Soleus Research Articles

Muscle structure assessment using synchrotron radiation X-ray micro-computed tomography in murine with cerebral ischemia.

Muscles are crucial for balance and walking, activities which depend specifically on the lower extremity muscles. Therefore, the evaluation of stroke induced atrophy and paralysis is essential; however, determining the extent of damage in the days after its occurrence remains challenging. In this study, we evaluated ischemic stroke-induced soleus muscle damage in gerbils using synchrotron radiation X-ray micro-computed tomography (SR-µCT), comparing a control group (n = 3), animals 7 days after stroke (7 d, n = 3), and animals 14 days after stroke (14 d, n = 3). The left muscle was paralyzed, whereas the right muscle was not. Subsequently, we quantified the assessment by segmenting the soleus muscle based on the extracellular space/matrix and fiber region to determine the degree of damage. The muscle fiber-to-extracellular space/matrix ratio were significantly damaged due to paralysis on the left side (control vs. 14 d, P = 0.040). Muscle area was significantly different at 14 d between the left and right sides (P = 0.010). Additionally, the left local fascicle surface area, thickness, global pennation angle, and local fascicle angle were significantly different between the control and 14 d groups (P = 0.002, P = 0.007, P = 0.005, and P = 0.014 respectively). These findings underscore the potential of post-stroke animal studies in improving rehabilitation treatment for the central nervous system by assessing the degree of muscle recovery.

Curcumin Mitigates Muscle Atrophy Potentially by Attenuating Calcium Signaling and Inflammation in a Spinal Nerve Ligation Model

Denervation-induced calcium/calmodulin-dependent protein kinase II (CaMKII) activation and inflammation can result in muscle atrophy. Curcumin and bisdemethoxycurcumin are well known to exhibit an anti-inflammatory effect. In addition, curcumin has been shown to attenuate CaMKII activation in neuronal cells. This study aimed to examine the effect of curcumin or bisdemethoxycurcumin on CaMKII activation, inflammation, and muscle cross-sectional area (CSA) in spinal nerve ligated rats. Sixteen female rats were assigned to sham (CON), spinal nerve ligation (SNL), SNL+ curcumin 100 mg/kg BW (100CUR), and SNL+ bisdemethoxycurcumin 50 mg/kg BW (50CMO) for 4 weeks. Ipsilateral (surgical) soleus and tibialis anterior (TA) muscles was stained for dystrophin to measure CSA. Ipsilateral and contralateral (non-surgical) plantaris muscles were analyzed for protein content for acetylcholine receptor (AChR), CaMKII, CaMKIIThr286, nuclear factor-κB (NF-κB), NF-κBSer536, and interleukin-1β (IL-1β) and normalized to α-tubulin and then CON. A significant (p < 0.050) group effect was observed for TA CSA where CON (11,082.25 ± 1617.68 μm2; p < 0.001) and 100CUR (9931.04 ± 2060.87 μm2; p = 0.018) were larger than SNL (4062.25 ± 151.86 μm2). In the ipsilateral plantaris, the SNL (4.49 ± 0.69) group had greater CaMKII activation compared to CON (1.00 ± 0.25; p = 0.010), 100CUR (1.12 ± 0.45; p = 0.017), and 50CMO (0.78 ± 0.19; p = 0.009). The ipsilateral plantaris (2.11 ± 0.66) had greater IL-1β protein content than the contralateral leg (0.65 ± 0.14; p = 0.041) in the SNL group. In plantaris, the SNL (1.65 ± 0.51) group had greater NF-κB activation compared to CON (1.00 ± 0.29; p = 0.021), 100CUR (0.61 ± 0.10; p = 0.003), 50CMO (0.77 ± 0.25; p = 0.009) groups. The observed reduction in Ca2+ signaling and inflammation in type II plantaris muscle fibers might reflect the changes within the type II TA muscle fibers which may contribute to the mitigation of TA mass loss with curcumin supplementation.

Sucla2 Knock-Out in Skeletal Muscle Yields Mouse Model of Mitochondrial Myopathy With Muscle Type-Specific Phenotypes.

Pathogenic variants in subunits of succinyl-CoA synthetase (SCS) are associated with mitochondrial encephalomyopathy in humans. SCS catalyses the conversion of succinyl-CoA to succinate coupled with substrate-level phosphorylation of either ADP or GDP in the TCA cycle. This report presents a muscle-specific conditional knock-out (KO) mouse model of Sucla2, the ADP-specific beta subunit of SCS, generating a novel invivo model of mitochondrial myopathy. The mouse model was generated using the Cre-Lox system, with the human skeletal actin (HSA) promoter driving Cre-recombination of a CRISPR-Cas9-generated Sucla2 floxed allele within skeletal muscle. Inactivation of Sucla2 was validated using RT-qPCR and western blot, and both enzyme activity and serum metabolites were quantified by mass spectrometry. To characterize the model invivo, whole-body phenotyping was conducted, with mice undergoing a panel of strength and locomotor behavioural assays. Additionally, exvivo contractility experiments were performed on the soleus (SOL) and extensor digitorum longus (EDL) muscles. SOL and EDL cryosections were also subject to imaging analyses to assess muscle fibre-specific phenotypes. Molecular validation confirmed 68% reduction of Sucla2 transcript within the mutant skeletal muscle (p < 0.001) and 95% functionally reduced SUCLA2 protein (p < 0.0001). By 3 weeks of age, Sucla2 KO mice were 44% the size of controls by body weight (p < 0.0001). Mutant mice also exhibited 34%-40% reduced grip strength (p < 0.01) and reduced spontaneous exercise, spending about 88% less cumulative time on a running wheel (p < 0.0001). Contractile function was also perturbed in a muscle-specific manner; although no genotype-specific deficiencies were seen in EDL function, SUCLA2-deficient SOL muscles generated 40% less specific tetanic force (p < 0.0001), alongside slower contraction and relaxation rates (p < 0.001). Similarly, a SOL-specific threefold increase in mitochondria (p < 0.0001) was observed, with qualitatively increased staining for both COX and SDH, and the proportion of Type 1 myosin heavy chain expressing fibres within the SOL was nearly doubled (95% increase, p < 0.0001) in the Sucla2 KO mice compared with that in controls. SUCLA2 loss within murine skeletal muscle yields a model of SCS-deficient mitochondrial myopathy with reduced body weight, muscle weakness and exercise intolerance. Physiological and morphological analyses of hindlimb muscles showed remarkable differences in exvivo function and cellular consequences between the EDL and SOL muscles, with SOL muscles significantly more impacted by Sucla2 inactivation. This novel model will provide an invaluable tool for investigations of muscle-specific and fibre type-specific pathogenic mechanisms to better understand SCS-deficient myopathy.

Comprehensive corrective exercise program improves ankle function in female athletes with limited weight-bearing ankle dorsiflexion: A randomized controlled trial.

Limited ankle dorsiflexion range of motion is one of the most important risk factors for lower limb injury, which changes the biomechanics and the neuromuscular control of the lower limb muscles. This study aims to test the effectiveness of a comprehensive corrective exercise program (CCEP) on the range of motion, proprioception, dynamic balance, and muscle activation in female athletes with limited weight-bearing lunge ankle dorsiflexion range of motion. 30 female athletes aged 15 to 25 years with dorsiflexion under 34° were randomized to two groups. The intervention group (n = 15) received eight weeks of CCEP including soft tissue mobilization, joint mobilization, stretching, and strengthening, and the control (n = 15) group did not receive any intervention. range of motion, proprioception, dynamic balance, and muscle activation were assessed before and after the intervention. The training group showed clinically acceptable and statistically significant changes in ankle dorsiflexion range of motion (ES = 0.714), balance (ES = 0.423), and proprioception (ES = 0.253; P < 0.05). There were significant changes in the activity of the tibialis anterior and soleus muscles in the dynamic overhead squat test (descending and ascending phases) and the activity of the medial gastrocnemius in the descending phase decreased significantly (P < 0.05). No significant change was observed in the activity of the peroneus longus muscle (P > 0.05). The findings show that CCEP appears to be beneficial in increasing dorsiflexion range of motion, proprioception, balance, and decreasing ankle muscle activity among individuals with limited ankle dorsiflexion. Improving the dorsiflexion range of motion may be promising for reducing ankle sprain injury.

The Effect of Geranylgeraniol and Ginger on Satellite Cells Myogenic State in Type 2 Diabetic Rats

Type 2 diabetes (T2D) is associated with increased inflammation and reactive oxygen species (ROS) in muscles, leading to basal satellite cell (SC) myogenic impairment (i.e., reduction in SC pool), which is critical for maintaining skeletal muscle mass. T2D may contribute to muscle atrophy, possibly due to reductions in the SC pool. Geranylgeraniol (GGOH) and ginger can reduce inflammation and enhance SC myogenesis in damaged muscles, thereby alleviating muscle atrophy; however, their effect on basal SC myogenic state and muscle mass in T2D rats is limited. Rats consumed a control diet (CON), high-fat diet with 35 mg/kg of streptozotocin (HFD), a HFD with 800 mg/kg body weight of GGOH (GG), or a HFD with 0.75% ginger root extract (GRE). In the eighth week, their soleus muscles were analyzed for Pax7, MyoD, and MSTN gene and protein expression, SC myogenic state, and muscle cross-sectional area (CSA). The HFD group had a significantly lower number of Pax7+/MyoD− and Pax7+/MSTN+ cells, less Pax7 and MyoD gene expression, and less MyoD and MSTN protein expression, with a smaller CSA than the CON group. Compared to the GG and GRE groups, the HFD group had a significantly lower number of Pax7+/MSTN+ cells, less MyoD protein expression, and smaller CSA. The GRE group also had a significantly lower number of Pax7−/MyoD+ and greater MSTN protein expression than the HFD group. Nevertheless, the CON group had a significantly greater number of Pax7+/MyoD− than the GG and GRE groups, and a greater number of Pax7−/MyoD+ cells than the GRE group with a larger CSA than the GG group. GGOH and ginger persevered muscle CSA, possibly through increased MyoD and the ability to maintain the SC pool in T2D rats.

Effects of Supervised Exercise Therapy on Muscle Function During Walking in Patients with Peripheral Artery Disease

Background: Although supervised exercise therapy (SET) is a primary treatment for peripheral artery disease (PAD), the current literature is limited regarding the mechanisms contributing to increased walking distances, including how lower extremity muscle function is altered after SET. This study aimed to investigate the effects of SET on lower extremity muscle function during walking in patients with PAD. Methods: Twelve patients with PAD participated in a 6-month SET program consisting of three weekly exercise sessions (a total of 72 sessions) and adhered to the American College of Sports Medicine’s (ACSM) recommendations. Each session started with a 5 min warm-up of mild walking and static stretching of upper and lower body muscles, followed by 50 min of intermitted exercise on a treadmill, and then finished with 5 min of cool-down activities similar to the warm-up. Each patient walked across a 10 m pathway with reflective markers on their lower limbs twice: before (baseline) and after six months of participation in SET (post-exercise). Marker coordinates and ground reaction forces were recorded and imported to OpenSim software (version 4.0) for gait simulations. Muscle force, muscle power, and metabolic rate were estimated from OpenSim and compared between the baseline and post-exercise. Results: The mean plantar flexor force was not altered after SET. However, individuals’ plantar flexor muscles demonstrated improvements in force production (lateral gastrocnemius: 75–80% of stance, Cohen’s d = 0.20–0.43; medial gastrocnemius: 65–85% of stance, Cohen’s d = 0.20–0.71; soleus: 90–95% of stance, Cohen’s d = 0.20–0.26). Furthermore, plantar flexor power increased (80–95% of stance, Cohen’s d = 0.20–0.39) and this was attributed to increased power in the lateral gastrocnemius (80–85% of stance, Cohen’s d = 0.20–0.47), medial gastrocnemius (80–90% of stance, Cohen’s d = 0.22–0.60), and soleus muscles (85–95% of stance, Cohen’s d = 0.20–0.49). Similarly, other muscle groups (knee extensors, knee flexors, hip abductors, hip adductors, hip extensors, and hip flexors) also exhibited force and power increases after SET. Additionally, force and power variances were significantly decreased in several muscle groups (plantar flexors, knee extensors, hip abductors, hip external rotators, hip extensors, and hip flexors). Total metabolic rate also increased during the stance period where muscle force and power were elevated after SET (early stance: 5–25%, Cohen’s d = 0.20–0.82; mid stance: 35–45%, Cohen’s d = 0.20–0.47; late stance: 75–80%, Cohen’s d = 0.20–0.36). Conclusions: Our results suggest that from a biomechanics perspective, muscle functions during walking are improved in patients with PAD after SET; however, the improvements were generally small and were not reflected by all muscle groups.

Stress Granule Assembly in Pulmonary Arterial Hypertension

The role of stress granules (SGs) in pulmonary arterial hypertension (PAH) is unknown. We hypothesized that SG formation contributes to abnormal vascular phenotypes, and cardiac and skeletal muscle dysfunction in PAH. Using the rat Sugen/hypoxia (SU/Hx) model of PAH, we demonstrate the formation of SG puncta and increased expression of SG proteins compared to control animals in lungs, right ventricles, and soleus muscles. Acetazolamide (ACTZ) treatment ameliorated the disease and reduced SG formation in all of these tissues. Primary pulmonary artery smooth muscle cells (PASMCs) from diseased animals had increased SG protein expression and SG number after acute oxidative stress and this was ameliorated by ACTZ. Pharmacologic inhibition of SG formation or genetic ablation of the SG assembly protein (G3BP1) altered the SU/Hx-PASMC phenotype by decreasing proliferation, increasing apoptosis and modulating synthetic and contractile marker expression. In human PAH lungs, we found increased SG puncta in pulmonary arteries compared to control lungs and in human PAH-PASMCs we found increased SGs after acute oxidative stress compared to healthy PASMCs. Genetic ablation of G3BP1 in human PAH-PASMCs resulted in a phenotypic switch to a less synthetic and more contractile phenotype. We conclude that increased SG formation in PASMCs and other tissues may contribute to PAH pathogenesis.

Kinetic changes of gait initiation in individuals with chronic ankle instability: A systematic review.

Gait initiation (GI) in individuals with chronic ankle instability (CAI) has shown differences in the center of pressure (COP) and muscular measures compared to healthy controls. Some studies reported that these alterations appeared when GI was with the affected leg, while others indicated that they occurred when GI was with the non-affected leg. This systematic review aimed to understand kinetic and muscular differences between individuals with CAI, healthy controls, and the affected and non-affected legs of individuals with CAI. PubMed, Science Direct, Web of Science, Google Scholar, and Scopus databases (1990-2023) were searched using the Population, Exposure, Comparator, and Outcome measure. The PRISMA guidelines were followed. The outcome measures were the peak and rate of COP displacement in the medial-lateral and anterior-posterior directions, and resultant plane during phases 1, 2, and 3 of COP trace during GI and the duration of each phase. The other measures included the onset time of the tibialis anterior and soleus muscle activity between individuals with CAI, healthy controls, and the affected and non-affected legs of the individuals with CAI. The studies' quality assessment was conducted based on the Strengthening the Reporting of Observational Studies in Epidemiology checklist. Five studies were included in the final evaluation. The results of included studies showed, individuals with CAI spent less time during phases 1 and 2, as well as a shorter peak of COP displacement in the lateral direction during phase 1 compared to healthy controls, regardless of whether the GI was with the affected or non-affected leg. Individuals with CAI have probably adopted a strategy involving adjusting the peak of COP displacement to manage internal sway while in a single-leg stance. Overall, there was no comprehensive conclusion about differences between the two legs in individuals with CAI.

Relationship Between the Amount of Change in Echo Intensity and Young’s Modulus of the Soleus Muscle After Ankle Fracture Surgery

Relationship Between the Amount of Change in Echo Intensity and Young’s Modulus of the Soleus Muscle After Ankle Fracture Surgery

Beneficial effects of the SGLT2 inhibition on oxidative skeletal muscle in infarcted rats

Abstract Introduction Metabolic, morphological, and biochemical changes in skeletal muscle are common in chronic heart failure patients. Despite the cardioprotective effects of sodium-glucose co-transporter 2 (SGLT2) inhibition in heart failure, the impact of this drug on skeletal muscle remains poorly understood. This study investigated the effects of SGLT2 inhibitor empagliflozin (EMPA) on the soleus muscle of rats with myocardial infarction (MI)-induced heart failure. Methods One week after MI induction, male Wistar rats were divided into Sham (n=10), Sham+EMPA (n=12), MI (n=10) and MI+EMPA (n=09) groups. EMPA was added to rat chow (5 mg/kg/day) for 12 weeks. Cardiac remodeling was evaluated by echocardiogram. Enzymatic activity and oxidative stress marker concentrations were assessed by spectrophotometry; protein expression was evaluated by Western blotting. After histological analysis of the left ventricle (LV), only rats with MI greater than 35% of total LV area were included in the study. Statistical analysis: ANOVA and Tukey or Kruskal–Wallis and Dunn and t test; p&amp;lt;0.05. Results Infarction size did not differ between groups. Infarcted groups had larger LV systolic and diastolic diameters, LV diastolic area (Sham 47±7.1; Sham+EMPA 48±4.5; MI 100±16.1*; MI+EMPA 85±10.1#† mm2; p&amp;lt;0.05: * vs Sham; # vs Sham+EMPA; † vs MI), and left atrial diameter (Sham 5.62±0.2; Sham+EMPA 5.65±0.3; MI 7.69±1.3*; MI+EMPA 6.85±0.9#† mm; p&amp;lt;0.05: * vs Sham; # vs Sham+EMPA; † vs MI); and lower LV posterior wall shortening velocity and ejection fraction than control groups. Echocardiographic changes were attenuated in MI+EMPA compared to MI. Other results are shown in Picture 1. Conclusion SGLT2 inhibitor empagliflozin attenuates infarction-induced cardiac remodeling in rats. Empagliflozin prevents increase in oxidative stress, modulates protein turnover by IGF-1 pathway, and attenuates morphological and biochemical changes in the soleus muscle of infarcted rats.

Decreased number of satellite cells-derived myonuclei in both fast- and slow-twitch muscles in HeyL-KO mice during voluntary running exercise

BackgroundSkeletal muscles possess unique abilities known as adaptation or plasticity. When exposed to external stimuli, such as mechanical loading, both myofiber size and myonuclear number increase. Muscle stem cells, also known as muscle satellite cells (MuSCs), play vital roles in these changes. HeyL, a direct target of Notch signaling, is crucial for efficient muscle hypertrophy because it ensures MuSC proliferation in surgically overloaded muscles by inhibiting the premature differentiation. However, it remains unclear whether HeyL is essential for MuSC expansion in physiologically exercised muscles. Additionally, the influence of myofiber type on the requirement for HeyL in MuSCs within exercised muscles remains unclear.MethodsWe used a voluntary wheel running model and HeyL-knockout mice to investigate the impact of HeyL deficiency on MuSC-derived myonuclei, MuSC behavior, muscle weight, myofiber size, and myofiber type in the running mice.ResultsThe number of new MuSC-derived myonuclei was significantly lower in both slow-twitch soleus and fast-twitch plantaris muscles from exercised HeyL-knockout mice than in control mice. However, expect for the frequency of Type IIb myofiber in plantaris muscle, exercised HeyL-knockout mice exhibited similar responses to control mice regarding myofiber size and type.ConclusionsHeyL expression is crucial for MuSC expansion during physiological exercise in both slow and fast muscles. The frequency of Type IIb myofiber in plantaris muscle of HeyL-knockout mice was not significantly reduced compared to that of control mice. However, the absence of HeyL did not affect the increased size and frequency of Type IIa myofiber in plantaris muscles. In this model, no detectable changes in myofiber size or type were observed in the soleus muscles of either control or HeyL-knockout mice. These findings imply that the requirement for MuSCs in the wheel-running model is difficult to observe due to the relatively low degree of hypertrophy compared to surgically overloaded models.

Effect of foot type on electromyography characteristics and synergy of lower limb muscles during running

The foot structure is associated with different running mechanics. The central nervous system is responsible for using the muscles through synergies during locomotion. The purpose of the present study was to examine the effect of foot structure on the electromyography factors and the synergy of the selected muscles of the lower extremity. Tibialis anterior, extensor digitorum longus, peroneus longus, soleus, biceps femoris, vastus lateralis, gastrocnemius lateralis and medialis muscles activity of 60 barefoot recreational runners with different foot structures was recorded while running at a speed of 3.3 m/s. Muscle activity was measured in the running cycle. Besides, muscle synergies were extracted using non-negative matrix factorization algorithm. The results showed that there were differences between groups with different foot type in muscle activity under different phases of running in some muscles. Furthermore, the findings indicated that the number of synergies was similar in different groups and the relative weight of muscles was not different across groups. In conclusion, despite the difference in muscle activity under different phases of the running cycle, muscle synergies are similar among the groups. This can indicate similar control by the central nervous system in runners with different arch structures while running and the observed changes in muscle activity can be attributed to the type of forces exerted on the body, the length-tension relationship, and changes in the direction of the lower limbs in people with different arch structures.

Nrf2 deficiency in muscle attenuates experimental autoimmune myositis-induced muscle weakness.

Idiopathic inflammatory myopathies (IIMs) are systemic autoimmune diseases characterised by muscle weakness. Although multiple physiological and pathological processes are associated with IIMs, T-lymphocyte infiltration into muscle plays a key role in the development and exacerbation of IIMs. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a key transcription factor that regulates inflammatory responses; therefore, muscle Nrf2 may serve an important role in the development of IIMs. In this study, we demonstrated that experimental autoimmune myositis (EAM) causes loss of muscle mass and function in oxidative and glycolytic muscles in C57BL/6 mice. EAM increased CD4+ and CD8+ T-lymphocyte infiltration, as well as interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α) mRNA expression in oxidative soleus and glycolytic extensor digitorum longus muscles, along with elevated chemokine mRNA levels (i.e. CCL3, CCL5, CXCL9, CXCL10 and CXCL16). IFN-γ and TNF-α treatments increased the mRNA expression levels of these chemokines in C2C12 myotubes. EAM also increased phosphorylated Nrf2 at Ser40 in soleus and glycolytic white vastus lateralis muscle. Although the expression of several chemokines was affected by Nrf2 activation following tert-butylhydroquinone treatment or Keap1 knockdown, CCL5 mRNA expression significantly increased in C2C12 myotubes and mouse skeletal muscle. Moreover, muscle-specific Nrf2 knockout in mice attenuates EAM-induced loss of muscle mass and function, which was associated with the inhibition of CCL5 mRNA expression, CD8+ T-lymphocyte infiltration and IFN-γ mRNA expression. Collectively, these findings reveal that regulating Nrf2 activity is a promising therapeutic approach for treating IIM-mediated muscle weakness. KEY POINTS: Experimental autoimmune myositis (EAM) causes loss of muscle mass and function. Loss of muscle mass and function in EAM were associated with increased chemokine mRNA expression (i.e. CCL3, CCL5, CXCL9, CXCL10 and CXCL16), T-lymphocyte infiltration and inflammatory cytokine mRNA expression (i.e. IFN-γ and TNF-α) in the skeletal muscle. EAM activated Nrf2 in muscle and increased Nrf2 activity in vivo and in vitro increased CCL5 mRNA expression. Muscle-specific Nrf2 knockout in mice attenuated EAM-induced muscle weakness by inhibiting CCL5 mRNA expression, CD8+ T-lymphocyte migration and IFN-γ mRNA expression in muscles. These results provide further evidence for the potential therapeutic targeting of Nrf2 to mitigate EAM-induced muscle weakness.

Key Anabolic Markers in Human M. Soleus after 21-Day Head-Down Tilt Bedrest

Prolonged bed rest can have a significant negative effect on skeletal muscle, leading to muscle wasting and reduced strength. This process can occur in as little as 10 days in healthy individuals, with the loss of muscle mass and strength being particularly pronounced during the first week of immobilization. Head-down tilt bed rest (HDT) is a method used to simulate the physiological changes that occur in weightlessness during spaceflight. This technique involves lying in bed with the head tilted downward. This paper is dedicated to the analysis of key anabolic markers of the soleus muscle during 21 days of HDT BR. The HDT BR experiment was conducted at the Institute of Biomedical Problems, Russian Academy of Sciences. Six healthy male volunteers, aged 25–35 years, were subjected to 21 days of strict bed rest with a tilt angle of –6°. A needle biopsy of the m.soleus was performed using the Bergström method before the start of HDT BR and on day 21 of HDT BR. The biopsy material was immediately frozen in liquid nitrogen for further Western blot and PCR analysis. Examination of mTORC1 substrates showed a significant decrease in p70 and 4EBP1 phosphorylation after HDT BR. We also observed a significant decrease in the phosphorylation of another ribosomal kinase, p90RSK, a significant increase in eEF2 phosphorylation and an increase in eEF2k mRNA expression. In addition, the phosphorylation of AMPK and its substrate ACC decreased after HDT BR. The data obtained in this work support the hypothesis that a decrease in protein synthesis, together with an increase in proteolysis, contributes to the development of human m. soleus atrophy after 21 days of HDT BR.

Metabolic pathways for removing reactive aldehydes are diminished in the skeletal muscle during heart failure.

Muscle wasting is a serious complication in heart failure patients. Oxidative stress and inflammation are implicated in the pathogenesis of muscle wasting. Oxidative stress leads to the formation of toxic lipid peroxidation products, such as 4-hydroxy-2-nonenal (HNE), which covalently bind with proteins and DNA and activate atrophic pathways. Whether the formation of lipid peroxidation products and metabolic pathways that remove these toxic products are affected during heart failure-associated skeletal muscle wasting has never been studied. Male C57BL/6J mice were subjected to sham and transverse aortic constriction (TAC) surgeries for 4, 8 or 14weeks. Different skeletal muscle beds were weighed, and the total cross-sectional area of the gastrocnemius muscle was measured via immunohistochemistry. Muscle function and muscle stiffness were measured by a grip strength meter and atomic force microscope, respectively. Atrophic and inflammatory marker levels were measured via qRT‒PCR. The levels of acrolein and HNE-protein adducts, aldehyde-removing enzymes, the histidyl dipeptide-synthesizing enzyme carnosine synthase (CARNS), and amino acid transporters in the gastrocnemius muscle were measured via Western blotting and qRT‒PCR. Histidyl dipeptides and histidyl dipeptide aldehyde conjugates in theGastrocnemius and soleus muscles were analyzed by LC/MS-MS. Body weight, gastrocnemius muscle and soleus muscle weights and the total cross-sectional area of the gastrocnemius musclewere decreased after 14weeks of TAC. Heart weight, cardiac function, grip strength and muscle stiffness were decreased in the TAC-operated mice. Expression of the atrophic and inflammatory markers Atrogin1 and TNF-α, respectively, was increased ~ 1.5-2fold in the gastrocnemius muscle after 14weeks of TAC (p < 0.05 and p = 0.004 vs sham). The formation of HNE and acrolein protein adducts was increased, and the expression of the aldehyde-removing enzyme aldehyde dehydrogenase (ALDH2) was decreased in the gastrocnemius muscle of TAC mice. Carnosine (sham: 5.76 ± 1.3 vs TAC: 4.72 ± 0.7nmol/mg tissue, p = 0.04) and total histidyl dipeptide levels (carnosine and anserine; sham: 11.97 ± 1.5 vs TAC: 10.13 ± 1.4nmol/mg tissue, p < 0.05) were decreased in the gastrocnemius muscle of TAC mice. Depletion of histidyl dipeptides diminished the aldehyde removal capacity of the atrophic gastrocnemius muscle. Furthermore, CARNS and TAUT protein expression were decreased in the atrophic gastrocnemius muscle. Our data reveals that reduced expression of ALDH2 and depletion of histidyl dipeptides in the gastrocnemius muscle during heart failure leads to the accumulation of toxic aldehydes and might contribute to muscle wasting.

Effects of altered contractile environment on muscle shape change in the triceps surae.

Skeletal muscles change shape when they contract. Current insights into the effects of shape change on muscle function have primarily come from experiments on isolated muscles operating at maximal activation levels. However, when muscles contract and change shape, the forces they apply onto surrounding muscles will also change. The impact of an altered contractile environment (i.e., mechanical behaviour of surrounding muscle) on muscle shape change, remains unknown. To address this, we altered the mechanical contributions of the gastrocnemii during isometric plantarflexion contractions [via changing knee angle] and determined if there were associated changes in how the muscles of the triceps surae bulged in thickness during a ramped contraction. We combined B-mode ultrasound imaging with surface electromyography to quantify the neuromechanical contributions of the medial (MG) and lateral gastrocnemius (MG) and soleus (SOL) muscles during isometric plantarflexion contractions. Our results demonstrated that at the same SOL activity levels, altering knee angle had no influence on the magnitude of muscle shape change (thickness) in the triceps surae muscles. We observed high levels of inter-individual variability in muscle bulging patterns, particularly in the knee flexed position, suggesting a complex relationship between muscle bulging and activation strategies in the triceps surae, which may be related to differences in muscle mechanical properties between participants or across muscles. Our findings highlight the dynamics of in vivo bulging interactions among muscles within the triceps surae and provide insights for future investigations into the impact of altered contractile environments on three-dimensional muscle deformations and force production.

Skeletal muscle fibre type-dependent effects of atorvastatin on the PI3K/Akt/mTOR signalling pathway and atrophy-related genes in rats

BackgroundOne of the probable causes of statin myotoxicity is an imbalance between protein synthesis and degradation. These processes are regulated by the PI3K/Akt/mTOR pathway and the ubiquitin‒proteasome system (UPS). The aim of this study was to assess whether the effects of atorvastatin on PI3K/Akt/mTOR pathway downstream proteins, the FoxO3a transcription factor and the UPS genes, i.e., MuRF-1 and MAFbx, depend on muscle fibre type.Methods and resultsAtorvastatin (50 mg/kg) was administered to Wistar rats. The levels of selected PI3K/Akt/mTOR pathway proteins were assayed via Western blotting, whereas MuRF-1, MAFbx and FoxO3a mRNA levels were measured using reverse transcription quantitative polymerase chain reaction (RT‒qPCR). Gomöri trichrome staining was performed to assess skeletal muscle pathology. A decrease in the P-Akt/Akt ratio was observed in the gastrocnemius muscle (MG), whereas an increase in the P-Akt/Akt ratio was observed in the soleus muscle (SOL). FoxO3a gene expression increased in the SOL and extensor digitorum longus (EDL) muscles. MuRF-1 gene expression increased in the MG, and MAFbx expression increased in the EDL. No histopathological changes were observed in any of the tested muscles.ConclusionsIn the absence of overt muscle damage, atorvastatin decreased the P-Akt/Akt ratio in the MG, indicating an increase in inactive Akt. Consistent with the decrease in Akt activation, rpS6 phosphorylation decreased. In SOL, atorvastatin increased the P-Akt/Akt ratio, indicating Akt activation. P-FoxO3a and the P-FoxO3a/FoxO3a ratio increased, suggesting that FoxO3a inactivation occurred. Moreover, in the SOL, atorvastatin did not affect the expression of atrophy-related genes. These findings indicate that atorvastatin has no adverse effect on the Akt pathway in the SOL. Our results showed that the effects of atorvastatin on the Akt signalling pathway and atrophy-related gene expression depend on muscle type.

Quantifying the Fascicular Changes in Recovered Achilles Tendon Patients Using Diffusion Magnetic Resonance Imaging and Tractography.

Regardless of the way of treatment, persistent deficits in calf muscles in recovered patients from Achilles tendon rupture (ATR) exist long-term postinjury. Studies on calf muscle changes mostly highlight morphological changes in the calf muscles and Achilles tendon. However, limited attention has been given to fascicular changes. Diffusion tensor imaging (DTI) can provide a better understanding of the characteristics and properties of tissues with organized microstructure. In the current study, we used DTI-derived indices (mean diffusivity (MD), fractional anisotropy (FA), and eigenvalues-λ 1, λ 2, and λ 3) and fiber tractography to better understand the soleus muscle after recovery from ATR by comparing the results of injured legs with healthy ones. Our findings suggest that the standard deviations of measured parameters (FA, MD, and eigenvalues) within the soleus muscle are better predictors of the changes associated with the ATR as compared to the control counterpart for the volumetric region of interest (ROI). Additionally, in four out of five participants, smaller tracts were observed in the injured leg compared to the healthy one, as evidenced by the fiber length distribution of the tracts. Altogether, this study demonstrates the feasibility of the DTI and fiber tractography approaches to quantify the fascicular changes in the individuals recovered from ATR.

HouShiHeiSan attenuates sarcopenia in middle cerebral artery occlusion (MCAO) rats

Ethnopharmacological relevancePhysical therapy is the main clinical treatment for limb symptoms after ischemic stroke, and there is a lack of reliable drug intervention programs. HouShiHeiSan (HS)comes from “Synopsis of the Golden Chamber”, where it is recorded: “seauelae of wind stroke and heaviness of limbs”, indicating this formulae is a promising opion for clinical practice. Aim of the studyThe aim of this study is to explore the therapeutic effect of HS on sarcopenia after ischemic stroke (ISS) by using the middle cerebral artery occlusion (MCAO) rats. Materials and methodsAfter 7 days of adaptive feeding Sprague-Dawley (SD) rats were randomly divided into sham and MCAO surgery groups. After MCAO operation, the agreement of the models was evaluated with a laser speckle instrument, and then, treatment groups were administered HS and related solvent. During the 7 days treatment period, the Zea-Longa score was used to assess the neural function, the treadmill for exercise capacity and traction instrument for grip strength. Besides, the physiological electrical signal system was used to record muscular electrical signals, while the muscle thickness was measured by ultrasound. After data acquisition on the 7th day after MCAO operation, the soleus muscle was dissected, and the indexes of length, weight of whole muscle tissue and cross-sectional area of muscular cells by H&E were recorded. Subsequently, mechanistic indicators were examined. MuRF1 and MAFbx expression was detected by immunohistochemistry (IHC). Furthermore, the expression level of more related indicators of muscular differentiation and cellular proterin balance, including mTOR, p-mTOR, AKT, p-AKT, p70s6k, p-p70s6, FOXO1, p-FOXO1, MyoD1, Myostatin, MuRF1 and MAFbx, were tested via Western blot. ResultsHS improved motor performance and promoted muscle regeneration in MCAO rats. In terms of motor ability, HS mixed with alcohol significantly improved the neurological function damage, reduce the weight loss, increase the running distance per unit time and increase the grip strength. The postoperative muscle electrical signal intensity increased, and muscle thickness, weight, and length were maintained. The HS with alcohol group significantly maintained the cross-sectional size of muscle cells and reduced the number of MyoD1 and myostatin-positive cells in the muscle tissue. It simultaneously promoted the expression of p-mTOR, p-AKT, p-p70s6k, and MyoD1 to promote the synthesis of muscle proteins and inhibited the expression of p-FOXO1, myostatin, MAFbx, and MuRF1 to reduce muscle protein degradation. ConclusionHS can enhance muscle protein synthesis and decrease protein breakdown by activating the AKT/mTOR/FOXO1 pathway, thereby preserving muscle health and enhancing motor performance following stroke in rats.

Antioxidant mito-TEMPO prevents the increase in tropomyosin oxidation and mitochondrial calcium accumulation under 7-day rat hindlimb suspension

After the first day of muscle disuse (unloading) mitochondria-derived ROS accumulate in the postural-tonic soleus muscle. It is known that excess of ROS can lead to the accumulation of intramitochondrial calcium and overload of mitochondria with calcium, can negatively affect mitochondrial function and fatigue resistance of soleus muscle.We assumed that the use of mitochondrial ROS scavenger mito-TEMPO will be able to prevent the unloading-induced disruption of mitochondrial functions and will help maintain soleus muscle fatigue resistance.To test this hypothesis, male rats were divided into 3 groups (n = 16 in each): vivarium control with placebo (C), 7-day hindlimb suspension with placebo (7HS) and 7-day hindlimb suspension with intraperitoneal administration of the mimetic superoxide dismutase mito-TEMPO at a dose of 1 mg/kg (7HSM). In the 7HS group, increased fatigue of the soleus muscle was found in the ex vivo test, accompanied with increased activity of ETC complex I and “leak” respiration, as well as a twofold increased content of oxidized tropomyosin (a marker of ROS level in tissues) and increase in intramitochondrial calcium compared to C. In 7HSM, the activity of ETC complex I and “leak” respiration had no significant differences from the control group, and the increase in intramitochondrial calcium and the content of oxidized tropomyosin was partially prevented, however, muscle fatigue was also significantly higher than in the control group. Thus, mitochondrial ROS under 7-day muscle unloading contribute to the accumulation of intramitochondrial calcium and oxidation of tropomyosin, but do not have a significant effect on soleus muscle function.