일회성 운동 및 회복기 흰쥐 골격근 내 Small HSP 및 HSP 70과 골격근 손상인자 발현 양상에 대한 연구

박대령 ( Dae Ryoung Park ),윤정수 ( Chung Su Yoon ),임채일 ( Chae Il Lim ),하태균 ( Tae Geun Ha ) 한국운동생리학회 2009 운동과학 Vol.18 No.3

일회성 운동 전· 후 식초음료 복용이 인체의 항염증 반응에 미치는 영향

박대령(Park, Dae-Ryoung),양민영(Yang, Min-Young),희뢰(Ji, Lei),윤정수(Yoon, Chung-Su) 한국체육과학회 2012 한국체육과학회지 Vol.21 No.6

The purpose of this study was to determine to the effect of vinegar drinks on blood inflammatory response during acute endurance exercise and recovery. We investigated the effect of vinegar drinks on serum CK, cellular immune cell, nitric oxide, CRP and IL-6 in human blood. For this, 20 healthy male was recruited and classified control group (CON, N=10) and vinegar drinks supplement group (VS, N=10). Both group carried out acute endurance running exercise (HRmax 70~80%), and Blood collected during the exercise and recovery period. The results of the experiment showed the VS group was significantly decreased Serum CK and immune cell activation (neutrophil, lymphocyte) and inflammatory cytokines (NO, CRP, IL-6)(p<.05). This effect might be brought to the decrease of DOMS after exercise. As results, we derived the conclusion that vinegar drinks supplement on exercise and recovery period is effective on decrease inflammatory response. However a downside of these experiments is that the reason and process for the direct evidence for the reduction of inflammatory response is still unknown.

무부하 자유수영 운동이 C57BL/6mouse 골격근의 PARP및 SIRT1 protein의 발현에 미치는 영향

박대령(Park, Dae-Ryoung),김정석(Kim, Jeong-Seok) 한국체육과학회 2014 한국체육과학회지 Vol.23 No.2

The aim of this study was to investigate effect of PARP inhibition to SIRT1 expression and NAD cyclases during skeletal muscle contraction. unloading swimming exercise and PARP inhibitor, PJ34, used for PARP inhibition during muscle contraction in vivo and in vitro. NAD cyclase related protein expression level were compared by western blotting. Our results demonstrated that PARP inhibition not influenced to SIRT1 and PGC-1α expression. This means no co-regulation effect between SIRT1 and PARP. These results indicated that exercise induced muscle damage mediated PARP activating signal not involved exercise induced thermogenesis through SIRT1 activation.

근수축에 의한 Nitric Oxide 생성과 미토콘드리아 생합성 조절 체계에서 CaMKII의 작용

박대령 ( Dae-ryoung Park ) 한국운동생리학회(구 한국운동과학회) 2016 운동과학 Vol.25 No.2

PURPOSE: This study was to investigate the effect of CaMKII mediated nitric oxide formation on skeletal muscle contraction induced mitochondria biogenesis. METHODS: To examine the effect of CaMKII inhibition on nitric oxide synthase regulation during muscle contraction or exercise, we designed in vivo and in vitro study. In vitro study was skeletal muscle primary study for identify relation between CaMKII and NOS and in vivo study was treadmill running study for NO formation regulation by nNOS or eNOS in tissue level. RESULTS: In skeletal muscle primary cell, nNOS inhibition by nNOS inhibitor or nNOS KD inhibited contraction induced NO formation or mitochondria biogenesis related factor upregulation including PGC-1α deacetylation or mitochondrial mRNA but not by eNOS inhibitor or eNOS KD. CaMKII inhibition by AIP attenuated contraction induced NO formation, nNOS phosphorylation and mitochondria biogenesis related factor upregulation including PGC-1α deacetylation or mitochondrial mRNA. However in vivo nNOS or eNOS KD both inhibited NO formation and mitochondria biogenesis related factor upregulation including PGC-1α deacetylation or mitochondrial mRNA during treadmill running. CONCLUSIONS: These results suggest that increase of CaMKII activity regulates NO formation during muscle contraction via regulation of nNOS phosphorylation and plays pivotal role in mitochondria biogenesis.

근수축시 해당작용에 의한 근형질 세망의 Ca²⁺ 변화가 미토콘드리아 Ca²⁺ 증가에 미치는 영향

박대령 ( Dae-ryoung Park ) 한국운동생리학회(구 한국운동과학회) 2017 운동과학 Vol.26 No.3

PURPOSE: This study was to investigate the Glycolysis mediated sarcoplasmic reticulum (SR) Ca²⁺ signal regulates mitochondria Ca²⁺ during skeletal muscle contraction by using glycolysis inhibitor. METHODS: To examine the effect of Glycolysis inhibitor on SR and mitochondria Ca²⁺ content, we used skeletal muscle fiber from gastrocnemius muscle. 2-deoxy glucose and 3-bromo pyruvate used as glycolysis inhibitor, it applied to electrically stimulated muscle contraction experiment. Intracellular Ca²⁺ content, SR, mitochondria Ca²⁺ level and mitochondria membrane potential (MMP) was detected by confocal microscope. Mitochondrial energy metabolism related enzyme, citric acid synthase activity also examined for mitochondrial function during the muscle contraction. RESULTS: Treatment of 2-DG and 3BP decreased the muscle contraction induced SR Ca²⁺ increase however the mitochondria Ca²⁺ level was increased by treatment of inhibitors and showed and overloading as compared with the control group. Glycolysis inhibitor and thapsigargin treatment showed a significant decrease in MPP of skeletal muscle cells compared to the control group. CS activity significantly decreased after pretreatment of glycolysis inhibitor during skeletal muscle contraction. These results suggest that regulation of mitochondrial Ca²⁺ levels by glycolysis is an important factor in mitochondrial energy production during skeletal muscle contraction CONCLUSIONS: These results suggest that mitochondria Ca²⁺ level can be regulated by SR Ca²⁺ level and glycolytic regulation of intraocular Ca²⁺ signal play pivotal role in regulation of mitochondria energy metabolism during the muscle contraction.

운동 빈도가 노화된 흰쥐 골격근 내 미토콘드리아 생합성 및 동화작용 신호경로 인자의 발현에 미치는 영향

박대령(Park, Dae-Ryoung),김정석(Kim, Jeong-Seok) 한국체육과학회 2014 한국체육과학회지 Vol.23 No.1

The purpose of this study was to investigate the effects of exercise frequency different on expression of mitochondrial biogenesis and anabolic signaling pathway factors in aged rats skeletal muscle. The aged rats in four groups exercises: young control (YC group, n=5), old control (OC group, n=5), old exercise 3days/week (0E3 group, n=7), old exercise 5days/week (OE5 group, n=6) on a treadmill run with an intensity of 21m/min, 10° intensity inclination five times for a week during 6 weeks. After exercise training, expression of the p-AMPK, PGC-1α, SIRT-1, HSP78 and IRS-1, GLUT-4, p-mTOR levels were increased significantly in OE5 groups. The major finding of this study was that OE5 groups contribute to enhance mitochondrial biogenesis and anabolic signaling pathway in aged skeletal muscle than in OE3 groups.

운동 빈도의 차이가 노화된 흰쥐 골격근 내 autophagy factors의 발현에 미치는 영향

박대령(Park, Dae-Ryoung),강윤석(Kang, Yun-Seok),김정석(Kim, Jeong-Seok) 한국체육과학회 2014 한국체육과학회지 Vol.23 No.5

The purpose of this sutyd was to investigate the effects of exercise frequency different on expression of autophagic markers in aged rats skeletal muscle. Divide into six groups: young control (YC group, n=5), young exercise 3days/week (YE3 group, n=7), young exercise 5days/week (YE5 group, n=7), old control (OC group, n=6), old exercise 3days/week (OE3 group, n=5), old exercise 5days/week (OE5 group, n=6) on a treadmill run with an intensity of 21m/min, 10° incline during 6 weeks. After exercise training, expression of the Atg5/12, Beclin 1, Atg7, Cathepsin L (CTSL) levels were increased significantly in both OE3 and OE5 groups. In contrast, LC3=Ⅱ and p62 levels were decreased significantly in the exercise groups in both OE3 and OE5. The major finding of this study was that OE5 groups contribute to enhance autophagic signaling pathway in aged skeletal muscle than in OE3 groups.

고강도 지구성 운동이 골격근내 p53, p-FoxO1 및 MuRF1 단백질의 발현에 미치는 영향

강윤석 ( Youn Seok Kang ),김정하 ( Jeong Ha Kim ),유성경 ( Seong Kyeong Yu ),박대령 ( Dae Ryoung Park ),김재철 ( Jae Cheol Kim ) 한국운동생리학회(구 한국운동과학회) 2011 운동과학 Vol.20 No.4

강윤석, 김정하, 유성경, 박대령, 김재철. 고강도 지구성 운동이 골격근내 p53, p-FoxO1 및 MuRF1 단백질의 발현에 미치는 영향. 운동과학. 제20권 제4호. 379-388. 2011. 고강도 지구력 운동(26 m/min, 10°, 15 min, 30 min, 45 min, 60 min, 90 min)을 하는 동안 쥐의 골격근내 p53, p-FoxO1 및 MuRF1 단백질의 발현을 규명하는데 연구의 목적이 있으며, 골격근내 단백질 발현은 western blotting 방법을 통해 분석하였다. p53 단백질의 발현의 경우 적색 및 백색 비복근 모두 운동 45분에 현저하게 증가하였다. p-FoxO1 단백질 발현의 경우적색 비복근에서는 운동 30분~45분의 증가한 반면에 백색 비복근에서는 운동 15분과 60분에서 높게 증가하는 것으로 나타났다. MuRF1 단백질 발현의 경우 운동 15분과 45분에서 증가한 반면에 백색 비복근에서는 운동 15분에서 높게 증가하는 것으로 나타났다. 이러한 결과는 고강도 지구력 운동을 하는 동안 p53, p-FoxO1 및 MuRF1 단백질이 근세포 손상에 대한 항상성을 유지하는데 중요하게 작용하고 있는 것으로 사료된다. Kang, Y. S., Kim, J. H., Yu, S. K., Park, D. R., Kim, J. C. The expression of p53, pFoxO-1 and MuRF1 protein in skeletal muscle during intensity endurance training. Exercise Science. 20(4): 379-388, 2011. The aim of this study was to investigate the expression of p53, p-FOXO-1, and MuRF-1 protein in the skeletal muscle of Sprague-Dawley rats during intensity endurance exercise (26 m/min, 10°, 15 min, 30 min, 45 min, 60 min, 90 min). The expression of p53, p-FoxO1, and MuRF1 protein in skeletal muscle by western blotting. The expression of p53, p-FoxO1, and MuRF1 protein significantly were increased at 15min to 30 min during intensity endurance exercise and then gradually returned to bofore exercise level. These finding suggest that p53, p-FoxO1, and MuRF1 protein plays a role in the maintennance of homeostasis in skeletal muscle during intensity endurance exercise.

반복적인 근 수축으로 인한 ROS 방어 체계의 향상이 Palmitate로 인한 ER stress 유발 및 미토콘드리아 칼슘에 미치는 효과

한진섭(ZhenXie Han),김상현(Kim, Shang-Hyun),박대령(Park, Dae-Ryoung) 한국체육과학회 2020 한국체육과학회지 Vol.29 No.1

PURPOSE: This study, it was intended to induce repetitive muscular dystrophy in skeletal muscle cells (C2C12) with continuous electro stimulation, check the improvement of the antioxidant system, and determine whether the ER stress by the palmitate can be controlled by iterative muscle contraction. METHODS: To examine the expression of catalase on palmitate-induced ER stress and mitochondria calcium level, 100uM palmitate was treated with C2C12 cell. Palmitate-induced reactive oxygen species(ROS) formation was measured by DCFDA fluorescence dye. ER Ca<SUP>2+</SUP> level was measured by using thapsigargin-induced ER Ca<SUP>2+</SUP> release method which was detected by Flou-4 am, Ca<SUP>2+</SUP> dye. palmitateinduced mitochondria calcium was measured by TMRM dye. ER stress protein PERK expression, phosphorylation, and ER Ca<SUP>2+</SUP> releasing channel related protein, RYR1 and p-RYR were analyzed by western blotting. All parameters were compared between control cell, palmitate - treated cell and palmitate and electro stimulation - treated cell. In addition, the catalase inhibitor, 3-Amino-1,2,4-triazole (3-AT), was treated in cells where the expression of the catalase was increased through electrical stimulation, and an experiment was conducted to establish the relationship between the catalase suppression and the ER stress. RESULTS: The one-week electro stimilation significantly increased the expression of the catalase in skeletal muscle cells and was shown to significantly inhibit the level of ER Ca<SUP>2+</SUP> by Palmitate. In addition, muscle contraction reduced of mitochondria Ca<SUP>2+</SUP> levels by palmitate was also shown to be inhibited, and the expression and phosphorylation of PERK / RYR1, an indicator were reduced. The result of processing 3-AT on cells that performed electro stimulation shows that the increase in ER stress indicators due to palmitate is significantly higher than that of electro stimulating cells. Phosphorylation of PERK / RYR also tends to increase. CONCLUSIONS: Repeated muscle contraction improves anti-oxidization capability in skeletal muscle, and activates the protective system of skeletal muscle related to metabolic disease. Palmitate increase the ROS of skeletal muscle cells and increase the ER stress. This study suggests that the incidence of catalase increases through skeletal muscle contraction an important role in defending the ER stress from palmitate processing, and suggests that the ROS defense system due to the Catalase is an important mechanism for suppressing ER stress and maintaining the mitochondria Ca<SUP>2+</SUP> level.

원문 : Dexamethasone 및 전기 자극 처치 시 근세포로부터 유발된 exosome의 항 근위축 및 근비대 관련 microRNA 및 단백질의 변화에 미치는 효과

김정석 ( Jeong Suk Kim ),박소영 ( So Young Park ),윤정수 ( Chung Su Yoon ),박대령 ( Dae Ryoung Park ) 한국운동생리학회(구 한국운동과학회) 2014 운동과학 Vol.23 No.4

본 연구의 목적은 골격근 세포에 dexamethasone (Dex) 및 전기 자극(ES) 처치 시 발생하는 microRNA(miR)가 각각의 자극에 따라 어떠한 profile을 가지게 되며, 이러한 exosome을 골격근 primary cell에 처치 시 근위축 및 근비대 관련 인자의 인산화가 어떻게 변하는지를 규명하기 위한 것이었다. C2C12 세포주에 Dex 및 ES를 처치하여 골격근 세포로부터 유리된 exosome의 miR의 profile을 분석하였으며, 이러한 과정에서 얻어진 exosome을 골격근 primary cell에 재처리하여 골격근 세포에서의 근위축 및 근비대 관련 인자의 인산화 변화를 관찰하였다. C2C12 세포주에 Dex 혹은 ES를 처치한 결과, 근위축 관련 miR인 miR-1, miR-206, miR-322와 근비대 관련 miR-23a의 발현 양상이 유의하게 차이나는 것을 확인하였다. 또한 이러한 세포에서 얻어진 exosome을 골격근 primary cell에 처치한 결과, Dex를 처치한 세포에서 얻어진exosome은 근위축 관련 인자인 FOXO와 MuRF의 인산화를 증가시키는 것에 비해 ES를 처치한 세포에서 얻어진 exosome은 근비대관련인자인 mTOR와 p70S6K의 인산화를 증가시키는 것으로 나타났다. 이러한 결과는 각각의 신호체계에 의해 발생되는 골격근내 exosome이 세포의 신호전달 체계에 중요한 역할을 하고 있음을 제시하고 있으며, exosome에 의한 miR의 전달이 근위축 및 근비대와 관련된 신호단백질의 인산화를 조절할 수 있음을 제시하고 있다. The purpose of this study was to examine the effect of dexamethasone (Dex) treatment and electrical stimulation (ES) on skeletal muscle cell derived exosome`s microRNA (miR) profile and its effect on skeletal muscle signal protein expression. Exosome obtained from two different C2C12 cell were treated with Dex or ES. Then we profiled exosome`s miR expression and atrophy related miR-1, miR-206, miR-322 were significantly increased with DEX however, hypertrophy related miR-23a was significantly increased only in the ES. Further we treated two different types of exosome to Dex treated skeletal muscle primary cell and two types of exosome differently regulated atrophy related protein (FOXO/MurF) and hypertrophy related protein (mTOR/p70S6K) phosphorylaton. In conclusion, the results indicate that atrophy or hypertrophy signal induces different exosome including different profile of miR and it plays important role in skeletal muscle atrophy and hypertrophy.