Structural muscle damage and muscle strength after incremental number of isometric and forced lengthening contractions

Summary Exercise-induced muscle damage is characterized by histological changes, like Z-line streaming, inflammatory response and decreased muscle function reflected in a prolonged decline in maximal isometric muscle strength after eccentric work. It is assumed that force decrement is mainly related to the amount of structural damage. However, the relationship between number of eccentric contractions, magnitude of structural damage and force decrement is not very well documented. Therefore we studied the effect of an increasing number of both isometric and eccentric (forced lengthening) contractions on histological parameters of muscle damage and maximal isometric force in an experimental in situ rat model. Tibialis anterior muscles of male Wistar rats were subjected to an increasing number of either isometric or eccentric contractions and were examined for histological markers of muscle damage. The present study shows that muscle damage increases progressively with the number of forced lengthening contractions. Maximal isometric torque was found to decline after both types of exercise. However, the decline after forced lengthening exercise was more pronounced. Only a weak relationship between percentage of histological muscle damage and isometric torque after forced lengthening contractions was found. The findings of the present study suggest that the decline in muscle force after eccentric exercise may partly be attributed to other factors than structural damage.     

Increased susceptibility of EDL muscles from mdx mice to damage induced by contractions with stretch

Summary Absence of dystrophin in mdx muscles may render the muscle more susceptible to damage when submitted to high stress levels. To test this, typically slow (soleus) and fast (EDL) limb muscles of dystrophic (mdx) and normal (C57BL/10) mice were submitted (in vitro) to a series of isometric contractions, followed by a series of contractions with stretches. Muscle injury was assessed by monitoring the force signal. Membrane damage was evaluated by bathing the muscle in Procion Red, a dye that does not penetrate intact fibres, and subsequent analysis by light microscopy. After isometric contractions, only a very small force drop (<3% of maximal isometric force) was observed which indicated that no injury had occurred in soleus and EDL muscles in either mdx or C57 strains. After contractions with a stretch, a force drop of 10% was observed in soleus muscles from both strains and in EDL muscles from C57 mice. However, in mdx mice EDL muscles displayed an irreversible force drop of 40–60%. Histological analysis of the muscles indicates that force drop is associated with membrane damage. These results show that EDL muscles from mdx mice are more vulnerable than their controls, supporting the structural role hypothesis for dystrophin. Furthermore, they suggest that contractions with stretches may contribute to the muscle damage and degeneration observed in DMD-patients.     

The disruption of myofibre structures in rat skeletal muscle after forced lengthening contractions

 Specific antibodies against structural proteins (actin, desmin, dystrophin, fibronectin) of muscle fibres were used to study the effect of forced lengthening contractions on muscle microarchitecture. Tibialis anterior (TA) muscle of male Wistar rats were subjected to 240 forced lengthening contractions. At consecutive time points (0, and 6 h, 2, 4, and 7 days) after stimulation, the TA muscle was excised for biochemical and histological assays. β-Glucuronidase activity, a quantitative indicator of muscle damage, showed increased values 2–7 days after the lengthening, peaking on day 4 (11.7-fold increase). A typical course of histopathological changes (myofibre swelling, necrosis and regeneration) was observed. In immunohistochemistry, the earliest abnormality observed was discontinuous dystrophin staining in some swollen fibres immediately after commencement of exercise, while at the same time no alterations occurred in the staining of the other antibodies studied. Six hours later, all the swollen fibres were uniformly desmin as well as dystrophin negative. The great majority, but not all, of the swollen fibres showed disorganized actin staining and intramyocellular localization of fibronectin. The early phase disruption of myofibre structures as measured in this study provides evidence of their central role following damage in skeletal muscle. These results suggest that the sequence of structural changes in the route to muscle fibre necrosis in injury induced by forced lengthening contraction originates in the disruption of the plasma membrane and the intermediate filament, which leads to disturbances in the myofibrillar system. Received: 26 January 1998 / Received after revision: 12 May 1998 / Accepted: 14 May 1998     

Metabolic cost of lengthening, isometric and shortening contractions in maximally stimulated rat skeletal muscle

AIM: The present study investigated the energy cost of lengthening, isometric and shortening contractions in rat muscle (n = 19). METHODS: With electrical stimulation the rat medial gastrocnemius muscle was maximally stimulated to perform 10 lengthening, isometric and shortening contractions (velocity 25 mm s(-1)) under experimental conditions (e.g. temperature, movement velocity) that resemble conditions in human movement. RESULTS: Mean +/- SD force-time-integral of the first contraction was significantly different between the three protocols, 2.4 +/- 0.2, 1.7 +/- 0.2 and 1.0 +/- 0.2 N s, respectively (P < 0.05). High-energy phosphate consumption was not significantly different between the three modes of exercise but a trend could be observed from lengthening (7.7 +/- 2.7 micromol approximately P muscle(-1)) to isometric (8.9 +/- 2.2 micromol approximately P muscle(-1)) to shortening contractions (10.4 +/- 1.6 micromol approximately P muscle(-1)). The ratio of high-energy phosphate consumption to force-time-integral was significantly lower for lengthening [0.3 +/- 0.1 micromol approximately P (N s)(-1)] and isometric [0.6 +/- 0.2 micromol approximately P (N s)(-1)] contractions compared with shortening [1.2 +/- 0.2 micromol approximately P (N s)(-1)] contractions (P < 0.05). CONCLUSION: The present results of maximally stimulated muscles are comparable with data in the literature for voluntary human exercise showing that the energy cost of force production during lengthening exercise is approximately 30% of that in shortening exercise. The present study suggests that this finding in humans probably does reflect intrinsic muscle properties rather than effects of differential recruitment and/or coactivation.     

Motor unit synchronisation is enhanced during slow lengthening contractions of a hand muscle

This study examined the strength of motor unit synchronisation based on time- and frequency-domain measures during postural, shortening and lengthening contractions of a hand muscle in young adults. Single motor unit activity was recorded with intramuscular electrodes in the left first dorsal interosseus muscle as the subject held the index finger at a constant position while supporting a light load for 2-5 min. The subject then performed slow (1.7 deg s⁻¹) shortening and lengthening contractions to lift and lower the load. The movement required subjects to perform 10-25 constant-velocity contractions with the index finger over a 10 deg range of motion by using 6 s shortening and lengthening contractions. Individual discharge times were obtained from 23 pairs of motor units in 14 subjects to assess the strength of motor unit synchronisation and coherence during the three tasks. The strength of motor unit synchronisation was approximately 50 % greater during the lengthening contractions compared with the postural and shortening contractions, and the width of the central synchronous peak in the cross-correlation histogram was ≈4 ms narrower during shortening contractions. These findings reveal that there is an increase in common input to motoneurones during lengthening contractions and a greater relative contribution of direct common inputs to motoneurones during shortening contractions compared with postural tasks. Furthermore, the amount of motor unit coherence in the low-frequency band (2-12 Hz) was reduced during shortening contractions compared with postural and lengthening contractions. These data indicate that the timing of inputs received by the motoneurones innervating the first dorsal interosseus of young adults differs during postural, shortening and lengthening contractions against a light load.     

Differences in stretch reflex responses of elbow flexor muscles during shortening, lengthening and isometric contractions

Stretch reflexes were evoked in elbow flexor muscles undergoing three different muscle contractions, i.e. isotonic shortening (SHO) and lengthening (LEN), and isometric (ISO) contractions. The intermuscle relationships for the magnitude of the stretch reflex component in the eletromyographic (EMG) activities of two main elbow flexor muscles, i.e. the biceps brachii (BB) and the brachioradialis (BRD), were compared among the three types of contractions. The subjects were requested to move their forearms sinusoidally (0.1?Hz) against a constant pre-load between elbow joint angles of 10° (0°?=?full extension) and 80° during SHO and LEN, and to keep an angle of 45° during the ISO. The perturbations were applied at the elbow angle of 45° in pseudo-random order. The EMG signals were rectified and averaged over a period of 100?ms before and 400?ms after the onset of the perturbation 40–50 times. From the ensemble averaged EMG waveform, the background activity (BGA), short (20–50?ms) and long latency (M2, 50–80, M3, 80–100?ms) reflex and voluntary activity (100–150?ms) components were measured. The results showed that both BGA and reflex EMG activity of the two elbow flexor muscles were markedly decreased during the lengthening contraction compared to the SHO and ISO contractions. Furthermore, the changes of reflex EMG components in the BRD muscle were more pronounced than those in the BB muscle, i.e. the ratios of M2 and M3 magnitudes between BRD and BB (BRD:BB) were significantly reduced during the LEN contractions. These results would suggest that the gain of long latency stretch reflex EMG activities in synergistic muscles might be modulated independently according to the model of muscle contraction.     

Effects of repeated lengthening contractions on skeletal muscle adaptations in female rats

We examined the adaptation of plantar flexor muscles of female rats to 6 weeks (5 days/week) of lengthening contractions. After repeated lengthening contractions, a decrease in myofiber area of gastrocnemius medialis (26%) was accompanied by an increase in extracellular matrix (ECM) (42%) and collagen content (30.9%) without changes in muscle mass. Decrease in myofiber area (13%) and muscle mass of soleus (19%) was associated with increased collagen content (28%) and ECM (15%). Relative number of soleus myofibers stained for fast myosin increased by 26%. For plantaris, increases in collagen content (32.3%), percent ECM (17%), and myofiber area (6%) were recorded. We also observed (1) increases (3.3%) in the collagen content of the Achilles tendon, (2) no change in the crosslink content of any of the tissues tested, and (3) no difference in the force-frequency relationship of the plantar flexor muscles. Substantial decreases in myofiber areas with increases in muscle connective tissue by 6 weeks of repeated lengthening contractions did not appear to result in isometric force loss.     

Tetrodotoxin-resistant contractions induced by electrical stimulation of bladder muscle from man, rabbit and rat

Isolated detrusor preparations from man, rabbit and rat were suspended in an organ bath and isometric tension was recorded. The preparations were stimulated electrically in the presence of Bay K8644 and nifedipine before and after neuronal blockade with tetrodotoxin. Transmural electrical stimulation produced frequency-dependent contractions in all preparations. Bay K8644 significantly increased and nifedipine decreased these contractions. TTX effectively suppressed the response to electrical field stimulation in all species. When Bay K8644 was added to TTX blocked preparations, the responses to electrical stimulation were partly restored in bladder strips from man and rat. No increase in response was seen in the rabbit preparations. However, if the extracellular K+-concentration was increased to 10 mM (which per se did not affect the response) Bay K8644 significantly increased the contractions. All responses elicited by electrical stimulation in the presence of TTX were abolished by nifedipine. It is concluded that if the bladder smooth muscle is exposed to factors that can increase its sensitivity to contractile agents, this may result in uncontrolled (unstable) bladder contractions. Such contractions may use the 'normal' transmitter substances, but may be triggered at a lower stimulus intensity than normal. As a non-specific increase in membrane excitability seems to be associated with an influx of calcium through voltage-sensitive calcium channels, calcium antagonists, together with agents specifically blocking relevant transmitter substances, would offer an effective therapy against the unstable bladder.     

Low-intensity eccentric contractions attenuate muscle damage induced by subsequent maximal eccentric exercise of the knee extensors in the elderly

This study investigated whether low-intensity eccentric contractions of the knee extensors would attenuate the magnitude of muscle damage induced by maximal eccentric exercise of the same muscle performed 7 days later using elderly individuals. Healthy older men (66.4 ± 4.6 years) were assigned to control or experimental (Exp) group (n = 13 per group). The control group performed six sets of ten maximal eccentric contractions (MaxECC) of the knee extensors of non-dominant leg. The Exp group performed six sets of ten low-intensity eccentric contractions of the knee extensors on a leg extension machine by lowering a weight of 10 % maximal voluntary isometric knee extension strength (10 %ECC) 7 days prior to MaxECC. Changes in maximal voluntary isokinetic concentric torque (MVC-CON), angle at peak torque, range of motion (ROM), upper thigh circumference, muscle soreness, plasma creatine kinase activity and myoglobin (Mb) concentration and B-mode ultrasound echo-intensity before and for 5 days after MaxECC were compared between groups by a mixed factor ANOVA. No significant changes in any variables were observed following 10 %ECC. Following MaxECC, all variables changed significantly, and changes in all variables except for angle at peak torque were significantly different between groups. MVC-CON and ROM decreased smaller and recovered faster (P < 0.05) for Exp than control group, and changes in other variables were smaller (P < 0.05) for Exp group compared with control group. These results suggest that preconditioning knee extensor muscles with low-intensity eccentric contractions was effective for attenuating muscle damage induced by subsequent MaxECC of the knee extensors for elderly individuals.     

Pulsed ultrasound therapy accelerates the recovery of skeletal muscle damage induced by Bothrops jararacussu venom

We studied the effect of pulsed ultrasound therapy (UST) and antibothropic polyvalent antivenom (PAV) on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group) received a perimuscular injection of venom (1 mg/kg) and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm², pulsed mode). Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg) partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively). Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively). The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.     

Growth induced by incremental static stretch in adult rabbit latissimus dorsi muscle

Incrementally applied static stretch over 3 weeks resulted in a 72 % increase in the weight of the in situ latissimus dorsi muscle in rabbits. True growth rather than tissue oedema was confirmed by increases in the protein content (130 %), the cross-sectional area of the type I fibres (30 %) and the muscle length (i.e. number of sarcomeres in series increased 25 %). Despite an increase in the proportion of fibres staining positive for the enzyme succinate dehydrogenase (SDH), the myosin ATPase stain showed no appreciable fibre type transformation. While total power output in the stretched muscle was unchanged, its maximum mass specific power output, as determined by oscillatory work loops, was decreased by 50 %. The cross-sectional area that was occupied by connective tissue increased from 15 to 19 % in the stretched muscles, with a concomitant increase in passive energy dissipation. Some incrementally stretched muscles were then allowed an additional 3 weeks of maintained stretch to determine whether the adaptive changes would be preserved or reversed. Previous gains in muscle weight, length and area of type I fibres all remained. In contrast, the connective tissue content and the passive properties returned to control values during this period.     

The mechanism of myometrial contractions induced by endothelin-1 in rat

Experiments were performed to characterize endothelin-1-induced contractions and the role of endothelin (ET) receptor subtypes in rat myometrium. The binding sites of [(125)I]-ET-1 were saturable with high affinity. Scatchard plot analysis revealed that ET-1 binding sites in the myometrium constituted a single population. The dissociation equilibrium constant (Kd) and the maximum binding sites (Bmax) were determined to be 48.9+/-3.0 pM and 1364.0+/-210.3 fmol/mg protein respectively. Specific [(125)I]-ET-1 binding was inhibited completely by unlabelled ET-1 and Ro 46-2005 (mixed-type ET receptor antagonist), but not fully (90.7+/-1.4%) by BQ 123 (a selective ETA receptor antagonist), and not at all by RES 701-1 (a selective ETB receptor antagonist). ET-1 induced myometrial contractions were composed of two types, an increase in resting tone and rhythmic contractions. These contractions were inhibited by BQ 123 and Ro 46-2005, but not by RES 701-1. ET-1-induced contractions were greatly reduced in Ca2+-free Krebs' solution. Nifedipine abolished the rhythmic contractions without affecting the increase in resting tone. These results suggest that ETA receptors are predominantly localized in rat myometrium and that excitation of ETA receptors evokes two types of contractions by increasing the cytoplasmic Ca2+ concentration.      

局部注射布比卡因建立大鼠骨骼肌损伤模型的组织形态学评价

BACKGROUND: The neurotoxicity and cardiotoxicity of bupivacaine have been reported frequently. However, the studies on bupivacaine-induced muscle toxicity are few.     

Reduced voluntary activation of human skeletal muscle during shortening and lengthening contractions in whole body hyperthermia

This study examined the effect of whole body hyperthermia on the voluntary activation of exercised and non-exercised skeletal muscle performing a series of lengthening and shortening contractions. Thirteen subjects exercised on a cycle ergometer at 60% of maximal oxygen consumption until voluntary exhaustion in ambient conditions of approximately 40 degrees C and 60% relative humidity. Before and immediately following the cycle protocol, subjects performed a series of 25 continuous isokinetic shortening and lengthening maximal voluntary contractions (MVCs) of the leg extensors and forearm flexors. Voluntary activation for shortening and lengthening contractions for the forearm and leg was assessed prior to and following the 25 MVCs by superimposing a paired electrical stimulus to the femoral nerve and the biceps brachii during additional MVCs. Exercise to exhaustion increased rectal temperature to 39.35+/-0.50 degrees C. Voluntary activation remained unchanged following the prehyperthermia endurance set of shortening and lengthening maximal contractions in both the forearm flexors and leg extensors. Similarly, voluntary activation remained at prehyperthermic levels for the single MVCs immediately following the cycle trial. However, by the time of completion of the posthyperthermia endurance contractions, voluntary activation had declined significantly by 5.87+/-7.56 and 8.46+/-9.26% in the shortening and lengthening phases, respectively, for the leg extensors but not for the forearm flexors. These results indicate that the central nervous system (CNS) reduces voluntary drive to skeletal muscle performing both shortening and lengthening contractions following exercise-induced hyperthermia. The reductions in voluntary activation were only observed following a series of dynamic movements, indicating that the CNS allows for initial and brief 're-activation' of skeletal muscle following exercise-induced hyperthermia.