In Vivo Metabolic Roles of G Proteins of the Gi Family Studied with Novel Mouse Models

G protein-coupled receptors (GPCRs) are the target of ~30-35% of all FDA-approved drugs. The individual members of the GPCR superfamily couple to one or more functional classes of heterotrimeric G proteins. The physiological outcome of activating a particular GPCR in vivo depends on the pattern of receptor distribution and the type of G proteins activated by the receptor. Based on the structural and functional properties of their α-subunits, heterotrimeric G proteins are subclassified into four major families: Gs, Gi/o, Gq/11, and G12/13. Recent studies with genetically engineered mice have yielded important novel insights into the metabolic roles of Gi/o-type G proteins. For example, recent data indicate that Gi signaling in pancreatic α-cells plays a key role in regulating glucagon release and whole body glucose homeostasis. Receptor-mediated activation of hepatic Gi signaling stimulates hepatic glucose production, suggesting that inhibition of hepatic Gi signaling could prove clinically useful to reduce pathologically elevated blood glucose levels. Activation of adipocyte Gi signaling reduces plasma free fatty acid levels, thus leading to improved insulin sensitivity in obese, glucose-intolerant mice. These new data suggest that Gi-coupled receptors that are enriched in metabolically important cell types represent potential targets for the development of novel drugs useful for the treatment of type 2 diabetes and related metabolic disorders.

Peak Fat Oxidation Rate Is Closely Associated With Plasma Free Fatty Acid Concentrations in Women; Similar to Men

Introduction: In men, whole body peak fat oxidation (PFO) determined by a graded exercise test is closely tied to plasma free fatty acid (FFA) availability. Men and women exhibit divergent metabolic responses to fasting and exercise, and it remains unknown how the combined fasting and exercise affect substrate utilization in women. We aimed to investigate this, hypothesizing that increased plasma FFA concentrations in women caused by fasting and repeated exercise will increase PFO during exercise. Then, that PFO would be higher in women compared with men (data from a previous study).Methods: On two separate days, 11 young endurance-trained women were investigated, either after an overnight fast (Fast) or 3.5 h after a standardized meal (Fed). On each day, a validated graded exercise protocol (GXT), used to establish PFO by indirect calorimetry, was performed four times separated by 3.5 h of bed rest both in the fasted (Fast) or fed (Fed) state.Results: Peak fat oxidation increased in the fasted state from 11 ± 3 (after an overnight fast, Fast 1) to 16 ± 3 (mean ± SD) mg/min/kg lean body mass (LBM) (after ~22 h fast, Fast 4), and this was highly associated with plasma FFA concentrations, which increased from 404 ± 203 (Fast 1) to 865 ± 210 μmol/L (Fast 4). No increase in PFO was found during the fed condition with repeated exercise. Compared with trained men from a former identical study, we found no sex differences in relative PFO (mg/min/kg LBM) between men and women, in spite of significant differences in plasma FFA concentrations during exercise after fasting.Conclusion: Peak fat oxidation increased with fasting and repeated exercise in trained women, but the relative PFO was similar in young trained men and women, despite major differences in plasma lipid concentrations during graded exercise.

Obesity progression causes liver steatosis and co-morbidities without apparent cardiac metabolic and functional decline

The goal of this study was to test if obesity progression can be a risk factor to alter cardiac metabolism and function along the time. Male Wistar rats were randomly divided to receive either chow diet (12.0% calories from fat) [C group] or high-fat diet (49.7% calories from fat) plus sucrose in the drinking water (100% from carbohydrate) [H group] for 6, 12 and 24 weeks. The Western diet significantly increased adiposity index of rats in all three experimental periods compared to C group. This was associated with increased plasma levels of insulin, resistin, leptin, glucose, triacylglycerol and decreased adiponectin, however, all variables were stable along the time except insulin and leptin. Plasma free fatty acid was only elevated with 24 weeks treatment. The obesity status resulted in hepatic steatosis progression in H group, while oxidative stress, hepatic inflammatory foci as well as TNF-α and IL-6 mRNA levels were not affected. There are no cardiac performance decline as well as metabolism cardiac changes in H group when compared with C. In conclusion, Western diet induced and promoted obesity, co-morbidities and hepatic steatosis progression while was not associated with apparent alterations of cardiac metabolism and function. These results suggest that obesity progression seems to affect the organs of distinct ways, and cardiac dysfunction is a question of time

Plasma Free Fatty Acid Concentration as a Modifiable Risk Factor for Metabolic Disease

Plasma free fatty acid (FFA) concentration is elevated in obesity, insulin resistance (IR), non-alcoholic fatty liver disease (NAFLD), type 2 diabetes (T2D), and related comorbidities such as cardiovascular disease (CVD). Furthermore, experimentally manipulating plasma FFA in the laboratory setting modulates metabolic markers of these disease processes. In this article, evidence is presented indicating that plasma FFA is a disease risk factor. Elevations of plasma FFA can promote ectopic lipid deposition, IR, as well as vascular and cardiac dysfunction. Typically, elevated plasma FFA results from accelerated adipose tissue lipolysis, caused by a high adipose tissue mass, adrenal hormones, or other physiological stressors. Reducing an individual’s postabsorptive and postprandial plasma FFA concentration is expected to improve health. Lifestyle change could provide a significant opportunity for plasma FFA reduction. Various factors can impact plasma FFA concentration, such as chronic restriction of dietary energy intake and weight loss, as well as exercise, sleep quality and quantity, and cigarette smoking. In this review, consideration is given to multiple factors which lead to plasma FFA elevation and subsequent disruption of metabolic health. From considering a variety of medical conditions and lifestyle factors, it becomes clear that plasma FFA concentration is a modifiable risk factor for metabolic disease.

Fasting hypoglycaemia secondary to carnitine deficiency: a late consequence of gastric bypass

Twelve years following gastric bypass surgery, a cachectic 69-year-old woman presented with both fasting and postprandial hypoglycaemia. Postprandial symptoms were relieved by dietary modification and acarbose, as is common in such cases. During a supervised fast, symptomatic hypoglycaemia occurred. Concurrent laboratory testing showed suppression of plasma insulin, c-peptide, proinsulin and insulin-like growth factor II. However, beta-hydroxybutyrate was also low, surprising given insulin deficiency. Elevated plasma free fatty acid (FFA) concentrations suggested that lipolysis was not impaired, making cachexia/malnutrition a less likely cause of hypoglycaemia. The apparent diagnosis was failure to counter-regulate—subsequent plasma carnitine measurements showed carnitine deficiency which presumably prevented FFA transport across mitochondrial membranes for ketogenesis. Repletion with high-dose oral carnitine supplements effected resolution of fasting hypoglycaemia.

Transient elevation of triacylglycerol content in the liver: a fundamental component of the acute response to exercise

Exercise is well appreciated as a therapeutic approach to improve health. While chronic exercise training can change metabolism, even a single exercise session can have significant effects upon metabolism. Responses of adipose tissue lipolysis and skeletal muscle triacylglycerol (TAG) utilization have been well-appreciated as components of the acute exercise response. However, there are other central components of the physiological response to be considered, as well. A robust and growing body of literature depicts a rapid responsiveness of hepatic TAG content to single bouts of exercise, and there is a remaining need to incorporate this information into our overall understanding of how exercise affects the liver. TAG content in the liver increases during an exercise session and can continue to rise for a few hours afterwards, followed by a fairly rapid return to baseline. We summarize evidence that rapid responsiveness of hepatic TAG content to metabolic stress is a fundamental component of the exercise response. Adipose tissue lipolysis and plasma free fatty acid concentration are likely the major metabolic controllers of enhanced lipid storage in the liver after each exercise bout, and we discuss nutritional impacts as well as health implications. While traditionally clinicians would be merely concerned with hepatic lipids in overnight-fasted, rested individuals, it is now apparent that the content of hepatic TAG fluctuates in response to metabolic challenges such as exercise, and these responses likely exert significant impacts on health and cellular homeostasis.

0295 Skeletal Muscle Diacylglycerol Accumulation and Impaired Insulin Sensitivity During Insufficient Sleep

Introduction Insufficient sleep impairs insulin sensitivity; however, the mechanism(s) by which this occurs are unknown. We previously reported an elevation in plasma free fatty acid concentration during insufficient sleep, suggesting dysregulated lipid metabolism. Lipid accumulation in muscle—specifically certain species of diacylglycerol (DAG)—is associated with impaired insulin sensitivity. We therefore tested the hypothesis that insufficient sleep leads to skeletal muscle DAG accumulation. Methods As part of an ongoing study, thirteen sedentary, healthy, lean adults (25.8±3.2y; 22.7±1.9kg/m2; 3F; mean±SD) participated in a controlled 6-day in-laboratory protocol with 9h in bed (habitual sleep) followed by 4 nights of 5h in bed (insufficient sleep), achieved by delaying bedtime by 4 hours. For one week prior to the study, participants maintained a 9h sleep schedule. Participants consumed energy balanced diets 3 days prior to and throughout the laboratory protocol. Insulin sensitivity was assessed using a hyperinsulinemic euglycemic clamp before and after insufficient sleep. Skeletal muscle biopsies of the vastus lateralis were taken immediately before each clamp. In a subset of subjects (n=10), quantitative lipidomic analyses using LC/MS/MS were performed on biopsied muscle tissue. Results Insulin sensitivity was impaired following insufficient sleep (10.7±1.5 vs 9.6±1.2 mg/kg/min, p<0.05, mean±SEM). There were no changes in skeletal muscle concentration of total triglycerides (TAGs), nor specific TAG species. However, insufficient sleep tended to increase skeletal muscle accumulation of total 1,2-DAGs (p=0.13) and significantly increased specific saturated species of 1,2-DAG, including Di-C18:0 DAG (p<0.05), previously implicated in insulin resistance. In contrast, 1,3-DAGs are not thought to impair insulin sensitivity and specific species were decreased or unchanged during insufficient sleep. Conclusion Preliminary findings suggest that skeletal muscle lipid accumulation of diacylglycerol species during insufficient sleep may be a contributing mechanism by which insufficient sleep dysregulates metabolic physiology. Support NIH K01DK110138, R03 DK118309, UL1 TR002535, and GCRC RR-00036