본 연구는 도시숲에서 산림치유 프로그램 활동이 도시에 거주하는 노인여성의 골격근량과 내장지방 량, 우울에 어떤 변화가 있는지 확인하기 위해 진행되었다. 연구대상자는 경기도 A 시에 거주하는 60세 이상의 노인여성으로 실험군 21명, 대조군 12명으로 총 33명으로 구성되었다. 산림치유 프로그 램은 주1회 2시간씩 총8회로 진행하였다. 프로그램 전후 체성분분석기(Inbody 720, Biospace, Korea)를 이용하여 골격근량과 내장지방량을 측정하였고, 우울척도는 GDS(Geriatric Depression Scale)를 이용하였다. 실험군 대상자의 나이는 평균 60.3±9.4였으며 대조군 나이는 평균63.6±5.8 이었다. 산림치유프로그램 참여 후 실험군 대상자의 골격근 변화는 참여 전 2.52에서 참여 후 2.47로 유의한 변화를 보였으나 대조군은1.64에서 1.99로 유의한 변화는 아니었다. 내장지방량의 변화는 실험군 대상자는 참여 전 34.27에서 참여 후 31.64로 유의한 변화를 보였으나 대조군은 35.02에서 33.18로 유의한 변화는 아니었다. 우울의 변화는 실험군에서 참여 전 5.10에서 참여 후 3.85로 유의한 변화를 보였다. 도시숲을 이용한 산림치유프로그램 활동이 노인여성의 만성질환의 요인이 되는 골 격근량과 내장지방량에 긍정적인 영향을 미쳤음이 검증되었고, 실험군의 우울감 역시 감소됨을 알 수 있었다. 규칙적이고 지속적인 산림치유 프로그램의 참여를 통해 노인의 신체적 건강과 우울 등 심리적 건강증진에 기여하는 하나의 기초자료로 제안하고자 한다.
이 연구의 목적은 근육 세포의 증식 배양을 위해 필요한 핵심 인자인 혈청이 혈청 대체물의 첨가에 의해 대체될 수 있는지를 분자 생물학적 측면에서 검증하는 것이다. 혈청 대체물이 첨가된 low serum (5% FBS) 기반의 promo cell 배지는 성장 배양액으로 사용되었고, 마우스 하지 골격근의 근육세포는 pre-plating (pp) 방법에 의해 분리되었다. Pre-plating 4의 세포는 작고 둥근 형태의 굴절성을 가진 Myoblast/satellite like cells의 형태가 관찰되었으며, 근육 줄기세포 전사인자들(Pax3/7, Myf5, Myod1)과 골격근 발달 전사인자(Myog)의 발현량이 섬유아세포와 비교하여 높게 나타났다. 따라서 그들을 Myoblast derived cells로 명명하고, 기본 세포주로 사용하였다. 분리된 MDCs는 5%, 10% 혹은 20% 혈청이 첨가된 배양액에서 2주간 배양되었다. 배양 6일째부터 대조군(5% 혈청)과 비교하여 20% 혈청은 세포 수가 증가하였으며, 양적 혈청 농도 의존성이 확인되었다. 증식 및 세포사멸 관련 유전자들은 대조군과 비교하였다. 배양 1주 차에 20% FBS군은 세포증식 촉진 유전자인 Myc 발현이 증가한 반면 pro-apoptosis 유전자인 Bax의 발현량이 감소했고, 2주 차에는 세포주기정지 인자인Cdkn1a의 감소와 Myc의 지속적인 증가와 Bax/Bcl의 감소가 나타났다. 각기 다른 FBS 처리농도에서 배양된 Myoblast derived cells을 동일한 Myotube유도 배양액에서 2주간 분화하였다. 대조군과 비교하여 20% FBS군은 Myod1, Myog, Myf6, Myh1의 유의적 증가가 1주부터 확인되었다. 결론적으로 혈청 대체 물질은 근육세포 배양액에서 혈청의 효과를 완벽히 모사할 수 없음이 증명되었고, 따라서 체외에서 상업적 목적의 근육 세포 대량 증식 및 분화를 위해서는 적절한 혈청 대체물 개발이 선행돼야 할 것으로 사료된다.
Amyotrophic lateral sclerosis (ALS) is progressive neurological disease that results in the death of motor neurons in the brain and spinal cord, leading to a decrease in skeletal muscle size and muscle weakness, wasting, or paralysis. Most research on ALS has focused on motor neuron death, and the underlying mechanisms are not well understood. This study examined the molecular mechanisms underlying muscle degeneration. We compared the protein and cytokine profiles of gastrocnemius muscle in ALS model hSOD1G93A mice at pre-symptomatic and symptomatic stages by western blotting. Pro-inflammatory factors including tumor necrosis factor-α, interleukin (IL)-1β and IL-6, and cluster of differentiation 11b were upregulated in the muscle of symptomatic as compared to pre-symptomatic mice. Additionally, the levels of oxidative stress-related proteins, heme oxygenase-1 and ferritin, were increased in muscle from symptomatic as compared to pre-symptomatic mice. We also observed increased autophagy dysfunction and metabolic dysregulation in the muscles of symptomatic hSOD1G93A as compared to non-Tg and pre-symptomatic hSOD1G93A mice, which was accompanied by upregulation of thrombospondin- 1, Prospero-related homeobox 1, glial fibrillary acidic protein, and DNA-damage-inducible 45α. Increased inflammation, oxidative stress, and autophagy contribute to motor neuron death and muscle atrophy in ALS, and the factors involved in these processes are potential therapeutic targets for treatment of this disease.
Inflammation is an important protective response mechanism that occurs against microbial invasion or injury. However, excessive inflammation may lead to cause of morbidity and mortality in diseases. The activated macrophages plays a vital role in inflammatory response by stimulation of lipopolysaccharide (LPS) and tumor necrosis factor-α (TNF-α). This activation further damages the host by inducing certain pro-inflammatory mediators such as nitric oxide (NO), interleukin-1β (IL-1β), interleukin- 6 (IL-6), TNF-α, inducible nitrous oxide (iNOS) and cyclooxygenase (COX-2). Flavonoids are bioactive compounds with potential effects as anti-cancer, anti-inflammation, anti-viral and anti-bacterial activities. Polymethoxyflavones (PMFs) are unique to citrus plants which are of specific interest owing to their biological effects that includes lipoprotein metabolism and anti-inflammatory activity. Sinensetin is one of the PMFs having five methoxy groups on the basic benzo-γ-pyrone skeleton with a carbonyl group at the C4 position. Sinensetin have been known for exerting various pharmacological activities including anti-angiogenesis, anti-diabetic and anti-inflammatory activities. However, there are no studies focused on the anti-inflammatory effects of sinensetin on skeletal muscle cells. In the present study, we investigated the antiinflammatory effect of flavonoids isolated from Sinensetin on the production of pro-inflammatory mediators mediated by nuclear factor-kappa B (NF-κB) by inhibition of signal transduction in LPS - induced L6 skeletal muscle cells.
Because the pig is a valuable candidate for a preclinical model of human cell therapy as well as an important food source, understanding a physiology of pig myogenic progenitors such as skeletal muscle satellite cells and myoblasts is required for cure of muscular diseases and improvement of meat production. For these reasons, we tried to isolate and culture the pig progenitor cells from skeletal muscle. Pig satellite cells, known as muscle stem cells, were isolated from biceps femoris of neonatal pigs by enzymatic digestion method. Muscle satellite cells are quiescent in uninjured muscle. Upon injury, they are activated into proliferating state, known as myoblasts, by growth factors and, in turn, differentiated forward to myocytes and myotubes. To trigger proliferation in vitro, the isolated satellite cells were cultured with epidermal growth factor (EGF) and dexamethasone (BMP4 inhibitor). As a result, the pig satellite cells were efficiently converted into proliferating myoblasts and stably maintained over an extended period. The myoblasts were confirmed by markers of PAX7, MYF5, and MYOD1. Our finding showed that modulating EGF and BMP4 signaling are essential for maintaining muscle stem cells. This culture method could be applied for a production of cultured meat and further basic research of muscle development.
This work was supported by the Korea Institute of Planning and Evaluation for Technology in food, agriculture, forestry, and fisheries (IPET) through the Development of High Value-Added Food Technology Program funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA; 118042-03-1-HD020).
To test the muscle cell specific gene expression, we examined the ability of human α-skeletal muscle actin (ACTA) promoter or human myoglobin (hMb) promoter to direct the expression of the GFP gene in both muscle and non-muscle cells, respectively. C2C12 cells, a mouse myoblast cell line, provide a powerful model to study skeletal muscle differentiation in vitro. We intended to use this cell line as a model for skeletal muscle-specific gene expression during myogenic differentiation from myoblast to myotubes. We compared marker gene expression profiles of proliferating and differentiated C2C12 cells using RT-PCR and fluorescent microscopy analysis. Also, we found that the expression of PCK1 gene under the control of ACTA promoter was proportionally increased as C2C12 differentiated into myotube form. PCK1 is involved in the regulation of gluconeogenesis. In previous research, transgenic mice with overexpressing PCK1 in skeletal muscle showed a greatly enhanced level of physical activity, which extends well into old age. This is due, in part, to an increased number of mitochondria and a high concentration of triglyceride in their skeletal muscles. These mice also had very little body fat, despite eating 60% more than controls. We also constructed a mesenchymal stem cell line and fetal fibroblast cell line for the experiments aiming to make transgenic animals in which the PCK1 gene is specifically expressed in muscle tissue. Accumulated knowledge of this approach could be applicable to a variety of related biological areas including transgenic animal research, gene function study, anti-aging study, etc.
This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Export Promotion Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (316002-5).
본 연구는 짚신나물 열수 추출물의 α-glucosidase 저해 활 성을 측정하고, 분화된 근육세포에서 glucose 이용과 인슐린 신호전달에 미치는 영향을 분석하였다. 짚신나물 열수 추출 물(10 ㎎/㎖)은 α-glucosidase 활성을 67% 저해하였으며, 같은 농도의 양성대조구인 acarbose(63%)와 유사한 저해 효과를 보였다. 짚신나물 열수 추출물이 α-glucosidase에 의한 단당 류 생성을 저해함으로 식사 후 혈당이 급격히 상승하는 것을 억제하는데 효과적인 소재로 이용 가능성을 확인하였다. 또한 근육세포에서 인슐린 저항성을 유발하기 위해 지방산(1 mM, palmitic acid)를 처리하였고, glucose의 세포내 유입이 감소 되는 것을 확인하였다. 지방산 처리 세포 모델에서 짚신나 물 열수 추출물(10 ㎍/㎖)은 glucose 이용을 유의적으로 회복 시켜 주었다. Normal 상태의 배양조건에서 근육세포의 포도 당 이용능은 짚신나물 열수 추출물(100 ㎍/㎖) 처리에 의해 유의적으로 증가하였다. 근육세포 내로 glucose 유입은 운반 단백질인 Glut4를 통해 이루어지며, 이것은 인슐린이 신호전 달을 통해 조절한다. 짚신나물 열수 추출물의 세포 내 glucose 이용 증가 효과는 인슐린 신호전달 관련 분자인 Akt 유전자 와 단백질 발현을 증가시킨 것과 관련되는 것으로 추정된다. 결론적으로, 짚신나물 열수 추출물은 소화기관에서의 탄수화 물 흡수 저해와 근육세포 내 glucose 이용 증가를 통해 혈당 조절 및 당 대사 개선에 긍정적인 영향을 미치고 있음을 확인 하였다.
X-ray diffraction studies have been made to investigate the effects of binding of ADP, ADP+Vi, ADP+AIF4, ADP+BeF3 on the structure of glycerinated rabbit skeletal muscle in the rigor state. Although these phosphate analogs are known to bind actively cycling myosin heads, it is not clear whether they can bind to the attached heads in the rigor muscle. We have found that these analogs can bind to the myosin heads attached to actin filaments in the rigor state. The present results indicate that (1) bound myosin heads altered their conformation in the proximal end toward the plane perpendicular to the fiber axis when MgADP bound to them, and (2) myosin heads were dissociated substantially (up to 50%) from actin filaments but still remained in the vicinity of actin filaments when MgADP and metallofluorides (AIF4 and BeF3) or vanadate bound to them. We detected new conformations of myosin heads attached to actin filaments when they had MgADP or ADP.Pi analogs. We report here these findings on the effects of MgADP and MgADP+phosphate analogs to the rigor crossbridges.
The aim of this study was to identify the effect of suspension unloading (SU) and electrical stimulation upon the development of neonatal muscular system. For this study, the neonatal rats were randomly divided into three groups: a control group, an experimental group I, and an experimental group II. The SU for experimental group I and II was applied from postnatal day (PD) 5 to PD 30. The electrical stimulation for soleus muscle of experimental group IIwas applied from PD 16 to PD 30 using neuromuscular electrical stimulation (NMES), which gave isometric contraction with 10 pps for 30 minutes twice a day. In order to observe the effect of SU and ES, this study observed myocyte enhancer factor 2C (MEF2C) and vascular endothelial growth factor (VEGF) immunoreactivity in the soleus muscles at PD 15 and PD 30. In addition, the motor behavior test was performed through footprint analysis at PD 30. The following is the result. At PD 15, the soleus muscles of experimental group Iand II had significantly lower MEF2C, VEGF immunoreactivity than the control group. It proved that microgravity conditions restricted the development of the skeletal muscle cells at PD 15. At PD 30, soleus muscles of the control group and experimental group II had significantly higher MEF2C, VEGF, immunoreactivity than experimental group I. It proved that the NMES facilitated the development of the skeletal muscle cells. At PD 30, it showed that SU caused the decrease in stride length of parameter of gait analysis and an increase in toe-out angle, and that the NMES decreased these variations. These results suggest that weight bearing during neonatal developmental period is essential for muscular development. They also reveal that NMES can encourage the development of muscular systems by fully supplementing the effect of weight bearing, which is an essential factor in the neonatal developmental process.
Since ultrasound has different reflections depending on components of organization, analysis of ultrasound images of skeletal muscle can offer both quantitative and qualitative reports as concerns skeletal muscle structure. This study is focused on the ultrasound method for evaluating the structural characteristics of skeletal muscle and also conducted to examine its practicality. After obtaining images of the elbow flexors from an ultrasound image device with 88 normal subjects whose ages were between twenty and seventy years old (44 men and 44 women), muscular density and white area index (WAI) which indicated structural characteristics of skeletal muscle were analyzed with gray scale analysis. The study examined correlations between subject's age and items which obtained from measuring ultrasound images and the differences in relations to sex and age. Muscular density and WAI had a high correlation with age and were significantly increased in men and women with greater age. The quantitative evaluation method of skeletal muscle structure which analyzed the ultrasound images has high practicality because it is a non-invasive method which complements physical therapy diagnosis and research methods and promotes functionality evaluation.
The purpose of this study was to investigate the treatment effect of three interventions on the disuse atrophy of rat hindlimb after two weeks suspension. Forty-eight 11~12 weeks old female Sprague-Dawley white rats were divided into four intervention groups: 1) suspension only (S; n=10), 2) intensive weight bearing treadmill (IWBT; n=10), 3) electrical stimulation (ES; n=9), 4) 2)+3) (ES/IWBT; n=9). Another 10 rats received no intervention or hindlimb suspension and served as controls (C). After the interventions, 1) the cross-sectional area (CSA), 2) the ratio of white muscle fiber composition (WMFC), 3) isometric tetanic tension (ITT), and 4) muscle weights (MWs) were measured from the four calf muscle specimens. The results were as follows: 1. In all intervention groups, the CSAs of medial and lateral gastrocnemius (MG LG), soleus (SOL), and flexor digitorum superficialis (FDS) decreased when compared to the control (C) group (p<.05). The CSA increased in FDS and LG for the IWBT group, in SOL for the ES/IWBT group compared to the S only group (p<.05). 2. The ratios of WMFC in MG, LG, SOL, and FDS increased compared to the C group for all interventions (p<.05). The ratios of WMFC decreased in SOL and LG for the IWBT group, in SOL for the ES/IWBT group compared to the S only group, and decreased in SOL for the ES/IWBT group compared to the IWBT group (p<.05). 3. The ITT in the MG, LG, SOL, and FDS decreased compared to the C group for all interventions (p<.05). The ITT increased in MG LG/FDS, SOL, and the whole calf muscles (WCMs) in the IWBT, ES and ES/IWBT groups compared to the S only group (p<.05). 4. The MWs in MG LG/FDS, SOL, WCMs decreased compared to the C group for all interventions (p<.05). The MWs increased in MG LG/FDS and WCMs for the IWBT group, in SOL for the ES group, and in SOL for the ES/IWBT group compared to the S only group (p<.05). 5. In atrophied muscles, the IWBT group showed the best recovery and the ES/IWBT and ES groups followed in decreasing order. The most susceptible muscle to disuse atrophy was the SOL. But conversely, it showed the best recovery in the ES/IWBT group. After two weeks of hindlimb suspension, the calf muscles of rats atrophied and their isometric tension decreased. These changes were best reversed by hindlimb-focused treadmill activity. The next best results were achieved by electrical stimulation combined with the treadmill followed by only electrical stimulation. These findings indicate that full weight bearing treadmill activity alone or in combination with electrical stimulation are effective treatments for non-weight bearing induced muscle atrophy. Further study of the effect of different intensities of electrical stimulation and variations in the duration period of full weight bearing treadmill activity on disuse atrophy is recommended.
The decrease of muscle power and muscle size between twenties and seventies was about 30% and 40% respectively. The loss of muscle mass by aging resulted in the decrease of muscle power. The loss of muscle mass was due to the decrease of number of Type I fiber and Type II fiber and size of each muscle fiber. The aging skeletal muscle didn't show the loss of glycolysis capacity but showed 20% decrease of the oxidative enzymes and muscle vascularization. The vigorous endurance exercise training with graded intensity played a role in the vascular proliferation, increase of activity of oxidative enzymes and improvement of max. The graded resistance exercise also played a role in the muscle hypertrophy and increase of muscle power, if it performed with adequate intensity and period. The exercise adaptation of aging skeletal muscle prevented it from sarcopenia, provided the activity of daily living with great effect and provided the aging related disease, that is Type II diabetes mellitus, arteriosclerosis, hypertension, osteoporosis and obesity, with great effect.
Astragalus membranaceus is a well known oriental medicinal herb. The polysaccharides of the aboveground parts (AMA) and the radix (AMR) of A. membranaceus are the most important functional constituents. Methods and Results: The aim of this study was to determine the effects of AMA and AMR on the oxidative damage induced in the skeletal muscle of rats subjected to exhaustive exercise. Sprague-Dawley rats were randomly divided into exercise and non-exercise groups; in the groups receiving the test compounds, AMA and AMR were administered orally for 30 days. Skeletal muscle samples were collected from each rat after running to exhaustion on a treadmill to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and the concentation of malondialdehyde (MDA). The antioxidant enzyme activities of SOD and GSH-Px of skeletal muscle of AMA- and AMR-treated groups were significantly higher than those of the control and commercial sports drink (SPD)-treated groups in exhaustive exercise rats. In addition, MDA concentrations in the skeletal muscle of the AMA- and AMR-treated groups were significantly lower than those of the control and SPD-treated groups. In the present study, the effects of AMA and AMR on exercise endurance capacity were also evaluated in mice subjected to a swimming exercise test. AMA and AMR supplementation prolonged the swimming time of mice and enhanced exercise endurance capacity. AMA and AMR possess the ability to retard and lower the production of blood lactate, and prevent the decrease of serum blood glucose. Conclusions: These results showed that, AMR and AMA exerted beneficial effect in mice, increasing the activity of the antioxidant systems and inhibiting oxidative stress induced by exhaustive exercise. The compounds improved exercise performance and showed anti-fatigue effects against exhaustive exercise.