Effect of treadmill exercise combined with bone marrow stromal cell transplantation on atrophy-related signaling pathway in the denervated soleus muscle

The purpose of this study was to investigate whether combination of low-intensity exercise with bone marrow stromal cell (BMSC) transplantation could regulate protein kinas B (Akt)- mammalian target of rapamycin (mTOR) and Wnt3a-β-catenin signaling pathways for prevention of soleus muscle atrophy after sciatic nerve injury (SNI). The experimental rats divided into 5 groups (n= 10): normal control group, SNI+sedentary group (SED), SNI+low-intensity treadmill exercise group (TEX), SNI+BMSC transplantation group (BMSC), SNI+TEX+BMSC transplantation group (TEX+BMSC). Sciatic nerve crush injury was applied into the middle of thigh twice for 1 min and 30 sec at interval. Low-intensity treadmill exercise was comprised of walking at a speed of 4 to 8 m/min for 30 min once a day. cultured BMSC at a density of 5× 106 in 50-μL phosphate-buffered saline was injected into the distal portion of the injured sciatic nerves. TEX+BMSC group dramatically upregulated expression levels of growth-associated protein-43 in the injured sciatic nerve at 2 weeks postinjury. Also, although Akt and mTOR signaling pathway significantly increased in TEX and BMSC groups than SED group, TEX+BMSC group showed more potent increment on this signaling in soleus muscle after SNI. Lastly, Wnt3a and the nuclear translocation of β-catenin and nuclear factor-kappa B in soleus were increased by SNI, but TEX+BMSC group significantly downregulated activity of this signaling pathway in the nuclear cell lysate of soleus muscle. Present findings provide new information that combination of low-intensity treadmill exercise might be effective therapeutic approach on restriction of soleus muscle atrophy after peripheral nerve injury.

Histomorphometric study of the soleus muscle under conditions of modeling of spinal cord contusion injury: experimental morphological study

Objective. To conduct a morphometric analysis of the soleus muscle of rats after moderate spinal cord contusion injury.Material and Methods. Experiments were performed on female Wistar rats aged 8–12 months, weighing 270–320 g. Animals of the experimental group (n = 25) underwent laminectomy at the T9 level under general anesthesia and modeling of spinal contusion injury of moderate severity. Intact rats constituted the control group (n = 10). Euthanasia was performed on the 5th, 15th, 30th, 60th, 90th, and 180th days of the experiment. Paraffin sections were stained with hematoxylin-eosin and Masson, the diameters of muscle fibers were determined by computer morphometry, and histograms of their distribution were obtained.Results. In the soleus muscle, the signs of reversible reparative processes prevailed in response to neurotrophic damage. It was evidenced by a local increase in the diversity of myocyte diameters and the loss of polygonality of their profiles, focal destruction of muscle fibers, activation of the connective tissue component, disorganization of some intramuscular nerve conductors, and vascular fibrosis of perimysium. Nevertheless, the histostructure of an intact muscle prevailed in the course of the experiment, which was confirmed by the data of morphometric analysis. All histograms of the distribution of the muscle fiber diameters are unimodal with a mode in the range of 30–41 μm. On the 180th day, the maximum myocyte diameters in the histogram of the left limb muscle belonged to the range of 21–30 μm, which was typical for histograms in the intact group.Conclusion. The nature of the plastic reorganization of the soleus muscle when neurotrophic control is impaired indicates compensatory regeneration of muscle tissue by the type of restitution, which opens up the possibility of predicting the rehabilitation period. It is advisable to take this into account when developing medical and social programs and therapeutic measures, where the most important role is played by superficial neuromuscular and functional electrical stimulation.

Corrigendum notice: Protective Effect of Interval Exercise Training with Different Intensity and Alpha-Lipoic Acid Supplement on Nav1.3 Protein in Soleus Muscle of Diabetic Rats

The article's abstract is not available.

Effects of Noninvasive Low-Intensity Focus Ultrasound Neuromodulation on Spinal Cord Neurocircuits In Vivo

Although neurocircuits can be activated by focused ultrasound stimulation, it is unclear whether this is also true for spinal cord neurocircuits. In this study, we used low-intensity focused ultrasound (LIFU) to stimulate lumbar 4–lumbar 5 (L4–L5) segments of the spinal cord of normal Sprague Dawley rats with a clapper. The activation of the spinal cord neurocircuits enhanced soleus muscle contraction as measured by electromyography (EMG). Neuronal activation and injury were assessed by EMG, western blotting (WB), immunofluorescence, hematoxylin and eosin (H&E) staining, Nissl staining, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), somatosensory evoked potentials (SEPs), motor evoked potentials (MEPs), and the Basso–Beattie–Bresnahan locomotor rating scale. When the LIFU intensity was more than 0.5 MPa, LIFU stimulation induced soleus muscle contraction and increased the EMG amplitudes ( P < 0.05 ) and the number of c-fos- and GAD65-positive cells ( P < 0.05 ). When the LIFU intensity was 3.0 MPa, the LIFU stimulation led to spinal cord damage and decreased SEP amplitudes for electrophysiological assessment ( P < 0.05 ); this resulted in coagulation necrosis, structural destruction, neuronal loss in the dorsal horn by H&E and Nissl staining, and increased expression of GFAP, IL-1β, TNF-α, and caspase-3 by IHC, ELISA, and WB ( P < 0.05 ). These results show that LIFU can activate spinal cord neurocircuits and that LIFU stimulation with an irradiation intensity ≤1.5 MPa is a safe neurostimulation method for the spinal cord.

HDAC4 Is Indispensable for Reduced Slow Myosin Expression at the Early Stage of Hindlimb Unloading in Rat Soleus Muscle

It is well known that reduced contractile activity of the main postural soleus muscle during long-term bedrest, immobilization, hindlimb unloading, and space flight leads to increased expression of fast isoforms and decreased expression of the slow isoform of myosin heavy chain (MyHC). The signaling cascade such as HDAC4/MEF2-D pathway is well-known to take part in regulating MyHC I gene expression. Earlier, we found a significant increase of HDAC4 in myonuclei due to AMPK dephosphorylation during 24 h of hindlimb unloading via hindlimb suspension (HU) and it had a significant impact on the expression of MyHC isoforms in rat soleus causing a decrease in MyHC I(β) pre-mRNA and mRNA expression as well as MyHC IIa mRNA expression. We hypothesized that dephosphorylated HDAC4 translocates into the nuclei and can lead to a reduced expression of slow MyHC. To test this hypothesis, Wistar rats were treated with HDAC4 inhibitor (Tasquinimod) for 7 days before HU as well as during 24 h of HU. We discovered that Tasquinimod treatment prevented a decrease in pre-mRNA expression of MyHC I. Furthermore, 24 h of hindlimb suspension resulted in HDAC4 nuclear accumulation of rat soleus but Tasquinimod pretreatment prevented this accumulation. The results of the study indicate that HDAC4 after 24 h of HU had a significant impact on the precursor MyHC I mRNA expression in rat soleus.

Electrical Stimulated Glut4 Signaling Attenuates Critical Illness-Associated Muscle Wasting

Background: Critical illness myopathy (CIM) is a debilitating condition characterized by the preferential loss of the motor protein myosin. CIM is a byproduct of critical care, attributed to impaired recovery, longterm complications, and mortality. CIM pathophysiology is complex, heterogeneous and remains incompletely understood, however loss of mechanical stimuli contributes to critical illness associated muscle atrophy and weakness. Passive mechanical loading (ML) and electrical stimulation (ES) therapies augment muscle mass and function. While having beneficial outcomes, the mechanistic underpinning of these therapies is less known. Therefore, here we aimed to assess the mechanism by which chronic supramaximal ES ameliorates CIM in a unique experimental rat model of critical care. Methods: Rats were subjected to 8 days critical care conditions entailing deep sedation, controlled mechanical ventilation, and immobilization with and without direct soleus ES. Muscle size and function were assessed at the single cell level. RNAseq and Western blotting were employed to understand the mechanisms driving ES muscle outcomes in CIM. Results: Following 8 days of controlled mechanical ventilation and immobilization, soleus muscle mass, Myosin:Actin ratio and single muscle fiber maximum force normalized to cross-sectional area (specific force) were reduced by 40-50% (p< 0.0001). ES significantly reduced the loss of soleus muscle fiber cross-sectional area (CSA) and Myosin:Actin ratio by approximately 30% (p< 0.05) yet failed to effect specific force. RNAseq pathway analysis revealed downregulation of insulin signaling in the soleus muscle following critical care and GLUT4 trafficking was reduced by 55% leading to an 85% reduction of muscle glycogen content (p< 0.01). ES promoted phosphofructokinase and insulin signaling pathways to control levels (p< 0.05), consistent with the maintenance of GLUT4 translocation and glycogen levels. AMPK, but not AKT, signaling pathway was stimulated following ES, where the downstream target TBC1D4 increased 3 logFC (p= 0.029) and AMPK-specific P-TBC1D4 levels were increased approximately 2-fold (p= 0.06). Reduction of muscle protein degradation rather than protein synthesis promoted soleus CSA, as ES reduced E3 ubiquitin proteins, Atrogin-1 (p= 0.006) and MuRF1 (p= 0.08) by approximately 50%, downstream of AMPK-FoxO3. Conclusions: ES maintained GLUT4 translocation through increased AMPK-TBC1D4 signaling leading to improved muscle glucose homeostasis. Soleus CSA and myosin content was promoted through reduced protein degradation via AMPK-FoxO3 E3 ligases, Atrogin-1 and MuRF1. These results demonstrate chronic supramaximal ES reduces critical care associated muscle wasting, preserved glucose signaling and reduced muscle protein degradation in CIM.

Protective effect of interval exercise training with different intensity and alpha-lipoic acid supplement on Nav1.3 protein in soleus muscle of diabetic rats

Introduction: Diabetes is a common metabolic disease, which leads to diabetic peripheral neuropathy. Peripheral neuron damage result in Nav1.3 elevations. Exercise training has beneficial role in diabetes management and peripheral neuropathy. Alpha lipoic acid (ALA) is a powerful biological antioxidant. However, the role of exercise training and ALA on Nav1.3 are not well understood. The aim of the present study was to investigate the effect of training with different intensity and Alpha lipoic acid supplement on soleus muscle Nav1.3 protein in rats with type 2 diabetes. Thirty-five male Wistar rats were randomly divided into seven groups: healthy control, diabetic, complementary diabetic, intensive exercise diabetic, moderate exercise diabetic, intensive exercise + supplemental diabetic, moderate exercise + complementary diabetic. Methods: In this experimental study, 35 male Wistar rats were randomly divided into seven groups: healthy control, diabetic (D), complementary (alpha lipoic acid) diabetic (ALA), diabetic high intensity training (HIT), diabetic moderate intensity training (MIT), diabetes HIT+ALA (ALA + HIT), diabetic MIT + ALA (ALA + MIT). Rats were diabetic by intra-peritoneal injection of STZ. The HIT and MIT protocols were performed five days a week for six weeks. HIIT included 10 bouts of four minutes (running at 85–90% of maximum speed) and MIT 13 bouts of four minutes (running at 65–70% of maximum speed). ALA was administered orally 20 mg/kg once a day by gavage. Nav1.3 protein levels were measured by immunohistochemistry method. Statistical operations were performed with SPSS version 16 software. One-way analysis of variance and Tukey were used to analyze the data. Results: The level of Nav1.3 increased significantly in diabetic group compared to the control (p≤0.0001). Moreover, HIT (p=0.0015), MIT p=0.0056), ALA+HIT (p≤0.0001) and ALA+MIT (p≤0.0001) decreased significantly Nav1.3 compared to the diabetic group. Conclusion: HIT and MIT can reduce the expression of NaV1.3 in soleus muscle in diabetic rats. ALA combined with exercise training can be more effective to reduce diabetic neuropathy.