Research Article| Volume 141, 155399, April 2023

Short-chain fatty acid production in accessible and inaccessible body pools as assessed by novel stable tracer pulse approach is reduced by aging independent of presence of COPD

Published:January 12, 2023DOI:


      • Previous methods to measure (intestinal) SCFA production might be inaccurate.
      • We assessed SCFA production with stable tracer pulse approach and compartmental modeling.
      • SCFA production and fluxes were reduced in aging but not affected by presence of COPD.
      • Non-compartmental SCFA production measurements underestimate production in inaccessible (intestinal) pool.



      Production rates of the short-chain fatty acids (SCFA) acetate, propionate, and butyrate, which are beneficial metabolites of the intestinal microbiota, are difficult to measure in humans due to inaccessibility of the intestine to perform measurements, and the high first-pass metabolism of SCFAs in colonocytes and liver. We developed a stable tracer pulse approach to estimate SCFA whole-body production (WBP) in the accessible pool representing the systemic circulation and interstitial fluid. Compartmental modeling of plasma enrichment data allowed us to additionally calculate SCFA kinetics and pool sizes in the inaccessible pool likely representing the intestine with microbiota. We also studied the effects of aging and the presence of Chronic Obstructive Pulmonary Disease (COPD) on SCFA kinetics.


      In this observational study, we designed a two-compartmental model to determine SCFA kinetics in 31 young (20–29 y) and 71 older (55–87 y) adults, as well as in 33 clinically stable patients with moderate to very severe COPD (mean (SD) FEV1, 46.5 (16.2)% of predicted). Participants received in the fasted state a pulse containing stable tracers of acetate, propionate, and butyrate intravenously and blood was sampled four times over a 30 min period. We measured tracer-tracee ratios by GC–MS and used parameters obtained from two-exponential curve fitting to calculate non-compartmental SCFA WBP and perform compartmental analysis. Statistics were done by ANCOVA.


      Acetate, propionate, and butyrate WBP and fluxes between the accessible and inaccessible pools were lower in older than young adults (all q < 0.0001). Moreover, older participants had lower acetate (q < 0.0001) and propionate (q = 0.019) production rates in the inaccessible pool as well as smaller sizes of the accessible and inaccessible acetate pools (both q < 0.0001) than young participants. WBP, compartmental SCFA kinetics, and pool sizes did not differ between COPD patients and older adults (all q > 0.05). Overall and independent of the group studied, calculated production rates in the inaccessible pool were on average 7 (acetate), 11 (propionate), and 16 (butyrate) times higher than non-compartmental WBP, and sizes of inaccessible pools were 24 (acetate), 31 (propionate), and 55 (butyrate) times higher than sizes of accessible pools (all p < 0.0001).


      Non-compartmental production measurements of SCFAs in the accessible pool (i.e. systemic circulation) substantially underestimate the SCFA production in the inaccessible pool, which likely represents the intestine with microbiota, as assessed by compartmental analysis.

      Graphical abstract


      BMI (body mass index), COPD (Chronic Obstructive Pulmonary Disease), FEV1 (forced expiratory volume in 1 s), Fij (tracee flux from compartment j to compartment i [μmol·min−1]), FVC (forced vital capacity), kij (fractional tracer exchange rate from compartment j to compartment i [min−1]), Qi (tracee pool size in compartment i [μmol]), qi (tracer pool size in compartment i [μmol]), SCFA (short-chain fatty acid), TTR (tracer-tracee ratio), Ui (endogenous production in compartment i [μmol·min−1]), WBP (whole-body production [μmol·min−1]), WbRa (whole-body rate of appearance [μmol·min−1])


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        • van der Hee B.
        • Wells J.M.
        Microbial regulation of host physiology by short-chain fatty acids.
        Trends Microbiol. 2021; 29: 700-712
        • Auger C.
        • Vinaik R.
        • Appanna V.D.
        • Jeschke M.G.
        Beyond mitochondria: alternative energy-producing pathways from all strata of life.
        Metabolism. 2021; 118154733
        • Markowiak-Kopeć P.
        • Śliżewska K.
        The effect of probiotics on the production of short-chain fatty acids by human intestinal microbiome.
        Nutrients. 2020; 12: 1107
        • Oliver A.
        • Chase A.B.
        • Weihe C.
        • Orchanian S.B.
        • Riedel S.F.
        • Hendrickson C.L.
        • et al.
        High-fiber, whole-food dietary intervention alters the human gut microbiome but not fecal short-chain fatty acids.
        mSystems. 2021; 6: e00115-e00121
        • Clauss M.
        • Gérard P.
        • Mosca A.
        • Leclerc M.
        Interplay between exercise and gut microbiome in the context of human health and performance.
        Front Nutr. 2021; 8: 637010
        • Boets E.
        • Deroover L.
        • Houben E.
        • Vermeulen K.
        • Gomand S.V.
        • Delcour J.A.
        • et al.
        Quantification of in vivo colonic short chain fatty acid production from inulin.
        Nutrients. 2015; 7: 8916-8929
        • Neis E.P.
        • Bloemen J.G.
        • Rensen S.S.
        • van der Vorst J.R.
        • van den Broek M.A.
        • Venema K.
        • et al.
        Effects of liver resection on hepatic short-chain fatty acid metabolism in humans.
        PLoS One. 2016; 11e0166161
        • Boets E.
        • Gomand S.V.
        • Deroover L.
        • Preston T.
        • Vermeulen K.
        • De Preter V.
        • et al.
        Systemic availability and metabolism of colonic-derived short-chain fatty acids in healthy subjects: a stable isotope study.
        J Physiol. 2017; 595: 541-555
        • den Besten G.
        • Lange K.
        • Havinga R.
        • van Dijk T.H.
        • Gerding A.
        • van Eunen K.
        • et al.
        Gut-derived short-chain fatty acids are vividly assimilated into host carbohydrates and lipids.
        Am J Physiol Gastrointest Liver Physiol. 2013; 305: G900-G910
        • Walter J.H.
        • Thompson G.N.
        • Leonard J.V.
        • Heatherington C.S.
        • Bartlett K.
        Measurement of propionate turnover in vivo using sodium [2H5]propionate and sodium [13C]propionate.
        Clin Chim Acta. 1989; 182: 141-150
        • Pouteau E.
        • Piloquet H.
        • Maugeais P.
        • Champ M.
        • Dumon H.
        • Nguyen P.
        • et al.
        Kinetic aspects of acetate metabolism in healthy humans using [1-13C] acetate.
        Am J Physiol. 1996; 271: E58-E64
        • Ten Have G.A.M.
        • Engelen M.
        • Wolfe R.R.
        • Deutz N.E.P.
        Phenylalanine isotope pulse method to measure effect of sepsis on protein breakdown and membrane transport in the pig.
        Am J Physiol Endocrinol Metab. 2017; 312: E519-E529
        • Deutz N.E.P.
        • Thaden J.J.
        • Ten Have G.A.M.
        • Walker D.K.
        • Engelen M.
        Metabolic phenotyping using kinetic measurements in young and older healthy adults.
        Metabolism. 2018; 78: 167-178
        • Ghane P.
        • Kirschner S.K.
        • Engelen M.P.K.J.
        • Ivanov I.
        • Braga-Neto U.M.
        • Deutz N.E.P.
        Compartmental data analysis to determine whole-body shortchain fatty acid production through the use of a pulse of stable tracers in humans.
        Curr Dev Nutr. 2018; 2 (Abst): 38
        • Mason A.
        • Engelen M.
        • Ivanov I.
        • Toffolo G.M.
        • Deutz N.E.P.
        A four-compartment compartmental model to assess net whole body protein breakdown using a pulse of phenylalanine and tyrosine stable isotopes in humans.
        Am J Physiol Endocrinol Metab. 2017; 313: E63-E74
        • Engelen M.P.
        • Ten Have G.A.
        • Thaden J.J.
        • Deutz N.E.
        New advances in stable tracer methods to assess whole-body protein and amino acid metabolism.
        Curr Opin Clin Nutr Metab Care. 2019; 22: 337-346
        • Salazar N.
        • Arboleya S.
        • Fernandez-Navarro T.
        • de Los Reyes-Gavilan C.G.
        • Gonzalez S.
        • Gueimonde M.
        Age-associated changes in gut microbiota and dietary components related with the immune system in adulthood and old age: a cross-sectional study.
        Nutrients. 2019; 11: 1765
        • Bowerman K.L.
        • Rehman S.F.
        • Vaughan A.
        • Lachner N.
        • Budden K.F.
        • Kim R.Y.
        • et al.
        Disease-associated gut microbiome and metabolome changes in patients with chronic obstructive pulmonary disease.
        Nat Commun. 2020; 11: 5886
        • Li N.
        • Dai Z.
        • Wang Z.
        • Deng Z.
        • Zhang J.
        • Pu J.
        • et al.
        Gut microbiota dysbiosis contributes to the development of chronic obstructive pulmonary disease.
        Respir Res. 2021; 22: 274
        • Kirschner S.K.
        • Deutz N.E.P.
        • Jonker R.
        • Olde Damink S.W.M.
        • Harrykissoon R.I.
        • Zachria A.J.
        • et al.
        Intestinal function is impaired in patients with chronic obstructive pulmonary disease.
        Clin Nutr. 2021; 40: 2270-2277
        • Cruthirds C.L.
        • Deutz N.E.P.
        • Ligthart-Melis G.C.
        • Simbo S.Y.
        • Engelen M.
        Walking exercise alters protein digestion, amino acid absorption, and whole body protein kinetics in older adults with and without COPD.
        J Appl Physiol. 2021; 130: 435-444
        • Ghosh T.S.
        • Shanahan F.
        • O'Toole P.W.
        The gut microbiome as a modulator of healthy ageing.
        Nat Rev Gastroenterol Hepatol. 2022; 19: 565-584
        • Vogelmeier C.F.
        • Criner G.J.
        • Martinez F.J.
        • Anzueto A.
        • Barnes P.J.
        • Bourbeau J.
        • et al.
        Global strategy for the diagnosis, management, and prevention of chronic obstructive lung disease 2017 report. GOLD executive summary.
        Am J Respir Crit Care Med. 2017; 195: 557-582
        • Charlson M.E.
        • Pompei P.
        • Ales K.L.
        • MacKenzie C.R.
        A new method of classifying prognostic comorbidity in longitudinal studies: development and validation.
        J Chronic Dis. 1987; 40: 373-383
        • Jonker R.
        • Deutz N.E.P.
        • Schols A.
        • Veley E.A.
        • Harrykissoon R.
        • Zachria A.J.
        • et al.
        Whole body protein anabolism in COPD patients and healthy older adults is not enhanced by adding either carbohydrates or leucine to a serving of protein.
        Clin Nutr. 2019; 38: 1684-1691
        • Mahler D.A.
        • Wells C.K.
        Evaluation of clinical methods for rating dyspnea.
        Chest. 1988; 93: 580-586
        • Abumrad N.N.
        • Rabin D.
        • Diamond M.P.
        • Lacy W.W.
        Use of a heated superficial hand vein as an alternative site for the measurement of amino acid concentrations and for the study of glucose and alanine kinetics in man.
        Metabolism. 1981; 30: 936-940
        • Santos-Marcos J.A.
        • Rangel-Zuniga O.A.
        • Jimenez-Lucena R.
        • Quintana-Navarro G.M.
        • Garcia-Carpintero S.
        • Malagon M.M.
        • et al.
        Influence of gender and menopausal status on gut microbiota.
        Maturitas. 2018; 116: 43-53
        • Kirschner S.
        • Deutz N.
        • Hermann C.
        • Engelen M.
        Plasma short-chain fatty acid (SCFA) concentrations are related to cognition and adipose tissue mass but not muscle function in older adults.
        Curr Dev Nutr. 2022; 6 (Abst): 33
        • Pouteau E.
        • Rochat F.
        • Jann A.
        • Meirim I.
        • Sanchez-Garcia J.-L.
        • Ornstein K.
        • et al.
        Chicory increases acetate turnover, but not propionate and butyrate peripheral turnovers in rats.
        Br J Nutr. 2008; 99: 287-296
        • Cobelli C.
        • Foster D.
        • Toffolo G.
        Tracer kinetics in biomedical research: from data to model.
        Springer Science & Business Media, 2007
        • Wolfe R.R.
        • Chinkes D.L.
        Chap4: calculation of substrate kinetics: multiple pool-models. Isotope tracers in metabolic research.
        in: Second ed. John Wiley & Sons, Inc., 2005: 51-76
        • Glickman M.E.
        • Rao S.R.
        • Schultz M.R.
        False discovery rate control is a recommended alternative to Bonferroni-type adjustments in health studies.
        J Clin Epidemiol. 2014; 67: 850-857
        • Cruz-Jentoft A.J.
        • Bahat G.
        • Bauer J.
        • Boirie Y.
        • Bruyere O.
        • Cederholm T.
        • et al.
        Sarcopenia: revised European consensus on definition and diagnosis.
        Age Ageing. 2019; 48: 16-31
        • Kirschner S.K.
        • Deutz N.E.P.
        • Rijnaarts I.
        • Smit T.J.
        • Larsen D.J.
        • Engelen M.P.K.J.
        Impaired intestinal function is associated with lower muscle and cognitive health and well-being in patients with congestive heart failure.
        Jpen-Parenter Enter. 2022; 46: 660-670
        • Matthews D.E.
        • Downey R.S.
        Measurement of urea kinetics in humans: a validation of stable isotope tracer methods.
        Am J Physiol. 1984; 246: E519-E527
        • Dong Y.
        • Chen L.
        • Gutin B.
        • Zhu H.
        Total, insoluble, and soluble dietary fiber intake and insulin resistance and blood pressure in adolescents.
        Eur J Clin Nutr. 2019; 73: 1172-1178
        • Kirschner S.K.
        • Ten Have G.A.
        • Engelen M.P.
        • Deutz N.E.
        Transorgan short-chain fatty acid fluxes in the fasted and postprandial state in the pig.
        Am J Physiol Endocrinol Metab. 2021; 321: E665-E673
        • Joppa P.
        • Tkacova R.
        • Franssen F.M.E.
        • Hanson C.
        • Rennard S.I.
        • Silverman E.K.
        • et al.
        Sarcopenic obesity, functional outcomes, and systemic inflammation in patients with chronic obstructive pulmonary disease.
        J Am Med Dir Assoc. 2016; 17: 712-718
        • Furutate R.
        • Ishii T.
        • Wakabayashi R.
        • Motegi T.
        • Yamada K.
        • Gemma A.
        • et al.
        Excessive visceral fat accumulation in advanced chronic obstructive pulmonary disease.
        Int J Chron Obstruct Pulmon Dis. 2011; 6: 423
        • Montandon S.A.
        • Jornayvaz F.R.
        Effects of antidiabetic drugs on gut microbiota composition.
        Genes. 2017; 8: 250
        • Sun C.
        • Wang Z.
        • Hu L.
        • Zhang X.
        • Chen J.
        • Yu Z.
        • et al.
        Targets of statins intervention in LDL-C metabolism: gut microbiota.
        Front Cardiovasc Med. 2022; : 9