Carrie G. Sharoff

AMP-activated protein kinase in skeletal muscle: From structure and localization to its role as a master regulator of cellular metabolism

Abstract.The AMP-activated protein kinase (AMPK) is a metabolite sensing serine/threonine kinase that has been termed the master regulator of cellular energy metabolism due to its numerous roles in the regulation of glucose, lipid, and protein metabolism. In this review, we first summarize the current literature on a number of important aspects of AMPK in skeletal muscle. These include the following: (1) the structural components of the three AMPK subunits (i.e. AMPKα, β, and γ), and their differential localization in response to stimulation in muscle; (2) the biochemical regulation of AMPK by AMP, protein phosphatases, and its three known upstream kinases, LKB1, Ca2+/calmodulin-dependent protein kinase kinase (CaMKK), and transforming growth factor-β-activated kinase 1 (TAK1); (3) the pharmacological agents that are currently available for the activation and inhibition of AMPK; (4) the physiological stimuli that activate AMPK in muscle; and (5) the metabolic processes that AMPK regulates in skeletal muscle.

Effects of exercise on energy-regulating hormones and appetite in men and women.

When previously sedentary men and women follow exercise training programs with ad libitum feeding, men lose body fat, but women do not. The purpose of this study was to evaluate whether this observation could be related to sex differences in the way energy-regulating hormones and appetite perception respond to exercise. Eighteen (9 men, 9 women) overweight/obese individuals completed four bouts of exercise with energy added to the baseline diet to maintain energy balance (BAL), and four bouts without energy added to induce energy deficit (DEF). Concentrations of acylated ghrelin, insulin, and leptin, as well as appetite ratings were measured in response to a meal after a no-exercise baseline and both exercise conditions. In men, acylated ghrelin area under the curve (AUC) was not different between conditions. In women, acylated ghrelin AUC was higher after DEF (+32%) and BAL (+25%), and the change from baseline was higher than men (P < 0.05). In men, insulin AUC was reduced (-17%) after DEF (P < 0.05), but not BAL. In women, insulin AUC was lower (P < 0.05) after DEF (-28%) and BAL (-15%). Leptin concentrations were not different across conditions in either sex. In men, but not in women, appetite was inhibited after BAL relative to DEF. The results indicate that, in women, exercise altered energy-regulating hormones in a direction expected to stimulate energy intake, regardless of energy status. In men, the response to exercise was abolished when energy balance was maintained. The data are consistent with the paradigm that mechanisms to maintain body fat are more effective in women.

Combining short-term metformin treatment and one bout of exercise does not increase insulin action in insulin-resistant individuals

Results from the Diabetes Prevention Program highlight the effectiveness of metformin or regular physical activity in the prevention of type 2 diabetes. Independently, metformin and exercise increase insulin sensitivity, but they have not been studied in combination. To assess the combined effects, insulin-resistant subjects (n = 9) matched for weight, body fat, and aerobic fitness were studied before any treatment (B), after 2-3 wk of 2,000 mg/day metformin (MET), and after metformin plus 40 min of exercise at 65% Vo(2peak) (MET + Ex). A second group (n = 7) was studied at baseline and after an identical bout of exercise with no metformin (Ex). Biopsies of the vastus lateralis were taken at B, after MET, immediately after MET + Ex (group 1), or immediately after Ex (group 2). Insulin sensitivity was assessed 4 h postexercise with a euglycemic hyperinsulinemic (40 mU.m(2).min(-1)) clamp enriched with [6,6-(2)H]glucose. Insulin sensitivity was 54% higher after Ex (P < 0.01), but there was no change with Met + Ex. Skeletal muscle AMPKalpha2 activity was elevated threefold (P < 0.01) after Ex, but there was no increase with MET + Ex. These findings suggest that the combination of short-term metformin treatment and an acute bout of exercise does not enhance insulin sensitivity, and the addition of metformin may attenuate the well-documented effects of exercise alone.

Effects of short-term exercise and energy surplus on hormones related to regulation of energy balance.

Energy surplus raises circulating concentrations of leptin and insulin while lowering plasma ghrelin. Exercise has the opposite effects. The purpose of this study was to determine whether exercise counters the hormonal effects of energy surplus independent of changes in energy balance. To do that, we assessed plasma concentrations of leptin, insulin, and ghrelin at baseline, after overfeeding, and after overfeeding plus exercise. Baseline (B) leptin and insulin concentrations and ghrelin area under the curve were measured during an oral glucose challenge in 9 healthy, active subjects (6 male, 3 female) after 2 days in energy balance without exercise. Measurements were repeated after subjects were overfed by +3213 +/- 849 kJ/d for 3 more sedentary days (OF). In the third condition, the same net energy surplus (+3125 +/- 933 kJ/d) was generated for 24 hours by doubling the overfeeding (+6284 +/- 1669 kJ/d) and countering it with a bout of exercise (expenditure = 3063 +/- 803 kJ); and measurements were made the next day (OF + EX). Compared with B, leptin went up (5.8 +/- 8.2 to 7.6 +/- 10.6 ng/mL) after OF, but was not significantly higher after OF + EX (7.1 +/- 10.2 ng/mL). Compared with B, insulin was +36% and +43% higher after OF and OF + EX, respectively. In contrast, ghrelin area under the curve did not change after OF but was significantly lower (-14%) than B or OF after OF + EX (indicating greater suppression). These data suggest that the effect of short-term exercise on fasting leptin and insulin depends on energy balance but the ghrelin response may be partially mediated by effects of exercise independent of energy balance.

Impact of Metformin on Peak Aerobic Capacity

Individually, exercise and the drug metformin have been shown to prevent or delay type 2 diabetes. Metformin mildly inhibits complex I of the electron transport system and may impact aerobic capacity in people exercising while taking metformin. The purpose of the study was to evaluate the effects of metformin on maximal aerobic capacity in healthy individuals without mitochondrial dysfunction. Seventeen healthy, normal-weight men (n=11) and women (n=6) participated in a double-blind, placebo-controlled, cross-over design. Peak aerobic capacity was measured twice using a continuous, incrementally graded protocol; once after 7-9 d of metformin (final dose=2000 mg/d) and once with placebo, with 1 week between tests. The order of the conditions was counterbalanced. Peak oxygen uptake (VO2 peak), heart rate (HR), ventilation (VE), respiratory exchange ratio (RER), rating of perceived exertion (RPE), and test duration were compared across conditions using paired t tests with the R statistical program. VO2 peak (-2.7%), peak heart rate (-2.0%), peak ventilation (-6.2%), peak RER (-3.0%), and exercise duration (-4.1%) were all reduced slightly, but significantly, with metformin (all p<0.05). There was no effect of metformin on RPE or ventilatory breakpoint. Correlations between the decrement in VO2 peak and any of the other outcome variables were weak (r2<0.20) and not significant. Short-term treatment with metformin has statistically significant, but physiologically subtle, effects that reduce key outcomes related to maximal exercise capacity. Whether this small but consistent effect is manifested in people with insulin resistance or diabetes who already have some degree of mitochondrial dysfunction remains to be determined.

The effect of carbohydrate availability following exercise on whole-body insulin action

One bout of exercise enhances insulin-stimulated glucose uptake (insulin action), but the effect is blunted by consumption of carbohydrate-containing food after exercise. The independent roles of energy and carbohydrate in mediating post-exercise insulin action have not been systematically evaluated in humans. The purpose of this study was to determine if varying carbohydrate availability, with energy intake held constant, mediates post-exercise insulin action. Ten young (21 +/- 2 y, overweight (body fat 37% +/- 3%) men and women completed 3 conditions in random order: (i) no-exercise (BASE), (ii) exercise with energy balance but carbohydrate deficit (C-DEF), and (iii) exercise with energy and carbohydrate balance (C-BAL). In the exercise conditions, subjects expended 30% of total daily energy expenditure on a cycle ergometer at 70% VO2 peak. Following exercise, subjects consumed a meal that replaced expended energy (~3000 kJ) and was either balanced (intake = expenditure) or deficient (-100 g) in carbohydrate. Twelve hours later, insulin action was measured by continuous infusion of glucose with stable isotope tracer (CIG-SIT). Changes in insulin action were evaluated using a one-way ANOVA with repeated measures. During CIG-SIT, non-oxidative glucose disposal (i.e., glucose storage) was higher in C-DEF than in BASE (27.2 +/- 3.2 vs. 16.9 +/- 3.5 micromol.L-1.kg-1.min-1, p < 0.05). Conversely, glucose oxidation was lower in C-DEF (8.6 +/- 1.3 micromol.L-1.kg-1.min-1) compared with C-BAL (12.2 +/- 1.2 micromol.L-1.kg-1.min-1), and BASE (17.1 +/- 2.2 micromol.L-1.kg-1.min-1), p < 0.05). Fasting fat oxidation was higher in C-DEF than in BASE (109.8 +/- 10.5 vs. 80.7 +/- 9.6 mg.min-1, p < 0.05). In C-DEF, enhanced insulin action was correlated with the magnitude of the carbohydrate deficit (r = 0.82, p < 0.01). Following exercise, re-feeding expended energy, but not carbohydrate, increased fasting fat oxidation, and shifted insulin-mediated glucose disposal toward increased storage and away from oxidation.

Effect of timing of energy and carbohydrate replacement on post-exercise insulin action

The nutritional environment surrounding an exercise bout modulates post-exercise insulin action. The purpose of this study was to determine how timing energy and carbohydrate replacement proximate to an exercise bout influences exercise-enhanced insulin action. To create an appropriate baseline, sensitivity to insulin was reduced in 9 healthy young men (n=6) and women (n=3) by 2 days of energy surplus and detraining. Then, insulin action (glucose uptake per unit plasma insulin) was assessed by stable isotope dilution during a continuous glucose infusion 12 h after a standardized meal under 4 conditions. In 3 conditions, the meal replaced the energy and carbohydrate expended during an exercise bout (62.9+/-2.8 min cycle ergometry at 65% VO2 peak followed by ten 30 s sprints). The meal was given before (Pre), immediately after (ImmPost), or 3 h after exercise (Delay). The 4th condition was a no-exercise control (Control). Data were analyzed using linear mixed-effects models with planned contrasts. Relative to Control, insulin action increased by 22% in Pre (p=0.05), 44% in ImmPost (p<0.01), and 19% in Delay (p=0.09). Non-oxidative disposal was higher, and oxidative disposal was lower in ImmPost relative to Control and Pre (p<0.05). Hepatic glucose production was suppressed by the infusion to a greater extent in Pre and Delay (76.9%+/-8.8% and 81.2%+/-4.7%) compared with ImmPost (64.7%+/-10.0%). A bout of exercise enhances insulin action even when expended energy and carbohydrate are replaced. Further, timing of energy and carbohydrate consumption subtly modulates the effectiveness of exercise to enhance insulin action.