Soleus Muscle Of Diabetic Rats Research Articles

Exercise ameliorates muscular excessive mitochondrial fission, insulin resistance and inflammation in diabetic rats via irisin/AMPK activation

This study aimed to investigate the effects of exercise on excessive mitochondrial fission, insulin resistance, and inflammation in the muscles of diabetic rats. The role of the irisin/AMPK pathway in regulating exercise effects was also determined. Thirty-two 8-week-old male Wistar rats were randomly divided into four groups (n = 8 per group): one control group (Con) and three experimental groups. Type 2 diabetes mellitus (T2DM) was induced in the experimental groups via a high-fat diet followed by a single intraperitoneal injection of streptozotocin (STZ) at a dosage of 30 mg/kg body weight. After T2DM induction, groups were assigned as sedentary (DM), subjected to 8 weeks of treadmill exercise training (Ex), or exercise training combined with 8-week cycloRGDyk treatment (ExRg). Upon completion of the last training session, all rats were euthanized and samples of fasting blood and soleus muscle were collected for analysis using ELISA, immunofluorescence, RT-qPCR, and Western blotting. Statistical differences between groups were analyzed using one-way ANOVA, and differences between two groups were assessed using t-tests. Our findings demonstrate that exercise training markedly ameliorated hyperglycaemia, hyperlipidaemia, and insulin resistance in diabetic rats (p < 0.05). It also mitigated the disarranged morphology and inflammation of skeletal muscle associated with T2DM (p < 0.05). Crucially, exercise training suppressed muscular excessive mitochondrial fission in the soleus muscle of diabetic rats (p < 0.05), and enhanced irisin and p-AMPK levels significantly (p < 0.05). However, exercise-induced irisin and p-AMPK expression were inhibited by cycloRGDyk treatment (p < 0.05). Furthermore, the administration of CycloRGDyk blocked the effects of exercise training in reducing excessive mitochondrial fission and inflammation in the soleus muscle of diabetic rats, as well as the positive effects of exercise training on improving hyperlipidemia and insulin sensitivity in diabetic rats (p < 0.05). These results indicate that regular exercise training effectively ameliorates insulin resistance and glucolipid metabolic dysfunction, and reduces inflammation in skeletal muscle. These benefits are partially mediated by reductions in mitochondrial fission through the irisin/AMPK signalling pathway.

Alleviative effects of α-lipoic acid on muscle atrophy via the modulation of TNF-α/JNK and PI3K/AKT pathways in high-fat diet and streptozotocin-induced type 2 diabetic rats.

Diabetes mellitus (DM) is often accompanied by clinical complications such as sarcopenia. Previous studies have indicated that oxidative stress and insulin resistance (IR) are highly associated with the pathogenesis of diabetic myopathy. α-lipoic acid (ALA), a potent biological antioxidant, exists abundantly in a variety of plants and vegetables. This study aimed to investigate the ameliorative effect of ALA on muscle atrophy in type 2 diabetic rats induced by high-fat diet feeding (HFD) plus streptozotocin (STZ) injection. The HFD/STZ-induced diabetic rats were orally administered 50, 100, or 200 mg/kg body weight ALA once a day for 13 weeks. The results showed that ALA at the tested concentrations significantly increased the soleus muscle mass and muscle fibers in diabetic rats. Proinflammatory cytokines, such as tumor necrosis factor (TNF)-α, were found to decrease in both the serum and muscle of ALA-treated diabetic rats. ALA significantly reduced the protein-expression levels of phosphorylated c-Jun N-terminal kinase (pJNK)/JNK, forkhead box O3 (FOXO3), and muscle ring-finger protein-1 (Murf1); whereas, it enhanced the protein-expression levels of phosphoinositide 3-kinase (PI3K), phosphorylated protein kinase B (pAKT)/AKT, myogenin determination gene D (MyoD), the mechanistic target of rapamycin (mTOR), and myosin heavy chain (MyHC) in the soleus muscle of diabetic rats. The results from this study suggested that ALA treatment may preserve soleus muscle mass, alleviate muscle atrophy by suppressing the TNF-α/JNK pathway, and ameliorate the PI3K/AKT pathway in HFD/STZ-induced type 2 diabetic rats.

Effects of two workload-matched high intensity interval training protocols on regulatory factors associated with mitochondrial biogenesis in the soleus muscle of diabetic rats.

Aims: High intensity interval training (HIIT) improves mitochondrial characteristics. This study compared the impact of two workload-matched high intensity interval training (HIIT) protocols with different work:recovery ratios on regulatory factors related to mitochondrial biogenesis in the soleus muscle of diabetic rats. Materials and methods: Twenty-four Wistar rats were randomly divided into four equal-sized groups: non-diabetic control, diabetic control (DC), diabetic with long recovery exercise [4–5 × 2-min running at 80%–90% of the maximum speed reached with 2-min of recovery at 40% of the maximum speed reached (DHIIT1:1)], and diabetic with short recovery exercise (5–6 × 2-min running at 80%–90% of the maximum speed reached with 1-min of recovery at 30% of the maximum speed reached [DHIIT2:1]). Both HIIT protocols were completed five times/week for 4 weeks while maintaining equal running distances in each session. Results: Gene and protein expressions of PGC-1α, p53, and citrate synthase of the muscles increased significantly following DHIIT1:1 and DHIIT2:1 compared to DC (p ˂ 0.05). Most parameters, except for PGC-1α protein (p = 0.597), were significantly higher in DHIIT2:1 than in DHIIT1:1 (p ˂ 0.05). Both DHIIT groups showed significant increases in maximum speed with larger increases in DHIIT2:1 compared with DHIIT1:1. Conclusion: Our findings indicate that both HIIT protocols can potently up-regulate gene and protein expression of PGC-1α, p53, and CS. However, DHIIT2:1 has superior effects compared with DHIIT1:1 in improving mitochondrial adaptive responses in diabetic rats.

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.

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.&#x0D; 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.&#x0D; 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.&#x0D; 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.

Effects of combined treatment with blood flow restriction and low-current electrical stimulation on capillary regression in the soleus muscle of diabetic rats

The combined treatment of blood flow restriction and low intensity electrical stimulation attenuated type 2 diabetes (T2D)-associated capillary regression in the skeletal muscles. The treatment inhibits the T2D-associated increase in antiangiogenic factors via inhibition of intramuscular chronic hypoxia; it can inhibit intramuscular chronic hypoxia by enhancing proangiogenic factors. These results suggest that the combined treatment may be an effective therapeutic intervention for the prevention of T2D-associated capillary regression in the skeletal muscles.

Investigation of the pronounced erythropoietin-induced reduction in hyperglycemia in type 1-like diabetic rats.

Erythropoietin (EPO) is known to stimulate erythropoiesis after binding with its specific receptor. In clinics, EPO is widely used in hemodialyzed patients with diabetes. However, changes in the expression of the erythropoietin receptor (EPOR) under diabetic conditions are still unclear. Therefore, we investigated EPOR expression both in vivo and in vitro. Streptozotocin-induced type 1-like diabetic rats (STZ rats) were used to evaluate the blood glucose-lowering effects of EPO. The expression and activity of the transducer and activator of transcription 3 (STAT3), the potential signaling molecule, was investigated in cultured rat skeletal myoblast (L6) cells incubated in high-glucose (HG) medium to mimic the in vivo changes. The EPO-induced reduction in hyperglycemia was more pronounced in diabetic rats. The increased EPOR expression in the soleus muscle of diabetic rats was reversed by the reduction in hyperglycemia. Glucose uptake was also increased in high-glucose (HG)-treated L6 cells. Western blotting results indicated that the EPO-induced hyperglycemic activity was enhanced mainly through an increase in EPOR expression. Increased EPOR expression was associated with the enhanced nuclear expression of STAT3 in HG-exposed L6 cells. In addition, treatment with siRNA specific to STAT3 reversed the increased expression of EPOR observed in these cells. Treatment with Stattic at a dose sufficient to inhibit STAT3 reduced the expression level of EPOR in STZ rats. In conclusion, the increased expression of EPOR by hyperglycemia is mainly associated with an augmented expression of nuclear STAT3, which was identified both in vivo and in vitro in the present study.

Potential mechanism of anti-diabetic activity of Picrorhiza kurroa

Picrorhiza kurroa Royle ex Benth. (Scrophulariaceae) is a traditional Ayurvedic herb known as Kutki. It is used as a remedy for diabetes by tribes of North Eastern Himalayan region of India. Present study was conducted to explore the mechanism of antidiabetic activity of standardized aqueous extract of Picrorhiza kurroa (PkE). PkE (100 and 200 mg/kg/day) was orally administered to streptozotocin induced diabetic rats, for 14 consecutive days. Plasma insulin levels were measured and pancreas of rat was subjected to histopathological investigations. Glucose transporter type 4 (GLUT-4) protein content in the total membrane fractions of soleus muscle was estimated by Western blot analysis. Plasma insulin level was significantly increased along with concomitant increase in GLUT-4 content of total membrane fractions of soleus muscle of diabetic rats treated with extract. There was evidence of regeneration of β-cell of pancreatic islets of PkE treated group in histopathological examinations. PkE increased the insulin-mediated translocation of GLUT-4 from cytosol to plasma membrane or increased GLUT-4 expression, which in turn facilitated glucose uptake by skeletal muscles in diabetic rats.

Treadmill exercise suppresses muscle cell apoptosis by increasing nerve growth factor levels and stimulating p-phosphatidylinositol 3-kinase activation in the soleus of diabetic rats

We investigated the effects of treadmill exercise performed regularly for 6 weeks on the levels of nerve growth factor (NGF), tyrosine kinase A and p75 receptors, phosphatidylinositol 3-kinase (PI3-K), mitogen-activated protein kinase/extracellular signal-regulated kinase (Erk) 1,2, cyclic AMP response element-binding protein (CREB), and caspase-3 in the soleus of rats with streptozotocin (STZ)-induced diabetes. Thirty-two male Sprague-Dawley rats were divided into the following four groups: (1) normal control group (NCG; n = 8), (2) normal exercise group (NEG; n = 8), (3) diabetes control group (DCG; n = 8), and (4) diabetes exercise group (DEG; n = 8). Diabetes was induced by intraperitoneal injection of STZ (55 mg/kg dissolved in 0.05 M citrate buffer, pH 4.5). Rats were subjected to treadmill exercise 5 days a week for 6 weeks. The protein level of NGF significantly increased in the NEG and DEG (p < 0.001), whereas the levels of tyrosine kinase A and p75 receptors significantly increased in the NEG (p < 0.001). The levels of t-PI3-K, p-PI3-K, and p-CREB, and the p-CREB/t-CREB ratio significantly increased in the NEG (p < 0.001, respectively). The p-PI3-K/t-PI3-K ratio significantly increased in the DEG (p < 0.001). The p-Erk1/t-Erk1 ratio significantly increased in the NEG (p < 0.001), whereas the p-Erk2/t-Erk2 ratio significantly decreased in the DCG and DEG (p < 0.001). The caspase-3 level significantly increased in the DCG compared with that in the DEG (p < 0.001). These results suggest that treadmill exercise increases NGF levels and accelerates p-PI3-K activation in order to suppress apoptotic cell death in the soleus muscle of diabetic rats.

The slow sarco/endoplasmic reticulum Ca2+ -ATPase declines independently of slow myosin in soleus muscle of diabetic rats.

The sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) isoforms are normally expressed in coordination with the corresponding myosin heavy chain (MyHC) isoforms in the fibers of skeletal muscle but this coordination is often disrupted in pathological conditions. In the streptozotocin-induced diabetes of rats (stz-rats), the soleus muscle showed peripheral neuropathy and the SERCA2a level decreased in type I (slow-oxidative) fibers compared to the control muscles, whereas the expression of the corresponding slow MyHC1 did not change. No difference was found at the mRNA and protein levels of SERCA and MyHC isoforms in the whole soleus, except that the level of the SERCA2a protein specifically declined in stz-rats compared to the controls. This shows that the coordinated expression of SERCA2a and MyHC1 is disrupted at the SERCA2a protein level in the diabetic soleus. The results are in line with previous observations that regulators of the Ca-homeostasis may adapt faster to type I diabetes than the contractile elements.

Lectins binding during alloxan-induced diabetes in rat soleus muscle

Membrane structural changes of soleus muscle of alloxan-diabetic rats were detected with a panel of six biotinylated lectins. Samples of muscles were obtained from normal and diabetic rats. The biotinylated lectins in staining were detected by avidin-peroxidase complex. Lectin stainning of soleus muscle cryostat sections from alloxan-diabetic rats were compared with similar cryostat sections from normal rats. All lectins were bounded to soleus muscle cell membranes of normal and diabetic rats except the Ulex europeus agglutinin I (UEA-I). UEA-I was not stained for cell membranes of soleus muscles of normal and diabetic rats. But intercellular areas of normal and diabetic soleus muscle were stained for UEA-I. Griffonia simplicifolia I (GS-I) lectin was bounded more strongly to cell membrane of diabetic rats than other lectins. Not only cell membranes but also cytoplasmic miyofibrills of diabetic muscle cells were stained by Griffonia simplicifolia (GS1) lectin. It was concluded that cell membranes of soleus muscle of diabetic rats were differently stained for lectins than those of normal rats. Our data can supplement on the use of lectins as probes for changes in carbohydrate-containing constituents of the alloxan-diabetic muscle cell membrane information obtained from normal and diabetic

Effect of Endurance Exercise on Myosin Heavy Chain Isoform Expression in Diabetic Rats with Peripheral Neuropathy

This study evaluated the effect of endurance exercise on myosin heavy chain (MHC) isoform expression in soleus muscle of diabetic rats with peripheral neuropathy. Male Sprague Dawley rats were randomly divided into four groups: control sedentary, diabetic sedentary, control exercise, and diabetic exercise. The exercised animals performed treadmill running five times per week. After 12 wks, electrophysiologic testing documented peripheral neuropathy in the diabetic rats. The soleus muscles were then excised and quick-frozen. Cross-sections were immunohistochemically stained for slow, fast, developmental, and neonatal MHCs. Fiber-type composition and fiber cross-sectional areas were then determined. The diabetic groups showed a significantly greater percentage of fast MHC than did the control groups, regardless of exercise status (diabetic sedentary, 22.6%; diabetic exercise, 25.2%; control sedentary, 13.5%; control exercise, 13.1%). The diabetics also showed a significantly lower percentage of slow-only MHC than controls (diabetic sedentary, 77.1%; diabetic exercise, 74.3%; control sedentary, 86.2%; control exercise, 86.1%). No differences in muscle fiber cross-sectional area existed between the groups. The exercised animals showed greater expression of developmental MHC than did the sedentary animals (diabetic sedentary, 1.6%; diabetic exercise, 3.8%; control sedentary, 0.8%; control exercise, 2.0%). The altered slow and fast MHC expression in the diabetic muscle is similar to MHC expression in several other conditions, including decreased neuromuscular activity and denervation. Mechanisms of this MHC expression shift are unknown. Chronic endurance training does not alter adult MHC expression in the diabetic animals. The developmental MHC expression is likely a manifestation of uphill treadmill running due to eccentric contractions in the soleus resulting in myofiber injury and regeneration.

Myosin heavy chain isoform immunolabelling in diabetic rats with peripheral neuropathy

This study evaluated mature and immature myosin heavy chain (MHC) isoform immunolocalisation in soleus muscle of diabetic rats with documented motor neuropathy. Sprague Dawley rats were assigned to one of three groups: control (C), diabetic with insulin (DI), or diabetic without insulin (DNI). Twelve weeks after diabetes induction, soleus muscles were excised and quick-frozen. Cross-sections were labelled immunohistochemically for slow, fast, developmental and neonatal MHC isoforms to determine fiber-type composition. Fiber cross-sectional areas were determined morphometrically. Results revealed that DNI and DI muscles contained greater percentages of myofibers positive for fast MHC compared with controls. DNI animals also showed a lower percentage of myofibers positive for slow MHC compared to the DI group. The number of fibers immunolabelled for developmental MHC isoforms was greater in DNI animals than in the other groups. The differences in slow and fast MHC-labelling appear to indicate a condition of altered neuromuscular activity affecting the diabetic muscles. The increase in developmental MHC-labelling in the DNI muscles could indicate myofiber regeneration or reinnervation that would be more pronounced in the DNI animals in context of their more severe neuropathy. Insulin appeared to influence muscle fiber cross-sectional area and possibly fiber-type grouping frequency; the potential mechanism for these effects was not elucidated.

The effects of peroxovanadate and peroxovanadyl on glucose metabolism in vivo and identification of signal transduction proteins involved in the mechanism of action in isolated soleus muscle

The insulin-like effects of peroxovanate (POV) and peroxovanadyl (PSV) on rates of lactate formation and glycogen synthesis were measured in isolated incubated soleus muscle preparations. In another experiment rats were made insulin deficient by streptozotocin injection and treated with POV and PSV (0.25 mM) administered in the drinking water and in the course of 7 days glycemia were determined. Also, signal transduction proteins ERK 1 and ERK 2 involved in the insulin signaling were measured in soleus muscle of diabetic rats treated with POV and PSV. Peroxides of vanadate and vanadyl significantly stimulated glucose utilization in soleus muscle preparations in vitro. The stimulation of glycogen synthesis and lactate formation by POV and PSV was similar to insulin stimuli. Rats treated with POV or PSV presented reduction of glycemia, food and fluid intake with amelioration of the diabetic state during the short period of treatment (7 days). POV and PSV modulated ERK1/2 phosphorilation and the insulin administration in these rats caused an addictive effect on phosphorilation state of these proteins.

Brain-Derived Neurotrophic Factor mRNA Levels Are Up-Regulated in Hindlimb Skeletal Muscle of Diabetic Rats: Effect of Denervation

In a previous study the levels of brain-derived neurotrophic factor (BDNF) mRNA were shown to be elevated in skeletal muscle of the diabetic rat compared with age-matched control animals. It was proposed that diabetes-induced changes in nerve function may initiate changes in nerve/muscle contact akin to those following denervation of target skeletal muscle. In this study hindlimb skeletal muscles were denervated by sciatic nerve crush or transection and the effect on BDNF mRNA levels in control and diabetic rats was observed using Northern blotting. Contralateral to the side of nerve injury, the levels of BDNF mRNA in soleus muscle of diabetic rats were higher than in controls (three- to sevenfold), as has been seen previously in diabetic rats without any axotomy. Sciatic nerve crush or transection, of 1 week or of 3 weeks duration, lowered the levels of BDNF mRNA by 50% in ipsilateral soleus muscle of diabetic rats. BDNF mRNA levels in contralateral gastrocnemius muscle were not similarly raised in diabetic rats compared with controls and nerve injury had no effect. In control animals, ipsilaterally, the BDNF mRNA levels of soleus muscle were raised approximately twofold at 1 week and were lowered by approximately 50% at 3 weeks following nerve injury. Neurotrophin-3 mRNA levels were reduced approximately 50% in soleus muscle of diabetic rats compared with control rats, and nerve injury had no significant effect. The specific up-regulation of BDNF mRNA in soleus muscle of diabetic rats is discussed in terms of a proposed diabetes-induced ischemia within hindlimb skeletal muscle, with a protective role for BDNF in muscle and/or nerve being introduced.