Measurement of de novo hepatic lipogenesis in humans using stable isotopes

M. K. Hellerstein, M. Christiansen, S. Kaempfer, C. Kletke, K. Wu, J. S. Reid, K. Mulligan, N. S. Hellerstein, C. H L Shackleton

Research output: Contribution to journalArticle

273 Citations (Scopus)

Abstract

Direct measurement of de novo lipogenesis has not previously been possible in humans. We measured de novo hepatic lipogenesis in normal men by means of stable isotopes and by combining the acetylated-xenobiotic probe technique with mass isotopomer analysis of secreted very low density lipoprotein-fatty acids (VLDL-FA). Sulfamethoxazole (SMX) was administered with [13C]acetate during an overnight fast followed by refeeding with intravenous glucose (7-10 mg/kg of weight per min), oral Ensure (7-10 mg of carbohydrate/kg of weight per min), or a high-carbohydrate mixed-meal breakfast (3.5 g of carbohydrate/kg of weight). Respiratory quotients remained < 1.0. High-performance liquid chromatography/mass spectrometry-determined enrichments in SMX-acetate attained stable plateau values, and hepatic acetyl-coenzyme A (CoA) dilution rate did not increase with refeeding (∼ 0.024 mmol/kg per min). The fraction of VLDL-palmitate derived from de novo lipogenesis was only 0.91±0.27% (fasted) and 1.64-1.97% (fed). For stearate, this was 0.37±0.08% and 0.47-0.64%. Precursor enrichments predicted from isotopomer ratios were close to measured SMX-acetate enrichments, indicating that SMX-acetate samples the true lipogenic acetyl-CoA pool. Stearate synthesis was less than palmitate and the two did not move in parallel. Estimated total VLDL-FA synthesis is < 500 mg/ day. Thus, de novo hepatic lipogenesis is a quantitatively minor pathway, consistent with gas exchange estimates; fatty acid futile cycling (oxidation/resynthesis) is not thermogenically significant; and synthesis rates of different nonessential fatty acids by human liver are not identical in nonoverfed normal men. The contribution and regulation of de novo lipogenesis in other settings can be studied using this technique.

Original languageEnglish (US)
Pages (from-to)1841-1852
Number of pages12
JournalJournal of Clinical Investigation
Volume87
Issue number5
StatePublished - May 1991

Fingerprint

Lipogenesis
Sulfamethoxazole
Isotopes
Acetates
Fatty Acids
Liver
Stearates
Acetyl Coenzyme A
VLDL Lipoproteins
Palmitates
Carbohydrates
Weights and Measures
Substrate Cycling
Breakfast
Xenobiotics
Meals
Mass Spectrometry
Gases
High Pressure Liquid Chromatography
Glucose

Keywords

  • Carbohydrate disposal
  • High-performance liquid chromatography/mass spectrometry
  • Isotopomers
  • Nonessential fatty acids
  • Precursor-product relationship
  • Xenobiotic probes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Hellerstein, M. K., Christiansen, M., Kaempfer, S., Kletke, C., Wu, K., Reid, J. S., ... Shackleton, C. H. L. (1991). Measurement of de novo hepatic lipogenesis in humans using stable isotopes. Journal of Clinical Investigation, 87(5), 1841-1852.

Measurement of de novo hepatic lipogenesis in humans using stable isotopes. / Hellerstein, M. K.; Christiansen, M.; Kaempfer, S.; Kletke, C.; Wu, K.; Reid, J. S.; Mulligan, K.; Hellerstein, N. S.; Shackleton, C. H L.

In: Journal of Clinical Investigation, Vol. 87, No. 5, 05.1991, p. 1841-1852.

Research output: Contribution to journalArticle

Hellerstein, MK, Christiansen, M, Kaempfer, S, Kletke, C, Wu, K, Reid, JS, Mulligan, K, Hellerstein, NS & Shackleton, CHL 1991, 'Measurement of de novo hepatic lipogenesis in humans using stable isotopes', Journal of Clinical Investigation, vol. 87, no. 5, pp. 1841-1852.
Hellerstein MK, Christiansen M, Kaempfer S, Kletke C, Wu K, Reid JS et al. Measurement of de novo hepatic lipogenesis in humans using stable isotopes. Journal of Clinical Investigation. 1991 May;87(5):1841-1852.
Hellerstein, M. K. ; Christiansen, M. ; Kaempfer, S. ; Kletke, C. ; Wu, K. ; Reid, J. S. ; Mulligan, K. ; Hellerstein, N. S. ; Shackleton, C. H L. / Measurement of de novo hepatic lipogenesis in humans using stable isotopes. In: Journal of Clinical Investigation. 1991 ; Vol. 87, No. 5. pp. 1841-1852.
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abstract = "Direct measurement of de novo lipogenesis has not previously been possible in humans. We measured de novo hepatic lipogenesis in normal men by means of stable isotopes and by combining the acetylated-xenobiotic probe technique with mass isotopomer analysis of secreted very low density lipoprotein-fatty acids (VLDL-FA). Sulfamethoxazole (SMX) was administered with [13C]acetate during an overnight fast followed by refeeding with intravenous glucose (7-10 mg/kg of weight per min), oral Ensure (7-10 mg of carbohydrate/kg of weight per min), or a high-carbohydrate mixed-meal breakfast (3.5 g of carbohydrate/kg of weight). Respiratory quotients remained < 1.0. High-performance liquid chromatography/mass spectrometry-determined enrichments in SMX-acetate attained stable plateau values, and hepatic acetyl-coenzyme A (CoA) dilution rate did not increase with refeeding (∼ 0.024 mmol/kg per min). The fraction of VLDL-palmitate derived from de novo lipogenesis was only 0.91±0.27{\%} (fasted) and 1.64-1.97{\%} (fed). For stearate, this was 0.37±0.08{\%} and 0.47-0.64{\%}. Precursor enrichments predicted from isotopomer ratios were close to measured SMX-acetate enrichments, indicating that SMX-acetate samples the true lipogenic acetyl-CoA pool. Stearate synthesis was less than palmitate and the two did not move in parallel. Estimated total VLDL-FA synthesis is < 500 mg/ day. Thus, de novo hepatic lipogenesis is a quantitatively minor pathway, consistent with gas exchange estimates; fatty acid futile cycling (oxidation/resynthesis) is not thermogenically significant; and synthesis rates of different nonessential fatty acids by human liver are not identical in nonoverfed normal men. The contribution and regulation of de novo lipogenesis in other settings can be studied using this technique.",
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