The purpose of this study was to determine the effect of periodic walking during hindlimb suspension on the mass, relative weight, fiber type distribution and cross-sectional area of Type I and II fibers in the developing Type II plantaris muscle. To examine the effectiveness of periodic walking on mass and fiber size, the hindlimbs of young female Wistar rats were suspended (HS group) and half of these rats walked on a treadmill for 45 min/day(15 min every 4 hours) at 5 meters/min at a 15 degree grade(HS-W group) After seven days of hindlimb suspension, the plantaris muscle wet weight was 28.40% significantly smaller(P<0.005) and relative plantaris muscle weight was 26.97% smaller compared with those of control rats(P<0.05). The plantaris muscle wet weight and the relative plantaris muscle weight increased by 46.60% and 49.23% respectively with periodic walking, moreover. the plantaris muscle wet weight and the relative plantaris muscle weight of the HS-W rats recovered to the level of the control rats. No change was observed in fiber type percentage of the developing plantaris muscle following one week of hindlimb suspension or periodic walking during hindlimb suspension. Type I and II fiber cross-sectional areas of the developing plantaris muscle were 42.51% and 43. 68% lower in the HS group than in the control group(p<0.0001), Type I and II fiber cross-sectional areas of the developing plantaris were 30.82% and 45.97% greater in the HS-W group than in the HS group(p<0.0001), whereas Type I and II fiber cross-sectional area of HS-W group were less than those of the control group(P<0.0001) The results suggest that periodic walking can attenuate developing plantaris muscle atrophy induced by hindlimb suspension.
Chronic or acute alcohol abuse often leads to liver injury associated with alcoholic hepatitis, liver fibrosis, cirrhosis, and liver cancer. In addition to the liver, alcohol abuse also induces a variety of other tissue injuries including pancreatitis, cardiomyopathy, neurotoxicity and muscle loss. Chronic skeletal muscle myopathy, independent of peripheral neuropathy, is well recognised in alcoholic patients. Several mechanisms may be involved in the pathogenesis of alcoholic myopathy. Ethanol is a potent inhibitor of muscle protein synthesis. Gastrocnemius and plantaris muscles are Type II fiber-predominant and usually considered representative of the musculature as a whole. Whereas, soleus muscle is Type I fiber predominant. Shihosogan-san is a traditional Korean medicine that is widely employed to treat indigestion and liver diseases. Muscle diseases are often related to liver diseases and conditions. We therefore tested the hypothesis that treatment with Shihosogan-san could ameliorate the ethanol-induced changes in muscle protein synthesis. Young male Sprague-Dawley rats were orally given 25% ethanol (5ml/kg, body weight) daily with Ethanol for 28 days. Normal group was similarly administrated with saline. In Shihosogan-san treated group, rats were orally administrated Shihosogan-san extract, and rats of EtOH group were given with the vehicle only. After 4 week, the morphology of gastrocnemius and plantaris muscles were assessed by hematoxylin and eosin staining. For comparative purposes, liver function was also investigated. The muscles from rats of EtOH group displayed a significant reduction in average cross section area compared to Normal group. Shihosogan-san treated group had increased fiber compared to the EtOH group. Moreover, Shihosogan-san treated group compared with EtOH group showed significantly decreased pro-apoptotic BAX expression and increased anti-apoptotic Bcl-2 expression. In conclusion, Shihosogan-san extract showed ameliorating effects on chronic alcohol toxicity in skeletal muscle.
Objectives Ethanol is a potent inhibitor of muscle protein synthesis. Muscle mass is regulated by the balance between rates of protein synthesis and protein breakdown. Both acute and chronic alcohol consumption inhibits synthesis to a greater extent than degradation. Protein synthesis is more intensely decreased in type II fibers than in type I fibers. Apoptosis has been shown to occur frequently in a variety of tissues in response to chronic alcohol feeding. Increased muscle fiber apoptosis has been shown in alcoholics with myopathy. Pueraria radix has been used for many disorders such as fevers, gastrointestinal disorders, muscle aches, allergies, respiratory problems, skin problems, high blood pressure, migraine headaches, lowering cholesterol and treating chronic alcoholism. We therefore tested the hypothesis that oral treatment with Pueraria radix could reduce the ethanol-induced muscle atrophy. Methods Young male Sprague-Dawley rats were orally given 25% ethanol (5 ml/kg, body weight) daily with Ethanol for 4 weeks. Normal group was similarly administrated with saline. The Rats of Pueraria radix treated group (EtOH+PR) were orally administrated Pueraria radix water extract, and rats of EtOH group were given with the vehicle only. After 4 week, the morphology of gastrocnemius and plantaris muscles were assessed by hematoxylin and eosin staining. The immunoreactivities of pre-apoptotic BAX and anti-apoptotic Bcl-2 proteins were also measured. Results The muscles from rats of EtOH group represented a significant reduction in average cross section area compared to Normal group. EtOH+PR group had increased fiber compared to the EtOH group. Moreover, to investigate the ethanol-induced muscular apoptosis, the immunohistochemical analysis of Bax and Bcl-2 was carried out. The treatment with Pueraria radix (EtOH+PR) significantly decreased BAX expression and increased Bcl-2 expression 4 weeks after ethanol administration when compared with Normal group. Conclusions These results suggest that Pueraria radix water extract has protective effects on chronic alcohol induced myopathy.
Objective : This experimental study was designed to investigate the effect of Chaenomelis fructus herbal acupuncture on the soleus muscle and tibialis anterior muscle of hindlimb-suspended rats. The measurement have been performed on the composition of type I, type IIa and type IIb muscle fibers in the soleus muscle and tibialis anterior muscie - histochemical study. Materials and Methods : Sprague-Dawley rats weighing about 250g were subjected to hindlimb suspension and divided into six groups; 1. Normal group: normai group 2. Control group: group of hindlimb suspension for 7 days 3. NT group: non-treatment group for 2 or 4 weeks after 7days of hindlimb suspension. 4. EX gtoup: running exercise group for 2 or 4 weeks after 7days of hindlimb suspension. 5. HA group: administration of Chaenomelis fructus herbal acupuncture during 2 or 4 weeks after 7 days of hindlimb suspension. 6. EXHA group: administration of Chaenomelis fructus herbal acupuncture and running exercise during 2 or 4 weeks after 7 days of hindlimb suspension. Results Results : 1. The composition of type I muscle fibers in the soleus muscle significantly increased in 2 or 4 weeks HA and EXHA groups more than control group, and the composition of type IIa muscle fibers in the soleus muscle significantly decreased in 2 or 4 weeks HA and EXHA goups more than control group. 2. The composition of type I muscie fibers in the tibialis anterior muscle significantly increased in 2 or 4 weeks HA and EXHA groups more than control group, and the composition of type IIb muscle fibers in the soleus muscle significantly decreased in 2 or 4 weeks HA and EXHA groups more than control group. 3. The size of type I muscle fibers in the soleus muscle significantly increased in 2 or 4 weeks HA and EXHA groups more than control group, and the size of type IIa muscle fibers in the soleus muscle significantly increased in 2 weeks EXHA group and 4 weeks HA and EXHA groups more than control group. 4. The size of type I, IIa muscle fibers in the tibialis anterior muscle significantly increased in 2 or 4weeks HA, EXHA and EX groups more than control group, and the size of type IIb muscle fibers significantly increased in 2 weeks EXHA group and 4 weeks HA, EXHA and EX groups more than control group. 5. The capiliaries number per fiber of muscle fibers in the soleus and tibialis anterior muscle showed no differences in all experimental groups more than control group. Conclusion : According to the above results, it is shown that Chaenomelis fructus herbal acupuncture could be effective in the treatment of muscle atrophy.
The purpose of this study was to determine the effect of intermittent low - intensity, short duration exercise during hindlimb suspension on the mass, relative weight, myofibrillar protein content, cross-sectional area of Type I and Type II fibers and SDH activity in Type II(plantaris) muscle. To examine the effectiveness of intermittent low-intensity, short duration exercise on mass, myofibrillar protein content and fiber size, the hindlimbs of adult female Wistar rats were suspended(HS) and half of these rats walked on a treadmill for 45 min/day(9 min every 2h) at 5m /min and a 15$^{\circ}$grade (HS-EX). Plantaris wet weight was 19.67% significantly smaller(p<0.005) and relative plantaris weight was 6.25% smaller compared with those of control rats following seven days of hindlimb suspension. Plantaris wet weight and relative plantaris weight increased by 27.66%, 12.22% each through intermit-tent exercise during hindlimb suspension(p<0.005, p<0.05), moreover, plantaris wet weight and relative plantaris weight of the HS-EX rats were similar to those of control rats. Soleus wet weight and relative soleus weight decreased significantly by 31% and 22.0% in the HS rats(p<0.05). Soleus wet weight and relative soleus weight increased by 10.41%, 25.64% respectively through intermittent ex-ercise during hindlimb suspension, furthermore, soleus wet weight and relative weight of the HS-EX rats were closer to those of control rats. Myofibrillar protein content of plantaris and soleus decreased significantly by 51.49%, 59.65% each, following seven days of hindlimb suspension (p<0.005) Myofibrillar protein content of plantaris and soleus increased by 51.79%, 75.47% each with significance through intermittent exercise during hindlimb suspension(p<0.005). Myofibrillar protein content of plantaris and soleus in HS-EX rats was smaller than that of control rats. No change was observed in fiber type percentage following 1 week of hindlimb suspension or exercise during hindlimb suspension. The type I fiber cross-sectional area of both soleus and plantaris muscle was 18.72% and 41.07% lower in the HS than that of the controls (p<0.05, p<.001), that of both muscles was 6.60% and 29. 3% greater in the HS-EX than that of the HS rats. HS plus intermittent low- intensity short duration exercise resulted in Type I fiber cross-sectional area closer to the controls. Type II fiber cross-sectional area of both plantaris and soleus muscle was 22.45% and 22.58% sl nailer in the HS than in the controls, that of both muschles in the HS-EX was 14.10%, 5.78% greater than HS. Intermittent exercise during hindlimb suspension resulted in Type I, II fiber cross-sectional area closer to the control value. There was no change in SDH activity following 1week of hindlimb suspension or exercise during hindlimb suspension in the plantaris muscle. The results suggest that intermittent low intensity short duration exercise can ameliorate Type II muscular atrophy Induced by hindlimb suspension.
Purpose: The balance between synthesis and degradation of proteins plays a critical role in the maintenance of skeletal muscle mass. Mitochondrial dysfunction has been closely associated with skeletal muscle atrophy caused by aging, cancer, and chemotherapy. Polygalacin D is a saponin derivative isolated from Platycodon grandiflorum (Jacq.) A. DC. This study aimed to investigate the effects of polygalacin D on myoblast differentiation and muscle atrophy in association with mitochondrial function in in vitro and in zebrafish models in vivo. Methods: C2C12 myoblasts were cultured in differentiation media containing different concentrations of polygalacin D, followed by the immunostaining of the myotubes with myosin heavy chain (MHC). The mRNA expression of markers related to myogenesis, muscle atrophy, and mitochondrial function was determined by real-time quantitative reverse transcription polymerase chain reaction. Wild type AB* zebrafish (Danio rerio) embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without polygalacin D, and immunostained to detect slow and fast types of muscle fibers. The Tg(Xla.Eef1a1:mitoEGFP) zebrafish expressing mitochondria-targeted green fluorescent protein was used to monitor mitochondrial morphology. Results: The exposure of C2C12 myotubes to 0.1 ng/mL of polygalacin D increased the formation of MHC-positive multinucleated myotubes (≥ 8 nuclei) compared with the control. Polygalacin D significantly increased the expression of MHC isoforms (Myh1, Myh2, Myh4, and Myh7) involved in myoblast differentiation while it decreased the expression of atrophic markers including muscle RING-finger protein-1 (MuRF1), mothers against decapentaplegic homolog (Smad)2, and Smad3. In addition, polygalacin D promoted peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α) expression and reduced the level of mitochondrial fission regulators such as dynamin-1-like protein (Drp1) and mitochondrial fission 1 (Fis1). In a zebrafish model of FOLFIRI-induced muscle atrophy, polygalacin D improved not only mitochondrial dysfunction but also slow and fast muscle fiber atrophy. Conclusion: These results demonstrated that polygalacin D promotes myogenesis and alleviates chemotherapy-induced muscle atrophy by improving mitochondrial function. Thus, polygalacin D could be useful as nutrition support to prevent and ameliorate muscle wasting and weakness.
A 3-year-old, spayed female Shih-tzu dog was presented due to acute vomiting, diarrhea, and generalized weakness. The dog had generalized weakness, increased respiratory rate, and respiratory muscle effect. Neurologic examination revealed appendicular muscular weakness and decreased in tone of the anal sphincter. Megaesophagus was confirmed by radiographic examinations. Other than type 2 fiber atrophy, no specific abnormalities were identified in histopathologic examinations of muscle biopsies from the left pelvic limb. Serum acetylcholine receptor (AChR) antibody titer was increased (0.78 nmol/L reference range, less than 0.6 nmol/L), confirming a diagnosis of acute fulminating myasthenia gravis. The dog dramatically responded to pyridostigmine bromide and had marked improvement in muscle strength, megaesophagus, and respiratory function. The dog has been successfully managed for 7 months after initial treatment.
The spinal cord is highly complex, consisting of a specialized neural network that comprised both neuronal and non-neuronal cells. Any kind of injury and/or insult to the spinal cord leads to a series of damaging events resulting in motor and/or sensory deficits below the level of injury. As a result, muscle paralysis (or paresis) leading to muscle atrophy or shrinking of the muscle along with changes in muscle fiber type, and contractile properties have been observed. Traditionally, histology had been used as a gold standard to characterize spinal cord injury (SCI)-induced adaptation in spinal cord and skeletal muscle. However, histology measurements is invasive and cannot be used for longitudinal analysis. Therefore, the use of conventional magnetic resonance imaging (MRI) is promoted to be used as an alternative non-invasive method, which allows the repeated measurements over time and secures the safety against radiation by using radiofrequency pulse. Currently, many of pathological changes and adaptations occurring after SCI can be measured by MRI methods, specifically 3-dimensional MRI with the advanced diffusion tensor imaging technique. Both techniques have shown to be sensitive in measuring morphological and structural changes in skeletal muscle and the spinal cord.
This study investigated how dietary fat affects muscle atrophy and lipid metabolism in various muscles during hindlimb immobilization in rats. Twenty-four male Sprague?Dawley rats had their left hindlimb immobilized and were divided into four groups by dietary fat content and composition. The contralateral hindlimb (control) was compared with the immobilized limb in all dietary groups. Rats (n = 6/group) were fed a 4% corn oil diet (CO), 2.6% corn oil + 1.4% fish oil diet (FO), 30% corn oil diet (HCO), or a 30% beef tallow diet (HBT)after their hind limbs were immobilized for 10 days. Data were collected for the gastrocnemius, plantaris and soleus muscles. Muscle atrophy was induced significantly after 10 days of hindlimb immobilization, resulting in significantly decreased muscle mass and total muscle protein content. The protein levels of peroxisome proliferator activated receptor ${\delta}$ (PPAR${\delta}$) in the plantaris, gastrocnemius, and soleus increased following hindlimb immobilization irrespective of dietary fat intake. Interestingly, the PPAR${\delta}$ mRNA level in the plantaris decreased significantly in all groups and that in the FO group was lower than that in the other groups. The soleus PPAR${\delta}$ mRNA level decreased significantly following hindlimb immobilization in the FO group only. Muscle carnitine palmitoyl transferase 1 (mCPT1) mRNA level was not affected by hindlimb immobilization. However, the mCPT1 mRNA level in the FO group was significantly lower in the plantaris but higher in the soleus than that in the other groups. The pyruvate dehydrogenase kinase 4 (PDK4) mRNA level in the plantaris decreased significantly, whereas that in the soleus increased significantly following hindlimb immobilization. The plantaris, but not soleus, PDK4 mRNA level was significantly higher in the FO group than that in the CO group. The increased PPAR${\delta}$ protein level following hindlimb immobilization may have suppressed triglyceride accumulation in muscles and different types of dietary fat may have differentially affected muscle atrophy according to muscle type. Our results suggest that ${\omega}$-3 polyunsaturated fatty acids may suppress muscle atrophy and lipid accumulation by positively affecting the expression level and activity of PPAR${\delta}$ and PPAR${\delta}$-related enzymes, which are supposed to play an important role in muscle lipid metabolism.
Background: The ginsenoside Rg1 has been shown to exert various pharmacological activities with health benefits. Previously, we have reported that Rg1 promoted myogenic differentiation and myotube growth in C2C12 myoblasts. In this study, the in vivo effect of Rg1 on fiber-type composition and oxidative metabolism in skeletal muscle was examined. Methods: To examine the effect of Rg1 on skeletal muscle, 3-month-old mice were treated with Rg1 for 5 weeks. To assess muscle strength, grip strength tests were performed, and the lower hind limb muscles were harvested, followed by various detailed analysis, such as histological staining, immunoblotting, immunostaining, and real-time quantitative reverse transcription polymerase chain reaction. In addition, to verify the in vivo data, primary myoblasts isolated from mice were treated with Rg1, and the Rg1 effect on myotube growth was examined by immunoblotting and immunostaining analysis. Results: Rg1 treatment increased the expression of myosin heavy chain isoforms characteristic for both oxidative and glycolytic muscle fibers; increased myofiber sizes were accompanied by enhanced muscle strength. Rg1 treatment also enhanced oxidative muscle metabolism with elevated oxidative phosphorylation proteins. Furthermore, Rg1-treated muscles exhibited increased levels of anabolic S6 kinase signaling. Conclusion: Rg1 improves muscle functionality via enhancing muscle gene expression and oxidative muscle metabolism in mice.
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