Sprint-interval training-induced alterations of Myosin heavy chain isoforms and enzyme activities in rat diaphragm: effect of normobaric hypoxia.

The purpose of this study was twofold: (i) to investigate if sprint-interval training (SIT) alters myosin heavy chain (MyHC) isoform composition and bioenergetic properties within the rat diaphragm, and (ii) to determine if mild normobaric hypoxia would enhance the effects of SIT-induced diaphragmatic adaptation. Male Wistar rats (8 weeks old) were randomly assigned to one of four groups (n = 7/group): (i) normoxic control (NC); (ii) normoxic training (NT); (iii) hypoxic control (HC); or (iv) hypoxic training (HT). The NT and HT groups were engaged in SIT (1 min sprint and 2-5 min rest, 6-10 sets/day, 5-6 days/week) on a treadmill for 9 weeks. Animals in the HC and HT groups were exposed to normobaric hypoxia (14.5% O(2)) during an SIT program from the 4th week of the training period. After completion of the training program, MyHC composition, citrate synthase (CS) activity, and lactate dehydrogenase (LDH) activity in the diaphragm and plantaris muscle were analyzed. An analysis of diaphragmatic MyHC composition demonstrated increased type IIa and decreased type IId/x for both training groups (P < 0.05), with the HT group producing greater changes than the NT group (P < 0.05). The plantaris muscle, however, showed increased Type IIa and IId/x and decreased Type IIb for both the NT and HT groups (P < 0.05). CS activity increased only for the training groups (P < 0.05), and this change was greater for the HT group in the diaphragm and for the NT group in the plantaris muscle (P < 0.05). Further, diaphragmatic LDH activity in HT was significantly lower (P < 0.05) than in HC and NT. These findings demonstrated that SIT could induce alterations in MyHC composition from fast to slow within type II isoforms and also improve the oxidative capacity in the diaphragm and plantaris muscles. It is of importance that our data revealed that SIT-induced diaphragmatic adaptations were enhanced when SIT was performed in normobaric hypoxia.

[Analysis of respiratory muscle structure and tumor necrosis and insulin-like growth factor expression in chronic obstructive pulmonary disease: are samples valid if obtained during thoracotomy performed because of localized pulmonary neoplasia?]. / Análisis estructual y expresión de los factores de necrosis tumoral y crecimiento insulina-like en los músculos respiratorios de pacientes con EPOS. Son válidas las muestras obtenidas en el curso de una toracotomía por neoplasia pulmonar localizada?

OBJECTIVE: Various methods have been used to obtain samples to study the structure of human respiratory muscles and the expression of diverse substances in them. Samples are most often obtained from autopsies, from muscle biopsies during thoracotomy performed because of a localized pulmonary lesion (TLL), and from ambulatory thoracoscopic biopsy in patients free of comorbidity (AT). The disadvantage of the first 2 of these methods lies in the possibility of interference from factors related to the patient's death in the first case or from the disease that necessitated surgery in the second. Although AT is free from the disadvantages of the other 2 methods, it is impossible to obtain samples of the diaphragm the principal respiratory muscle with this procedure. The objective of this study was to analyze the fibrous structure of the external intercostal muscle of patients with chronic obstructive pulmonary disease and to quantify the expression of the principal inflammatory cytokine tumor necrosis factor alpha (TNF-alpha)- and of insulin-like growth factor (IGF-1) in the same muscle, comparing the results obtained with TLL and AT samples. METHODS: Prospective and consecutive samples were taken of the external intercostal muscle (fifth space, anterior axillary line) in 15 patients with chronic obstructive pulmonary disease (mean [SD] age 66 [6] years; forced expiratory volume in 1 second 49% [9%] of predicted; PaO2 75 [9] mm Hg). Samples were taken during TLL (8 patients, all with pulmonary neoplasms but carefully selected in order to rule out systemic effects) or TA (7 patients). Patients with serious comorbidity were excluded from the second group. Samples were processed for structural analysis of fibers (immunohistochemical and enzymatic histochemical) and genetic expression of TNF-alpha and IGF-1 (real-time polymerase chain reaction). RESULTS: No differences in the structure of fibers were found between the 2 groups. No differences were observed in the expression of TNF-alpha or IGF-1. CONCLUSIONS: Using rigorous criteria, the TLL method appears to be suitable for studying the structural characteristics and expression of inflammatory cytokines and growth factors in the external intercostal muscle. Moreover, it can also be inferred that TLL is probably also useful for obtaining samples of the diaphragm, a muscle which cannot currently be sampled by any alternative method.

Effects of the beta(2)-agonist clenbuterol on respiratory and limb muscles of weaning rats.

The aim of this study was to analyze the effects of chronic administration of the beta(2)-agonist clenbuterol (1.5 mg x kg(-1) x day(-1) for 4 wk in the drinking water) on respiratory (diaphragm and parasternal intercostal) and hindlimb (tibialis and soleus) muscles in young rats during postnatal development (21 to 49 postnatal days). The treatment resulted in very little stimulation of muscle growth. Significant slow-to-fast transitions in the expression of myosin heavy chain isoforms and significant increases in the myofibrillar ATPase activity were found in the diaphragm and soleus, whereas tibialis anterior and intercostal muscles did not show any significant fiber-type alteration. Decrease of oxidative enzyme activities and increase of glycolytic enzyme activities were also observed. It is concluded that whereas the growth stimulation is age dependent and only detectable in adult rats, the fiber-type transformation is also present in weaning rats and particularly evident in the soleus and diaphragm. The fiber-type transformation caused by clenbuterol might lead to an enhancement of contractile performance and also to a reduced resistance to fatigue.

Hypoxemia-induced modification of troponin I and T in canine diaphragm.

Impaired muscle function (fatigue) may result, in part, from modification of contractile proteins due to inadequate O(2) delivery. We hypothesized that severe hypoxemia would modify skeletal troponin I (TnI) and T (TnT), two regulatory contractile proteins, in respiratory muscles. Severe isocapnic hypoxemia (arterial partial pressure of O(2) of approximately 25 Torr) in six pentobarbital sodium-anesthetized spontaneously breathing dogs increased respiratory frequency and electromyographic activity of the diaphragm and internal and external obliques, with death occurring after 131-285 min. Western blot analysis revealed proteolysis of TnI and TnT, 17.5- and 28-kDa fragments, respectively, and higher molecular mass covalent complexes, one of which (42 kDa) contained TnI (or some fragment of it) and probably TnT in the costal and crural diaphragms but not the intercostal or abdominal muscles. These modifications of myofibrillar proteins may provide a molecular basis for contractile dysfunction, including respiratory failure, under conditions of limited O(2) delivery.

Role of beta-adrenergic mechanisms in exercise training-induced metabolic changes in respiratory and locomotor muscle.

To test the hypothesis that beta-adrenergic stimulation is required for the normal increase in oxidative capacity of respiratory and locomotor skeletal muscle in response to exercise training, we examined the effects of beta-blockade on muscle oxidative capacity in trained and sedentary rats. Thirty-four female adult Sprague-Dawley rats were randomly divided into four experimental groups: 1) trained+propranolol (TP); 2) trained + sham injection (TS); 3) sedentary + propranolol (SP); and 4) sedentary + sham injection (SS). Training increased (p < 0.05) citrate synthase (CS) activity in the plantaris (+29%) and costal diaphragm (+12%) of TS animals compared to SS animals. In contrast, training did not (p > 0.05) increase costal diaphragm CS activity in TP animals compared to the SS group. Further, although training increased (p < 0.05) plantaris CS activity in the TP group (+18%) compared to the SP group, the training-induced increase in muscle CS activity was 11% lower (p < 0.05) than observed in TS animals. Collectively, these results suggest that beta-adrenergic mechanisms may play a role in the normal training-induced increase in oxidative capacity in both respiratory and locomotor skeletal muscles.