Metabolic and functional adaptation of the diaphragm to training with resistive loads.

To study the metabolic and functional changes that occur during training with inspiratory flow resistive loads, a chronically instrumented unanesthetized sheep preparation was used. Sheep were exposed to resistances ranging from 50 to 100 cmH2O.l-1.s, for 2-4 h/day, 5-6 days/wk, for a total of 3 wk. Load intensity was adjusted to maintain arterial Po2 (PaO2) above 60 Torr and arterial PCO2 (PaCO2) below 45 Torr. Training produced significant (P less than 0.05) increases in citrate synthase, 3-hydroxyacyl-CoA dehydrogenase, and cytochrome oxidase in the costal and crural diaphragm of the trained sheep (n = 9) compared with control sheep (n = 7). Phosphofructokinase did not increase. In the quadriceps, citrate synthase, 3-hydroxyacyl-CoA dehydrogenase, and phosphofructokinase did not change with training but cytochrome oxidase increased significantly (P less than 0.01). Function of the diaphragm was assessed in a subset of five sheep exposed to the same severe load 1 wk before and 2 days after the final training session. After training, sheep had a lower PaCO2 (10-40%), generated a higher transdiaphragmatic pressure (20-40%), and could sustain this level of transdiaphragmatic pressure for 0.5-2 h longer. The respiratory duty cycle was 10-15% lower, whereas minute ventilation and tidal volume were 20-30% higher in the posttraining test. We conclude that 1) training with inspiratory flow resistive loads improves the performance of the respiratory neuromuscular system and 2) the shift in enzyme profile of the diaphragm is at least in part responsible for this improvement.

Ventilatory response to fatiguing and nonfatiguing resistive loads in awake sheep.

To study the changes in ventilation induced by inspiratory flow-resistive (IFR) loads, we applied moderate and severe IFR loads in chronically instrumented and awake sheep. We measured inspired minute ventilation (VI), ventilatory pattern [inspiratory time (TI), expiratory time (TE), respiratory cycle time (TT), tidal volume (VT), mean inspiratory flow (VT/TI), and respiratory duty cycle (TI/TT)], transdiaphragmatic pressure (Pdi), functional residual capacity (FRC), blood gas tensions, and recorded diaphragmatic electromyogram. With both moderate and severe loads, Pdi, TI, and TI/TT increased, TE, TT, VT, VT/TI, and VI decreased, and hypercapnia ensued. FRC did not change significantly with moderate loads but decreased by 30-40% with severe loads. With severe loads, arterial PCO2 (PaCO2) stabilized at approximately 60 Torr within 10-15 min and rose further to levels exceeding 80 Torr when Pdi dropped. This was associated with a lengthening in TE and a decrease in breathing frequency, VI, and TI/TT. We conclude that 1) timing and volume responses to IFR loads are not sufficient to prevent alveolar hypoventilation, 2) with severe loads the considerable increase in Pdi, TI/TT, and PaCO2 may reduce respiratory muscle endurance, and 3) the changes in ventilation associated with neuromuscular fatigue occur after the drop in Pdi. We believe that these ventilatory changes are dictated by the mechanical capability of the respiratory muscles or induced by a decrease in central neural output to these muscles or both.

O2 metabolism of the sheep diaphragm during flow resistive loaded breathing.

To determine whether O2 availability limited diaphragmatic performance, we subjected unanesthetized sheep to severe (n = 11) and moderate (n = 3) inspiratory flow resistive loads and studied the phrenic venous effluent. We measured transdiaphragmatic pressure (Pdi), systemic arterial and phrenic venous blood gas tensions, and lactate and pyruvate concentrations. In four sheep with severe loads, we measured O2 saturation (SO2), O2 content, and hemoglobin. We found that with severe loads Pdi increased to 74.7 +/- 6.0 cmH2O by 40 min of loading, remained stable for 20-30 more min, then slowly decreased. In every sheep, arterial PCO2 increased when Pdi decreased. With moderate loads Pdi increased to and maintained levels of 40-55 cmH2O. With both loads, venous PO2, SO2, and O2 content decreased initially and then increased, so that the arteriovenous difference in O2 content decreased as loading continued. Hemoglobin increased slowly in three of four sheep. There were no appreciable changes in arterial or venous lactate and pyruvate during loading or recovery. We conclude that the changes in venous PO2, SO2, and O2 content may be the result of changes in hemoglobin, blood flow to the diaphragm, or limitation of O2 diffusion. Our data do not support the hypothesis that in sheep subjected to inspiratory flow resistive loads O2 availability limits diaphragmatic performance.

Adaptation of respiratory muscles to acute and chronic stress. Considerations on energy and fuels.

Although the majority of patients with acute or chronic pulmonary disease and respiratory failure do not have respiratory muscle disease or dysfunction at the outset of their clinical condition, most if not all do not have a normal respiratory musculature at the time of respiratory failure, especially those who have had a chronic course. Indeed, a large number of alterations in the muscle structure and metabolism occur as a result of a chronic load. Most of these changes are compensatory, but some may be maladaptive and deleterious to function. In addition, the compensatory mechanisms can be limited, and it is now believed that respiratory failure is at least in part exaggerated, or initiated, by the failure of the respiratory musculature. Therefore, the failure of these muscles can lead to hypoventilation, apnea, poor gas exchange, and clinically serious cardiovascular consequences. Although research efforts have increased in the past several years, there is still a great deal to learn about the function of the respiratory musculature under stress. Some of the notable questions are: What is the relation between inspiratory muscles (for example, the diaphragm and the intercostals) and the muscles of the airways during loaded breathing and in the presence of respiratory failure? How does the differential respiratory output to the various motor neuron pools change and evolve during the chronic loaded breathing? What kind of compensatory mechanisms (for example, humoral, biochemical, neural, or mechanical) can the respiratory muscles use to preserve function? What is the relation between the nutritional status of the individual and its effect on the function of these muscles? and What effect(s) do the changes associated with chronic pulmonary disease (for example, hypoxia, hypercapnea) have on respiratory muscle metabolism and function? This is an exciting area of research that has enormous potential for clinical applicability, and we believe that we are just at the very beginning.

Respiratory muscle response to load and glycogen content in type I and II fibers.

To study the relation between the response of respiratory muscle to inspiratory loads and glycogen content, we subjected unanesthetized sheep to moderate and severe inspiratory flow resistive (IFR) loads. Only severe IFR loads eventually led to a decrease in transdiaphragmatic pressure (Pdi) and a concomitant rise in PaCO2. Respiratory and nonrespiratory skeletal muscle samples were obtained at necropsy. Glycogen content was determined biochemically in muscle homogenates. Frozen sections were stained with periodic acid-Schiff (PAS) for glycogen and fibers were typed using myosin ATPase stain. Fibers were categorized as full, intermediate, or devoid of glycogen by a subjective scoring system of PAS staining intensity. We found that glycogen content decreased in the costal and crural diaphragm and in the intercostal muscles as the duration of moderate IFR loaded breathing was increased. With severe loads glycogen content decreased significantly, reaching about 40 and 22% of control levels in the costal and crural diaphragm, respectively (P less than 0.01). In addition, with severe IFR loads, a statistically significant proportion of both type I and type II muscle fibers was depleted of glycogen when compared with that of controls (P less than 0.05), but more type II fibers were depleted than type I fibers (50 vs 23%). These data indicate that in sheep subjected to IFR loads: (1) glycogen content in the respiratory muscles decreases as the severity and duration of loaded breathing increases and (2) respiratory muscle fatigue occurs at a time when considerable glycogen is still present in type I fibers in the diaphragm.