Fig 1: HO-1 is induced by aerobic exercise(A) HO-1 expression in WT and Hpx-/- mice over time in plantaris muscle after an acute aerobic exercise bout. Data represent mean ± SEM. *p < 0.05, ***p < 0.001 versus baseline; #p < 0.05, ##p < 0.01, ###p < 0.001 versus Hpx-/- baseline; n = 4–6/time point.(B–E) HO-1 mRNA and protein expression with representative immunoblots in the plantaris muscle and heart, respectively, after 6 weeks of aerobic exercise training in WT mice. Data represent mean ± SEM. *p < 0.05, **p < 0.01; n = 6/group.(F) Representative image of primary skeletal muscle myotube cells.(G and H) Hemopexin and HO-1 expression at different concentrations of heme in myotube cells.(I) Lipid peroxidation levels in myotubes after 4 h of treatment with heme (50 µM).(J) Myotubes silenced to Hpx (siHpx) and HO-1 (siHO-1) were evaluated for viability after treatment with heme (50 µM) and hemopexin (1 µM).Data represent mean ± SEM. *p < 0.05, **p < 0.01 versus control.
Fig 2: Effects of low (LCR) and high (HCR) intrinsic running capacity on heme processing(A and B) Serum heme and Hpx levels comparing LCR and HCR rats.(C and D) Hpx and HO-1 expression in plantaris skeletal muscle.(E–H) Hpx and HO-1 expression in liver and heart, respectively, comparing LCR and HCR animals.(I) Gene expression in plantaris muscle comparing LCR and HCR rats.Results are expressed as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001; n = 10/group.
Fig 3: Trauma injury releases free heme.(A) Levels of free heme in the plasma from healthy volunteers and trauma patients on day 1 (n = 6/group). (B and C) Levels of free heme (n = 3–10/group) and hemopexin (Hpx; n = 3–4/group) in the plasma after liver crush injury in mice. (D) Hmox1 mRNA expression levels in the injured left liver lobes and the uninjured right liver lobes after crush injury (normalized to beta-actin; n = 3–5/group). (E) HO-1 protein levels in the naive (n = 2) and injured liver lobes after crush injury (normalized to vinculin; n = 4/group). Western blot images are shown below. (F and G) Mean fluorescent intensity (MFI) of HO-1 in blood cells (WBC; n = 3/group) and BM cells 24 hours after liver crush injury (n = 4/group). (H) Levels of free heme in the plasma after liver crush injury in Hpx–/– mice (n = 3-4/group). (I) Fold change of S. aureus CFU in the BALF 24 hours after infection in C57BL/6 mice and Hpx–/– mice, both with liver crush injury performed 4 hours before the infection (n = 14/group). (J) Fold change of S. aureus CFU in the lungs 24 hours after infection in C57BL/6 mice with infection only (n = 11), trauma followed by infection (n = 7), and trauma followed by Hpx treatment and then infection (n = 3). All data are presented as mean ± SEM. Statistical analyses for A, F, G, and I were performed by unpaired 2-tailed Student’s t test. All other P values were calculated by 1-way ANOVA with post hoc Tukey’s test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig 4: Aerobic exercise promotes circulating heme release and upregulates hemopexin (Hpx) levels(A–D) Serum CK, LDH, heme, and Hpx levels in WT and Hpx-/- mice at different time points after acute running exercise on a treadmill. Data represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 versus WT baseline; #p < 0.05 versus Hpx-/- baseline; +p < 0.05, +++p < 0.001 WT versus Hpx-/-; n = 4–6/time point.(E–H) Serum heme and Hpx levels in the serum, heart, and plantaris muscle after 6 weeks of exercise training in WT (WTTR) mice.(I–K) Serum heme levels in Hpx-/- mice and running capacity in WT and Hpx-/- mice before and after 6 weeks of exercise training.(L) Flvcr2 gene expression in plantaris skeletal muscle before and after 6 weeks of aerobic exercise training in WT and Hpx-/- mice.Data represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001; n = 6/group.
Supplier Page from Novus Biologicals, a Bio-Techne Brand for Mouse Hemopexin ELISA Kit (Colorimetric)