Effects of β-hydroxy-β-methylbutyrate free acid and cold water immersion on expression of CR3 and MIP-1β following resistance exercise

Am J Physiol Regul Integr Comp Physiol. 2014 Apr 1;306(7):R483-9. doi: 10.1152/ajpregu.00542.2013. Epub 2014 Feb 5.


The inflammatory response to muscle-damaging exercise requires monocyte mobilization and adhesion. Complement receptor type 3 (CR3) and macrophage inflammatory protein (MIP)-1β enables monocyte recruitment, adhesion, and subsequent infiltration into damaged muscle tissue. The purpose of this study was to examine the effects of cold water immersion (CWI) and/or β-hydroxy-β-methylbutyrate free acid (HMB-FA) on CR3 expression and MIP-1β concentration after four sets of up to 10 repetitions of squat, dead lift, and split squat exercises at 70-80% 1-repetition maximum. Thirty-nine resistance-trained men (22.2 ± 2.5 yr) were randomly divided into four groups: 1) placebo (PL), 2) HMB-FA, 3) HMB-FA-CWI, and 4) PL-CWI. The HMB-FA groups ingested 3 g/day, and CWI groups were submersed into 10-12°C water for 10 min after exercise. Blood was sampled at baseline (PRE), immediately post- (IP), 30 min post- (30P), 24 h post- (24P), and 48 h post (48P)-exercise. Circulating MIP-1β was assayed and CR3 expression on CD14+ monocytes was measured by flow cytometry. Without treatment, CR3 expression significantly elevated at 30P compared with other time points (P = 0.030-0.047). HMB-FA significantly elevated the percentage of monocytes expressing CR3 between IP and 24P (P = 0.046) and between IP and 48P (P = 0.046). No time effect was observed for MIP-1β concentration. The recovery modalities showed to attenuate the rise in CR3 following exercise. Additionally, supplementation with HMB-FA significantly elevated the percentage of monocytes expressing CR3 during recovery. Although the time course that inflammatory responses are most beneficial remains to be determined, recovery modalities may alter immune cell mobilization and adhesion mechanisms during tissue recovery.

Keywords: CD11b; macrophage-1 antigen, supplement; muscle damage; recovery.

Publication types

  • Randomized Controlled Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Chemokine CCL4 / blood*
  • Cold Temperature*
  • Double-Blind Method
  • Florida
  • Humans
  • Immersion*
  • Macrophage-1 Antigen / blood
  • Macrophage-1 Antigen / drug effects*
  • Male
  • Monocytes / drug effects*
  • Monocytes / immunology
  • Monocytes / metabolism
  • Muscle Contraction*
  • Muscle, Skeletal / drug effects*
  • Muscle, Skeletal / immunology
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / pathology
  • Recovery of Function
  • Resistance Training*
  • Time Factors
  • Valerates / pharmacology*
  • Water*
  • Young Adult


  • CCL4 protein, human
  • Chemokine CCL4
  • Macrophage-1 Antigen
  • Valerates
  • Water
  • beta-hydroxyisovaleric acid