Peroxisome proliferator-activated receptor-gamma coactivator-1alpha overexpression increases lipid oxidation in myocytes from extremely obese individuals

Leslie A Consitt, Jill A Bell, Timothy R Koves, Deborah M Muoio, Matthew W Hulver, Kimberly R Haynie, G Lynis Dohm, Joseph A Houmard, Leslie A Consitt, Jill A Bell, Timothy R Koves, Deborah M Muoio, Matthew W Hulver, Kimberly R Haynie, G Lynis Dohm, Joseph A Houmard

Abstract

Objective: To determine whether the obesity-related decrement in fatty acid oxidation (FAO) in primary human skeletal muscle cells (HSkMC) is linked with lower mitochondrial content and whether this deficit could be corrected via overexpression of peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha).

Research design and methods: FAO was studied in HSkMC from lean (BMI 22.4 +/- 0.9 kg/m(2); N = 12) and extremely obese (45.3 +/- 1.4 kg/m(2); N = 9) subjects. Recombinant adenovirus was used to increase HSkMC PGC-1alpha expression (3.5- and 8.0-fold), followed by assessment of mitochondrial content (mtDNA and cytochrome C oxidase IV [COXIV]), complete ((14)CO(2) production from labeled oleate), and incomplete (acid soluble metabolites [ASM]) FAO, and glycerolipid synthesis.

Results: Obesity was associated with a 30% decrease (P < 0.05) in complete FAO, which was accompanied by higher relative rates of incomplete FAO ([(14)C]ASM production/(14)CO(2)), increased partitioning of fatty acid toward storage, and lower (P < 0.05) mtDNA (-27%), COXIV (-35%), and mitochondrial transcription factor (mtTFA) (-43%) protein levels. PGC-1alpha overexpression increased (P < 0.05) FAO, mtDNA, COXIV, mtTFA, and fatty acid incorporation into triacylglycerol in both lean and obese groups. Perturbations in FAO, triacylglycerol synthesis, mtDNA, COXIV, and mtTFA in obese compared with lean HSkMC persisted despite PGC-1alpha overexpression. When adjusted for mtDNA and COXIV content, FAO was equivalent between lean and obese groups.

Conclusion: Reduced mitochondrial content is related to impaired FAO in HSkMC derived from obese individuals. Increasing PGC-1alpha protein levels did not correct the obesity-related absolute reduction in FAO or mtDNA content, implicating mechanisms other than PGC-1alpha abundance.

Figures

FIG. 1.
FIG. 1.
Ad-PGC-1α overexpression in cultured myotubes (HSkMC) from lean and obese donors. A: PGC-1α protein content in no-virus controls (NVC), Ad-β-gal controls, and Ad-PGC-1α–treated HSkMC. PGC-1α protein content increased dose dependently in HSkMC from lean and obese donors. B: mtTFA and COXIV protein content increased dose dependently with increasing PGC-1α viral titer in HSkMC.
FIG. 2.
FIG. 2.
Effect of PGC-1α overexpression on FAO and oxidation efficiency in HSkMC from lean (■) and obese (□) donors. HSkMC cultured from lean (n = 12) and obese (n = 9) donors was treated with either low- or high-dose recombinant Ad-β-gal or PGC-1α and incubated with either 100 μmol/l (A, C, and E) or 500 (B, D, and F) μmol/l [14C] oleate. Complete FAO was measured from 14C-labeled incorporation into CO2 (A and B). Total FAO (C and D) was measured as the sum of 14C-labeled incorporation into CO2 and 14C-labeled incorporation into ASMs, with ASM serving as an index of incomplete FAO. Oxidation efficiency was determined as the ratio of ASM to complete FAO, represented as ASM/CO2 (E and F), with higher values indicative of reduced efficiency. Data are expressed as means ± SE and significant differences denoted at the P ≤ 0.05 level. *Significant difference between lean and obese for that treatment. †Significant main effect comparing control (β-gal) and PGC-1 α overexpression (Ad-PGC-1α) at the respective adenoviral doses. ‡Significant difference between the high and low Ad-PGC-1α doses. §Significant increase in lean subjects with PGC-1α overexpression compared with control at the respective adenoviral doses.
FIG. 3.
FIG. 3.
Effect of PGC-1α overexpression on fatty acid incorporation in HSkMC from lean (■) and obese (□) donors. HSkMC cultured from lean (n = 8) and obese (n = 8) donors were incubated with either 100 μmol/l (A, C, and E) or 500 μmol/l (B, D, and F) [14C] oleate, and 14C-labeled incorporation into glycerolipid (A and B), TAG (C and D), and DAG (E and F) was determined. Data are expressed as means ± SE and significant differences denoted at the P ≤ 0.05 level. *Significant difference between lean and obese for that treatment. †Significant main effect comparing control (β-gal) and PGC-1α overexpression (Ad-PGC-1α) at the respective adenoviral doses. ‡Significant difference between the high and low Ad-PGC-1α doses.
FIG. 4.
FIG. 4.
PGC-1α overexpression does not normalize lipid partitioning rates between HSkMC from lean (■) and obese (□) donors. The partitioning of fatty acids between oxidative and storage pathways was evaluated by dividing the rate of oleate esterified into glycerolipid by the rate completely oxidized in response to either 100 μmol/l (A) or 500 μmol/l (B) [14C] oleate in HSkMC from lean (n = 8) and obese (n = 8) donors. Data are means ± SE and significant differences denoted at the P ≤ 0.05 level. *Significant difference between lean and obese for that treatment. †Significant main effect comparing control (β-gal) and PGC-1α overexpression (Ad-PGC-1α) at the respective adenoviral doses. ‡Significant difference between the high and low Ad-PGC-1α doses.
FIG. 5.
FIG. 5.
PGC-1α increases mtDNA (A), COXIV protein (B), and mtTFA protein (C) in HSkMC from lean (■) and obese (□) donors. mtDNA, n = 9 for lean and obese; COXIV and mtTFA, n = 8 for lean and obese. Data are expressed as means ± SE and significant differences denoted at the P ≤ 0.05 level. *Significant difference between lean and obese for that treatment. †Significant main effect comparing control (β-gal) and PGC-1α overexpression (Ad-PGC-1α) at the respective adenoviral doses. ‡Significant difference between the high and low Ad-PGC-1α doses. NVC, no-virus control.
FIG. 6.
FIG. 6.
FAO does not differ between HSkMC from lean (■) and obese (□) donors when normalized to indexes of mitochondrial content. FAO normalization to indexes of mitochondrial content were evaluated by dividing the rate of complete oleate oxidation from 14C-labeled incorporation into CO2 (FAO) by mtDNA copy number per diploid nuclear genome (A and B) or by COXIV protein expression in arbitrary units (C and D) under both 100 umol/l (A and C) and 500 umol/l (B and D) oleate conditions. FAO/mtDNA, n = 9 for lean and obese; FAO/COXIV, n = 8 for lean and obese. *Significant difference between lean and obese for that treatment. †Significant main effect comparing control (β-gal) and PGC-1α overexpression (Ad-PGC-1α) at the respective adenoviral doses.

References

    1. Kelley DE, Goodpaster BH: Effects of physical activity on insulin action and glucose tolerance in obesity. Med Sci Sports Exerc 1999; 31: S619–S623
    1. Kim JY, Hickner RC, Cortright RL, Dohm GL, Houmard JA: Lipid oxidation is reduced in obese human skeletal muscle. Am J Physiol Endocrinol Metab 2000; 279: E1039–E1044
    1. Hulver MW, Berggren JR, Cortright RN, Dudek RW, Thompson RP, Pories WJ, MacDonald KG, Cline GW, Shulman GI, Dohm GL, Houmard JA: Skeletal muscle lipid metabolism with obesity. Am J Physiol Endocrinol Metab 2003; 284: E741–E747
    1. Guesbeck NR, Hickey MS, MacDonald KG, Pories WJ, Harper I, Ravussin E, Dohm GL, Houmard JA: Substrate utilization during exercise in formerly morbidly obese women. J Appl Physiol 2001; 90: 1007–1012
    1. Thyfault JP, Kraus RM, Hickner RC, Howell AW, Wolfe RR, Dohm GL: Impaired plasma fatty acid oxidation in extremely obese women. Am J Physiol Endocrinol Metab 2004; 287: E1076–E1081
    1. Holloway GP, Thrush AB, Heigenhauser GJ, Tandon NN, Dyck DJ, Bonen A, Spriet LL: Skeletal muscle mitochondrial FAT/CD36 content and palmitate oxidation are not decreased in obese women. Am J Physiol Endocrinol Metab 2007; 292: E1782–E1789
    1. Ritov VB, Menshikova EV, He J, Ferrell RE, Goodpaster BH, Kelley DE: Deficiency of subsarcolemmal mitochondria in obesity and type 2 diabetes. Diabetes 2005; 54: 8–14
    1. Kelley DE, He J, Menshikova EV, Ritov VB: Dysfunction of mitochondria in human skeletal muscle in type 2 diabetes. Diabetes 2002; 51: 2944–2950
    1. Simoneau JA, Veerkamp JH, Turcotte LP, Kelley DE: Markers of capacity to utilize fatty acids in human skeletal muscle: relation to insulin resistance and obesity and effects of weight loss. Faseb J 1999; 13: 2051–2060
    1. Goodyear LJ, Giorgino F, Balon TW, Condorelli G, Smith RJ: Effects of contractile activity on tyrosine phosphoproteins and PI 3-kinase activity in rat skeletal muscle. Am J Physiol 1995; 268: E987–E995
    1. Hulver MW, Berggren JR, Carper MJ, Miyazaki M, Ntambi JM, Hoffman EP, Thyfault JP, Stevens R, Dohm GL, Houmard JA, Muoio DM: Elevated stearoyl-CoA desaturase-1 expression in skeletal muscle contributes to abnormal fatty acid partitioning in obese humans. Cell Metab 2005; 2: 251–261
    1. Malenfant P, Joanisse DR, Theriault R, Goodpaster BH, Kelley DE, Simoneau JA: Fat content in individual muscle fibers of lean and obese subjects. Int J Obes Relat Metab Disord 2001; 25: 1316–1321
    1. Berggren JR, Boyle KE, Chapman WH, Houmard JA: Skeletal muscle lipid oxidation and obesity: influence of weight loss and exercise. Am J Physiol Endocrinol Metab 2008; 294: E726–E732
    1. Rohas LM, St-Pierre J, Uldry M, Jager S, Handschin C, Spiegelman BM: A fundamental system of cellular energy homeostasis regulated by PGC-1alpha. Proc Natl Acad Sci U S A 2007; 104: 7933–7938
    1. Benton CR, Nickerson JG, Lally J, Han XX, Holloway GP, Glatz JF, Luiken JJ, Graham TE, Heikkila JJ, Bonen A: Modest PGC-1alpha overexpression in muscle in vivo is sufficient to increase insulin sensitivity and palmitate oxidation in subsarcolemmal, not intermyofibrillar, mitochondria. J Biol Chem 2008; 283: 4228–4240
    1. Baar K, Wende AR, Jones TE, Marison M, Nolte LA, Chen M, Kelly DP, Holloszy JO: Adaptations of skeletal muscle to exercise: rapid increase in the transcriptional coactivator PGC-1. Faseb J 2002; 16: 1879–1886
    1. Koves TR, Li P, An J, Akimoto T, Slentz D, Ilkayeva O, Dohm GL, Yan Z, Newgard CB, Muoio DM: Peroxisome proliferator-activated receptor-gamma co-activator 1alpha-mediated metabolic remodeling of skeletal myocytes mimics exercise training and reverses lipid-induced mitochondrial inefficiency. J Biol Chem 2005; 280: 33588–33598
    1. Berggren JR, Tanner CJ, Houmard JA: Primary cell cultures in the study of human muscle metabolism. Exerc Sport Sci Rev 2007; 35: 56–61
    1. Muoio DM, Way JM, Tanner CJ, Winegar DA, Kliewer SA, Houmard JA, Kraus WE, Dohm GL: Peroxisome proliferator–activated receptor-α regulates fatty acid utilization in primary human skeletal muscle cells. Diabetes 2002; 51: 901–909
    1. Russell AP, Feilchenfeldt J, Schreiber S, Praz M, Crettenand A, Gobelet C, Meier CA, Bell DR, Kralli A, Giacobino JP, Deriaz O: Endurance training in humans leads to fiber type-specific increases in levels of peroxisome proliferator–activated receptor-γ coactivator-1 and peroxisome proliferator–activated receptor-α in skeletal muscle. Diabetes 2003; 52: 2874–2881
    1. Miller FJ, Rosenfeldt FL, Zhang C, Linnane AW, Nagley P: Precise determination of mitochondrial DNA copy number in human skeletal and cardiac muscle by a PCR-based assay: lack of change of copy number with age. Nucleic Acid Res 2003; 31: e61.
    1. Menshikova EV, Ritov VB, Toledo FG, Ferrell RE, Goodpaster BH, Kelley DE: Effects of weight loss and physical activity on skeletal muscle mitochondrial function in obesity. Am J Physiol Endocrinol Metab 2005; 288: E818–E825
    1. Menshikova EV, Ritov VB, Fairfull L, Ferrell RE, Kelley DE, Goodpaster BH: Effects of exercise on mitochondrial content and function in aging human skeletal muscle. J Gerontol A Biol Sci Med Sci 2006; 61: 534–540
    1. Szuhai K, Ouweland J, Dirks R, Lemaitre M, Truffert J, Janssen G, Tanke H, Holme E, Maassen J, Raap A: Simultaneous A8344G heteroplasmy and mitochondrial DNA copy number quantification in myoclonus epilepsy and ragged-red fibers (MERRF) syndrome by a multiplex molecular beacon based real-time fluorescence PCR. Nucleic Acid Res 2001; 29: E13.
    1. Ukropcova B, McNeil M, Sereda O, de Jonge L, Xie H, Bray GA, Smith SR: Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor. J Clin Invest 2005; 115: 1934–1941
    1. Holloway GP, Bonen A, Spriet LL: Regulation of skeletal muscle mitochondrial fatty acid metabolism in lean and obese individuals. Am J Clin Nutr 2009; 89: 455S–462S
    1. Tanner CJ, Barakat HA, Dohm GL, Pories WJ, MacDonald KG, Cunningham PR, Swanson MS, Houmard JA: Muscle fiber type is associated with obesity and weight loss. Am J Physiol Endocrinol Metab 2002; 282: E1191–E1196
    1. Gaster M, Rustan AC, Aas V, Beck-Nielsen H: Reduced lipid oxidation in skeletal muscle from type 2 diabetic subjects may be of genetic origin: evidence from cultured myotubes. Diabetes 2004; 53: 542–548
    1. Gleyzer N, Vercauteren K, Scarpulla RC: Control of mitochondrial transcription specificity factors (TFB1M and TFB2M) by nuclear respiratory factors (NRF-1 and NRF-2) and PGC-1 family coactivators. Mol Cell Biol 2005; 25: 1354–1366
    1. Kelly DP, Scarpulla RC: Transcriptional regulatory circuits controlling mitochondrial biogenesis and function. Genes Dev 2004; 18: 357–368
    1. Lehman JJ, Barger PM, Kovacs A, Saffitz JE, Medeiros DM, Kelly DP: Peroxisome proliferator-activated receptor gamma coactivator-1 promotes cardiac mitochondrial biogenesis. J Clin Invest 2000; 106: 847–856
    1. Puigserver P, Wu Z, Park CW, Graves R, Wright M, Spiegelman BM: A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis. Cell 1998; 92: 829–839
    1. Shadel GS, Clayton DA: Mitochondrial transcription initiation: variation and conservation. J Biol Chem 1993; 268: 16083–16086
    1. Hood DA: Mechanisms of exercise-induced mitochondrial biogenesis in skeletal muscle. Appl Physiol Nutr Metab 2009; 34: 465–472
    1. Itani SI, Ruderman NB, Schmieder F, Boden G: Lipid-induced insulin resistance in human muscle is associated with changes in diacylglycerol, protein kinase C, and IκB-α. Diabetes 2002; 51: 2005–2011
    1. Koves TR, Ussher JR, Noland RC, Slentz D, Mosedale M, Ilkayeva O, Bain J, Stevens R, Dyck JR, Newgard CB, Lopaschuk GD, Muoio DM: Mitochondrial overload and incomplete fatty acid oxidation contribute to skeletal muscle insulin resistance. Cell Metab 2008; 7: 45–56
    1. Choi CS, Befroy DE, Codella R, Kim S, Reznick RM, Hwang YJ, Liu ZX, Lee HY, Distefano A, Samuel VT, Zhang D, Cline GW, Handschin C, Lin J, Petersen KF, Spiegelman BM, Shulman GI: Paradoxical effects of increased expression of PGC-1alpha on muscle mitochondrial function and insulin-stimulated muscle glucose metabolism. Proc Natl Acad Sci U S A 2008; 105: 19926–19931

Source: PubMed

스폰서 및 공동 작업자

건강 상태

약물 개입

3
구독하다