Skip to main content

β-Hydroxy β-methylbutyrate free acid alters cortisol responses, but not myofibrillar proteolysis, during a 24-h fast

  • Grant M. Tinsley (a1), Amy H. Givan (a1), Austin J. Graybeal (a1), Michael I. Villarreal (a1) and Austin G. Cross (a1)...

This study was a randomised, double-blind, placebo-controlled cross-over trial examining the effects of β-hydroxy β-methylbutyrate free acid (HMB-FA) supplementation on muscle protein breakdown, cortisol, testosterone and resting energy expenditure (REE) during acute fasting. Conditions consisted of supplementation with 3 g/d HMB-FA or placebo during a 3-d meat-free diet followed by a 24-h fast. Urine was collected before and during the 24-h fast for analysis of 3-methylhistidine:creatinine ratio (3MH:CR). Salivary cortisol, testosterone, their ratio (T:C), and the cortisol awakening response were assessed. ANOVA was used to analyse all dependent variables, and linear mixed models were used to confirm the absence of carryover effects. Eleven participants (six females, five males) completed the study. Urinary HMB concentrations confirmed compliance with supplementation. 3MH:CR was unaffected by fasting and supplementation, but the cortisol awakening response differed between conditions. In both conditions, cortisol increased from awakening to 30 min post-awakening (P=0·01). Cortisol was reduced from 30 to 45 min post-awakening with HMB-FA (−32 %, d=−1·0, P=0·04), but not placebo (PL) (−6 %, d=−0·2, P=0·14). In males, T:C increased from 0 to 24 h of fasting with HMB-FA (+162 %, d=3·0, P=0·001), but not placebo (+13 %, d=0·4, P=0·60), due to reductions in cortisol. REE was higher at 24 h of fasting than 16 h of fasting independent of supplementation (+4·0 %, d=0·3, P=0·04). In conclusion, HMB-FA may affect cortisol responses, but not myofibrillar proteolysis, during acute 24-h fasting.

Corresponding author
* Corresponding author: G. M. Tinsley, email
Hide All
1. Dulloo, AG, Jacquet, J, Miles-Chan, JL, et al. (2017) Passive and active roles of fat-free mass in the control of energy intake and body composition regulation. Eur J Clin Nutr 71, 353357.
2. Dulloo, AG (2017) Collateral fattening: when a deficit in lean body mass drives overeating. Obesity 25, 277279.
3. Ravussin, E, Burnand, B, Schutz, Y, et al. (1982) Twenty-four-hour energy expenditure and resting metabolic rate in obese, moderately obese, and control subjects. Am J Clin Nutr 35, 566573.
4. Varady, KA (2011) Intermittent versus daily calorie restriction: which diet regimen is more effective for weight loss? Obes Rev 12, e593e601.
5. Tinsley, GM & Bounty, PML (2015) Effects of intermittent fasting on body composition and clinical health markers in humans. Nutr Rev 73, 661674.
6. Trepanowski, JF, Kroeger, CM, Barnosky, A, et al. (2017) Effect of alternate-day fasting on weight loss, weight maintenance, and cardioprotection among metabolically healthy obese adults: a randomized clinical trial. JAMA Intern Med 177, 930938.
7. Tinsley, GM & Horne, BD (2018) Intermittent fasting and cardiovascular disease: current evidence and unresolved questions. Future Cardiol 14, 4754.
8. Rennie, MJ, Edwards, RHT, Halliday, D, et al. (1982) Muscle protein synthesis measured by stable isotope techniques in man: the effects of feeding and fasting. Clin Sci 63, 519.
9. Tsalikian, E, Howard, C, Gerich, JE, et al. (1984) Increased leucine flux in short-term fasted human subjects: evidence for increased proteolysis. Am J Physiol 247, E323E327.
10. Moro, T, Tinsley, G, Bianco, A, et al. (2016) Effects of eight weeks of time-restricted feeding (16/8) on basal metabolism, maximal strength, body composition, inflammation, and cardiovascular risk factors in resistance-trained males. J Transl Med 14, 290.
11. Tinsley, GM, Forsse, JS, Butler, NK, et al. (2017) Time-restricted feeding in young men performing resistance training: a randomized controlled trial. Eur J Sport Sci 17, 200207.
12. Lee, IM, Shiroma, EJ, Lobelo, F, et al. (2012) Effect of physical inactivity on major non-communicable diseases worldwide: an analysis of burden of disease and life expectancy. Lancet 380, 219229.
13. Smith, HJ, Mukerji, P & Tisdale, MJ (2005) Attenuation of proteasome-induced proteolysis in skeletal muscle by {beta}-hydroxy-{beta}-methylbutyrate in cancer-induced muscle loss. Cancer Res 65, 277283.
14. Holecek, M, Muthny, T, Kovarik, M, et al. (2009) Effect of beta-hydroxy-beta-methylbutyrate (HMB) on protein metabolism in whole body and in selected tissues. Food Chem Toxicol 47, 255259.
15. Hao, Y, Jackson, JR, Wang, Y, et al. (2011) beta-Hydroxy-beta-methylbutyrate reduces myonuclear apoptosis during recovery from hind limb suspension-induced muscle fiber atrophy in aged rats. Am J Physiol Regul Integr Comp Physiol 301, R701R715.
16. Shirato, M, Tsuchiya, Y, Sato, T, et al. (2016) Effects of combined β-hydroxy-β-methylbutyrate (HMB) and whey protein ingestion on symptoms of eccentric exercise-induced muscle damage. J Int Soc Sports Nutr 13, 7.
17. Wilson, JM, Kim, JS, Lee, SR, et al. (2009) Acute and timing effects of beta-hydroxy-beta-methylbutyrate (HMB) on indirect markers of skeletal muscle damage. Nutr Metab (Lond) 6, 6.
18. Nissen, S, Sharp, R, Ray, M, et al. (1996) Effect of leucine metabolite β-hydroxy-β-methylbutyrate on muscle metabolism during resistance-exercise training. J Appl Physiol 81, 20952104.
19. Soeters, MR, Soeters, PB, Schooneman, MG, et al. (2012) Adaptive reciprocity of lipid and glucose metabolism in human short-term starvation. Am J Physiol 303, E1397E1407.
20. Rojdmark, S (1987) Influence of short-term fasting on the pituitary–testicular axis in normal men. Hormone Res 25, 140146.
21. Zauner, C, Schneeweiss, B, Kranz, A, et al. (2000) Resting energy expenditure in short-term starvation is increased as a result of an increase in serum norepinephrine. Am J Clin Nutr 71, 15111515.
22. Mansell, PI, Fellows, IW & Macdonald, IA (1990) Enhanced thermogenic response to epinephrine after 48-h starvation in humans. Am J Physiol 258, R87R93.
23. Klein, S, Sakurai, Y, Romijn, JA, et al. (1993) Progressive alterations in lipid and glucose metabolism during short-term fasting in young adult men. Am J Physiol 265, E801E806.
24. Norrelund, H, Nair, KS, Jorgensen, JO, et al. (2001) The protein-retaining effects of growth hormone during fasting involve inhibition of muscle-protein breakdown. Diabetes 50, 96104.
25. Elia, M, Carter, A, Bacon, S, et al. (1980) The effect of 3-methylhistidine in food on its urinary excretion in man. Clin Sci 59, 509511.
26. Elia, M, Carter, A, Bacon, S, et al. (1981) Clinical usefulness of urinary 3-methylhistidine excretion in indicating muscle protein breakdown. Br Med J 282, 351354.
27. US Department of Agriculture (2016) SuperTracker . Washington, DC: US Department of Agriculture.
28. Nissen, S, Van Koevering, M & Webb, D (1990) Analysis of beta-hydroxy-beta-methyl butyrate in plasma by gas chromatography and mass spectrometry. Anal Biochem 188, 1719.
29. Rathmacher, JA, Link, GA, Flakoll, PJ, et al. (1992) Gas chromatographic/mass spectrometric analysis of stable isotopes of 3-methylhistidine in biological fluids: application to plasma kinetics in vivo . Biol Mass Spectrom 21, 560566.
30. Salimetrics (2015) Saliva Collection Handbook. Carlsbad, CA: Salimetrics.
31. Stalder, T, Kirschbaum, C, Kudielka, BM, et al. (2016) Assessment of the cortisol awakening response: Expert consensus guidelines. Psychoneuroendocrinology 63, 414432.
32. Dorn, LD, Lucke, JF, Loucks, TL, et al. (2007) Salivary cortisol reflects serum cortisol: analysis of circadian profiles. Ann Clin Biochem 44, 281284.
33. Vining, RF, McGinley, RA, Maksvytis, JJ, et al. (1983) Salivary cortisol: a better measure of adrenal cortical function than serum cortisol. Ann Clin Biochem 20, 329335.
34. Vining, RF & McGinley, RA (1987) The measurement of hormones in saliva: possibilities and pitfalls. J Steroid Biochem 27, 8194.
35. Nahoul, K, Rao, LV & Scholler, R (1986) Saliva testosterone time-course response to hCG in adult normal men. Comparison with plasma levels. J Steroid Biochem 24, 10111015.
36. Vittek, J, L’Hommedieu, DG, Gordon, GG, et al. (1985) Direct radioimmunoassay (RIA) of salivary testosterone: correlation with free and total serum testosterone. Life Sci 37, 711716.
37. Salimetrics, L (2017) Testosterone testing in saliva & salivary testosterone research., (accessed March 2017).
38. Salimetrics, L (2017) Cortisol levels in saliva & salivary cortisol research - salimetrics. (accessed March 2017).
39. Compher, C, Frankenfield, D, Keim, N, et al. (2006) Best practice methods to apply to measurement of resting metabolic rate in adults: a systematic review. J Am Diet Assoc 106, 881903.
40. Paris, JJ, Franco, C, Sodano, R, et al. (2010) Sex differences in salivary cortisol in response to acute stressors among healthy participants, in recreational or pathological gamblers, and in those with posttraumatic stress disorder. Hormones Behav 57, 3545.
41. IBM (2017) Using linear mixed models to analyze a crossover trial. IBM Knowledge Center. (accessed June 2017).
42. Benedict, FG, Goodall, HW, Ash, JE, et al. (1915) A Study of Prolonged Fasting. Washington, DC: Carnegie Institution of Washington.
43. Nair, KS, Woolf, PD, Welle, SL, et al. (1987) Leucine, glucose, and energy metabolism after 3 days of fasting in healthy human subjects. Am J Clin Nutr 46, 557562.
44. Pozefsky, T, Tancredi, RG, Moxley, RT, et al. (1976) Effects of brief starvation on muscle amino acid metabolism in nonobese man. J Clin Invest 57, 444449.
45. Fryburg, DA, Barrett, EJ, Louard, RJ, et al. (1990) Effect of starvation on human muscle protein metabolism and its response to insulin. Am J Physiol 259, E477E482.
46. Vendelbo, MH, Moller, AB, Christensen, B, et al. (2014) Fasting increases human skeletal muscle net phenylalanine release and this is associated with decreased mTOR signaling. PLOS ONE 9, e102031.
47. Cahill, GF Jr (1970) Starvation in man. N Engl J Med 282, 668675.
48. Saudek, CD & Felig, P (1976) The metabolic events of starvation. Am J Med 60, 117126.
49. Henson, LC & Heber, D (1983) Whole body protein breakdown rates and hormonal adaptation in fasted obese subjects. J Clin Endocrinol Metab 57, 316319.
50. Giesecke, K, Magnusson, I, Ahlberg, M, et al. (1989) Protein and amino acid metabolism during early starvation as reflected by excretion of urea and methylhistidines. Metabolism 38, 11961200.
51. Seashore, JH, Huszar, G & Davis, EM (1981) Urinary 3-methylhistidine/creatinine ratio as a clinical tool: Correlation between 3-methylhistidine excretion and metabolic and clinical states in healthy and stressed premature infants. Metabolism 30, 959969.
52. Young, VR, Haverberg, LN, Bilmazes, C, et al. (1973) Potential use of 3-methylhistidine excretion as an index of progressive reduction in muscle protein catabolism during starvation. Metabolism 22, 14291436.
53. Lowry, SF, Horowitz, GD, Jeevanandam, M, et al. (1985) Whole-body protein breakdown and 3-methylhistidine excretion during brief fasting, starvation, and intravenous repletion in man. Ann Surg 202, 2127.
54. Hector, AJ, McGlory, C, Damas, F, et al. (2018) Pronounced energy restriction with elevated protein intake results in no change in proteolysis and reductions in skeletal muscle protein synthesis that are mitigated by resistance exercise. FASEB J 32, 265275.
55. Urhausen, A, Gabriel, H & Kindermann, W (1995) Blood hormones as markers of training stress and overtraining. Sports Med 20, 251276.
56. Lee, DY, Kim, E & Choi, MH (2015) Technical and clinical aspects of cortisol as a biochemical marker of chronic stress. BMB Rep 48, 209216.
57. Kelly, DM & Jones, TH (2013) Testosterone: a metabolic hormone in health and disease. The J Endocrinol 217, R25R45.
58. Chennaoui, M, Desgorces, F, Drogou, C, et al. (2009) Effects of Ramadan fasting on physical performance and metabolic, hormonal, and inflammatory parameters in middle-distance runners. Appl Physiol Nutr Metab 34, 587594.
59. Wilson, JM, Lowery, RP, Joy, JM, et al. (2013) β-Hydroxy-β-methylbutyrate free acid reduces markers of exercise-induced muscle damage and improves recovery in resistance-trained men. Br J Nutr 110, 538544.
60. Durkalec-Michalski, K & Jeszka, J (2016) The effect of β-hydroxy-β-methylbutyrate on aerobic capacity and body composition in trained athletes. J Strength Cond Res 30, 26172626.
61. Portal, S, Zadik, Z, Rabinowitz, J, et al. (2011) The effect of HMB supplementation on body composition, fitness, hormonal and inflammatory mediators in elite adolescent volleyball players: a prospective randomized, double-blind, placebo-controlled study. Eur J Appl Physiol 111, 22612269.
62. Hoffman, JR, Cooper, J, Wendell, M, et al. (2004) Effects of beta-hydroxy beta-methylbutyrate on power performance and indices of muscle damage and stress during high-intensity training. J Strength Cond Res 18, 747752.
63. Durkalec-Michalski, K & Jeszka, J (2015) The efficacy of a beta-hydroxy-beta-methylbutyrate supplementation on physical capacity, body composition and biochemical markers in elite rowers: a randomised, double-blind, placebo-controlled crossover study. J Int Soc Sports Nutr 12, 31.
64. Crowe, MJ, O’Connor, DM & Lukins, JE (2003) The effects of beta-hydroxy-beta-methylbutyrate (HMB) and HMB/creatine supplementation on indices of health in highly trained athletes. Int J Sport Nutr Exerc Metab 13, 184197.
65. Wilson, JM, Lowery, RP, Joy, JM, et al. (2014) The effects of 12 weeks of beta-hydroxy-beta-methylbutyrate free acid supplementation on muscle mass, strength, and power in resistance-trained individuals: a randomized, double-blind, placebo-controlled study. Eur J Appl Physiol 114, 12171227.
66. Asadi, A, Arazi, H & Suzuki, K (2017) Effects of β-hydroxy-β-methylbutyrate-free acid supplementation on strength, power and hormonal adaptations following resistance training. Nutrients 9, 1316.
67. Chrousos, GP (2009) Stress and disorders of the stress system. Nat Rev Endocrinol 5, 374381.
68. Wijngaarden, MA, Bakker, LE, van der Zon, GC, et al. (2014) Regulation of skeletal muscle energy/nutrient-sensing pathways during metabolic adaptation to fasting in healthy humans. Am J Physiol Endocrinol Metab 307, E885E895.
69. Yonamine, CY, Teixeira, SS, Campello, RS, et al. (2014) Beta hydroxy beta methylbutyrate supplementation impairs peripheral insulin sensitivity in healthy sedentary Wistar rats. Acta Physiologica 212, 6274.
70. Nunes, EA, Goncalves-Neto, LM, Ferreira, FB, et al. (2013) Glucose intolerance induced by glucocorticoid excess is further impaired by co-administration with beta-hydroxy-beta-methylbutyrate in rats. Appl Physiol Nutr Metab 38, 11371146.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

British Journal of Nutrition
  • ISSN: 0007-1145
  • EISSN: 1475-2662
  • URL: /core/journals/british-journal-of-nutrition
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Full text views

Total number of HTML views: 6
Total number of PDF views: 25 *
Loading metrics...

Abstract views

Total abstract views: 217 *
Loading metrics...

* Views captured on Cambridge Core between 6th March 2018 - 20th March 2018. This data will be updated every 24 hours.