Glucose kinetics differ between women and men, during and after exercise

Tracy J. Horton, Gary K. Grunwald, Jennifer Lavely and W. Troy Donahoo

J Appl Physiol 100:1883-1894, 2006. ; doi: 10.1152/japplphysiol.01431.2005 You might find this additional info useful... This article cites 61 articles, 45 of which you can access for free at: http://jap.physiology.org/content/100/6/1883.full#ref-list-1 This article has been cited by 6 other HighWire-hosted articles: http://jap.physiology.org/content/100/6/1883#cited-by Updated information and services including high resolution figures, can be found at: http://jap.physiology.org/content/100/6/1883.full Additional material and information about Journal of Applied Physiology can be found at: http://www.the-aps.org/publications/jappl This information is current as of January 5, 2013.
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Journal of Applied Physiology publishes original papers that deal with diverse area of research in applied physiology, especially those papers emphasizing adaptive and integrative mechanisms. It is published 12 times a year (monthly) by the American Physiological Society, 9650 Rockville Pike, Bethesda MD 20814-3991. Copyright 2006 the American Physiological Society. ISSN: 8750-7587, ESSN: 1522-1601. Visit our website at http://www.the-aps.org/.

J Appl Physiol 100: 18831894, 2006; doi:10.1152/japplphysiol.01431.2005.

Glucose kinetics differ between women and men, during and after exercise
Tracy J. Horton,1,3 Gary K. Grunwald,2 Jennifer Lavely,1 and W. Troy Donahoo4
1

Section of Nutrition, Department of Pediatrics, and 2Department of Preventive Medicine and Biostatistics, University of Colorado at Denver and Health Sciences Center, Denver; 3Department of Food Science and Human Nutrition, Colorado State University, Fort Collins; and 4Department of Preventive Medicine, Kaiser Permanente, Denver, Colorado

Submitted 14 November 2005; accepted in nal form 16 January 2006

Horton, Tracy J., Gary K. Grunwald, Jennifer Lavely, and W. Troy Donahoo. Glucose kinetics differ between women and men, during and after exercise. J Appl Physiol 100: 18831894, 2006; doi:10.1152/japplphysiol.01431.2005.As exercise can improve the regulation of glucose and carbohydrate metabolism, it is important to establish biological factors, such as sex, that may inuence these outcomes. Glucose kinetics, therefore, were compared between women and men at rest, during exercise, and postexercise. It was hypothesized that glucose ux would be signicantly lower in women than men during both the exercise and postexercise periods. Subjects included normal weight, healthy, eumenorrehic women and men, matched for habitual activity level and maximal oxygen uptake per kilogram lean body mass. Testing occurred following 3 days of diet control, with no exercise the day before. Subjects were tested in the overnight-fasted condition with women studied in the midluteal phase of the menstrual cycle. Resting (120 min), exercise (85% lactate threshold, 90 min), and postexercise (180 min) measurements of glucose ux and substrate metabolism were made. During exercise, women had a signicantly lower rate of glucose appearance (Ra) (P 0.001) and disappearance (Rd) (P 0.002) compared with men. Maximal values were achieved at 90 min of exercise for both glucose Ra (mean SE: 22.8 1.12 mol kg body wt 1 min 1 women and 33.6 1.79 mol kg body wt 1 min 1 men) and glucose Rd (23.2 1.26 and 34.1 1.71 mol kg body wt 1 min 1, respectively). Exercise epinephrine concentration was signicantly lower in women compared with men (P 0.02), as was the increment in glucagon from rest to exercise (P 0.04). During the postexercise period, glucose Ra and Rd were also signicantly lower in women vs. men (P 0.001), with differences diminishing over time. In conclusion, circulating blood glucose ux was signicantly lower during 90 min of moderate exercise, and immediately postexercise, in women compared with men. Sex differences in the glucagon increase to exercise, and/or the epinephrine levels during exercise, may play a role in determining these sex differences in exercise glucose turnover. sex; carbohydrate metabolism; catecholamines; glucagon
HABITUAL EXERCISE FORMS THE cornerstone of chronic disease prevention and treatment, including conditions characterized by the dysregulation of carbohydrate (CHO) and glucose metabolism, such as diabetes and obesity. Understanding the physiological basis for the benecial effects of exercise, along with elucidation of biological factors that affect the metabolic response to exercise is, therefore, of high importance. One biological factor that has recently been shown to affect exercise metabolism is sex (6, 10, 12, 18 20, 25, 40, 45, 46, 50, 51, 53). Determining male and female differences in the metabolic effects of exercise on glucose and CHO metabolism, therefore, is an important step in the delineation of biological factors that may impact the physiological benets of exercise.

With respect to whole body CHO metabolism, lean, eumenorrheic women have generally been observed to oxidize proportionally less CHO, and more lipid, compared with their male counterparts (2, 19, 25, 40, 51, 54), although not all studies report this (48, 50). Observations of a sex difference in exercise CHO oxidation have been made with exercise of mild to moderately high intensity [40 75% maximum O2 consump tion (VO2 max)], where both circulating blood glucose and intramuscular glycogen contribute to total CHO oxidation. The lesser relative CHO oxidation in women vs. men during mild to moderately high-intensity exercise could be due to a lower utilization of circulating blood glucose and/or muscle glycogen. Furthermore, sex differences in the use of these CHO sources could occur, even without differences in whole body CHO oxidation. In the few studies that have utilized stable isotope techniques to determine sex differences in exercise glucose ux, within the same study, both a similar (6, 48) and a lower (46) rate of exercise glucose turnover have been reported in women compared with men. The direct measurement of muscle glycogen changes from pre- to postexercise also provides conicting data with regard to sex differences, with both no difference (46) and a lower muscle glycogen utilization (51, 53) being reported in women compared with men. Factors related to differences in study design likely explain these inconclusive data. Nevertheless, whether or not sex differences in glucose turnover occur during exercise, and the contribution of blood glucose to total CHO utilization relative to muscle glycogen, warrants further investigation. The healthful effects of exercise are not limited to the increased energy expenditure and substrate utilization during the exercise bout itself. Indeed, the benecial effects of exercise with respect to substrate metabolism may be equally, if not more, important in the postexercise period. This is a time when factors come in to play to reestablish metabolic homeostasis and to replete fuel stores, especially hepatic and muscle glycogen, that have been utilized during exercise (33). Insulin action has been observed to increase by 4 h after exercise has ended (14, 38, 43, 44), and this effect may be maintained for up to 48 h (37). Concurrently, there is a decrease in CHO oxidation and a reciprocal increase in fat oxidation (25, 55). These factors promote glycogen repletion and the reestablishment of resting glucose homeostasis in the postexercise period. Sexbased differences in postexercise substrate metabolism are an area that has received little attention. Indeed, any potential sex differences in glucose and overall nutrient and hormone metabolism during exercise could result in sex differences in metabolism postexercise. For example, if men were to deplete
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Address for reprint requests and other correspondence: T. J. Horton, Sect. of Nutrition, Box C225, Univ. of Colorado Health Sciences Center, 4200 East 9th Ave., Denver, CO 80262 (e-mail: tracy.horton@uchsc.edu). http://www. jap.org

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muscle glycogen stores more than women during exercise, as some data would suggest (51, 53), this could increase glucose uptake more in men than women during the postexercise period, as lower muscle glycogen levels are associated with increased glucose uptake (42, 43). What may also be important is the impact of depleted glycogen stores, and changes in insulin action, on the partitioning of ingested nutrients between storage and oxidation during the postexercise period (7, 32). It could be hypothesized that a lesser reliance on blood glucose as a fuel source during exercise, and a greater utilization of muscle glycogen, and/or greater changes in postexercise insulin action, would result in more ingested CHO being directed toward storage vs. oxidation, thus increasing fat oxidation and decreasing net fat storage. Our laboratory has previously shown that, when subjects continue to fast for 2 h after 2 h of exercise at 40% VO2 max, whole body substrate oxidation does not differ between men and women (25). This does not, however, exclude the possibility that there are sex differences in glucose production and disposal, or between partitioning of glucose uptake between storage (glycogen repletion) and oxidation, especially after a meal. Hence, understanding potential sex-based differences in CHO metabolism during exercise and postexercise is relevant to our overall understanding of the regulation of glucose and nutrient metabolism. To further elucidate sex effects on exercise metabolism, the present study aimed to determine glucose kinetics, nutrient oxidation, and substrate and hormone responses to moderateintensity exercise of 90-min duration, and during the immediate 3 h postexercise in women relative to men. Care was taken to match the level of habitual activity, and VO2 max relative to lean body mass (LBM), between the sexes. In contrast to most previous investigations, women were studied in the midluteal phase of the menstrual cycle, rather than the follicular phase of the menstrual cycle, and steady-state exercise intensity was determined based on each individuals lactate threshold (LT).
METHODS

Table 1. Subject characteristics

Women Men

Age, yr Height, m Body weight, kg BMI, kg/m2 Body fat, % LBM, kg VO2max ml/min ml kg 1 min 1 ml kg LBM 1 min 1 LT ml/min % of VO2max Exercise activity, h/wk 3.9 METS (light activity) 48.9 METS (moderate activity) 8.9 METS (intense activity) Study day sex steroid hormone concentrations Estradiol, pg/ml Progesterone, ng/ml

34.0 1.66 56.9 20.5 22.4 41.6

6.3 0.09b 7.7a 1.6d 3.9a 5.2a

33.8 1.81 73.3 22.4 12.8 60.4

6.2 0.06 7.5 1.5 3.5 6.1

2,672 389a 47.1 4.6e 64.4 6.4 1,552 225c 58.3 5.4 0.08 0.13 3.85 2.22 3.01 2.37 155 152 (108 30)* 10.4 3.3 (10.2 3.4)*

3,921 512 53.8 7.4 65.1 7.5 2,287 504 57.9 6.6 0 0 3.82 2.71 3.45 2.11 30 8 0.7 0.2

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Values are means SD. BMI, body mass index; LBM, lean body mass (i.e., fat-free mass bone mass); VO2max, maximal oxygen uptake; LT, lactate threshold; METS, metabolic equivalents where 1 MET the oxygen consumption at rest. *Excludes one subject whose estradiol levels were unusually high (580 pg/ml) with normal progesterone (12.2 ng/ml); thus an analytical error was suspected. Women lower than men: a P 0.0001, b P 0.0003, c P 0.0004, d P 0.01, e P 0.02.

Subjects Lean, healthy women and men (20 45 yr) were recruited for the study. Female subjects were required to have a regular menstrual cycle ( 11 cycles over the past year), and all subjects were habitually active with men and women closely matched for activity level (Table 1). Medical exclusions included past or present history of cardiovascular disease, high blood pressure, diabetes, any hormonal imbalance or metabolic abnormality, and use of oral contraceptives or other hormones. A total of 24 subjects (11 women, 13 men) took part in the study. One woman was excluded, based on her study day hormone concentrations, indicating she was not in the luteal phase of her menstrual cycle. One man was excluded due to an extended delay between his prestudy VO2 max test and main study day testing, along with a signicant decrease in activity level (reported after the fact), which brought into question the accuracy of the relative intensity of the exercise on the main study day. This gave a total of 10 women and 12 men who completed the study. Ten of the men and women were pair-matched based on habitual activity level and VO2 max per kilogram LBM per minute. The remaining two men were typical of the group average; therefore, their data were included in the study analysis. Subject characteristics are shown in Table 1. The study protocol was approved by the University of Colorado Committee Institutional Review Board for the Protection of Human Subjects. All subjects read and signed an informed consent form before admission into the study.
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Preliminary assessments. A health and physical examination was completed on all subjects, including blood and urine analysis, to conrm that there was no medical reason for exclusion. Resting metabolic rate (RMR) was measured using indirect calorimetry via a metabolic cart system (Sensormedics 2900, Sensormedics, Yorba Linda, CA). Oxygen consumption and carbon dioxide production were used to calculate metabolic rate (62). This RMR value was used to determine energy intake of subjects during the period of prestudy diet control. Body composition was determined via dual-energy X-ray absorptiometry (Lunar, Madison, WI) (41). VO2 max was determined using a graded exercise test on a cycle ergometer (Lode Medical Technology, Groningen, The Netherlands). Subjects cycled at a constant revolutions per minute (70 90), while the resistance was gradually increased until volitional exhaustion. Workload commenced at 2550 W (depending on sex and habitual activity level) and was increased by 25 W every 2 min. To ensure maximum effort was achieved, two of the following criteria had to be fullled: whole body respiratory quotient 1.1, achieved maximum heart rate within 5% of the age-predicted maximum, and/or an increase in oxygen consumption in response to the nal workload of 2.0 ml kg body wt 1 min 1. For determination of LT, a retrograde dorsal hand intravenous (IV) catheter was placed, and the heated hand technique (36) was used to arterialize the blood. At the end of each workload stage (every 2 min), 0.5 ml of blood was drawn, placed in 8% perchloric acid (1.5 ml), vortexed, and placed on ice. Tubes were pre- and postweighed, and lactate concentrations were measured and adjusted for dilution. The log of the lactate concentration was plotted against the log of the oxygen consumption at the time of sampling. A two-line regression model was used to describe the two phases of the lactate accumulation. The point of intersection of these two lines was taken as the LT (59). Measurement of VO2 max, body composition, and RMR was performed in the luteal phase of the menstrual cycle for female subjects. Prestudy diet and exercise control. Subjects were fed a controlled diet for 3 days before the study day. All food was prepared by the General Clinical Research Center (GCRC) diet kitchen at the University of Colorado, and subjects were required to consume breakfast in the GCRC with other food prepared to take away. No other food was permitted, and
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subjects were required to consume all the food given. The only optional part of the diet were two food modules (840 kJ each, same composition as the overall diet), one or both of which the subjects could eat if they were hungry. The diet composition was 25% fat, 15% protein, and 60% CHO, and initial energy intake was calculated at 1.6 1.8 RMR based on subjects habitual activity level. This period of diet control helped minimize uctuations in energy balance, as suggested by body weight (BW) changes of 2%. Subjects were allowed to follow their usual activity routine for the rst 2 days of the diet, and on the last day they refrained from any planned exercise (energy intake, 1.6 RMR). Testing was, therefore, performed at least 36 h after the last exercise bout. Study Days Subjects stayed overnight on the GCRC the evening before the study. Between 1900 and 2000, subjects consumed their evening meal and had a light snack at 2200. After this, subjects remained fasted until the end of the study the following day. The study involved measurement of resting, exercise, and postexercise glucose kinetics, as well as respiratory gas exchange. Conrmation of menstrual cycle phase in women was based on serum estrogen and progesterone ( 2.5 ng/ml) levels measured on the day of the study (Table 1). Determination of glucose kinetics and circulating substrates and hormones. On the study day, IV catheter placement occurred between 6:45 and 7:30 AM. An infusion IV was placed in an antecubital vein for delivery of the stable isotope. In the contralateral arm, a sampling catheter was placed retrograde fashion into a dorsal hand vein or, if necessary, in a wrist vein. The heated hand technique (36) was used to obtain arterialized blood samples. Basal blood samples were taken, for determination of background enrichment of glucose, before the start of the tracer infusion at 0800 (time 0 at rest). A primed (17.6 mol/kg), constant (0.2 mol kg 1 min 1) infusion of [6,6-2H2]glucose began and continued for 120 min at rest. Subjects remained semirecumbent in bed over this time, and four blood samples were taken over the nal 30 min of rest (time t 45, 35, 25, and 15 relative to the start of exercise) for the determination of resting glucose kinetics, substrate, and hormone concentrations. Subjects were then transferred to an electronically braked stationary bike (Lode Medical Technology) and began the exercise period, which included a 5-min warm-up, followed by 90 min of steady-state exercise at 85% of each individuals LT. At the start of the exercise (warm-up), the glucose isotope infusion rate was increased twofold to minimize changes in isotope enrichment (63). Blood samples were drawn at t 10, 20, 30, 45, 60, 75, and 90 min of steady-state exercise. After completion of the exercise bout, subjects moved into bed and remained resting for 3 h postexercise. The isotope infusion rate was decreased to preexercise levels. Blood samples were drawn at t 110, 120, 135, 150, 180, 210, 240, and 270 min during the postexercise period. Respiratory gas exchange. In the 30 min before blood sampling at rest, a 15- to 20-min measurement of respiratory gas exchange was made via indirect calorimetry (Sensormedics 2900, Sensormedics, Yorba Linda, CA). During exercise, 3 20-min measurements of respiratory gas exchange were performed every 30 min. CHO and fat oxidation were calculated from the volume of O2 consumed and volume of CO2 expired after correcting for protein oxidation (26, 62). Protein oxidation within each period (rest, exercise, and postexercise) was estimated from urinary nitrogen excretion. Determination of circulating hormone and substrate levels. Two to three milliliters of blood were added to EDTA tubes for the measurement of tracer enrichment and plasma substrate concentrations. Samples were immediately placed on ice and spun, and plasma was separated. Approximately 0.5 ml of whole blood was added to a preweighed tube containing 1.5 ml of iced perchloric acid (8%). After vortexing, the tube was postweighed and spun to separate the supernatant. This was used to measure blood lactate. Two and one-half milliliters of whole blood were added to 40 l of preservative (EGTA 3.6 mg plus glutathione 2.4 mg in distilled water), for plasma catecholamine determinations. Blood for glucagon measurement (2 ml) was added to tubes containing EDTA plus
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500 kallikrein-inhibitor units of aprotinin. Samples were immediately placed on ice and spun. Approximately 7 ml of whole blood were allowed to clot, and the serum was separated off after spinning. This was used for determination of the remaining hormone and substrate concentrations. All plasma, serum, and supernatant samples were stored at 70C until analysis. Catecholamines were determined in duplicate by highperformance liquid chromatography with electrochemical detection [intra-assay coefcients of variation (CVs) 6.2% epinephrine (Epi), 4.9% norepinephrine (Norepi)] (8). Radioimmunoassays were used to determine serum insulin (Kabi Pharmacia, Piscataway, NJ), cortisol, progesterone, estradiol (Diagnostic Products, Los Angeles, CA), and glucagon (Linco Research, St. Louis, MO). Samples were run in duplicate with intra-assay CVs of 10, 6.7, 8.8, 7.5, and 9.4%, respectively. Blood lactate (Sigma Diagnostics, St. Louis, MO) was run in duplicate with intra-assay CV of 4.2%. Lactate concentrations and plasma catecholamines (Epi and Norepi) were measured on samples drawn at t 35, 15, 10, 20, 30, 45, 60, 75, 90, 110, 120, 150, 210, and 270 min. Glucagon, cortisol, and insulin were measured on samples drawn at t 15, 30, 60, 75, and 90 min. Postexercise, insulin was measured at t 120, 150, 180, 210, 240, and 270 min, whereas glucagon and cortisol were measured at t 120, 150, 210, and 270 min. Estradiol and progesterone were measured on samples drawn at t 45 min only. Determination of glucose isotope enrichment and concentration. This was measured via gas chromatography-mass spectrometry (GC-MS; GC model 6890 and 5973N, Agilent, Palo Alto, CA). First, plasma samples were thawed and 80 g of [U-13C]glucose added to act as an internal standard. The pentacetate derivative of glucose was then generated as follows. Samples were deproteinized with iced ethanol, and the supernatant was dried in a Speedvac at 50C for 2.5 h. Samples were then derivatized using 100 l of acetic anhydride-pyridine solution (1:1) and heating for 30 min at 100C. Ethyl acetate (100 l) was then added, and the samples were vortexed and then transferred to GC-MS vials for analysis. Injector temperature of the GC-MS was set at 250C, and initial oven temperature was set at 150C. The column used was an Agilent HP-5MS 0.25 mm 30 m with a 0.25-mm lm thickness. Oven temperature was increased 30C/min until a nal temperature of 250C was achieved. Helium was used as the carrier gas with a 20:1 ml/min pulsed split injection ratio. Transfer line temperature was set at 280C, source temperature at 250C, and quadruple temperature at 150C. Methane chemical ionization (63) was used to monitor selective ions with mass-to-charge ratios of 331 (M 0 from natural glucose), 333 (M 2 from [6,6-2H2]glucose), and 337 (M 6 from the [U-13C]glucose, internal standard). Natural glucose standards from 10 to 200 mg/dl were prepared and spiked with 80 g [U-13C]glucose to enable the generation of a calibration curve for determining the concentration of natural glucose. The calibration curve was constructed by comparing the known ratio of glucose/[U-13C]glucose to the measured area ratio of 331:337. Natural glucose concentration in the samples was then determined by measuring the sample 331:337 area ratio and using the linear equation obtained from the calibration curve to calculate the natural glucose concentration in milligrams per deciliter. Calculations. Glucose moles percent excess (MPE) and concentration data were spline tted to remove noise introduced by analytical and sampling errors (56). Glucose rates of appearance (Ra) and disappearance (Rd) were then calculated using the Steele equation, as modied for use with stable isotopes (47, 63). Ra F pV C2 Rd Ra C1 /2 E2 E1 / t2 E2 E1 /2 pV C2 C1 / t2 t1 t1 F

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where Ra is for the tracee ( mol/min), F is infusion rate of tracer ( mol kg 1 min 1), pV is effective volume of tracee distribution (100 ml/kg BW) (47), t1 is time 1 of sampling, t2 is time 2 of sampling, C1 is tracee concentration at t1, C2 is tracee concentration at t2, E1 is
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plasma MPE at t1, and E2 is plasma MPE at t2. For exercise substrate kinetics, no value was calculated for t 10 min; rather the t 10 min glucose concentration and MPE values were used to calculate the t 20 min value only. This was due to the 30-min duration of time between the nal t 15 min resting blood draw and the t 10 min exercise blood sample (the 30 min include the 5-min warm up), and the change in infusion rate part way through (infusion rate increasing at t 0 of steady-state exercise), which present suboptimal conditions for reliable tracer kinetic calculations. Data Analysis Subject characteristics were compared using an unpaired t-test. For each dependent variable, a general linear multivariate model (30), estimated using SAS PROC MIXED (SAS Institute, Cary, NC), was used to account for correlation between repeated measurements on the same subject. This model allowed different variances at the different measurement times, as well as different variances for men and women. This analysis provides a valid handling of the missing values that occurred randomly across subjects and study phase due to occasional problems with blood sampling. Differences in time courses between men and women were examined, as were interactions between time and sex. Comparisons of particular interest (e.g., men vs. women at a given time, differences between times, trends over time, and differences between men and women in the changes between two times) were estimated and tested using contrasts within the multivariate model. Comparisons between periods were made by averaging the values during the period for each subject and again using a multivariate model to account for correlations between periods on the same subject. Several variables (lactate, Epi, Norepi, and estrogen) were log transformed for the signicance tests in the multivariate model to account for skewness in the distributions. However, results are reported in the original units for easier interpretation. Results are presented as means SE. For glucose kinetic data, results are expressed both in terms of BW and LBM. The latter mode of expression adjusts for differences in the most metabolic active tissue mass between men and women and is consistent with the matching of men and women for VO2 max/kg LBM. For glucose disposal, glycogen utilization, and CHO oxidation during exercise, data were again expressed relative to LBM, as this parameter is the primary determinant of metabolic rate and glucose disposal during exercise. This effectively adjusts for exercise differences in energy expenditure between men and women. In addition, data were expressed relative to leg lean mass, which would be the major energy-consuming tissue mass during cycle exercise.
RESULTS

Table 3. Metabolic rate and respiratory exchange ratio

Women Men

Rest n MR, kJ/min MR, kJ kg LBM 1 h RER* Nonprotein RER* n MR, kJ/min MR, kJ kg LBM 1 h Total EE, kJ/90 min RER Nonprotein RER n MR, kJ/min MR, kJ kg LBM 1 h RER* Nonprotein RER* 4.26 6.15 0.754 0.741 10 0.19a 0.13b 0.012 0.016 10 1.3a,e 1.4e 119 0.006e 0.006e 5.10 5.14 0.749 0.726 11* 0.14 0.13 0.009 0.015 12 2.7e 2.3e 245a 0.009e 0.009e

Exercise 29.3 42.4 2,636 0.865 0.866 10 4.61 6.65 0.708 0.684 44.1 43.7 3,966 0.880 0.883 11* 5.67 5.70 0.698 0.667

Postexercise

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0.21a,c 0.15c 0.007c,d 0.007c,d

0.15c 0.09b,c 0.009c,d 0.012c,d

Values are means SE; n, no. of subjects. MR, metabolic rate; RER, respiratory gas exchange ratio; EE, energy expenditure. *n 11 for men (data excluded for one subject due to error in the dilution mode gas exchange data). a Women lower than men: P 0.002 rest, P 0.0002 exercise, P 0.0004 postexercise. b Women greater than men: P 0.0001. c Signicantly different from rest: these values range from P 0.0001 to 0.03. d Signicantly different from exercise: P 0.0001. e Signicantly different from rest and postexercise: P 0.0001.

Table 2 shows the exercise parameters for women and men. The relative intensity of the exercise was almost identical in women and men: 51% of VO2 max in both sexes, and 87 and 88% of LT, respectively. Energy Expenditure and Whole Body Substrate Oxidation The average metabolic rate and respiratory exchange ratio (RER) for rest, exercise, and postexercise are shown in Table 3. The expected increase in metabolic rate and RER between Table 2. Exercise parameters
Exercise VO2 n Heart Rate, beats/min Workload, W ml kg LBM 1 min
1

% of VO2max

% of LT

Women Men

10 12

130 4 126 5

89 5 133 9

32.6 1.1 33.4 1.8

50.8 1.3 51.1 1.7

87.2 0.9 88.3 0.7

Values are means

SE; n, no. of subjects. VO2, oxygen consumption. J Appl Physiol VOL

rest and exercise was observed. In addition, metabolic rate postexercise was signicantly elevated compared with preexercise (P 0.03 for women, P 0.0001 for men), whereas RER postexercise was signicantly reduced compared with preexercise (P 0.006 for women, P 0.0003 for men). Metabolic rate, relative to LBM, was not different between men and women during exercise, but it was signicantly greater in women vs. men at rest and postexercise (P 0.0001). No signicant sex differences were observed in RER, or nonprotein RER, for any study phase. During exercise, absolute rates of protein and CHO oxidation were lower in women vs. men (P 0.0001 and P 0.0023, respectively, Table 4), with a nearly signicant difference in absolute fat oxidation (P 0.06). Relative nutrient oxidation, however, was not different between the sexes, whether data were expressed as a percentage of total energy expended (Table 4), or relative to LBM (data not shown). It is noteworthy that the nonprotein RER postexercise was below 0.70. This meant that it would have been erroneous to use the postexercise respiratory gas exchange data to estimate substrate oxidation. Results suggest that, as subjects continued to fast in the postexercise period (after the 2-h rest period followed by 90-min exercise), there was signicant production of CO2 and/or utilization of O2, due to metabolic processes unrelated to direct nutrient oxidation. Glucose kinetics. Figure 1 shows the MPE (%) for glucose at rest, during the 90-min exercise, and over the 3 h postexercise. Although values tended to drift downward in men during exercise, large changes in isotope enrichment were avoided by doubling the isotope infusion rate at the onset of exercise and decreasing this postexercise. At rest, glucose turnover did not
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Table 4. Whole body nutrient oxidation

Women g/min % of EE g/min Men % of EE

Rest n CHO oxidation Fat oxidation Protein oxidation n CHO oxidation Fat oxidation Protein oxidation 10 0.022 0.011 9.1 5.0 0.073 0.006 67.7 4.9 0.050 0.004 23.3 1.5 Exercise 10 0.956 0.061 57.0 2.0 0.299 0.016 40.6 2.0 0.037 0.005 2.4 0.3 12 1.543 0.101 61.9 3.0 0.399 0.047 35.2 2.9 0.064 0.006 2.9 0.3 11* 0.014 0.012 4.8 4.1 0.085 0.005 65.9 4.8 0.076 0.005 29.3 1.9

Values are means SE; n, no. of subjects. *n 11 for men (data excluded for one subject due to error in the dilution mode gas exchange data). CHO, carbohydrate. Women lower than men: P 0.0002, P 0.0003, P 0.002.

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differ between women and men, whether data were expressed relative to BW (Fig. 2) or LBM (Fig. 3). Average glucose Ra at rest was 12.5 0.5 mol kg BW 1 min 1 in women and 12.6 0.5 mol kg BW 1 min 1 in men, while glucose Rd equaled 12.4 0.6 and 12.8 0.7 mol kg BW 1 min 1, respectively. During exercise, glucose Ra and Rd increased over time, but less so in women than in men (sex time interaction, P 0.0001 for data expressed relative to BW or LBM). In addition, the average glucose Ra and Rd during exercise, expressed per kilogram BW, were signicantly lower in women vs. men (P 0.001, glucose Ra and P 0.002, glucose Rd), with maximal rates occurring at 90 min of exercise for both glucose Ra (women 22.8 1.1 mol kg BW 1 min 1, men 33.6 1.8 mol kg BW 1 min 1) and glucose Rd (23.2 1.3 and 34.1 1.7 mol kg BW 1 min 1, respectively). Average exercise glucose Ra, per kilogram LBM, was also signicantly lower in women vs. men (P 0.04), whereas average glucose Rd per kilogram LBM tended to be lower (P 0.065) due to smaller differences in the rst 45 min of the exercise. Sex differences in glucose kinetics were maintained in the postexercise period. There was a signicant sex time interaction (P 0.0003 for glucose Ra

Fig. 2. Glucose rate of appearance (Ra; A) and glucose rate of disappearance (Rd; B) (per kg body weight) at rest ( 35 to 15 min), during cycle exercise (0 90 min), and postexercise (90 270 min). Values are means SE. A: signicant effect of time (P 0.0001) and sex time interaction (P 0.0001) for exercise and postexercise periods. Signicantly higher average glucose Ra in men vs. women during exercise (P 0.001) and postexercise (P 0.001). B: signicant sex time interaction (P 0.0002) for exercise and postexercise periods. Signicantly higher average glucose Rd in men vs. women during exercise (P 0.002) and postexercise (P 0.001).

Fig. 1. Glucose tracer enrichment, expressed as mole percent excess (MPE). Values are means SE. J Appl Physiol VOL

and Rd expressed relative to BW or LBM) due to a more immediate vs. gradual decline in postexercise glucose turnover in women compared with men. Furthermore, the average glucose Ra and Rd postexercise were signicantly lower in women vs. men, whether data were expressed relative to BW (P 0.001, Ra and Rd) or LBM (P 0.03 and P 0.05 for Ra and Rd, respectively). Estimated glucose and glycogen oxidation during exercise. It was assumed that all glucose uptake during exercise was directed toward oxidation, i.e., a maximal estimate. Thus estimated glycogen oxidation total CHO oxidation glucose oxidation. Table 5 shows results expressed in absolute terms and relative to measures of LBM and total CHO oxidawww.jap.org

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Fig. 3. Glucose rate of appearance (Ra; A) and glucose rate of disappearance (Rd; B) (per kg lean body mass) at rest ( 35 to 15 min), during cycle exercise (0 90 min), and postexercise (90 270 min). Values are means SE. A: signicant sex time interaction during exercise and postexercise (P 0.0002). Signicantly higher average glucose Ra in men vs. women during exercise (P 0.04) and postexercise (P 0.03). B: signicant sex time interaction during exercise (P 0.0001) and postexercise (P 0.0003). Higher average glucose Ra in men vs. women during exercise (P 0.065) and postexercise (P 0.05).

tion. Estimated blood glucose oxidation was signicantly lower in women vs. men in absolute terms (P 0.0001), when expressed relative to total LBM (P 0.04) or leg lean mass (P 0.02), but was not different when expressed as a percentage of total CHO oxidation. In contrast, estimated glycogen oxidation was only signicantly lower in absolute terms (P 0.003) in women vs. men. Circulating substrate concentrations. Figure 4 shows the changes in glucose and lactate concentrations during the study. Glucose concentration preexercise was signicantly lower in women vs. men (P 0.009), but it increased with the onset of exercise to achieve a similar level as that in men (P 0.59). There was a gradual decline in glucose concentration during exercise (P 0.0001 for decrease within each sex). Postexercise glucose levels fell initially in women, more so than in men, but the average glucose levels postexercise were not different between the sexes (P 0.12). In both women and men, postexercise glucose levels were signicantly lower compared with initial resting levels (P 0.0001). Lactate concentration showed the expected changes with exercise and postexercise (Fig. 4B). Average exercise lactate levels were signicantly lower in women vs. men (P 0.02), and the decrease in the average lactate concentration from exercise to postexercise was signicantly lower in women vs. men (P 0.04). Circulating hormone concentrations. Insulin concentration is shown in Fig. 5. There was a small but marginally signicant decline in insulin level with exercise (P 0.03 for men, P 0.09 for women). No sex differences in the pattern of insulin response, nor insulin concentrations during any study phase, were observed. Epi and Norepi changes throughout the study are shown in Fig. 6. Women had a signicantly lower Epi concentration compared with men at rest (P 0.05; 27 3 vs. 40 5 pg/ml, respectively) with no difference in Norepi concentrations. During exercise, there was a time course effect for Epi due to levels signicantly increasing in both women (P 0.01) and men (P 0.0001). Average Epi levels during exercise, however, were signicantly lower in women vs. men (P 0.02, average values, 78 6 vs. 128 9 pg/ml), and the magnitude of the sex difference in Epi concentration during exercise increased over time with the concentration at 90 min in women being one-half that observed in men. Postexercise, Epi was slightly lower in women vs. men (P 0.05; average 25 1 vs. 33 2 pg/ml). With respect to Norepi, the time course of changes during exercise differed between the sexes (P 0.0004), with men demonstrating a signicant increase (P

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Table 5. Estimated blood glucose and muscle glycogen oxidation during exercise
g/min g kg LBM
1

min

g kg leg LBM

min

% of Total CHOox

Women (n 10) Glucose oxidation* Glycogen oxidation* Men (n 12) Glucose oxidation* Glycogen oxidation*

0.225 0.009a 0.731 0.060b 0.388 0.019 1.155 0.104

0.0055 0.0003d 0.0175 0.0012 0.0064 0.0003 0.0191 0.0017

0.0157 0.008c 0.0500 0.0031 0.0190 0.001 0.0562 0.0049

24.3 1.5 75.7 1.5 26.4 2.2 73.6 2.2

Values are means SE; n, no. of subjects. % of total CHOox, percentage of total carbohydrate oxidation provided by glucose or glycogen oxidation. *Estimated values based on the assumption that 1) all glucose uptake during exercise is directed toward oxidation (thus this represents a maximal estimate); and 2) all of the difference between total carbohydrate oxidation and estimated glucose oxidation is due to glycogen oxidation. Women lower than men: a P 0.0001, b P 0.003, c P 0.02, d P 0.05. J Appl Physiol VOL
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0.002) and women demonstrating a signicant decrease (P 0.003). Postexercise, there was also a sex time interaction (P 0.04) due to a more gradual vs. immediate decline in Norepi in women compared with men. Average circulating Norepi levels were not signicantly different between men and women during any study phase. Figure 7 shows the glucagon and cortisol concentrations throughout the study. At rest, glucagon concentration was signicantly higher in women vs. men (P 0.002). Glucagon increased signicantly during exercise (P 0.0001 for men and P 0.013 for women), but glucagon concentrations were no longer signicantly different between the sexes. Thus the increment in glucagon from rest to exercise (average value) was signicantly less in women compared with men (P 0.04). There was no signicant sex difference in glucagon concentration postexercise, but the change in glucagon concentration from rest to postexercise showed a signicant sex difference (P 0.02) due to the glucagon staying somewhat

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Fig. 5. Insulin concentration at rest ( 15 min), during cycle exercise (0 90 min), and postexercise (90 270 min). Values are means SE. Signicant decline over time during exercise in men (P 0.03) and near signicant decline in women (P 0.09).

elevated in men, whereas it declined almost to resting levels in women. Although resting cortisol levels were lower in women vs. men (P 0.01), the magnitude of increase with exercise was not different between the sexes, nor was the decline after exercise (P 0.12 for both). There was a signicant decrease in cortisol over the postexercise period in both men and women (P 0.0001).
DISCUSSION

Fig. 4. A: glucose concentrations at rest ( 45 to 15 min), during cycle exercise (0 90 min), and postexercise (90 270 min). Signicant effect of time during exercise (P 0.0001 for men and women). Signicantly lower blood glucose in women vs. men at rest (P 0.009). B: lactate concentrations at rest (time t 15 average of t 35 and 15 min measures), during cycle exercise (0 90 min), and postexercise (90 270 min). Signicant effect of time during exercise (P 0.0007 for men and women) and postexercise. Signicantly lower average lactate concentration in women vs. men during exercise (P 0.02) and signicantly lower decrease in lactate concentration from exercise to postexercise (P 0.04). Values are means SE. J Appl Physiol VOL

The main nding from this study was that eumenorrheic women, studied in the luteal phase of the menstrual cycle, had a signicantly lower exercise glucose turnover compared with men, and this was maintained for the initial ( 2 h) postexercise period. Data suggest a greater exercise glucose clearance in men vs. women, as the higher rate of exercise glucose ux in men was observed at the same level of glycemia as women. Of the hormones measured, only the increase in Epi during exercise mirrored the sex differences in exercise glucose kinetics, with levels being signicantly lower in women vs. men. Notably, however, there was also a lesser increment in circulating glucagon levels from rest to exercise in women compared with men. Results from the current investigation agree with data from the study of Roepstorff et al. (46), which also reported a lower glucose turnover in highly trained women vs. men, during 90-min cycle exercise at 58% of VO2 max. Both studies also observed no sex difference in relative CHO utilization and no difference in glycogen utilization. It appears that certain similarities in protocol design, including carefully matching women and men in terms of VO2 max per kilogram LBM and habitual training status, stringent prestudy diet and exercise control, as well as studying subjects in the overnight-fasted state, resulted in a consensus between the results from the two studies. As exercising above or below the LT can signicantly affect glucose kinetics (9) and whole body substrate oxidation (9, 10), it is noteworthy that the agreement between the two studies occurred, despite differences in the method of calculating steady-state exercise intensity, relative to LT in the
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current investigation ( 87% LT, equivalent to 51% VO2 max) O2 max (58%) in the study of Roepstorff et al. and relative to V (46). It is likely, however, that the subjects in the study of Roepstorff et al. exercised below their LT, as these subject were a relatively homogeneous group of highly trained athletes. Potential hormonal factors that may have played a role in the observed sex differences in exercise glucose kinetics include the glucagon response and the catecholamine increase, in particular Epi, during exercise. Glucagon is an important regulator of exercise glucose production, especially when insulin levels remain very low, as observed during exercise in the present study (57). Although women had higher absolute concentrations of glucagon than men, the increase in exercise

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Fig. 7. Glucagon (A) and cortisol (B) concentrations at rest ( 15 min), during cycle exercise (0 90 min), and postexercise (90 270 min). Values are means SE. A: signicant increase during exercise (P 0.0001 for men, P 0.013 women). Signicantly higher average glucagon concentration at rest in women vs. men (P 0.002) and signicantly lower increment in average glucagon from rest to exercise (P 0.04) and lesser difference in the change from rest to postexercise (P 0.02). B: signicant effect of time postexercise (P 0.0001). Signicantly lower cortisol concentration at rest in women vs. men (P 0.01).

Fig. 6. Epinephrine (A) and norepinephrine (B) concentrations at rest (t 15 average of t 35 and 15 min measures), during cycle exercise (0 90 min), and postexercise (90 270 min). Values are means SE. A: signicantly greater average concentrations during exercise than at rest or postexercise (P 0.0001 for women and men). Signicant increase during exercise (P 0.0001 for men, P 0.01 for women). Signicantly lower average epinephrine concentration at rest (P 0.05), during exercise (P 0.02), and postexercise (P 0.05) in women vs. men. B: signicantly greater average concentrations during exercise than at rest or postexercise (P 0.0001 for men and women). Signicant increase during exercise for men (P 0.002), and signicant decrease during exercise for women (P 0.003). Signicant sex time interaction postexercise (P 0.04). J Appl Physiol VOL

glucagon level was signicantly lower in women. It has been shown that the increment in glucagon level from rest to exercise is a major determinant of the increase in glucose release during exercise (23, 58). Assuming no sex difference in hepatic glucagon extraction, then the lesser glucagon increase in women vs. men from rest to exercise could have been driving the sex differences in glucose release and, consequently, glucose utilization. Interestingly, resting glucose production was not different between men and women, despite the higher systemic glucagon levels in women, suggesting the systemic glucagon concentration per se may not relate to glucose production in the same way in men and women. The fact that glucose levels remained higher than preexercise basal levels in women throughout exercise suggests a better defense of circulating blood glucose in women, whereas the decline in
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men relative to rest may have stimulated a greater glucagon secretion during exercise. Interestingly, systemic Norepi levels showed a signicantly different pattern of response in women vs. men, decreasing after an initial increase in women vs. a continuous increase in men. This may also have played into the increase in glucagon release in men or could, in of itself, have played a role in the differential exercise glucose production between the sexes. The lower Epi increase in women vs. men is consistent with previous observations (6, 12, 19, 25, 51). As Epi can play some role in glucose production during exercise (28, 34), albeit to a lesser extent than glucagon, the lower response in women could have contributed to their lower glucose production compared with men. Elevated Epi has also been shown to decrease glucose uptake (61). Such an effect would predict a lower glucose Rd in men, as they had much higher Epi levels than women. The exercise Epi response does not, therefore, explain any of the sex difference in glucose Rd, and other factors must be overriding the effects of Epi on glucose uptake. Epi can signicantly increase muscle glycogenolysis (17, 39, 60). Estimated (muscle) glycogen utilization was signicantly lower in women vs. men in absolute terms but not when expressed relative to LBM of leg lean mass. Epi levels may, therefore, be more related to the absolute utilization of glycogen rather than the relative. It needs to be borne in mind, however, that the glycogen utilization value is purely an estimate dependent on the assumption of complete oxidation of all glucose that is taken up, complete oxidation of all glycogen that is broken down, and no net lactate production. Any of these scenarios may not be entirely true, to the extent that blood-derived and/or glycogen-derived glucose could be shunted through glycolysis to generate energy and lactate. Measurement of muscle glycogen changes from muscle biopsies is devoid of such assumptions and provides a direct measure of net glycogen utilization. With the use of this technique, no sex difference in net muscle glycogen utilization has been reported (46, 52), along with no sex difference in Epi levels (46). In contrast, a lower muscle glycogen decrease has been observed in women compared with men (90 100 min of running at 65% VO2 max), along with a lower Epi response (51). Thus the Epi response may relate more to muscle glycogen utilization in men and women, but the role it plays in determining sex differences in exercise glucose turnover requires further investigation. It is notable that women maintained the same level of glycemia during exercise as men, despite a lower glucose ux. This indicates that, in response to exercise, men increased glucose clearance more than women. Reasons for this are not apparent from the current data but may be related to factors controlling glucose uptake at the muscle. Interestingly, men had signicantly greater lactate levels during exercise than women, despite both groups exercising at the same percentage of LT. One possible reason for this could be a greater lactate production in men as a result of increased glycolysis. Indeed, there are data to suggest that men have a more glycolytic enzyme prole at the muscle relative to women (22). Thus the greater glucose clearance in men may have contributed to a greater glycolysis, meaning that not all the glucose that was taken up was directly oxidized. Alternatively, the lower lactate level during exercise in women could be due to a greater lactate clearance, and this could be related to testing of women in the luteal phase of the menstrual cycle. Previous studies have
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reported lower circulating lactate concentrations during steadystate exercise in the luteal vs. follicular phase of the menstrual cycle (3, 21, 27, 29, 64), whereas LT itself, determined by the break point in lactate level rather than the absolute lactate concentration, is not different between phases of the menstrual cycle (13, 16). It is possible that the prevailing sex-steroid environment either directly or indirectly played a role in determining the sex differences in exercise glucose kinetics. Exogenous estradiol administration has been consistently shown to reduce exercise glucose turnover (5, 11, 49), whereas progesterone appears to have no additional, or conversely antagonistic, effect (11). There is inconsistency regarding whether or not glucose kinetics are affected by menstrual cycle uctuations in estradiol and progesterone, with both no difference (3, 24) and a greater glucose ux (4, 64) being observed in the follicular vs. luteal phase. Reasons for these conicting data may be related to differences in the level of circulating estrogen in female subjects and/or the demands placed on glucose production (24). In the present investigation, women were tested in the midluteal phase of the menstrual cycle, whereas, in the study of Roepstorff et al. (46), women were studied, on average, at day 9 of the follicular phase of the menstrual cycle. Despite what would have been very different progesterone levels, estradiol would have been elevated in women in the two studies, and both studies observed a signicantly lower glucose turnover relative to men, thus potentially supporting a role for estradiol in determining the sex difference in exercise glucose kinetics. An indirect effect of the sex steroids could occur via interaction with the endocrine response, through a decrease or increase in other hormone release or through an enhancement or suppression of the action of glucoregulatory hormones. In the present study, we observed a signicantly lower increase in glucagon from rest to exercise in women vs. men. In a previous study (unpublished data), we also observed a greater exercise increase in plasma glucagon with exercise (120 min of 40% VO2 max exercise) in men (57 3.5 to 80 6.9 pg/ml) compared with women (51 3.9 and 59 3.7 pg/ml). It is notable that, despite a similarly reduced increment in glucagon level with exercise in women tested in the present study and our previous study, the absolute glucagon concentrations were much higher in women in the present study. This may be due to the phase of the menstrual cycle in which the two groups of women were tested: midluteal phase in the present study and follicular phase in the previous study. Greater glucagon levels have been observed in the luteal vs. follicular phase of the menstrual cycle (24, 31), and progesterone receptors are present on the -cells of the pancreas (15), suggesting progesterone may increase tonic glucagon secretion. Nevertheless, it appears that, although progesterone may affect basal glucagon level, it does not affect the response to exercise. The lower exercise glucagon response in women may, therefore, be due to a greater estrogen level in women, a higher testosterone level in men, or may be unrelated to the circulating sex steroid environment per se. Data presented here are the rst to report on postexercise differences in glucose kinetics between men and women. Mirroring what was observed during exercise, women had a signicantly lower glucose turnover than men. This was mainly due to a more immediate decrease in glucose ux at the end of exercise in women, whereas in men it took a little longer
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to reestablish basal glucose homeostasis. The higher glucose release in men postexercise may have been facilitated by the maintenance of an elevated glucagon level compared with preexercise, basal levels. By 120 min postexercise, however, glucose ux was similar between the sexes. The lower glucose concentration in women vs. men during the initial postexercise period appeared to be due to the early fall in glucose Ra exceeding the fall in Rd. This was insufcient to trigger any form of counterregulatory response, however. With respect to postexercise glycogen resynthesis, we had hoped to obtain an estimate of this from the difference between total CHO oxidation and tracer-determined glucose disposal. As presented in the results, however, the postexercise nonprotein RER was below 0.70, invalidating the use of the gas exchange data for the estimation of fat and CHO oxidation. The extended period of fasting, along with the 90-min exercise, likely introduced metabolic processes, including ketogenesis and gluconeogenesis (not balanced by oxidation) that would have incurred metabolic gas exchange unrelated to direct substrate oxidation. Thus we were not able to estimate if there were sex differences in nonoxidative glucose disposal (NOGD) postexercise. One might speculate, however, given the fact that our laboratory has previously observed no difference in postexercise CHO and lipid oxidation between the sexes under fasted conditions (25), a large part of the greater glucose Rd in men in the initial 2 h postexercise would have been due to a greater NOGD. It is worth considering why other studies have not observed a signicant sex difference in exercise glucose kinetics. In the study of Ruby et al. (48), shorter duration exercise (25 min at 70% LT immediately followed by 25 min at 90% LT) was employed. We currently observed the greatest difference in glucose kinetics between men and women after 45 min of continuous exercise at 87% LT. Testing of subjects 4.5 h after a high-CHO snack and at different times of day (meal consumption likely occurring for later testing) may have masked the potential sex differences in glucose turnover in the study of Carter et al. (6). Food consumption would have somewhat increased hepatic glycogen stores, which would otherwise be signicantly depleted by an overnight fast: the test condition of the current and previous study observing lower exercise glucose kinetics in women (46). It is possible, therefore, that the sex difference in exercise blood glucose production (and utilization) may only be manifest in the setting of low hepatic glycogen stores, and differences in glucose kinetics may only become apparent when hepatic stores are additionally depleted with 45 min of moderate exercise. Further support for this comes from two other studies. In a retrospective comparison of exercise glucose kinetics in women, to that of previously tested men (19), no difference in exercise glucose kinetics (45 65% VO2 max, 60 min) was observed with subjects tested 3.5 h after a high-CHO snack. In another study of the neuroendocrine response to exercise (80% LT, 90 min) in women vs. men, no difference in exercise glucose production or utilization was observed when similar euglycemia during exercise was maintained via exogenous glucose infusion (12). Such an infusion would relieve the need for endogenous glucose release, in the face of increased glucose utilization, and again could mask any potential sex differences in exercise glucose kinetics. These data together might suggest that, in the face of limited hepatic glycogen stores, the restraint observed in glucose
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release in women may be the cause of the lower glucose utilization, as opposed to the converse. As both gluconeogenesis and glycogenolysis contribute to glucose production, data might suggest that women are less able to increase the gluconeogenic contribution to glucose production in the face of low-hepatic glycogen stores and increased glucose utilization. Lower gluconeogenesis in women vs. men could be due to the lower increment in glucagon in women vs. men during exercise and/or a lower precursor delivery and/or extraction. In animal models, the female sex steroids can also decrease gluconeogenesis and glucose production at rest (1, 35). Given the importance of glucose production in terms of the regulation of normal glucose homeostasis, the mechanisms resulting in the sex differences in glucose production with exercise and the consequences with respect to glucose utilization are areas that require further investigation. The current investigation illustrates that sex-based differences in exercise and postexercise glucose metabolism can occur independently of differences in whole body nutrient oxidation. Specically, lean, healthy women had a signicantly lower glucose production and utilization during and up to 2 h after exercise compared with lean, healthy men. Furthermore, differences in the hormonal response to exercise, specically the glucagon response and to a lesser extent the Epi response, appear to play a role in determining these sex differences in exercise and postexercise glucose kinetics. Current data delineate sex differences in healthy individuals and form a basis from which comparisons can be made to populations characterized by the dysregulation of glucose metabolism, specically individuals with diabetes and/or obesity. As exercise can be benecial in the management of aberrant glucose metabolism in these populations, whether the extent of such effects is similar in men and women, and how this is hormonally mediated is of relevance. Understanding the physiological impact of sex on these factors will ultimately assist with the development of the most effective and appropriate exercise programs in men and women and enable realistic goals to be set regarding the nature and extent of anticipated benets.
ACKNOWLEDGMENTS The authors thank all of the subjects who volunteered for the study for their time and cooperation. We thank the GCRC nursing, dietary, and laboratory staff for valuable assistance, as well as the Mass Spectrometry and Energy Balance Core Laboratories of the Colorado Nutrition Research Unit for important provision of testing and analytical services. GRANTS This investigation was supported by National Institutes of Health Grants HL-59331, HL-04226, DK-48520, and by Public Health Services Research Grant M01 RR-00051 from the Division of Research Resources. REFERENCES 1. Ahmed-Sorour H and Bailey CJ. Role of ovarian hormones in the long-term control of glucose homeostasis, glycogen formation and gluconeogenesis. Ann Nutr Metab 25: 208 212, 1981. 2. Blatchford FK, Knowlton RG, and Schneider DA. Plasma FFA responses to prolonged walking in untrained men and women. Eur J Appl Physiol 53: 343347, 1985. 3. Braun B, Mawson JT, Muza SR, Dominick SB, Brooks GA, Horning MA, Rock PB, Moore LG, Mazzeo RS, Ezeji-Okoye SC, and Buttereld GE. Women at altitude: carbohydrate utilization during exercise at 4,300 m. J Appl Physiol 88: 246 256, 2000. 4. Campbell SE, Angus DJ, and Febbraio MA. Glucose kinetics and exercise performance during phases of the menstrual cycle: effect of www.jap.org

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SEX DIFFERENCES IN EXERCISE GLUCOSE KINETICS glucose ingestion. Am J Physiol Endocrinol Metab 281: E817E825, 2001. Carter S, McKenzie S, Mourtzakis M, Mahoney DJ, and Tarnoplosky MA. Short-term 17 -estradiol decreases glucose Ra but not whole body metabolism during endurance exercise. J Appl Physiol 90: 139 146, 2001. Carter SL, Rennie C, and Tarnopolsky MA. Substrate utilization during endurance exercise in men and women after endurance training. Am J Physiol Endocrinol Metab 280: E898 E907, 2001. Casey A, Mann R, Banister K, Fox J, Morris P, Macdonald IA, and Greenhaff PL. Effect of carbohydrate ingestion on glycogen resynthesis in human liver and skeletal muscle, measured by 13C MRS. Am J Physiol Endocrinol Metab 278: E65E75, 2000. Causon RM, Carruthers M, and Rodnight R. Assay of plasma catecholamines by liquid chromatography with electrochemical detection. Anal Biochem 116: 223226, 1981. Coggan AR, Kohrt WM, Spina RJ, Kirwan JP, Bier DM, and Holloszy JO. Plasma glucose kinetics during exercise in subjects with high and low lactate thresholds. J Appl Physiol 73: 18731880, 1992. Coyle EF, Coggan AR, Hopper MK, and Walters TJ. Determinants of endurance in well-trained cyclists. J Appl Physiol 64: 26222630, 1988. DEon TM, Sharoff C, Chipkin SR, Grow D, Ruby BC, and Braun B. Regulation of exercise carbohydrate metabolism by estrogen and progesterone in women. Am J Physiol Endocrinol Metab 283: E1046 E1055, 2002. Davis SN, Galassetti P, Wasserman DH, and Tate D. Effects of gender on neuroendocrine and metabolic counterregulatory responses to exercise in normal man. J Clin Endocrinol Metab 85: 224 230, 2000. Dean TM, Perreault L, Mazzeo RS, and Horton TJ. No effect of menstrual cycle phase on lactate threshold. J Appl Physiol 95: 25372543, 2003. DeFronzo RA, Ferrannini E, Sato Y, Felig P, and Wahren J. Synergistic interaction between exercise and insulin on peripheral glucose uptake. J Clin Invest 68: 1468 1474, 1981. Doglioni C, Gambacorta M, Zamboni G, Coggi G, and Viale G. Immunocytochemical localization of progesterone receptors in endocrinecells of the human pancreas. Am J Pathol 137: 999 1005, 1990. Dombovy ML, Bonekat HW, Williams TJ, and Staats BA. Exercise performance and ventilatory response in the menstrual-cycle. Med Sci Sports Exerc 19: 111117, 1987. Febbraio MA, Lambert DL, Starkie RL, Proietto J, and Hargreaves M. Effect of epinephrine on muscle glycogenolysis during exercise in trained men. J Appl Physiol 84: 465 470, 1998. Friedlander AL, Casazza GA, Horning MA, Buddinger TF, and Brooks GA. Effects of exercise intensity and training on lipid metabolism in young women. Am J Physiol Endocrinol Metab 275: E853E863, 1998. Friedlander AL, Casazza GA, Horning MA, Huie MJ, Piacentini MF, Trimmer JK, and Brooks GA. Training-induced alterations of carbohydrate metabolism in women: women respond differently from men. J Appl Physiol 85: 11751186, 1998. Friedlander AL, Casazza GA, Horning MA, Usaj A, and Brooks GA. Endurance training increases fatty acid turnover, but not fat oxidation, in young men. J Appl Physiol 86: 20972105, 1999. Galliven EA, Singh A, Michelson D, Bina S, Gold PW, and Deuster PA. Hormonal and metabolic responses to exercise across time of day and menstrual cycle phase. J Appl Physiol 83: 18221831, 1997. Green HJ, Fraser IG, and Ranney DA. Male and female differences in enzyme activities of energy metabolism in vastus lateralis muscle. J Neurol Sci 65: 323331, 1984. Hirsch IB, Marker JC, Smith LJ, Spina RJ, Parvin CA, Holloszy JO, and Cryer PE. Insulin and glucagon in prevention of hypoglycemia during exercise in humans. Am J Physiol Endocrinol Metab 260: E695 E704, 1991. Horton TJ, Miller EK, Glueck D, and Tench K. No effect of menstrual cycle phase on glucose kinetics and fuel oxidation during moderateintensity exercise. Am J Physiol Endocrinol Metab 282: E752E762, 2002. Horton TJ, Pagliassotti MJ, Hobbs K, and Hill JO. Fuel metabolism in men and women during and after long-duration exercise. J Appl Physiol 85: 18231832, 1998. Jequier E, Acheson K, and Schutz Y. Assessment of energy expenditure and fuel utilization in man. Annu Rev Nutr 7: 187208, 1987. Jurkowski JEH, Jones NL, Toews CJ, and Sutton JR. Effects of menstrual cycle on blood lactate, O2 delivery, and performance during exercise. J Appl Physiol 51: 14931499, 1981. J Appl Physiol VOL

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8. 9. 10. 11.

12. 13. 14. 15. 16. 17. 18. 19.

20. 21. 22. 23.

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