Impact of ageing on female metabolic flexibility: a cross-sectional pilot study in over-60 active women

Background Ageing influences the metabolic flexibility, albeit the physical status could determine this relationship. This cross-sectional study aims to describe and analyse the metabolic flexibility/inflexibility in a group of active older women, together with the impact of ageing and physical status on their oxidation rates and maximal fat oxidation (MFO). Methods Fifteen volunteers (69.00±6.97 years) from 24 women, completed an incremental cycling test until the second ventilatory threshold. Intensity increased 10W each 3min–15sec, starting at 30W. Gas exchange, heart rate, rate of perceived effort, pain scale and muscle power were registered, together with lactate. VO2 and VCO2 were considered for Fat and CHO oxidation (FATox & CHOox; Frayn’s equation) at the intensities 60%, 80% and 100% from the peak of power in the test (P100). Psychophysiological parameters were compared at MFO/FATmax and P100, together with the main correlation analyses, with and without P100 and VO2 as covariates. Results FATox was low at MFO (0.13; 95%CI [0.09-0.17] g·min−1·kg; 5.61 [3.59-7.63] g·min−1·kg FFM), with a shifting down and leftward of a short oxidation-rate curves. CHOox and FATox were both low for a reduced power with age (77.14±18.58 W & 39.29±9.17 W at P100 and MFO respectively), pointing to metabolic inflexibility in older women despite being active. Notwithstanding, the negative correlation between age and MFO (r=-0.54, p=0.04; R2=0.29) disappeared when normalized with P100 (r=-0.17, p=0.53), which was in turn strongly and negatively associated to age (r=-0.85, p<0.005; R2=0.72). P100 was also positive and moderately associated to MFO (r=0.71, p=0.01; R2=0.50). Conclusions Despite the inflexibility with age, physical status (i.e., larger muscular power) suggest a key role in the preservation of the metabolic health with aging in active women.

Bioactive Foods Decrease Liver and Brain Alterations Induced by a High-Fat-Sucrose Diet through Restoration of Gut Microbiota and Antioxidant Enzymes

Obesity is associated with cognitive deficit and liver alterations; however, it remains unclear whether a combination of functional foods could reverse cognitive damage and to what extent it would be associated with changes in gut microbiota and liver. With this aim, male Wistar rats were fed a high-fat-5%sucrose diet (HFS) for 4 mo. And were then fed for 1 mo. with bioactive foods. At the end of this period, liver, serum, feces, intestine, and brain samples were taken. Body composition, energy expenditure, LPS, hormones, intraperitoneal glucose tolerance test, behavioral tests, and gut microbiota were evaluated. We showed that male rats fed high-fat-sucrose diet developed gut microbiota dysbiosis, increased in body fat, decreased antioxidant activity, decreased brain neuropeptide Y, increased the number of astrocytes and activated microglia, along with reduced spine density associated with deficits in working memory. Ingestion of a combination of nopal, soy protein, curcumin, and chia seed oil (bioactive foods) for three months was associated with an increase in a cluster of bacteria with anti-inflammatory capacity, a decrease in serum LPS levels and an increase in serum eicosapentaenoic acid (EPA) with neuroprotective properties. In the liver, ingestion of bioactive food significantly increased antioxidant enzymes, decreased lipogenesis, reduced inflammation mediated by the TLR4-TNFα pathway along with a decrease in body fat, glucose intolerance, and metabolic inflexibility. Finally, neuroinflammation in the brain was reduced and working memory improved. Our study demonstrates that consumption of bioactive foods was associated with reduced liver, brain, and gut microbiota alterations in obese rats.

PPARα, δ and FOXO1 Gene Silencing Overturns Palmitate-Induced Inhibition of Pyruvate Oxidation Differentially in C2C12 Myotubes

The molecular mechanisms by which free fatty acids (FFA) inhibit muscle glucose oxidation is still elusive. We recently showed that C2C12 myotubes treated with palmitate (PAL) presented with greater protein expression levels of PDK4 and transcription factors PPARα and PPARδ and lower p-FOXO/t-FOXO protein ratios when compared to control. This was complemented with the hallmarks of metabolic inflexibility (MI), i.e., reduced rates of glucose uptake, PDC activity and maximal pyruvate-derived ATP production rates (MAPR). However, the relative contribution of these transcription factors to the increase in PDK4 and reduced glucose oxidation could not be established. Therefore, by using a similar myotube model, a series of individual siRNA gene silencing experiments, validated at transcriptional and translation levels, were performed in conjunction with measurements of glucose uptake, PDC activity, MAPR and concentrations of metabolites reflecting PDC flux (lactate and acetylcarnitine). Gene silencing of PPARα, δ and FOXO1 individually reduced PAL-mediated inhibition of PDC activity and increased glucose uptake, albeit by different mechanisms as only PPARδ and FOXO1 silencing markedly reduced PDK4 protein content. Additionally, PPARα and FOXO1 silencing, but not PPARδ, increased MAPR with PAL. PPARδ silencing also decreased FOXO1 protein. Since FOXO1 silencing did not alter PPARδ protein, this suggests that FOXO1 might be a PPARδ downstream target. In summary, this study suggests that the molecular mechanisms by which PAL reduces PDC-mediated glucose-derived pyruvate oxidation in muscle occur primarily through increased PPARδ and FOXO1 mediated increases in PDK4 protein expression and secondarily through PPARα mediated allosteric inhibition of PDC flux. Furthermore, since PPARδ seems to control FOXO1 expression, this may reflect an important role for PPARδ in preventing glucose oxidation under conditions of increased lipid availability.

Simulating Metabolic Flexibility in Low Energy Expenditure Conditions Using Genome-Scale Metabolic Models

Metabolic flexibility is the ability of an organism to adapt its energy source based on nutrient availability and energy requirements. In humans, this ability has been linked to cardio-metabolic health and healthy aging. Genome-scale metabolic models have been employed to simulate metabolic flexibility by computing the Respiratory Quotient (RQ), which is defined as the ratio of carbon dioxide produced to oxygen consumed, and varies between values of 0.7 for pure fat metabolism and 1.0 for pure carbohydrate metabolism. While the nutritional determinants of metabolic flexibility are known, the role of low energy expenditure and sedentary behavior in the development of metabolic inflexibility is less studied. In this study, we present a new description of metabolic flexibility in genome-scale metabolic models which accounts for energy expenditure, and we study the interactions between physical activity and nutrition in a set of patient-derived models of skeletal muscle metabolism in older adults. The simulations show that fuel choice is sensitive to ATP consumption rate in all models tested. The ability to adapt fuel utilization to energy demands is an intrinsic property of the metabolic network.

Metabolic flexibility during sleep

Known as metabolic flexibility, oxidized substrate is selected in response to changes in the nutritional state. Sleep imposes an extended duration of fasting, and oxidized substrates during sleep were assumed to progressively shift from carbohydrate to fat, thereby gradually decreasing the respiratory quotient (RQ). Contrary to this assumption, whole-room indirect calorimetry with improved time resolution revealed that RQ re-ascended prior to awakening, and nadir of RQ in non-obese young adults occurred earlier in women than men after bedtime. The transient decrease in RQ during sleep was blunted in metabolically inflexible men with smaller amplitude of diurnal rhythm in RQ. Similarly, the effect of 10 years difference in age on RQ became significant during sleep; the decrease in RQ during sleep was blunted in older subjects. Inter-individual difference in RQ become apparent during sleep, and it might serve as a window to gain insight into the early-stage pathogenesis of metabolic inflexibility.

Metabolic Response to Submaximal and Maximal Exercise in People with Severe Obesity, Prediabetes, and Diabetes

<b><i>Introduction:</i></b> Metabolic adaptations to maximal physical exercise in people with obesity (PwO) are scarcely described. This cross-sectional study evaluates the metabolic response to exercise via the respiratory exchange ratio (RER) in PwO and different degrees of glycemic control. <b><i>Methods:</i></b> Eighty-five PwO (body mass index 46.0 [39.0–54.0] kg/m²), that is, 32 normoglycemic (Ob-N), 25 prediabetic (Ob-preDM), and 28 diabetic (Ob-T2DM) subjects and 18 healthy subjects performed an incremental, maximal cardiopulmonary exercise test. The RER was measured at rest (RERrest) and at peak exercise (RERpeak). <b><i>Results:</i></b> RERpeak was significantly higher in healthy subjects than that in PwO. Among those, RERpeak was significantly higher in Ob-N than that in Ob-preDM and Ob-T2DM (1.20 [1.15–1.27] vs. 1.18 [1.10–1.22] <i>p</i> = 0.04 and vs. 1.14 [1.10–1.18] <i>p</i> &#x3c; 0.001, respectively). Accordingly, ΔRER (RERpeak-RERrest) was lower in Ob-preDM and Ob-T2DM than that in Ob-N (0.32 [0.26–0.39] <i>p</i> = 0.04 and 0.29 [0.24–0.36] <i>p</i> &#x3c; 0.001 vs. 0.38 [0.32–0.43], respectively), while no significant difference was found in ΔRER between Ob-preDM and Ob-T2DM and not even between Ob-N and healthy subjects. Moreover, ΔRER in PwO correlated with glucose area under curve (<i>p</i> = 0.002). <b><i>Conclusions:</i></b> PwO demonstrate restricted metabolic response during maximal exercise. Particularly, those with prediabetes already show metabolic inflexibility during exercise, similarly to those with type 2 diabetes. These findings also suggest a potential role of cardiopulmonary exercise testing in detecting early metabolic alterations in PwO.

Cardiac-Specific Overexpression of ERRγ in Mice Induces Severe Heart Dysfunction and Early Lethality

Proper cardiac function depends on the coordinated expression of multiple gene networks related to fuel utilization and mitochondrial ATP production, heart contraction, and ion transport. Key transcriptional regulators that regulate these gene networks have been identified. Among them, estrogen-related receptors (ERRs) have emerged as crucial modulators of cardiac function by regulating cellular metabolism and contraction machinery. Consistent with this role, lack of ERRα or ERRγ results in cardiac derangements that lead to functional maladaptation in response to increased workload. Interestingly, metabolic inflexibility associated with diabetic cardiomyopathy has been recently associated with increased mitochondrial fatty acid oxidation and expression of ERRγ, suggesting that sustained expression of this nuclear receptor could result in a cardiac pathogenic outcome. Here, we describe the generation of mice with cardiac-specific overexpression of ERRγ, which die at young ages due to heart failure. ERRγ transgenic mice show signs of dilated cardiomyopathy associated with cardiomyocyte hypertrophy, increased cell death, and fibrosis. Our results suggest that ERRγ could play a role in mediating cardiac pathogenic responses.

HIV-1 Viral Protein R Couples Metabolic inflexibility with White Adipose Tissue Thermogenesis

Persons living with HIV (PLWH) manifest chronic disorders of brown and white adipose tissues that lead to diabetes and metabolic syndrome. The mechanisms that link viral factors to defective adipose tissue function and abnormal energy balance in PLWH remain incompletely understood. Here, we explored how the HIV accessory protein viral protein R (Vpr) contributes to adaptive thermogenesis in two mouse models and human adipose tissues. Uncoupling protein 1 (UCP1) gene expression was strongly increased in subcutaneous white adipose tissue (WAT) biopsies from PLWH and in subcutaneous WAT of the Vpr mice, with near-equivalent mRNA copy number. Histology and functional studies confirmed beige transformation in subcutaneous but not visceral WAT in the Vpr mice. Measurements of energy balance indicated Vpr mice displayed metabolic inflexibility and could not shift efficiently from carbohydrate to fat metabolism during day-night cycles. Furthermore, Vpr mice showed a marked inability to defend body temperature when exposed to 4^oC. Importantly, Vpr couples higher tissue catecholamine levels with UCP1 expression independent of β-adrenergic receptors. Our data reveal surprising deficits of adaptive thermogenesis that drive metabolic inefficiency in HIV-1 Vpr mouse models, providing an expanded role for viral factors in the pathogenesis of metabolic disorders in PLWH.