling V, Miles JM. Insulin regu-lation of lipolysis in nondiabetic and IDDM subjects. Diabe-tes. 1989;38:1595–601.

41. Rosdahl H, Lind L, Millgard J, Lithell H, Ungerstedt U,Henriksson J. Effect of physiological hyperinsulinemia onblood flow and interstitial glucose concentration in humanskeletal muscle and adipose tissue studied by microdialysis.Diabetes. 1998;47:1296–301.

42. Wajchenberg BL, Bosco A, Marone MM, et al. Estimationof body fat and lean tissue distribution by dual energy X-rayabsorptiometry and abdominal body fat evaluation by com-puted tomography in Cushing’s disease. J Clin EndocrinolMetab. 1995;80:2791–4.

43. Summers LK, Arner P, Ilic V, Clark ML, Humphreys SM,Frayn KN. Adipose tissue metabolism in the postprandialperiod: microdialysis and arteriovenous techniques compared.Am J Physiol. 1998;274:E651–E5.

44. Marco J, Calle C, Roman D, Diaz-Fierros M, VillanuevaML, Valverde I. Hyperglucagonism induced by glucocorti-coid treatment in man. N Engl J Med. 1973;288:128–31.

45. Dinneen S, Alzaid A, Miles J, Rizza R. Metabolic effects ofthe nocturnal rise in cortisol on carbohydrate metabolism innormal humans. J Clin Invest. 1993;92:2283–90.

46. Gravholt CH, Moller N, Jensen MD, Christiansen JS,Schmitz O. Physiological levels of glucagon do not influencelipolysis in abdominal adipose tissue as assessed by microdi-alysis. J Clin Endocrinol Metab. 2001;86:2085–9.

47. Jensen MD, Heiling VJ, Miles JM. Effects of glucagon onfree fatty acid metabolism in humans. J Clin EndocrinolMetab. 1991;72:308–15.

48. Caruso M, Divertie GD, Jensen MD, Miles JM. Lack ofeffect of hyperglycemia on lipolysis in humans. Am J Physiol.1990;259:E542–E7.

49. Cersosimo E, Coppack S, Jensen M. Lack of effect ofhyperglycemia on lipolysis in humans. Am J Physiol. 1993;265:E821–E4.

50. McMahon M, Gerich J, Rizza R. Effects of glucocorti-coids on carbohydrate metabolism. Diabetes Metab Rev. 1988;4:17–30.

51. Tappy L, Randin D, Vollenweider P, et al. Mechanisms ofdexamethasone-induced insulin resistance in healthy humans.J Clin Endocrinol Metab. 1994;79:1063–9.

52. Coderre L, Srivastava AK, Chiasson JL. Effect ofhypercorticism on regulation of skeletal muscle glyco-gen metabolism by epinephrine. Am J Physiol. 1992;262:E434 –E9.



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