Hyperinsulinemia and skeletal muscle fatty acid trafficking.

We hypothesized that insulin alters plasma free fatty acid (FFA) trafficking into intramyocellular (im) long-chain acylcarnitines (imLCAC) and triglycerides (imTG). Overnight-fasted adults (n = 41) received intravenous infusions of [U-¹³C]palmitate (0400-0900 h) and [U-¹³C]oleate (0800-1400 h) to label imTG and imLCAC. A euglycemic-hyperinsulinemic (1.0 mU·kg fat-free mass⁻¹·min⁻¹) clamp (0800-1400 h) and two muscle biopsies (0900 h, 1400 h) were performed. The patterns of [U-¹³C]palmitate incorporation into imTG-palmitate and palmitoylcarnitine were similar to those we reported in overnight postabsorptive adults (saline control); the intramyocellular palmitoylcarnitine enrichment was not different from and correlated with imTG-palmitate enrichment for both the morning (r = 0.38, P = 0.02) and afternoon (r = 0.44, P = 0.006) biopsy samples. Plasma FFA concentrations, flux, and the incorporation of plasma oleate into imTG-oleate during hyperinsulinemia were ~1/10th of that observed in the previous saline control studies (P < 0.001). At the time of the second biopsy, the enrichment in oleoylcarnitine was <25% of that in imTG-oleate and was not correlated with imTG-oleate enrichment. The intramyocellular nonesterified fatty acid-palmitate-to-imTG-palmitate enrichment ratio was greater (P < 0.05) in women than men, suggesting that sex differences in intramyocellular palmitate trafficking may occur under hyperinsulinemic conditions. We conclude that plasma FFA trafficking into imTG during hyperinsulinemia is markedly suppressed, and these newly incorporated FFA fatty acids do not readily enter the LCAC preoxidative pools. Hyperinsulinemia does not seem to inhibit the entry of fatty acids from imTG pools that were labeled under fasting conditions, possibly reflecting the presence of two distinct imTG pools that are differentially regulated by insulin.

Asian Indians have enhanced skeletal muscle mitochondrial capacity to produce ATP in association with severe insulin resistance.

OBJECTIVE: Type 2 diabetes has become a global epidemic, and Asian Indians have a higher susceptibility to diabetes than Europeans. We investigated whether Indians had any metabolic differences compared with Northern European Americans that may render them more susceptible to diabetes. RESEARCH DESIGN AND METHODS: We studied 13 diabetic Indians, 13 nondiabetic Indians, and 13 nondiabetic Northern European Americans who were matched for age, BMI, and sex. The primary comparisons were insulin sensitivity by hyperinsulinemic-euglycemic clamp and skeletal muscle mitochondrial capacity for oxidative phosphorylation (OXPHOS) by measuring mitochondrial DNA copy number (mtDNA), OXPHOS gene transcripts, citrate synthase activity, and maximal mitochondrial ATP production rate (MAPR). Other factors that may cause insulin resistance were also measured. RESULTS: The glucose infusion rates required to maintain identical glucose levels during the similar insulin infusion rates were substantially lower in diabetic Indians than in the nondiabetic participants (P < 0.001), and they were lower in nondiabetic Indians than in nondiabetic Northern European Americans (P < 0.002). mtDNA (P < 0.02), OXPHOS gene transcripts (P < 0.01), citrate synthase, and MAPR (P < 0.03) were higher in Indians irrespective of their diabetic status. Intramuscular triglyceride, C-reactive protein, interleukin-6, and tumor necrosis factor-alpha concentrations were higher, whereas adiponectin concentrations were lower in diabetic Indians. CONCLUSIONS: Despite being more insulin resistant, diabetic Indians had similar muscle OXPHOS capacity as nondiabetic Indians, demonstrating that diabetes per se does not cause mitochondrial dysfunction. Indians irrespective of their diabetic status had higher OXPHOS capacity than Northern European Americans, although Indians were substantially more insulin resistant, indicating a dissociation between mitochondrial dysfunction and insulin resistance.

Skeletal muscle mitochondrial functions, mitochondrial DNA copy numbers, and gene transcript profiles in type 2 diabetic and nondiabetic subjects at equal levels of low or high insulin and euglycemia.

We investigated whether previously reported muscle mitochondrial dysfunction and altered gene transcript levels in type 2 diabetes might be secondary to abnormal blood glucose and insulin levels rather than an intrinsic defect of type 2 diabetes. A total of 13 type 2 diabetic and 17 nondiabetic subjects were studied on two separate occasions while maintaining similar insulin and glucose levels in both groups by 7-h infusions of somatostatin, low- or high-dose insulin (0.25 and 1.5 mU/kg of fat-free mass per min, respectively), and glucose. Muscle mitochondrial DNA abundance was not different between type 2 diabetic and nondiabetic subjects at both insulin levels, but the majority of transcripts in muscle that are involved mitochondrial functions were expressed at lower levels in type 2 diabetes at low levels of insulin. However, several gene transcripts that are specifically involved in the electron transport chain were expressed at higher levels in type 2 diabetic patients. After the low-dose insulin infusion, which achieved postabsorptive insulin levels, the muscle mitochondrial ATP production rate (MAPR) was not different between type 2 diabetic and nondiabetic subjects. However, increasing insulin to postprandial levels increased the MAPR in nondiabetic subjects but not in type 2 diabetic patients. The lack of MAPR increment in response to high-dose insulin in type 2 diabetic patients occurred in association with reduced glucose disposal and expression of peroxisome proliferator-activated receptor-gamma coactivator 1alpha, citrate synthase, and cytochrome c oxidase I. In conclusion, the current data supports that muscle mitochondrial dysfunction in type 2 diabetes is not an intrinsic defect, but instead a functional defect related to impaired response to insulin.

Evidence for increased and insulin-resistant lipolysis in skeletal muscle of high-fat-fed rats.

The metabolic and isotopic profiles of glycerol in skeletal muscle were examined using awake, fasted lean and high-fat-induced obese rats, and hyperinsulinemic-euglycemic clamp was performed to assess the effect of insulin. During the clamp, Intralipid (no heparin; Fresnius Kabi Clayton, Clayton, NC), free fatty acids, glycerol, and glucose were coinfused to maintain their respective basal plasma levels in both groups. At steady-state, [U-(14)C]glycerol was infused intravenously for 120 minutes followed by muscle biopsy. The classical phenotypic characteristics of obesity, namely, reduced insulin-stimulated glucose uptake, a failure to suppress systemic lipolysis by insulin, and elevated plasma fatty acid concentration, were observed in the obese rats. Novel observations showed that in the basal state, the isotopic specific activity (S.A.) of glycerol (dpm/nmol) in gastrocnemius (0.03 v 0.12), soleus (0.05 v 0.12), and tibialis anterior (0.03 v 0.12) was significantly lower (all P <.003) in obese than in lean rats despite similar concentrations, indicating an active basal intramyocellular lipolysis. In addition, the lipolysis appeared resistant to insulin because the suppression of muscle glycerol during the clamp was 8%, 12%, and 8% in obese compared to 67%, 71%, and 63% in the lean control for gastrocnemius (P =.001), soleus (P =.007), and tibialis anterior (P =.004), respectively. The active intracellular lipolysis likely disturbs metabolic functions that may contribute to insulin resistance.

Determination of skeletal muscle triglyceride synthesis using a single muscle biopsy.

A novel stable isotopic technique for the determination of triglyceride synthesis in skeletal muscle by using a single muscle biopsy has been developed and evaluated in rats. In previous studies using (13)C-tracers, muscle triglyceride synthesis is usually determined using at least 2 biopsies, the first of which serves as the baseline sample for the measurement of natural (13)C abundance. In the present studies, the baseline biopsy has been eliminated by making the use of the isotopic information of a nontraced fatty acid in the muscle triglyceride pool. This is based on the fact that the source and, hence, the natural (13)C abundance of fatty acids in the same triglyceride pool is similar. To demonstrate and validate the method, a series of rat studies have been conducted to have established that (1) the natural (13)C abundance of 4 major fatty acids in the muscle triglyceride pool is similar; (2) there are no (13)C-label exchanges between fatty acids in the lipid pool; and (3) the incorporation of (13)C-palmitate into muscle triglycerides determined using this technique favorably compared with that determined by the traditional method. This approach makes stable isotope studies possible in which more than 1 muscle biopsy is difficult or impossible. Therefore, it has the potential to facilitate investigation of triglyceride metabolism in the skeletal muscle.