지방의 종류가 다른 식이의 섭취가 하지고정 흰 쥐의 근 섬유별 근 위축과 PPAR${\delta}$ 활성에 미치는 영향

이호욱,박미나,이연숙,Lee, Ho-Uk,Park, Mi-Na,Lee, Yeon-Sook 한국영양학회 2011 Journal of Nutrition and Health Vol.44 No.5

본 연구는 지방의 종류와 함량이 다른 식이의 섭취가 하지 고정 흰 쥐의 근육 내 화학 조성, PPAR${\delta}$ 및 관련 효소의 발현에 미치는 영향을 근육 종류에 따라 알아 보기 위해 수행되었다. 연구결과를 요약하면 다음과 같다. 1) 10일 간의 하지고정과 식이 중재에 의해 실험동물의 체중, 식이섭취량 및 혈청 지방산에는 유의적인 차이가 없었다. 2) 10일 간의 하지고정에 따라 근육 양은 하지 근육 종류에 따라, 각각 soleus 에서 약 57%, plantaris에서 약 39% 그리고 gastrocnemius에서 약 38%로 달리 감소되었다 (p < 0.01). 특히 soleus에서 가장 많은 감소 현상을 보였다. 이러한 감소 경향은 식이 지방의 종류 및 함량에 영향을 받지 않았다. 3) 10일 간의 하지고정에 따라 근육 내 조성 변화의 경우, gastrocnemius에서 분석한 근육 총 단백질량은 정상 다리(contralateral)와 비교하여 하지고정에 따라 유의적으로 감소 (CO, 41%; FO, 40%; HCO, 37%; HBT, 38%)하였다. Plantaris, gastrocnemius에서 분석한 근육 내 중성지방량의 경우에도, 정상다리와 비교하여 하지고정에 따라 plantaris(CO, 39%; FO, 27%; HCO, 38%; HBT, 35%)와 gastrocnemius(CO, 38%; FO, 34%; HCO, 33%; HBT, 34%) 모두에서 유의적으로 감소하였다. 그러나, 근 단백질량과 중성지방량의 감소 현상은 모두 식이군 간의 차이는 나타나지 않았다. 4) PPAR${\delta}$ 단백질은 하지고정에 의해 plantaris에서는 유의적으로 증가 (p < 0.05)하였고, gastrocnemius, soleus에서는 증가하는 경향을 나타내었다. PPAR${\delta}$ 단백질과는 반대로 PPAR${\delta}$ mRNA는 하지고정에 의해 plantaris에서는 유의적으로 감소 하였고, soleus에서는 FO군만 PPAR${\delta}$ mRNA 발현 양이 유의적으로 감소하였다. PPAR${\alpha}$의 경우 하지고정에 따라 두 근육에서 모두 영향을 받지 않았다. mCPT1 mRNA는 하지고정에 따라 soleus와 plantaris 모두에서 변화가 없었지만, PDK4 mRNA는 하지고정에 따라 plantaris에서 유의적으로 감소하였고, 반대로 soleus에서는 유의적으로 증가하였다. 5) PPAR${\delta}$ 단백질은 세 근육의 FO군에서 모두 크게 증가하는 경향을 보였지만, 식이에 따른 유의적인 차이를 보이지는 않았다. PPAR${\delta}$ mRNA의 경우, plantaris의 CO군과 비교해 FO군에서 mRNA 발현이 유의적으로 낮은 수준이었고, 다른 군에서는 유의적인 차이가 없었다. Soleus의 PPAR${\delta}$ mRNA는 CO군과 비교해 FO, HCO 그리고 HBT군 모두에서 발현 양이 유의적으로 낮은 수준을 보였다. 반면, PPAR${\alpha}$의 mRNA 발현은 두 근육에서 모두 식이에 의한 영향을 받지 않았다. mCPT1 mRNA 발현의 경우, plantaris에서는 CO군과 비교해 FO군에서 발현 양이 유의적으로 낮은 수준을 보였지만, 고지방 식이 군에서는 유의적인 차이가 없었다. 반대로, soleus에서는 CO군과 비교해 FO군에서 발현이 유의적으로 높은 수준을 보였지만, 고지방 식이 군에서는 CO군과 비교해 유의적인 차이는 없었다. PDK4 mRNA의 경우, plantaris에서는 CO군과 비교해 FO군의 mRNA 발현 양이 유의적으로 높은 수준을 보였다. 고지방 식이 군에서는 CO군과 비교해 발현 양의 유의적인 차이가 없었다. 한편, soleus에서는 FO군에서 발현이 낮은 경향을 보였을 뿐, 식이에 따른 mRNA 발현에는 모든 군에서 유의적인 차이가 없었다. 본 연구에서는 10일간의 하지고정에 의해서 모든 근육에서 유의적인 근 위축이 유발되었고, 이에 따라 근육 양의 감소와 함께 근육 총 단백질의 유의적인 감소가 나 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.

Activation of mTOR for the Loss of Skeletal Muscle in a Hindlimb-Suspended Rat Model

유영민,박지형,서동현,Sinae Eom,정영진,김택중,한태영,김한성 한국정밀공학회 2015 International Journal of Precision Engineering and Vol.16 No.5

The hindlimb-suspended or unloading rodent model was developed to mimic the microgravity of a spaceflight environment and has been used extensively to study the physiological responses to various aspects of musculoskeletal loading and unloading. This study investigated a compensating response to skeletal muscle loss in a hindlimb-suspended model that involves up-regulation of mTOR phosphorylation at four weeks. Body weight and muscle volume significantly decreased over four weeks in the hindlimb-suspended group compared with the sham control. Expressions of p-mTOR, raptor protein, p-p70S6K, p-Akt, and p-ERK, anti- or pro-apoptotic p53, Bcl-2, Bcl-XL, Bax, and Bak proteins were significantly increased in the hindlimb-suspended group compared to the sham control at four weeks. These results indicate that p-mTOR and p-Akt/p-ERK proteins might be up-regulated to compensate for the loss of skeletal muscle in the hindlimb-suspended model. Expressions of p-AMPK, IRS1, and IRS2 proteins were significantly increased, but that of p-eIF2α was significantly reduced in the hindlimb-suspended group compared to the sham control at four weeks. Our results suggest that the loss of skeletal muscle in a hindlimb-suspended model induces activation of p-mTOR and p-Akt/p-ERK proteins, and that these signaling pathways may preserve protein synthesis and cell growth in skeletal muscles of hindlimb-suspended animals.

목과약침(木瓜藥鍼)이 후지체공(後肢滯空)으로 유발(誘發)된 근위축(筋萎縮) 흰쥐의 근섬유(筋纖維) 변화(變化)에 미치는 영향(影響)

전철기,남상수,박동석,Chun, Chul-Ki,Nam, Sang-Soo,Park, Dong-Suk 대한침구의학회 2001 대한침구의학회지 Vol.18 No.1

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.

Effects of Intermittent Sciatic Nerve Stimulation on the Soleus and Medial Gastrocnemius Muscle Atrophy in Hindlimb Suspended Rats

Park, Byung-Rim,Cho, Jung-Shick,Kim, Min-Sun,Chun, Sang-Woo The Korean Physiological Society 1992 대한생리학회지 Vol.26 No.2

The present study was designed to evaluate effects of intermittent electrical stimulation of the sciatic nerve on the atrophic response of antigravity muscles, such as the soleus (slow m.) and medial gastrocnemius (fast m.) muscles. Rats (Sprague-Dawley, 245-255g) were subjected to a hindlimb suspension and divided into three groups : one was with hindlimb suspension (MS) and another with hindlimb suspension plus intermittent electrical stimulation of the sciatic nerve (HS ES). Control group (CONT) was kept free without strain of the hindlimb. After 7 days of hindlimb suspension, the soleus and medial gastrocnemius muscles were cut at their insertion sites, and were then connected to the force transducer to observe their mechanical properties. Optimal pulse width and frequency of electrical stimulation were 0.2ms, 20Hz for the soleus muscle and 0.3ms, 40Hz for the medial gastrocnemius muscle under supramaximal stimulation. Body weight and circumference of the hindlimb were significantly decreased in HS and HS-ES groups compared with the control group. In HS-ES group, however, the weight of the soleus muscle was not different from that in the control group while the weight of the medial gastrocnemius muscle was lower than that in the control group. In HS group, mechanical properties of muscle contraction including contraction time, half relaxation time, twitch tension, tetanic tension, and fatigue index of both muscles were significantly decreased compared with the control group except for twitch tension and tetanic tension of medial gastrocnemius muscle. The degree of atrophy of the soleus muscle in HS group was more prominent than that of the medial gastrocnemius muscle. Twitch tension and fatigue index of the soleus muscle and fatigue index of the medial gastrocnemius muscle in HS-ES group were not different from those of the control group. While mechanical properties of the soleus muscle examined were all significantly increased in HS-ES group compared with HS group, only contraction time and fatigue index of the medial gastrocnemius muscle were significantly increased in HS-ES group. These data indicate that intermittent electrical stimulation may be useful in prevention of muscle atrophy.

AHCC Supplementation Attenuates Muscle Atrophy via Akt Activation in Hindlimb-suspended Rat

Young-ju Song,Sang-Hoon Bae,Jin-Young Park,Hye-Jin Lee,My-Young Lee,Seung-Jea Lee,Young-Ki Min,Jeong-Beom Lee 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.3

We investigated the inhibitory effect of active hexose correlated compound (AHCC) on muscle atrophy in hindlimb-suspended rats. Twenty-four six-week-old male Sprague-Dawley rats were randomly divided into three groups: the control sedentary group (CS, n = 8), the hindlimb-suspended group (HS, n = 8), and the hindlimbsuspended and AHCC-supplemented group (HSA, n = 8). Hindlimb suspension and AHCC supplementation were performed for two weeks. The HSA group was treated with AHCC (1 g/1 kg of body weight (BW)) orally in 0.3 mL of PBS solution, while the HS group received the vehicle (PBS solution) only. After two weeks, the cross-sectional area (CSA) of the HS and HSA groups decreased by approximately 36% (p < 0.05) and 19%, respectively, compared to the CS group. In addition, myonuclear numbers of the HS and HSA groups and the extensor digitorum longus (EDL) muscle weight of the HS group decreased 30% (p < 0.05) and 18%, respectively, compared to the CS group. AHCC supplementation increased the phosphorylation of pAkt/Akt in the HSA group compared to the HS group (p < 0.05). Furthermore, Fbx32 and MuRF1 protein expression in the HSA group recovered to the level of the CS group. Based on these results, AHCC supplementation may have a positive role in the prevention of muscle atrophy via Akt activation in hindlimb-suspended rats.

냉동보관 제대혈 신생혈관 유도성세포배양과 허혈성 하지 쥐모델에서의 치료효과연구

김성환 ( Sung Whan Kim ),손영덕 ( Young Doug Sohn ),변기현 ( Ki Hyun Byun ) 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4

Currently, human bone marrow was thought to be the main source of stem cells. However, bone marrow derived stem cell is a limited source of cells for allograft transplantation, because it is not easy to collect bone marrow and they also have the possibility of viral infection. Therefore, as an alternative stem cell source, human umbilical cord blood(hUCB) could overcome the limitations of bone marrow derived stem cells. However, it has been difficult to isolate mesenchymal stem cells(MSCs) from cryopreserved hUCB due to the containment of rare populations of mesenchymal progenitor cells(MPCs). We tried to establish the culture system for angiogenic cells isolation from cryopreserved hUCB. As a result, we successfully could culture angiogenic cells that have endothelial specific surface markers and the capacity for differentiation potential into endothelial cells. In addition, transplantation of these cells into ischemic hindlimb mice showed that increased blood flow and higher limb salvage rate in the hindlimb ischemia. Immunohistochemistry results also demonstrated that hUCB derived cells are detected in the arterial walls of the ischemic hindlimb. Therefore, it is suggested that transplantation of cultured angiogenic cells derived from cryopreserved hUCB may be a novel and useful therapeutic armament for similar ischemic diseases.

부분 진동이 하지현수에 의한 골다공증 예방에 미치는 효과

엄시내(Sinae Eom),박지형(Ji Hyung Park),서동현(Dong-Hyun Seo),김한성(Han Sung Kim) Korean Society for Precision Engineering 2014 한국정밀공학회지 Vol.31 No.8

The aim of this study was to assess the effects of artificial unloading induced by hindlimb suspension on the trabecular bone in tibiae. Twenty four 12-week-old Sprague-Dawley rats were assigned to 3 groups, namely, the control group (CON, n = 8), the hindlimb-suspended group (HLS, n = 8) and HLS with partial vibration group (HLSPV, n = 8). After 4 weeks, compared with CON group, HLS group had significantly greater decreases on BMD, BV/TV, Tb.N, Conn.Dn and increase on Tb.Sp (p < 0.05). However, there were no significant differences in BMD and the other micro structural parameters of tibial trabecular bones between CON and HLSPV (p > 0.05). These results implied that partial vibration might inhibit the bone loss induced by hindlimb suspension. Furthermore, we could expect to apply partial vibration system in space environment, to prevent bone loss in astronauts.

The anti-microbial peptide SR-0379 stimulates human endothelial progenitor cell-mediated repair of peripheral artery diseases

( Tae Wook Lee ),( Soon Chul Heo ),( Yang Woo Kwon ),( Gyu Tae Park ),( Jung Won Yoon ),( Seung-chul Kim ),( Il Ho Jang ),( Jae Ho Kim ) 생화학분자생물학회(구 한국생화학분자생물학회) 2017 BMB Reports Vol.50 No.10

Ischemia is a serious disease, characterized by an inadequate blood supply to an organ or part of the body. In the present study, we evaluated the effects of the anti-microbial peptide SR-0379 on the stem cell-mediated therapy of ischemic diseases. The migratory and tube-forming abilities of human endothelial progenitor cells (EPCs) were enhanced by treatment with SR-0379 in vitro. Intramuscular administration of SR-0379 into a murine ischemic hindlimb significantly enhanced blood perfusion, decreased tissue necrosis, and increased the number of blood vessels in the ischemic muscle. Moreover, co-administration of SR-0379 with EPCs stimulated blood perfusion in an ischemic hindlimb more than intramuscular injection with either SR-0379 or EPCs alone. This enhanced blood perfusion was accompanied by a significant increase in the number of CD31- and α-SMA-positive blood vessels in ischemic hindlimb. These results suggest that SR-0379 is a potential drug candidate for potentiating EPC-mediated therapy of ischemic diseases. [BMB Reports 2017; 50(10): 504-509]

주기적인 운동이 뒷다리 부유에 의해 유발된 쥐의 뒷다리근 위축경감에 미치는 영향

최명애,신기수 대한기초간호자연과학회 1999 Journal of korean biological nursing science Vol.1 No.1

The purpose of this study was to determine the effect of periodic low-intensity exercise during hindlimb suspension on the mass, relative weight, myofibrillar protein content in soleus, plantaris and gastrocnemius muscles. To examine the effectiveness of periodic low-intensity exercise on mass, and myofibrillar protein content of hindlimb muscles, adult female Wistar rats were suspended(HS) and half of these rats walked on a treadmill for 45min/day(15 min every 4h) at 5m/min and a 15˚ grade(HS-EX). Soleus wet weight was 33.51% significantly smaller(p<0.005) and relative soleus weight of hindlimb suspended rats was 31.96% smaller(p<0.005) compared with those of control rats following seven days of hindlimb suspension. Plantaris wet weight was 7.5% smaller(p<0.01) and relative plantaris weight was 11.83% smaller(p<0.05) compared with those of control rats following seven days of hindlimb suspension. Gastrocnemius wet weight was 11.31% significantly smaller(p<0.005) and relative gastrocnemius weight was 17.13% significantly smaller(p<0.005) compared with those of control rats following seven days of hindlimb suspension. Soleus wet weight while increased by relative soleus weight increased by 25.13%, 27.59% each through periodic low intensity exercise during hindlimb suspension(p<0.05, p<0.05). Plantaris wet weight and relative plantaris weight increased by 1.04%, 10.98%(p<0.05) each, and gastrocnemius wet weight and relative gastrocnemius weight increased by 1.98%, 12.02%(p<0.05) each through periodic low intensity exercise during hindlimb suspension. Wet weight of soleus, plantaris and gastrocnemius in HS-EX rats did not recover to control level. Myofibrillar protein content of soleus, plantaris and gastrocnemius was 48.24%, 40.85% and 37.33% significantly smaller(p<0.005) respectively compared with those of control rats following seven days of hindlimb suspension. Myofibrillar protein content of soleus, plantaris and gastrocnemius increased by 40.68%, 25.07% and 17.93%(p<0.005) each through periodic low intensity exercise during hindlimb suspension. Myofibrillar protein content of soleus, plantaris, and gastrocnemius in HS-EX rats did not recover to control level. The results suggest that periodic low intensity exercise can attenuate hindlimb muscle atrophy induced by hindlimb suspension.

전기자극이 흰쥐 위측근의 수축반응에 미치는 영향

김동희,김유섭,전향원 대한스포츠의학회 1994 대한스포츠의학회지 Vol.12 No.2

The effect of hindlimb suspension on the mechanical properties of contraction of the hindlimb muscles and the effect of the intermittent electrical stimulation on the atrophic response of antigravity muscles were evaluated. Muscle weight, contraction time (CT), half relaxation time(HRT), maximum twitch tension(Pt), maximum tetanic tension(Po), and fatigue index(FI), were investigated. Sprague-Dawley rats weighting 230-250g were subjected to hindlimb suspension and divided into six groups: the first was hindlimb suspension (HS) group, the second hindlimb suspension with 20Hz electrical stimulation (HS-ES 20), the third hindlimb suspension with 40Hz electrical stimulation (HS-ES 40), the fourth spontaneous recovery after hindlimb suspension (PHS-SR), the fifth electrical stimulation during recovery period (PHS-ES), and the last the control group. The results from the statistical analysis of these data were summarized as follows; 1. Weight of soleus and medial gastrocnemius muscles were decreased significantly by the 7 days hindlimb suspension, and the decrease of soleus muscle weight in the HS-ES 20 group and the decrease of medial gastrocnemius muscle weight in the HS-ES 20 and the HS-ES 40 group were less severe than those of the HS group. It proved that the hindlimb suspension with electrical stimulation might be useful in preventing the effect of muscle atrophy. 2. Muscle weight of the PHS-SR group and the PHS-ES group were not retrieved in the 7 days recovery period without hindlimb suspension but were superior in the recovery of muscle atrophy. The soleus muscle weight of the electrical stimulation group was similar to that of the control group, however, the medial gastrocnemius muscle weight shows no difference in the 3 groups. 3. CT, HRT, Pt and Po of the medial gastrocnemius muscle of the HS group decreased prominently, and the 20Hz and the 40HZ stimulation showed more efficient effect than the 20Hz in Po and FI of that muscle. 4. In the change of mechanical properties of the soleus muscle after the recovery period without the strain of the hindlimb in the PHS-ES group, the electrical stimulation was effective for preventing muscle atrophy. Especially GRT and FI of the muscle were retrievd similarly to those of the control group. Recovery of CT, HRT, Pt and Po of the medial gastrocnemius muscle were similar to those of the control group but FI of the PHS-SR and the PHS-ES group was decreased. Mechanical properties of the soleus muscle contraction were different from those of the medial gastrocnemius muscle. The changes of mechanical properties were more serious in the soleus than in the medial gastrocnemius muscle. Electrical stimulation during the recovery period or during the hindlimb suspension showed more prominent effect in prevention of recovery of muscle atrophy. In conclusion, the results of these data indicate that the intermittent electrical stimulation, especially after the appearance of muscle atrophy, may be useful in preventing muscle atrophy, and that the optimal pulse width and the frequency of electrical stimulation were 0.2ms and 20Hzfor the soleus muscle and 0.3ms and 40Hz for the medial gastrocnemius muscle, under the supramaximal stimulation.