Effects of short vs. prolonged mechanical ventilation on antioxidant systems in piglet diaphragm.

OBJECTIVE: Prolonged controlled mechanical ventilation (MV) is known to induce diaphragmatic oxidative stress that seems to be an important factor reducing force-generating capacity. To better understand the cellular mechanisms involved, this work examined the effect of short vs. prolonged MV on antioxidant defense in the diaphragm. DESIGN AND SETTING: Prospective, randomized, controlled animal study in a university laboratory. METHODS: Eleven piglets (15-20 kg) were assigned to one of two groups: a long-MV group (n=6) ventilated for 3 days or a short-MV group (n=5) ventilated for 3 h. Force frequency curves of the transdiaphragmatic pressure (Pdi) were obtained in vivo by phrenic nerve pacing. Oxidative stress was evaluated by thiobarbituric reactive substance (TBARs) content and the enzymatic antioxidant activity of both superoxide dismutase (SOD) and glutathione peroxidase (GPx) in samples of diaphragm. RESULTS: Pdi decreased in the long-MV group by 30-35% over the 3 days at all frequencies compared to the short-MV group. Diaphragm TBARs content was significantly higher and SOD activity lower in long-MV animals than in short-MV animals after 72 h. GPx activity tended to be lower in diaphragms from long-MV animals, but this difference was not significant. CONCLUSIONS: This study shows that 3 days of MV in piglets is associated with a decrease in antioxidant activity which could emphasize oxidative stress and both contribute to the diaphragm dysfunction caused by MV.

Changes in respiratory muscle endurance during puberty.

The evaluation of respiratory muscle endurance provides clinically useful information on muscle function, especially in children with respiratory and neuromuscular diseases. However, endurance may be lower in young children than in older children because of the major physical changes of puberty. We thus compared respiratory muscle endurance in 15 healthy pre- and peripubertal children (S1-S2/P1 group) and 14 healthy children near the end of the pubertal process (S4P4 group). All performed a respiratory muscle endurance test threshold load fixed at 50% of the individual maximal inspiratory pressure; (Pi(max)), using a standardized method with a controlled breathing pattern. No significant difference was found between groups for Pi(max). The mean endurance time limit for the S1-S2/P1 group was 138 +/- 20 sec. The S4P4 group was able to breathe with the threshold valve for more than 20 min (1,200 sec) without task failure, except for one girl (385 sec). This study shows that inspiratory muscle endurance is significantly lower in children in early puberty compared to children at the end of the pubertal process. If the underlying mechanisms are not well-known, the present study revealed that if we use the same inspiratory load in prepubertal children as in adults during clinical testing, we are likely to underestimate the susceptibility to task failure of their respiratory muscles. To define a fatigue threshold for the respiratory muscles, as a function of age, thus appears clinically important in further studies, particularly for the management of children with respiratory diseases.

The effect of respiratory muscle training with CO2 breathing on cellular adaptation of mdx mouse diaphragm.

The aim of our study was to investigate the cellular mechanisms induced by hypercapnic stimulation of ventilation, during 6 weeks/30 min per day, in 10 mdx and 8 C57BL10 mice (10+/-0.2 months old). Ten mdx and eight C57BL10 mice served as control group. This respiratory training increases in vitro maximal tetanic tension of the diaphragm only in mdx mice. Western blot analysis of diaphragm showed: (1) an over-expression of alpha-dystrobrevin in mdx and C57BL10 training group compared to control group (8100+/-710 versus 6100+/-520 and 2800+/-400 versus 2200+/-250 arbitrary units); (2) a decrease in utrophin expression only in mdx training group compared to control group (2100+/-320 versus 3100+/-125 arbitrary units). Daily respiratory muscle training in mdx mice, induces a beneficial effect on diaphragm strength, with an over-expression of alpha-dystrobrevin. Further studies are needed to determine if, in absence of dystrophin, the over-expression of alpha-dystrobrevin could be interpreted as a possible pathway to improve function of dystrophic muscle.

Effect of age on Hsp72 expression in leukocytes of healthy active people.

Aging mammalians show reduced expression and induction of cytoprotective heat shock proteins in response to physiological stresses. Physical training can increase Hsp72 expression in young and old animals, but whether same adaptations can be observed in old people remains unknown. We hypothesized that the maintenance of physical activity during aging should preserve Hsp72 expression in leukocytes of elderly people. Intracellular and surface Hsp72 (the inducible form of Hsp70) expression in leukocytes as well as apoptotic and necrotic leukocytes were measured by flow cytometry at rest and after maximal incremental test on treadmill in the following groups: 8 young subjects (25.3+/-0.6 year, G25), 12 sexagenarians (66.2+/-1 year, G65) and 9 octogenarians (82.2+/-1.2 year, G85), all physically active. Protein and lipid oxidation markers were also measured at rest and post-exercise. Results showed significant lower basal percentages of Hsp72-positive lymphocytes in G85 compared to G25. At rest, lower mean fluorescence intensity in Hsp72-positive monocytes was measured in G65 and G85 compared to G25, and in granulocytes from G85 compared to G25. Maximal exercise test induced systemic oxidative-stress in the three groups but did not induce any increase in apoptotic or necrotic cells. We observed a significant increase in the percentage of Hsp72-positive lymphocytes from G85. This study showed that maintaining physical activity during aging can preserve the ability to induce Hsp72 in response to physiological stress.

Does oxidative stress alter quadriceps endurance in chronic obstructive pulmonary disease?

The role of exercise-induced oxidative stress in the reduced quadriceps endurance of chronic obstructive pulmonary disease (COPD) patients has never been shown. We conducted a randomized, double-blind, and crossover study in which nine severe patients performed localized dynamic quadriceps endurance tests at 40% of maximal strength after oral treatment with the antioxidant, N-acetylcysteine (NAC), and placebo. Venous blood was sampled before, immediately after exercise, and 6 hours later. Endurance time improved by 25% after NAC treatment compared with placebo (p < 0.05). Superoxide anion (oxidant) release by stimulated phagocytes decreased after treatment (p < 0.05). No change in the antioxidant system was observed. Lipid peroxidation, an index of oxidative stress, was significantly increased 6 hours after exercise in the placebo condition (p < 0.05) but not after treatment. Advanced oxidized protein products, another index of oxidative stress, were also increased 6 hours after exercise by 139 +/- 27% in the placebo condition but only by 54 +/- 19% after treatment (p < 0.05). This study shows that NAC treatment in COPD reduced basal disturbance in the prooxidant system, improved endurance time, and prevented exercise-induced oxidative stress. Oxidative stress thus seems to be implicated in the reduced quadriceps endurance of patients with COPD.

Exercise-induced quadriceps oxidative stress and peripheral muscle dysfunction in patients with chronic obstructive pulmonary disease.

Exercise-induced muscle oxidative stress may be involved in the myopathy associated with chronic obstructive pulmonary disease (COPD). This study was designed to look at whether local exercise induces muscle oxidative stress and whether this oxidative stress may be associated with the reduced muscle endurance in patients with COPD. Quadriceps endurance was measured in 12 patients with COPD (FEV1 = 0.96 +/- 0.14 SEM) and 10 healthy sedentary subjects by repeated knee extensions of the dominant leg. Biopsies of the vastus lateralis muscle were obtained before and 48 hours after exercise. Muscle oxidative stress was measured by lipid peroxidation and oxidized proteins. Muscle antioxidant was evaluated by peroxidase glutathion activity. Quadriceps endurance was significantly reduced in patients with COPD when compared with the healthy control subjects (p < 0.01). Forty-eight hours postexercise, only patients with COPD had a significant increase in muscle lipid peroxidation (p < 0.05) and oxidized proteins (p < 0.05), whereas increased peroxidase glutathion activity was only observed in control subjects (p < 0.05). Both increases in muscle lipid peroxidation and oxidized proteins were significantly and inversely correlated with quadriceps endurance capacity in COPD (p < 0.05). In summary, local exercise induced muscle oxidative stress in patients with COPD, whereas it failed to raise antioxidant activity. In these individuals, muscle oxidative stress was associated with a reduced quadriceps endurance.