Short-term repeated treadmill exercise more increases cell proliferation and brain-derived neurotrophic factor expression in the rat hippocampus

정지헌 경희대학교 대학원 2011 국내박사

Physical exercise enhances cell proliferation and/or neurogenesis in the hippocampal dentate gyrus. Brain-derived neurotrophic factor (BDNF) is implicated in the modulation of neurotransmitters turnover, neurite genesis, and neuronal survival. BDNF binds to the tyrosine kinas B (TrK). In this study, we compared the effects of one bout of long-duration of treadmill exercise and repeated short-duration of treadmill exercise on cell proliferation and expressions of BDNF and TrKB in the hippocampus. Adult male Sprague-Dawley rats, weighing 200 ± 5 g, were randomly divided into three groups (n = 8 in each group): control group, one hour-one bout-treadmill exercise group, and ten minutes-six bouts-treadmill exercise group. The rats in one hour-one bout-treadmill exercise group was forced to run on a motorized treadmill for 60 min once a day (at 16:00), and the rats in ten minutes-six bouts-treadmill exercise group forced to run on a treadmill once 10 minutes per six times for a day (at 10:00, 11:00, 14:00, 15:00, 19:00, 20:00). This treadmill exercise was continued for 7 consecutive days. The rats in control group were left without running. Both types of treadmill exercise increased cell proliferation and enhanced expressions of BDNF and TrkB in the hippocampus. Cell proliferation and BDNF expression was more potently increased by ten minutes-six bouts treadmill exercise than one hour-one bout treadmill exercise. These results showed that repeated bouts of short-duration treadmill exercise may achieve more useful results for their brain functions than one-bout of long-duration treadmill exercise. 운동은 해마 치상회에서 신경세포의 생성을 증가시킨다. 뇌 유래 신경영양 인자 (brain-derived neurotrophic factor, BDNF)는 신경전달 물질의 분비, 신경 성장, 그리고 신경의 생존 등에 관여하며, tyrosine kinas B (TrK)에 결합하게 된다. 본 연구에서, 일회의 긴 시간 운동과 여러 번의 짧은 시간의 운동이 해마에서 신경세포 생성과 뇌 유래 신경영양 인자와 TrKB의 발현에 미치는 영향을 비교하였다. 무게 200 ± 5 g의 Sprague-Dawley계 흰쥐를 대조군, 하루 1시간 1회 운동군, 하루 10분씩 6회 운동군의 세 군으로 분류하였다 (각 군 = 8마리). 하루 1시간 1회 운동군은 16시에 매일 한 시간씩 트레드밀 운동을 시행하였다. 하루 10분씩 6회 운동군은 10시, 11시, 14시, 15시, 19시, 20시에 각 10분씩 하루 6번 트레드밀 운동을 시행하였다. 이 운동은 7일간 지속하였다. 두 가지 형태의 트레드밀 운동 모두 해마에서 신경세포 생성과 뇌 유래 신경영양 인자, 그리고 TrkB의 발현을 증가시켰다 신경세포 생성과 뇌 유래 신경영양 인자는 하루 10분씩 6회 운동군에서 더욱 증가되었다. 본 실험의 결과 긴 시간의 일회 운동보다 짧은 시간 여러 번 운동이 뇌기능의 향상에 좋은 결과를 미침을 알 수 있었다. 중심단어: 트레드밀 운동, 뇌유래 신경영양 인자, 세포 생성, 해마, 흰쥐

Spectral Analysis of Heart Rate Variability during Treadmill Exercise at Various Speeds and Grades

김기홍 Kyungpook National Univ. 1996 국내석사

본 연구는 다양한 속도 및 경사도의 treadmill 운동중 심박수 변이성을 스팩트럼 분석하여 순환계 조절기전을 밝힘을 목적으로 하였다. 20-26세의 남자 대학생 33명을 대상으로 하여 속도 4.02, 5.47, 6.76 및 8.05 km/h와 경사도 0, 4, 8, 12, 16 및 20%를 조합한 treadmill 운동을 실시하였다. 각 주행시 treadmill의 속도는 고정한 채 3분마다 경사도를 증가시켰으며, 심전도, 호흡 및 보행 동작을 A/D 변환기를 통해 컴퓨터 디스크에 연속 기록하였다. Fast Fourier transform 법에 따라 심박수 변이성의 power spectrum을 얻고, 이것으로부터 총 파워와 초저주파(VLF: 0-0.04 Hz), 저주파 (LF: 0.04-0.15 Hz) 및 고주파 (HF: 0.15-1.00 Hz) 영역에 대한 파워를 구하였다. 심박수는 안정시 74.4 土 2.1 beats/min였고 treadmill 운동중 속도와 경사도가 증가함에 따라 196.7 土 5.0 beats/min까지 점차적으로 증가하였다. 심박수 변이성의 총 파워는 안정시 35.0 土 6.7 (beats/min)²였고 운동중 1.9 土 0.3 (beats/min)²까지 점차적인 감소를 보였다. %VLF 파워는 안정시 34.5 土 3.7 %였고 최대 운동부하시 감소를 보인 것 외에는 운동중 유의한 변화를 보이지 않았다. %LF 파워는 안정시 44.1 土 3.0 %였고 심박수가 135 beats/min를 초과하는 운동중 4.5 土 1.0 %까지 점차적인 감소를 보였다. %HF 파워는 안정시 21.4 土 2.9 %였고 심박수가 165 beats/min를 초과하는 강한 운동중 87.1 ± 6.7 %까지 점차적인 증가를 보였다. 고주파 영역의 peak 주파수는 안정시 0.200 土 0.018 Hz였고 심박수가 135 beats/min를 초과하는 운동중 0.909 土 0.048 Hz까지 증가하였다. 호흡수는 안정시 18.0 土 1.5 breaths/min였고 운동중 53.0 土 3.7 breaths/min까지 유의하게 증가하였다. Treadmill 운동중 보수 (stride frequency)는 속도가 증가함에 따라 4.02 km/h시 55.6 土 0.9 steps/min로부터 8.05 km/h시 81.2 土 0.6 steps/min까지 증가하였다. 이상을 종합하면 심박수 변이성의 총 파워는 운동강도가 증가함에 따라 부교감 신경 활동도의 감퇴로 인해 점차적으로 감소하였다. 높은 강도의 운동시 속도와 경사도가 증가함에 따라 점차적으로 %LF 파워는 감소하였고 %HF 파워 및 그 peak 주파수는 증가하였으며, 이것은 각각 순환계 조절기전의 재조정과 호흡활동 및 호흡수의 증가에 기인하는 것으로 생각된다. This study was aimed to elucidate the relationship between combinations of treadmill speed-grade and heart rate variability by spectral analysis. Thirty-three untrained male college students aged 20-26 yr were employed to exercise on a treadmill using 4 speeds (4.02, 5.47, 6.76 and 8.05 km/h) and 6 grades (0, 4, 8, 12, 16 and 20%). A fixed speed was selected for each session with the grade increased every 3 min. The electrocardiogram, respiration and the stepping activity were continuously recorded through an A/D converter system on the computer disk. Power spectra of heart rate variability (HRV) were obtained by use of a fast Fourier transform algorithm. The frequency domain was divided into 3 bands: VLF (0-0.04 Hz), LF (0.04-0.15 Hz) and HF (0.15-1.00 Hz). Heart rate was 74.4 ± 2.1 beats/min at rest and showed a steady increase during treadmill exercise with increasing speed and grade up to 196.7 ± 5.0 beats/min. Total power of HRV was 35.0 ± 6.7 (beats/min)² at rest and progressively decreased during exercise down to 1.9 ± 0.3 (beats/min)². The %VLF power of HRV was 34.5 t 3.7 % at rest and showed no significant change during exercise except for a decrease observed at the highest intensities of exercise. The %LF power was 44.1 ± 3.0 % at rest and showed a progressive decrease down to 4.5 ± 1.0 % during those stages of exercise where heart rate was over 135 beats/min. The %HF power was 21.4 ± 2.9 % at rest and showed a progressive increase up to 87.1 ± 6.7 % during higher intensity exercise where heart rate was over 165 beats/min. Peak frequency of HF band was 0.200 ± 0.018 Hz at rest and was shifted to higher frequencies up to 0.909 ± 0.048 Hz at heart rates greater than 135 beats/min. Respiratory frequency was 18.0 ± 1.5 breaths/min at rest and significantly increased during exercise up to 53.0 ± 3.7 breaths/min. Stride frequency during treadmill exercise showed an increasing tendency with increasing speed from 55.6 ± 0.9 steps/min at 4.02 km/h to 81.2 ± 0.6 at 8.05 km/h. It was concluded that total power of HRV decreased progressively with increasing exercise intensity due to the withdrawal of parasympathetic activity. At higher exercise intensities, %LF power decreased and %HF power increased with its peak frequency shifted to higher values in a progressive mode with increasing speed and grade, reflecting a readjustment in the cardiovascular system and the increased respiration and its rate, respectively.

Impact of treadmill exercise after lymph node dissection followed by radiotherapy in a rodent model

김상아 울산대학교 대학원 2023 국내석사

Background and Purpose: Lymphedema is a chronic condition caused by radiotherapy and surgery, and exercise has been reported to have a positive effect on this condition. However, exercise-induced changes in the lymphatic system and skin after lymph node dissection and radiotherapy are not well established. Therefore, the aim of this study was to examine alterations in lymphatic drainage pathway, skin thickness, and the extent of fibrosis in rats with lymph node dissection and radiotherapy after treadmill exercise. Materials and Methods: 12 Sprague-Dawley rats were divided into exercise and control groups of 6 rats each, and lymphedema was induced by popliteal and inguinal lymph node dissection followed by irradiation. The exercise group underwent treadmill exercise for 30 minutes per day, five days a week for four weeks. The hind limbs were imaged using indocyanine green (ICG) lymphography every other week during the exercise period to visualize lymphatic function, while ankle thickness was measured weekly to assess changes in the extent of swelling. Histopathological analysis was conducted at the end of the exercise period to examine dermal thickness, lymphatic vessel density, and the degree of fibrosis. Results: Exercise after surgery and irradiation resulted in improved lymphatic fluid retention. At week 3, the ICG lymphography showed that the group that performed exercise had a higher occurrence of linear and splash patterns. In addition, Ankle thickness was consistently lower in the exercise group, but there was exclusively significant difference between exercise and control groups at week 4 (p= .016). The group that underwent exercise showed a thinner epidermis and dermis compared to the control group (p = .041 and .002 each), as well as a lower percentage of collagen area (p = .002) and a higher lymph vessel density (p = .002). Conclusion: The results show that exercise may have a positive effect on the pathology of the lymphatic system after treatment that may induce lymphedema, including surgery and radiotherapy.

The effect of the whole body vibration exercise on mechanism of the Alzheimer in the APPsw/PS2m transgenic mouse : 전신진동이 운동의 효과로서 APPsw/PS2m 형질전환 마우스의 알츠하이머 기전에 미치는 영향

권기천 한국체육대학교 일반대학원 2024 국내박사

The effect of the whole body vibration exercise on mechanism of the Alzheimer in the APPsw/PS2m transgenic mouse The purpose of this study was to determine the effect of 12-week whole-body vibration exercise on Alzheimer's disease compared to treadmill exercise. First, 24-month-old Alzheimer's disease mice were divided into a comparative group (NonTg, n=6), an Alzheimer's group (Tg, n=6), a treadmill exercise group (TTE, n=6), and a whole-body vibration exercise group (TVE, n=6). Treadmill exercise was performed for 5 days and 12 weeks at a speed of 8m/min for 5 minutes, 10m/min for the next 5 minutes, and 12m/min for the last 20 minutes, while whole-body vibration exercise was performed for 5 minutes at 4Hz, 6Hz 5 minutes, and 8Hz 20 minutes for 5 days and 12 weeks. As a result, cognitive function showed positive results. In body composition, only the whole-body vibration exercise group decreased weight and fat, and there was no difference in cortisol, a stress-related factor. As a result of analyzing the Amyloid beta-related indicators, Amyloid beta was reduced in the exercise group, and in APP and BACE1, all three groups increased compared to the NonTg group. As a result of analyzing autophagy-related indicators, Beclin1, BNIP3, and P62 increased in all three groups compared to the NonTg group, and the TVE group increased in LC3. LAMP2 also increased in three groups than in the NonTg group. The Anabolic indicators AKT, mTOR, and P70S6K increased in three groups compared to the NonTg group, and in p-AKT, p-mTOR, and p-P70S6K, they increased in the TTE TV group. In the case of Apoptosis, the TTE and TVE groups in Case 3 decreased compared to the Tg groups. In conclusion, it may not be effective in treating diseases in patients with Alzheimer's disease, but in terms of auxiliary exercise, treatment assistance, and disease prevention, whole-body vibration exercise can have the same effect as aerobic exercise. Keyword : Alzheimer’s disease, Treadmill exercise, Whole body vibration, Amyloid beta, Autophagy

허혈성 뇌졸중 질환을 유발한 실험동물의 뇌에서 대식세포이동저해인자의 발현 정도와 신경재생 간의 상관관계 : Correlation between neuronal regeneration and expression levels of macrophage migration inhibitory factor (MIF) after ischemic stroke in the rat bra

박채리 울산대학교 일반대학원 2016 국내석사

국문요약 목적: 본 연구의 목적은 뇌졸중 질환 동물모델에서 뇌 손상 이후 유산소 운동으로 인한 뇌신경 보호 효과를 확인하고 신경 세포 재생과 관련된 뇌 유래 신경 영양 인자 (BDNF) 발현 정도에 따르는 대식세포 이동 저해 인자 (MIF)의 발현양의 변화를 관찰함으로써 MIF가 뇌신경 세포 회복과 상관 관계가 있는지를 알아보고자 한다. 방법: 실험 군은 뇌졸중 유발 후 운동한 군 (Ex, n = 10), 운동하지 않은 군 (St, n = 10), 정상 상태에서 운동한 군 (Con-Ex, n = 10), 정상 상태에서 운동하지 않은 군 (Con-St, n = 10)으로 각각 분류한다. 뇌졸중 질환 유발을 위해서 실험 동물에서 중간 대뇌 동맥 결찰술(MCAO)을 시행하고 60분 후에 재관류 하였다 (n = 20). 운동 군은 수술 후1일째부터 5일에 걸쳐 하루에 30분씩 20m/s의 속도로 트레드밀 운동을 시행한다. 재 관류 후 각각 24 시간, 7 일 후에 MRI촬영과DTI측정 및 분석을 수행한다. (n = 40). 행동 실험은 뇌졸중 유발 이후 각각24 시간 후, 7 일 후에 평가한다. 조직학적 연구수행을 위해 뇌졸중 유발 이후 8 일째 실험동물에서 뇌조직을 적출한다. MIF와 BDNF의 발현 정도의 변화를 관찰하기 위해서 면역 조직 염색을 수행한다. 결과: 행동실험에서 중대뇌동맥 결찰술 7일 후, 뇌졸중 유발모델에서 운동한 군이 운동을 하지 않은 군에 비해 현저한 신경운동학적으로 향상을 보였다. BDNF 발현양은 뇌졸중 유발 모델의 penumbra region에서 운동한 군이 운동하지 않은 군에 비해서 통계적으로 유의미하게 현저한 증가를 보였고 MIF발현양은 penumbra와 core region 모두에서 통계적으로 유의미하게 현저한 증가를 보였다. 또한 중대뇌동맥 결찰술 후 운동한 군의 penumbra region에서 rFA값이 현저하게 증가하는 것을 확인하였다. BDNF와 MIF 각각에서 rFA값 사이의 상관성에 대한 통계 분석 결과 유의미한 양적 상관관계를 이루고 있음을 확인하였다. 결론: 유산소 운동은 뇌경색에서 피질척수로의 재생에 도움을 주고 신경학적 운동 기능을 향상시킨다. 뇌졸중 발생 이후 유산소 운동을 했을 때MIF와 BDNF 발현양이 증가하였고 이는 MIF가 신경재생에 중요한 역할을 하는 요소임을 시사한다. 중심단어: 허혈성 뇌졸중, 대식세포 이동 저해 인자, 유산소 운동, 신경 세포 재생 ABSTRACT Objective: The purpose of this study was to investigate the neuroprotective effects of exercise on brain damage and the expression levels of the macrophage migration inhibitory factor (MIF) and brain-derived neurotrophic factor (BDNF) in treadmill-exercise-induced neuronal regeneration in the ischemic penumbra of rat brains, and whether MIF changes correlated with reduced brain injury induced by treadmill exercise, in rats after cerebral ischemia. Method: Rats were assigned to one of four groups: ischemia and exercise group (Ex , n=10) , ischemia and sedentary group (St , n=10) , sham-surgery and exercise group (Con-Ex, n=10), sham-surgery and sedentary group (Con-St, n=10). Rats in the ischemia groups underwent middle cerebral artery occlusion (MCAO) for 60 minutes (n=40). Rats ran on a treadmill for 30 min once a day at a speed of 20m/min for 5days a week. Magnetic resonance imaging (MRI) and diffusion weighted image (DTI) were then acquired 24h, 7days after reperfusion (n=20). The total infarction volume was measured in T2WI at 24h, 7days after reperfusion. Behavioral test was evaluated in a 24h, 7days post-infarction. Rats were killed at 7 days after the operation. Then histological study was performed to observe the change of expression level of MIF and BDNF by immunohistochemistry. Result: To compare a behavioral ability between exercised and sedentary rat group, MCAO models with 7day after surgery showed a statistical significant difference (p < 0.05) between the treadmill exercise and sedentary state. BDNF and MIF expression level between exercised and sedentary groups in penumbra region, in the MCAO models showed a statistical significant difference (p<0.05) between the treadmill exercise and sedentary state. For a changes of relative fractional anisotropy in peri-infarct region, MACO models with 7day after surgery showed a statistical difference (p<0.05) between the treadmill exercise and sedentary state. Correlation between immunohistochemistry (IHC) and relative fractional anisotropy (rFA), were showed a statistical significant correlation (p<0.05) which is a positive correlation. Conclusion: Treadmill exercise helps a regeneration of the corticospinal tract and reduced ameliorated motor function. Moreover, treadmill exercise induced increasing expression levels of MIF and BDNF after ischemic stroke. These imply that MIF is an important role for the neuronal regeneration. Key Word: ischemic stroke, macrophage migration inhibitory factor (MIF), neuronal regeneration, treadmill exercise.

MPTP로 유발한 파킨슨병 생쥐 모델에서의 운동 효과 검증 : Effects of treadmill exercise in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine lesioned mouse model

백미나 가천대학교 대학원 2014 국내박사

Parkinson’s disease (PD) is a progressive neurodegenerative disease accompanied by motor dysfunctions. Exercise leads to promote neuroplasticity in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of PD. We evaluated the MPTP lesioned mouse models and investigated the effect of treadmill exercise using several motor behavioral tests and microPET scan. C57BL/6 mice were divided into four groups: (1) Saline, (2) Saline+Exercise (Saline+Ex), (3) MPTP, and (4) MPTP+Exercise (MPTP+Ex). Mice were pre-trained on rotarod prior to MPTP lesioning. At 5 days after MPTP lesioning, mice were tested rotarod to evaluate the MPTP lesioned models. And then exercise was initiated 5 days after MPTP lesioning and continued for 6 weeks. After the 5 days of MPTP lesioning, MPTP treated mice were significantly reduced the rotarod performance. At the end of 6 weeks exercise, exercised group showed significantly increased rotarod performance compared to non-exercise mice. Other motor behavioral tests including wire hanging test, vertical grid test and gait test have been shown significant results in exercised mice. Also, exercise led to change the DA-D2R expression. In saline group, exercised group showed significantly increased DA-D2R binding potential (BP). By contrast, exercised MPTP group mice showed different results. In the present study, we demonstrated that exercise improves motor function in MPTP lesioned mouse.

Effects of aerobic and resistance exercise training on leukemia inhibitory factor in skeletal muscle of aging mice

소병훈 서울대학교 대학원 2017 국내석사

Introduction Aging is an inevitable phenomenon in all living creature, deteriorating the body composition and physical activity capacity leading to mortality. Specially, age-related loss of muscle mass and strength are defined as Sarcopenia. Leukemia inhibitory factor, LIF, is not only known as a newly discovered myokine but also, an important player in skeletal muscle hypertrophy and regeneration following exercise and injury status. However, there is no study that reports a change of LIF expression in a chronic and regular exercise. As such, there was no relating study the expression of LIF during aging. Therefore, the purpose of this study is to investigate 12 weeks of resistance and aerobic exercise on expression of LIF in skeletal muscle of aging mice. Methods Twenty 19 months old male C57BL/6 mice were randomly assigned to each of the following groups: OLD-CON, sedentary aging mice group (n=6); OLD-AEX, 12 weeks of treadmill exercise group (n=7), OLD-REX, 12 weeks of resistance ladder climbing exercise group (n=7). Following 1 week of adaptation, OLD-AEX was performed for 12 weeks of forced treadmill exercise 3 days per week, and OLD-REX was performed for 12 weeks of resistance ladder climbing 3 days per week. During the intervention period, food intake was measured every week and grip strength and hanging tests were measured every two weeks. Body composition was measured by DXA (Dual Energy X-ray Absorptiometry) and LIF protein level was detected by using ELISA in each of muscle, including soleus (SOL), gastrocnemius (GAS), tibialis anterior (TA), and extensor digitorum longus (EDL). Western blotting analysis were performed to measure muscle pro-inflammatory factor, tumor necrosis factor-alpha (TNF- α) and interleukin-1 beta (IL-1β) in soleus muscle. Moreover, physical activity test was measured at the end of period. Results There was no significant difference between groups in the body composition and weekly body weight change. The change of grip strength was significantly increased in OLE-AEX compare to OLD-CON. Muscle cross sectional area was significantly increased in OLD-AEX and OLD-REX compare to OLD-CON. The free weight hanging test was no significant difference between each groups, but weighted bearing hanging test was significantly increased in OLD-AEX compare to OLD-CON. Also, physical activity test was significantly increased in both of OLD-AEX and OLD-REX compare to OLD-CON. Both of OLD-AEX and OLD-REX LIF protein levels in SOL, EDL, and GAS were significantly decreased compare to OLD-CON. However, TA LIF protein level was significantly decreased in OLD-REX. In addition, there was a negative correlation between SOL LIF protein level and hindlimb lean mass. TNF- α and IL-1β protein expression in soleus muscle were no significant difference between each groups. Negative correlation was found between soleus muscle LIF protein level and hindlimb lean mass. Conclusion 12 weeks of aerobic and resistance exercise training increased the grip strength, muscle cross sectional area, weighted hanging time, and physical activity. However, 12 weeks of aerobic and resistance exercise training decreased LIF protein levels in hindlimb muscle, and there was a negative correlation between soleus muscle LIF protein level and hinblimb lean mass.

The Effects of trunk mobility exercise and treadmill training on balance and functional gait ability of idiopathic with Parkinson’s disease

Hye-seong Jo 청주대학교 대학원 2022 국내석사

This study aimed to investigate the effects of integrating treadmill training on trunk mobility exercises on balance and walking function in patients with Parkinson’s disease (PD) . This single-participant experimental study used an A-B-C design: (A) baseline, (B) intervention-1 (trunk mobility exercise) , and (C) intervention-2 (trunk mobility exercise and treadmill training) . Outcome measures included the Unified Parkinson’s Disease Rating Scale Part III, 10-m walk test (10MWT) , timed up-and-go (TUG) test, Berg Balance Scale, and Dynamic Gait Index. No intervention was performed during the baseline phase (A) , and trunk mobility exercise focusing on big trunk and limb motions was performed during the intervention-1 phase (B) . The intervention-2 phase (C) included treadmill training in addition to trunk mobility exercise. Significantly improved scores were found for all parameters after the intervention-1 phase (p<0.05) , and these gains improved further after the intervention-2 phase (p<0.05) . Moreover, in most participants, individual 10MWT and TUG test scores showed clinical improvement after the intervention-2 phase, compared with those in the baseline phase, except for six and three participants, respectively. These findings suggest that trunk mobility exercise is advantageous for enhancing balance and walking function in patients with PD, with more benefits in integrating treadmill training. 본 연구는 몸통 가동성 운동과 트레드밀 훈련을 병합하여 중재하였을 때 파킨슨병 환자의 균형 및 보행 능력에 미치는 영향에 대해 알아보기 위하여 시행되었다. 14명 (남자:6명, 여자:8명) 의 파킨슨병 환자가 이 연구에 참여하였고, 단일화된 파킨슨병 계측척도 중 운동기능 검사, 10미터 보행검사, 동적보행지수, 일어나 걸어가기 검사, 버그 균형 척도로 평가 측정 수행되었다. 단일사례연구로 ABC 설계를 사용하였고 A는 기초선, B는 몸통 가동성 운동, C는 몸통 가동성 운동과 트레드밀 훈련 기간이다. 본 연구에서 모든 평가 값에서 유의미하게 개선된 점수가 확인이 되었으며, 몸통 가동성 운동만 적용한 것 또한 효과가 있었으나 트레드밀 훈련을 병합하여 적용한 것이 더 좋은 효과를 미쳤다. 단일사례연구로 대상자 수가 적었기 때문에 일반화하기 위해서는 대규모 무작위 대조군 연구가 필요할 것으로 사료된다.

The effect of treadmill exercise on skeletal muscle autophagy of obese rats induced by high-fat diet

조도근 한국체육대학교 대학원 2017 국내석사

(The) effect of exercise intensity on HSP expression and apoptosis in colon cancer model

조은효 서울대학교 대학원 2012 국내박사

Cancer is a result of aberrant regulation of apoptosis. Apoptosis is a programmed cell death, it is essential for the normal development of organisms to play a critical role in removing unwanted and potentially dangerous cells (tumor cells or cells infected by virus). HSP (Heat Shock Protein) is one of the quintessential regulator of apoptosis. It has strong cyto-protective effects and behaves as molecular chaperones for other cellular proteins. But in cancer, HSP has been correlated with tumor progression by inhibiting apoptotic pathway. Exercise is one of the stressor inducing overexpression of HSPs, thus it protects against inappropriate apoptosis of aging cell or normal cells. But the protective role of exercise-induced HSPs in tumor cells is unknown so far. Therefore the present study is to investigate the accurate biological mechanisms in the association between exercise and expression patterns of heat shock protein (HSP) and apoptosis in mouse colon cancer cell based on exercise intensity. Firstly, to identify the protective effect of exercise the protocol is as follows. ICR (n=20) mice were divided into 4 groups as follows: (1) control group (n=5, Con), (2) Low-intensity exercise group (n=5, LIE), (3) Moderate-intensity exercise group (n=5, MIE), (4) High-intensity exercise group (n=5, HIE). To induce colon cancer we injected one AOM (10mg/kg) and administrated 3 cycles of DSS solution. Exercise groups were subjected to exercise at a different speed on a treadmill for 12 weeks before and after injection (10, 18, 27 m/min, 30 min, 5 days/week, 0% grade). We performed TUNEL assay for apoptosis in colon tissue and expression level of HSP70 was analyzed using IHC staining. Results show that apoptosis in cancer cell in exercise groups (LIE, MIE) were significantly increased (p<.01), but the HSP70 expression in exercise groups was not markedly increased as compared with control group (p<.05). Secondly, to identify the treatment effect of exercise the protocol is as follows. ICR (n=29) mice were divided into 4 groups as follows: (1) control group (n=8, Con), (2) Low-intensity exercise group (n=6, LIE), (3) Moderate-intensity exercise group (n=7, MIE), (4) High-intensity exercise group (n=8, HIE). All mice were induced colon cancer with the same method, then the mice of exercise groups ran on a treadmill for 12 weeks only after tumor evaluation. We counted the number of polyps and the level of caspase-3, p53 and HSP70 which were analyzed using Real time RT-PCR and Western blot. Results show that the number of polyps was decreased (p<.05, P<.01) in exercise groups (MIE, HIE). Caspase-3, proapoptotic protein, was significantly increased (p<.001, p<.001. P<.01) in all exercise groups (LIE, MIE, HIE). Also, p53, tumor suppresser gene was markedly increased (p<.001, p<.05) in exercise groups (LIE, MIE). But the HSP70 expression in exercise groups were not significantly increased as compared with control group (p<.05). These results suggest that there was no overexpression of HSPs by exercise in tumor cell, thus the protective role of exercise-induced HSPs did not occurred. However exercise reduced the number of polyps in colon cancer and induced overexpression of proapoptotic proteins with the different mechanisms, regular treadmill exercise may be helpful to prevent and treat colon cancer. 암의 발병은 부적절한 세포사멸 (apoptosis)에 의한 세포의 생성과 사멸의 불균형과 관련이 있다. 세포사멸은 세포 내에 내재된 프로그램에 의한 세포의 죽음이며, 유기체로부터 불필요한 해로운 세포(암 세포 또는 바이러스에 감염된 세포)를 제거하여 신체 각 기관의 정상적인 발달에 기여한다. 세포사멸을 억제하는 물질로는 열충격단백질 (heat shock proteins; HSPs)이 있으며, 이는 변성된 다른 효소 단백질과 결합하여 적절한 접힘 (folding)을 유지시켜 세포의 손상을 최소화하는 분자적 샤페론 (molecular chaperone) 기능을 하는 것으로 알려져 있다. 이러한 HSPs는 운동에 의해 민감하게 반응하며, 그 발현량이 유의하게 증가하여 노화된 세포, 혹은 정상 세포의 부적절한 사멸을 억제시키는 것으로 보고되고 있다. 그러나 이러한 운동 유발성 HSPs 의 효과는 세포사멸 경로가 활성화되어야 하는 암세포에서는 어떠한 역할을 하는지 알려져 있지 않다. 따라서 본 연구의 목적은 장기간의 강도별 유산소성 운동(tredmill exercise)이 대장암 세포에서 HSP 발현양상과 세포사멸에 미치는 영향을 규명하는데 있다. 첫째, 운동이 암의 예방에 미치는 영향을 확인하기 위한 실험은 다음과같다. ICR (n=20) 쥐를 다음과 같이 4 그룹으로 나누었다: (1) 통제군 (n=5, Con), (2) 저강도 운동 그룹 (n=5, LIE), (3) 중강도 운동 그룹 (n=5, MIE), (4) 고강도 운동 그룹 (n=5, HIE). 대장암을 유발시키기 위하여 대장암유발 물질인 AOM (Azoxymethane, 10mg/kg)을 1회 주사하고, 3주 간격으로 식용수에 DSS (Dextran Sodium Sulfate)용액을 희석하여 총 3회에 걸쳐 제공하였다. 운동 그룹들은 AOM 주사 전 후로 12주 동안 각각 다른 강도의 트레드밀 운동을 실시하였다 (10, 18, 27 m/min, 30 min, 5 days/week, 0% grade). 대장 조직에서 세포사멸을 확인하기 위해 TUNEL assay, HSP70 발현량을 확인하기 위해 IHC staining을 실시하였다. 결과는 다음과 같다. 암세포의 세포사멸은 통제군에 비해 저, 중강도 운동군에서 유의하게 증가하였으나 (p<.01), HSP70 발현은 통제군에 비해 운동군에서 유의하게 증가하지 않았다 (p>.05). 둘째, 운동이 암의 치료에 미치는 영향을 확인하기 위한 실험은 다음과 같다. ICR (n=29) 쥐를 다음과 같이 4 그룹으로 나누었다: (1) 통제군 (n=8, Con), (2) 저강도 운동 그룹 (n=6, LIE), (3) 중강도 운동 그룹 (n=7, MIE), (4) 고강도 운동 그룹 (n=8, HIE). 같은 방법으로 대장암을 유발시켰으며, 첫번째 실험과 다르게 운동 그룹은 암이 형성된 후 12주 동안만 운동을 실시하였다. 운동의 효과를 확인하기 위해 폴립 수를 확인하였고, caspase-3, p53 그리고 HSP70의 발현량은 Real time RT-PCR 과 Western blot으로 분석하였다. 결과는 다음과 같다. 폴립의 수는 중, 고강도 운동군에서 통제군에 비해 유의하게 감소하였다 (p<.05, p<.01). 세포사멸을 유도하는 caspase-3는 모든 운동군에서 통제군에 비해 유의하게 증가하였다 (p<.001, p<.001. p<.01). 또한 암 억제유전자이며 세포사멸을 일으키는 유전자인 p53은 저, 중강도 운동군에서 증가하였다 (p<.001, p<.05). 그러나 HSP70 발현은 통제군과 운동군 사이에 유의한 차이가 나타나지 않았다 (p>.05). 본 연구 결과 암세포에서는 HSPs이 운동에 의해 과발현 되지 않았으며, 따라서 운동유발성 HSPs의 세포보호 효과가 암세포에서는 작용하지 않는 것으로 나타났다. 그와는 다른 기전으로 운동은 대장암에서 폴립의 형성을 억제시키고, 암세포의 사멸을 유도하는 인자들을 증가시킴으로써, 장기간 트레드밀 유산소 운동은 대장암의 예방과 치료에 효과가 있는 것으로 사료된다.