ATP-sensitive K+ channel blocker glibenclamide and diaphragm fatigue during normoxia and hypoxia

The role of ATP-sensitiveK⁺ channels in skeletal musclecontractile performance is controversial: blockers of these channels have been found to not alter, accelerate, or attenuate fatigue. Thepresent study reexamined whether glibenclamide affects contractile performance during repetitive contraction. Experiments systematically assessed the effects of stimulation paradigm, temperature, and presenceof hypoxia and in addition compared intertrain with intratrain fatigue.Adult rat diaphragm muscle strips were studied in vitro. At 37°Cand normoxia, glibenclamide did not significantly affect any measure offatigue during continuous 5- or 100-Hz or intermittent 20-Hzstimulation but progressively prolonged relaxation time during 20-Hzstimulation. At 20°C and normoxia, neither force nor relaxationrate was affected significantly by glibenclamide during 20-Hzstimulation. At 37°C and hypoxia, glibenclamide did notsignificantly affect fatigue at 5-Hz or intertrain fatigue during 20-Hzstimulation but reduced intratrain fatigue and prolonged relaxationtime during 20-Hz stimulation. These findings indicate that, althoughATP-sensitive K⁺ channels may beactivated during repetitive contraction, their activation has only amodest effect on the rate of fatigue development.

     

Effects of modulation of nitric oxide on rat diaphragm isotonic contractility during hypoxia.

Nitric oxide (NO) is essential for optimal myofilament function of the rat diaphragm in vitro during active shortening. Little is known about the role of NO in muscle contraction under hypoxic conditions. Hypoxia might increase the NO synthase (NOS) activity within the rat diaphragm. We hypothesized that NO plays a protective role in isotonic contractile and fatigue properties during hypoxia in vitro. The effects of the NOS inhibitor N(G)-monomethyl-l-arginine (l-NMMA), the NO scavenger hemoglobin, and the NO donor spermine NONOate on shortening velocity, power generation, and isotonic fatigability during hypoxia were evaluated (Po(2) approximately 7 kPa). l-NMMA and hemoglobin slowed the shortening velocity, depressed power generation, and increased isotonic fatigability during hypoxia. The effects of l-NMMA were prevented by coadministration with the NOS substrate l-arginine. Spermine NONOate did not alter isotonic contractile and fatigue properties during hypoxia. These results indicate that endogenous NO is needed for optimal muscle contraction of the rat diaphragm in vitro during hypoxia.     

Effects of respiratory muscle unloading on exercise-induced diaphragm fatigue.

We previously compared the effects of increased respiratory muscle work during whole body exercise and at rest on diaphragmatic fatigue and showed that the amount of diaphragmatic force output required to cause fatigue was reduced significantly during exercise (Babcock et al., J Appl Physiol 78: 1710, 1995). In this study, we use positive-pressure proportional assist ventilation (PAV) to unload the respiratory muscles during exercise to determine the effects of respiratory muscle work, per se, on exercise-induced diaphragmatic fatigue. After 8-13 min of exercise to exhaustion under control conditions at 80-85% maximal oxygen consumption, bilateral phrenic nerve stimulation using single-twitch stimuli (1 Hz) and paired stimuli (10-100 Hz) showed that diaphragmatic pressure was reduced by 20-30% for up to 60 min after exercise. Usage of PAV during heavy exercise reduced the work of breathing by 40-50% and oxygen consumption by 10-15% below control. PAV prevented exercise-induced diaphragmatic fatigue as determined by bilateral phrenic nerve stimulation at all frequencies and times postexercise. Our study has confirmed that high- and low-frequency diaphragmatic fatigue result from heavy-intensity whole body exercise to exhaustion; furthermore, the data show that the workload endured by the respiratory muscles is a critical determinant of this exercise-induced diaphragmatic fatigue.     

Changes in diaphragm motor unit EMG during fatigue

Fatigue-related changes in the waveform and root-mean-square (rms) values of evoked motor unit electromyographic (EMG) responses were studied in the right sternocostal region of the cat diaphragm. Motor units were isolated by microdissection and stimulation of C5 ventral root filaments and then classified as fast-twitch fatigable (FF), fast-twitch fatigue intermediate (FInt), fast-twitch fatigue resistant (FR), or slow-twitch (S) based on standard physiological criteria. The evoked EMG responses of S and FR units showed very little change during the fatigue test. The evoked EMG waveform and rms values of FF and FInt units displayed variable changes during the fatigue test. When changes were observed, they typically included a prolongation of the EMG waveform, a decrease in peak amplitude, and a decrease in rms value. The changes in EMG amplitude and rms values were not correlated. In more fatigable units, the decrease in force during the fatigue test generally exceeded the decrease in EMG rms values. Changes in the evoked force and EMG responses of multiple units innervated by C5 or C6 ventral roots were also examined during the fatigue test. The decrease in diaphragm force during the fatigue test closely matched the force decline predicted by the proportionate contribution of different motor unit types. However, the observed reduction in diaphragm EMG rms values during the fatigue test exceeded that predicted based on the aggregate contribution of different motor unit types. It was concluded that changes in EMG do not reflect the extent of diaphragm fatigue.     

Effects of hypoxia on metabolic rate of conscious adult cats during cold exposure

Oxygen consumption (VO2) and shivering movements were recorded in adult, conscious cats in a thermoneutral (24-27 degrees C) and in a cold (3-8 degrees C) environment during normoxia, hypoxia, or hyperoxia for 55 min. In the cold environment, VO2 correlated with shivering index (SI) under conditions of normoxia or ambient hypoxia (FIO2 = 0.12). During normoxia, VO2 was 63% higher in the cold than the thermoneutral environment. Ambient hypoxia acutely reduced VO2 in cold and thermoneutral environments, the decrement being greater for the former than the latter. Similarly, the variation in VO2 for unit change in SI was greater in hypoxia than normoxic conditions, suggesting that hypoxia influenced nonshivering as well as shivering components of cold-induced VO2. Hypoxia induced by CO (FICO = 0.002) also reduced VO2 and SI, a result that is consistent with previous results indicating that carotid body chemoreceptors do not mediate the suppression of shivering by ambient hypoxia. Hyperoxia increased VO2 and SI in the cold, and the effects of both hypoxia and hyperoxia in the cold were antagonized by increasing FICO2 to 0.03. The results demonstrate that hypoxia suppresses VO2 in the cold by reducing the intensity of shivering and, probably, by an action on metabolic rate that is unrelated to cold-induced calorigenesis.     

Muscle shortening increases sensitivity of fatigue to severe hypoxia in canine diaphragm.

The effects of inspired O2 on diaphragm tension development during fatigue were assessed using isovelocity (n = 6) and isometric (n = 6) muscle contractions performed during a series of exposures to moderate hypoxia [fraction of inspired O2 (FIO2) = 0.13], hyperoxia (FIO2 = 1), and severe hypoxia (FIO2 = 0.09). Muscle strips were created in situ from the canine diaphragm, attached to a linear ergometer, and electrically stimulated (30 Hz) to contract (contraction = 1.5 s/relaxation = 2 s) from optimal muscle length (Lo = 8.9 cm). Isovelocity contractions shortened to 0.70 Lo, resulting in a mean power output of 210 mW/cm2. Fatigue trials of 35 min duration were performed while inspired O2 was sequentially changed between the experimental mixtures and normoxia (FIO2 = 0.21) for 5-min periods. In this series, severe hypoxia consistently decreased isovelocity tension development by an average of 0.1 kg/cm2 (P less than 0.05), which was followed by a recovery of tension (P less than 0.05) on return to normoxia. These responses were not consistently observed in isometric trials. Neither isovelocity nor isometric tension development was influenced by moderate hypoxia or hyperoxia. These results demonstrate that the in situ diaphragm is relatively insensitive to rapid changes in O2 supply over a broad range and that the tension development of the shortening diaphragm appears to be more susceptible to severe hypoxia during fatigue. This may reflect a difference in either the metabolic or blood flow characteristics of shortening contractions of the diaphragm.     

Effects of DAP on diaphragm force and fatigue, including fatigue due to neurotransmission failure

Van Lunteren, Erik, and Michelle Moyer. Effects of DAPon diaphragm force and fatigue, including fatigue due toneurotransmission failure. J. Appl.Physiol. 81(5): 2214-2220, 1996.Among theaminopyridines, 3,4-diaminopyridine (DAP) is a more effectiveK⁺ channel blocker than is4-aminopyridine (4-AP), and, furthermore, DAP enhances neuromusculartransmission. Because 4-AP improves muscle contractility, wehypothesized that DAP would also increase force and, in addition,ameliorate fatigue and improve the neurotransmission failure componentof fatigue. Rat diaphragm strips were studied in vitro (37°C). Infield-stimulated muscle, 0.3 mM DAP significantly increased diaphragmtwitch force, prolonged contraction time, and shifted theforce-frequency relationship to the left without altering peak tetanicforce, resulting in increased force at stimulation frequencies 50 Hz.During 20-Hz intermittent stimulation, DAP increased diaphragm peakforce compared with control during a 150-s fatigue run and,furthermore, significantly improved maintenance of intratrain force.The relative contribution of neurotransmission failure to fatigue wasestimated by comparing the force generated by phrenic nerve-stimulatedmuscles with that generated by curare-treated field-stimulated muscles.DAP significantly increased force in nerve-stimulated muscles and, inaddition, reduced the neurotransmission failure contribution todiaphragm fatigue. Thus DAP increases muscle force atlow-to-intermediate stimulation frequencies, improves overall force andintratrain fatigue during 20-Hz intermittent stimulation, and reducesneurotransmission failure.

     

Changes in relaxation rate with diaphragmatic fatigue in humans

Maximum relaxation rate (MRR) and the time constant of relaxation (tau) of transdiaphragmatic pressure (Pdi) was measured in four male subjects and compared with the high-to-low frequency ratio (H/L) of the diaphragmatic electromyogram (EMG) as a predictor of diaphragmatic fatigue. Pdi and inspiratory time-to-total breath duration ratios (TI/TT) were varied, and TT and tidal volume were held constant; inspiratory resistances were used to increase Pdi. Studies were performed at various tension-time indices (TTdi = Pdi/Pdimax X TI/TT). Base-line MRR/Pdi was 0.0100 +/- 0.0004 (SE) ms-1, and baseline tau was 53.2 +/- 3.2 ms. At TTdi greater than 0.20, MRR and H/L decreased and tau increased, with maximum changes at the highest TTdi. At TTdi less than 0.20, there was no change in H/L, MRR, or tau. The time course of changes in H/L correlated with those of MRR and tau under fatiguing conditions. In this experimental setting, change in relaxation rate was as useful a predictor of diaphragmatic fatigue as fall in H/L of the diaphragmatic EMG.     

Effect of N-acetylcysteine on diaphragm fatigue

It has recently been postulated that diaphragm fatigue may be due, at least in part, to a form of low-grade injury to subcellular organelles. Moreover, several studies have shown that thiol-containing compounds can protect cardiac and striated skeletal muscle organelles from the deleterious effects of a number of physiological stresses. The purpose of the present study was to determine whether pretreatment with N-acetylcysteine (NAC), a thiol-containing compound, would attenuate the rate of development of diaphragmatic fatigue. Studies were performed with the use of an in situ rabbit diaphragm strip preparation that permitted direct and continuous measurement of diaphragm tension development. Diaphragm fatigue was induced by rhythmically stimulating strips to contract at 30/min (20-Hz trains) for 20 min. The diaphragm force-frequency relationship (10-, 20-, 50-, and 100-Hz stimuli) was assessed immediately before and after fatigue trials and then again 20 min into the period of recovery. Half the animals were treated with intravenous NAC before fatigue, whereas the remaining animals were given intravenous saline. The rate of development of fatigue was markedly greater in saline-treated control than in NAC-treated animals, with reductions in tension of 55 +/- 3 and 34 +/- 3%, respectively, in these two groups of animals over 20 min (P less than 0.001). Although rhythmic stimulation resulted in a downward shift in the force-frequency relationship in both NAC- and saline-treated animals, the magnitude of this shift was substantially greater in saline-treated animals (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of progressive hypoxia on parasternal, costal, and crural diaphragm activation

The distribution of motor drive to the costal and crural diaphragm and parasternal intercostal muscles was evaluated during progressive isocapnic hypoxia in anesthetized dogs. Bipolar stainless steel wire electrodes were placed unilaterally into the costal and crural portions of the diaphragm and into the parasternal intercostal muscle in the second or third intercostal space. Both peak and rate of rise of electromyographic activity of each chest wall muscle increased in curvilinear fashion in response to progressive hypoxia. Both crural and parasternal intercostal responses, however, were greater than those of the costal diaphragm. The onset of crural activation preceded that of the costal portion of the diaphragm and parasternal intercostal muscle activation. Despite differences in the degree of activation among the various chest wall muscles, the rate of increase in activation for any given muscle was linearly related to the rate of increases for the other two. This suggests that respiratory drive during progressive hypoxia increases in fixed proportion to the different chest wall inspiratory muscles. Our findings lend further support to the concept that the costal and crural diaphragm are governed by separate neural control mechanisms and, therefore, may be considered separate muscles.     

Effects of hypoxic hypoxia on O2 uptake and heart rate kinetics during heavy exercise

Engelen, Marielle, Janos Porszasz, Marshall Riley, KarlmanWasserman, Kazuhira Maehara, and Thomas J. Barstow. Effects ofhypoxic hypoxia on O₂ uptake andheart rate kinetics during heavy exercise. J. Appl.Physiol. 81(6): 2500-2508, 1996.It is unclearwhether hypoxia alters the kinetics ofO₂ uptake(O₂) during heavy exercise[above the lactic acidosis threshold (LAT)] and how thesealterations might be linked to the rise in blood lactate. Eight healthyvolunteers performed transitions from unloaded cycling to the sameabsolute heavy work rate for 8 min while breathing one of threeinspired O₂ concentrations: 21%(room air), 15% (mild hypoxia), and 12% (moderate hypoxia). Breathing12% O₂ slowed the time constantbut did not affect the amplitude of the primary rise inO₂ (period of first2-3 min of exercise) and had no significant effect on either thetime constant or the amplitude of the slowO₂ component (beginning2-3 min into exercise). Baseline heart rate was elevated inproportion to the severity of the hypoxia, but the amplitude andkinetics of increase during exercise and in recovery were unaffected bylevel of inspired O₂.We conclude that the predominant effect of hypoxia during heavyexercise is on the early energetics as a slowed time constant forO₂ and an additionalanaerobic contribution. However, the sum total of the processesrepresenting the slow component of O₂ is unaffected.

     

Respiratory sensation and pattern of respiratory muscle activation during diaphragm fatigue

We have examined the relationship between respiratory effort sensation (modified Borg scale) and amplitude of the integrated surface electromyogram of the diaphragm (Edi, esophageal electrode), rib cage muscles (Erc), and sternomastoid muscle (Esm) during the development of diaphragm fatigue in five normal subjects. Three conditions were studied: run A: transdiaphragmatic pressure (Pdi), 65% Pdimax; esophageal pressure (Pes), 60% Pesmax; run B: Pdi, 50% Pdimax; Pes, 60% Pesmax; and run C: Pdi, 50% Pdimax; Pes, 20% Pesmax. During all runs there was a progressive rise in sensation, which was greater in runs A and B than in run C (P less than 0.05, analysis of variance). There was no difference between runs A and B. At the end of run C subjects did not report a maximal Borg score despite their inability to generate the target Pdi. The increase in sensory score with fatigue correlated highly with Esm/Esmmax and with Erc/Ercmax. There was no correlation between sensory score and Edi/Edimax. We conclude that the increase in respiratory effort sensation that accompanies diaphragm fatigue is not due to perception of increased diaphragmatic activation. It may reflect increased overall respiratory motor output not directed to the diaphragm.     

Torque and mechanomyogram correlations during muscle relaxation: Effects of fatigue and time-course of recovery

To assess the validity and reliability of the mechanomyogram (MMG) as a tool to investigate the fatigue-induced changes in the muscle during relaxation, the torque and MMG signals from the gastrocnemius medialis muscle of 23 participants were recorded during tetanic electrically-elicited contractions before and immediately after fatigue, as well as at min 2 and 7 of recovery. The peak torque (pT), contraction time (CT) and relaxation time (RT), and the acceleration of force development (d2RFD) and relaxation (d2RFR) were calculated. The slope and τ of force relaxation were also determined. MMG peak-to-peak was assessed during contraction (MMG p–p) and relaxation (R-MMG p–p). After fatigue, pT, d2RFD, d2RFR, slope, MMG p–p and R-MMG p–p decreased significantly, while CT, RT and τ increased (P < 0.05 for all comparisons), remaining altered throughout the entire recovery period. R-MMG p–p correlated with pT, MMG p–p, slope, τ and d2RFR both before and after fatigue. Reliability measurements always ranged from high to very high. In conclusion, MMG may represent a valid and reliable index to monitor the fatigue-induced changes in muscle mechanical behavior, and could be therefore considered an effective alternative to the force signal, also during relaxation.