A. G. Goachet

Effect of a 120 km endurance race on plasma and muscular neutrophil elastase and myeloperoxidase concentrations in horses.

REASONS FOR PERFORMING STUDY Intense physical exercise can induce the degranulation of neutrophils leading to an increase in plasma concentration of the neutrophil marker enzymes myeloperoxidase (MPO) and elastase (ELT). These enzymes have pro-oxidative and pro-inflammatory properties and may play a role in the exercised-induced muscular damage. OBJECTIVES To measure MPO and ELT concentrations in plasma and muscles of endurance horses and to correlate them to the extent of exercise-induced muscular damage. METHODS Seven endurance horses qualified on 120 km races were tested in this study. Neutrophil count, serum creatine kinase (CK), plasmatic and muscular MPO and ELT concentrations were measured before and 2 h after a 120 km endurance race. RESULTS The race produced a significant increase of neutrophils, CK, and plasma MPO and ELT levels. A significant correlation was observed between the MPO and ELT values in plasma (r(2) = 0.92, P < 0.01) and in muscles (r(2) = 0.89, P < 0.01) while plasmatic concentrations of MPO and ELT were not significantly correlated to muscular ones. An increase of mean concentrations (± s.e.) of MPO (T0: 9.85 ± 3.9, T1: 228.9 ± 95.9 ng/mg proteins) and ELT (T0: 8.4 ± 2.4, T1: 74.5 ± 39.7 ng/mg proteins) in the muscles were observed after the race. Interestingly, the individual data showed large differences between the horses. Muscular MPO and ELT concentrations were significantly correlated to plasma CK levels. The coefficient of correlation (r(2)) was 0.69 (P < 0.01) for MPO and 0.66 (P < 0.01) for ELT, respectively. CONCLUSIONS Results underline the possible role of MPO and ELT in exercise-induced muscular damage. POTENTIAL RELEVANCE Further studies should investigate the effect of exercise type and intensity, as well as the role of the training state on MPO and ELT involvement in muscular damage. The assessment of the intensity of exercise-induced neutrophilic degranulation may have a potential role in the monitoring of the athletic career.

Alterations in mitochondrial respiratory function in response to endurance training and endurance racing

REASONS FOR PERFORMING STUDY Limited information exists about the muscle mitochondrial respiratory function changes that occur in horses during an endurance season. OBJECTIVES To determine effects of training and racing on muscle oxidative phosphorylation (OXPHOS) and electron transport system (ETS) capacities in horses with high resolution respirometry (HRR). METHODS Mitochondrial respiration was measured in microbiopsies taken from the triceps brachii (tb) and gluteus medius (gm) muscles in 8 endurance horses (7 purebred Arabians and 1 crossbred Arabian) before training (T0), after two 10 week training periods (T1, T2) and after 2 CEI** endurance races (R1, R2). Muscle OXPHOS capacity was determined using 2 titration protocols without (SUIT 1) or with pyruvate (SUIT 2) as substrate. Electrons enter at the level of Complex I, Complex II or both complexes simultaneously (Complexes I+II). Muscle ETS capacity was obtained by uncoupling Complexes I+II sustained respiration. RESULTS T1 improved OXPHOS and ETS capacities in the tb as demonstrated by the significant increase of oxygen fluxes vs. T0 (Complex I: +67%; ETS: +37%). Training improved only OXPHOS in the gm (Complex I: +34%). Among horses that completed the race, a significant decrease in OXPHOS (Complex I: ∼ -35%) and ETS (-22%) capacities was found in the tb with SUIT 2 indicating a reduced aerobic glycolysis. Significant correlations between CK activities and changes in OXPHOS were found suggesting a relationship between exercise-induced muscle damage and depression of mitochondrial respiration. CONCLUSIONS For the first time, OXPHOS and ETS capacities in equine muscle at different steps of an endurance season have been determined by HRR. Significant alterations in mitochondrial respiratory function in response to endurance training and endurance racing have been observed although these changes appeared to be muscle group specific.

Altered systolic left ventricular function in horses completing a long distance endurance race.

REASONS FOR PERFORMING STUDY It is unknown whether or not exercise-induced cardiac fatigue (EICF), as demonstrated in human athletes performing long duration exercise, occurs in endurance horses. OBJECTIVE To examine the effects of a long distance endurance race on left ventricular systolic function in horses. METHODS Echocardiography was performed before, and after, a 2 or 3 star international endurance race (106-132 km) in 11 horses. Systolic (s) and diastolic (d) interventricular and left ventricular free wall thickness (IVS and LVFW, respectively), left ventricular, left atrial and aortic internal diameter (LVID, LA and Ao, respectively), fractional shortening (FS) and ejection fraction (EF) were measured by echocardiography. Heart rate (HR), peak flow velocity (Vmax), flow velocity integral (FVI), ejection time (ET), pre-ejection period (PEP), velocity of circumferential fibre shortening (Vcf), stroke volume (SV) and cardiac output (CO) were measured from aortic Doppler wave recordings. RESULTS After the race, LVIDd, Ao, LA, EF, FS, FVI, SV, ET and ET indexed for HR were significantly lower and IVSd, LVFWd, HR, PEP, PEP/ET and Vcf significantly higher as compared with prerace values. Pre- to post exercise changes in those parameters were not significantly correlated with changes in HR or in LVIDd. CONCLUSIONS Results suggest that EICF, with a decrease in left ventricular systolic function, could occur post exercise in horses performing long duration endurance races. However, a multanecus effect of altered preload and heart rate on the studied variables cannot be discounted.

Specific immuno-extraction followed by enzymatic detection (SIEFED) of myeloperoxidase and mitochondrial complex I in muscular microbiopsies: preliminary results in endurance horses

REASONS FOR PERFORMING STUDY Intense exercise in horses induces an increase of creatine kinase (CK) and stimulation of neutrophils which release the strong oxidant enzyme, myeloperoxidase (MPO) into the blood. It is not known whether active MPO is found in equine muscles and whether oxidant activity of neutrophils could affect muscular tissues and mitochondrial activity. OBJECTIVES Specific immuno-extraction followed by enzymatic detection (SIEFED) methods will be employed for the first time to assess both the normal range of MPO and mitochondrial complex I (MCI) activities in equine muscular microbiopsies and to study the variation of these activities induced by endurance races. MATERIALS AND METHODS Forty-six microbiopsies were taken from 8 endurance Arabian horses (age: 10 ± 2 years) in the triceps brachii (n = 23) or in the gluteus medius muscle (n = 23). Myeloperoxidase and MCI activities were measured in muscle extracts by enzyme immunocapture assays or SIEFED methods. Further, 7 endurance horses were sampled in the triceps brachii muscle before (T0) and after (T1) a 120 km endurance race (mean speed: 15.4 ± 1.4 km/h). RESULTS The 46 microbiopsies from 8 horses revealed mean values for active MPO concentration and MCI activity of 21 ± 14 ng/mg proteins and 0.0172 ± 0.0066 mOD/min/μg proteins, respectively. No significant difference was observed between the 2 muscles. In 3 out of the 7 horses sampled after exercise, the 120 km endurance race induced a severe increase of muscle MPO activity (+118 ± 45% vs. T0), a large decrease of MCI activity (-63 ± 18% vs. T0) associated with a high mean plasma CK level (4642 ± 658 iu/l). In the 4 remaining horses, the 120 km endurance race did not modify the MPO and MCI activities and moderately increased the plasma CK level. CONCLUSIONS Preliminary observations showed a possible link between MPO activity and mitochondrial functions.