OBJECTIVE: Long-chain fatty acids (LCFAs) contribute to metabolic homeostasis in part via gene regulation. This study's objective was to identify novel LCFA target genes in human skeletal muscle cells (myotubes). RESEARCH DESIGN AND METHODS: In vitro methods included culture and treatment of human myotubes and C2C12 cells, gene array analysis, real-time RT-PCR, Western blotting, ELISA, chromatin immunoprecipitation, and RNA interference. Human subjects (two cohorts) were characterized by oral glucose tolerance test, hyperinsulinemic-euglycemic clamp, magnetic resonance imaging and spectroscopy, and standard blood analyses (glucose, insulin, C-peptide, and plasma lipids). RESULTS: We show here that ANGPTL4 (encoding angiopoietin-like protein 4) represents a prominent LCFA-responsive gene in human myotubes. LCFA activated peroxisome proliferator-activated receptor (PPAR)-delta, but not PPAR-alpha or -gamma, and pharmacological activation of PPAR-delta markedly induced ANGPTL4 production and secretion. In C2C12 myocytes, knockdown of PPARD, but not of PPARG, blocked LCFA-mediated ANGPTL4 induction, and LCFA treatment resulted in PPAR-delta recruitment to the ANGPTL4 gene. In addition, pharmacological PPAR-delta activation induced LIPE (encoding hormone-sensitive lipase), and this response crucially depended on ANGPTL4, as revealed by ANGPTL4 knockdown. In a human cohort of 108 thoroughly phenotyped subjects, plasma ANGPTL4 positively correlated with fasting nonesterified fatty acids (P = 0.0036) and adipose tissue lipolysis (P = 0.0012). Moreover, in 38 myotube donors, plasma ANGPTL4 levels and adipose tissue lipolysis in vivo were reflected by basal myotube ANGPTL4 expression in vitro (P = 0.02, both). CONCLUSIONS: ANGPTL4 is produced by human myotubes in response to LCFA via PPAR-delta, and muscle-derived ANGPTL4 seems to be of systemic relevance in humans.
PURPOSE: To evaluate whether techniques of high field magnetic resonance imaging may be used to characterize embryonic tissue during proliferation and differentiation. MATERIALS AND METHODS: Thirteen chicken embryos with incubation times between 5 days and 16 days have been measured in a small animal magnetic resonance imager (ClinScan, Bruker) at 7 Tesla using the built-in resonator. T1, T2-, and magnetization transfer imaging was performed using fast spin-echo with inversion recovery, half acquisition single shot turbo spin-echo, and spoiled gradient-echo sequences with and without off-resonance pulse, respectively. T1, T2, and magnetization transfer ratio (MTR) maps were calculated on a pixel-by-pixel basis. RESULTS: T1-, T2-, and MTR maps showed good image quality allowing for delineation of embryonic organs. During embryonic development, a decrease of T1 and T2 relaxation times was found, whereas, embryonic tissue typically showed an increase of magnetization transfer, for example, liver properties at day 5: T1 = 2431 +/- 163 ms, T2 = 122 +/- 12 ms, MTR = 9.2 +/- 4.2%; liver properties at day 16: T1 = 1763 +/- 89 ms, T2 = 71 +/- 4 ms, MTR = 16.9 +/- 2.2%. CONCLUSION: Embryonic tissues show changing relaxation and magnetization transfer properties during development, therefore, high field MRI seems suitable for characterization of tissue replacement derived from embryonic stem cells.
CONTEXT: Liver fat predicts insulin resistance in humans. So far, there is not much information on genetic determinants of liver fat. Hepatic lipase is a liver-specific enzyme that regulates lipid metabolism. OBJECTIVE: First, our object was to investigate whether the functional -514C>T polymorphism of the hepatic lipase gene is associated with liver fat content and with insulin sensitivity. Second, because this polymorphism displays gene-nutrient interactions, we assessed gene-gene interactions with the Pro12Ala polymorphism of the peroxisome proliferator-activated receptor-gamma(2) on liver fat content and insulin sensitivity. DESIGN AND METHODS: Cross-sectional data from a total of 1070 nondiabetic subjects were analyzed. Insulin sensitivity was estimated from a 75-g oral glucose tolerance test. A subgroup of 115 subjects underwent measurements of liver fat. RESULTS: The -514C>T polymorphism of the hepatic lipase gene was associated with higher liver fat content (P = 0.005) and lower insulin sensitivity (P = 0.02), both after adjustment for age, gender, and percentage of body fat. This was independent of serum adiponectin concentrations (P = 0.01 and 0.03). However, there was an interaction of the -514C>T polymorphism with the Pro12Ala variant on liver fat (P = 0.09) and insulin sensitivity (P = 0.01). Subjects carrying the -514C>T polymorphism had higher liver fat content and were insulin resistant only before the background of the Pro/Pro genotype of the Pro12Ala polymorphism. CONCLUSIONS: The -514C>T polymorphism of the hepatic lipase gene is associated with higher liver fat content and lower whole-body insulin sensitivity. However, these effects are modulated by the common Pro12Ala polymorphism in peroxisome proliferator-activated receptor-gamma(2). These findings may be relevant for intervention strategies to prevent increase in liver fat content and insulin resistance.