지구성 운동 수행에 따른 obese Zucker rat 골격근의 GLUT-4, v-와 t-SNARE 발현의 변화

김철현 ( Cheol Hyun Kim ),임예현 ( Yea Hyun Leem ),김문희 ( Moon Hee Kim ),조준용 ( Joon Yong Cho ),이규성 ( Kyu Sung Lee ),방상식 ( Sang Sik Bang ),장석암 ( Seok Am Jang ) 한국스포츠정책과학원(구 한국스포츠개발원) 2003 체육과학연구 Vol.14 No.4

인슐린 자극에 의한 골격근 GLUT-4의 세포막으로의 전좌(translocation)는 골격근 세포의 글루코스 섭취 능력을 향상시킨다. 특히, GLUT-4의 전좌는 특이적인 액포(vesicle)의 결합 단백질인 v-SNARE(vamp-2)가 표적 세포막에 위치한 상동성이 큰 t-SNARE(syntaxin-4)단백질을 특이적으로 인식하고 상호 작용하여 GLUT-4를 세포막 표면으로 가역적으로 전좌시켜 글루코스 섭취 능력을 증가시킨다. 따라서 이 연구에서는 지구성 운동수행에 따른 골격근의 GLUT-4 작용 기전과 글루코스 섭취 능력을 규명하는데 그 목적이 있다. 이를 위해 이 연구에서는 비만 실험모델 동물인 obese Zucker rat(fa/fa)를 활용하여 운동집단과 비교집단으로 구분한 후 8주간 실험을 실시하였다. 실험 결과, 지구성 운동수행에 따라 obese Zucker rat의 골격근(가자미근과 족저근) 내 인슐린 조절(insulin regulated)에 의한 글루코스 운반 단백질인 GLUT-4와 막 결합 단백질인 v-SNARE(vamp-2), t-SNARE(syntaxin-4)단백질의 발현이 증가한 것으로 나타났으며, In vitro 수준에서 2-deoxy-D-glucose(2-DG)수준을 측정하여 골격근 형태별(가자미근, 족저근) 글루코스 섭취능력을 분석한 결과, 운동집단이 비교집단에 비해 현저하게 높은 것으로 나타났다. 이러한 결과는 지구성 운동이 골격근의 v-SNARE(vamp-2)와 t-SNARE(syntaxin-4)의 상호작용을 향상시키고 GLUT-4 단백질의 전좌를 활성화시켜 글루코스 섭취 능력을 극대화시키는 것으로 나타났다. Insulin stimulation of skeletal muscle results in the translocation of GLUT4 from an intra cellular location to the plasma membrane; this translocation is defective in insulin resistance. Studies have suggested an important role for synaptobrevin and syntaxin homologues in this event, particularly the v-soluble N-ethylmaleimide attachment protein receptors (SNAREs) cellubrevin and vesicle-associated membrane protein-2 (V AMP-2) and the t-SNARE syntaxin-4, but the expression of these proteins has not been studied in insulin resistant tissues. Therefore, we examined SNARE protein content in skeletal muscle from obese Zucker rats and determined the effect of exercise on these proteins. GLUT4 levels in skeletal muscle from exercise were elevated compared with control group. V AMP-2, and syntaxin 4 protein levels were elevated in skeletal muscle from obese Zucker rats compared with controls. Restoration of normoglycemia and normo insulinemia in exercised obese Zucker rats improved V AMP-2, and syntaxin 4 protein to levels approaching normal in lean animals. These data show that elevated v- and t-SNARE protein levels are associated with insulin resistance in skeletal muscle and that these increase may be achieved by exercise concomitant with a restoration of glycemic control. In summary, we have demonstrated that v- and t- SNARE concentration as well as muscle GLUT4 protein and mRNA can increase significantly after prolonged exercise confirmed that this increase in GLUT4 protein is controlled at both the pre-translational and translational levels. The observation that improved glucose uptake suggests that the increased expression of GLUT4 protein expression can serve as a molecular yardstick for exercise-dependent glycogen deposition.

감금 스트레스에 의한 불안증 및 우울증 유사 행동에 대한 운동의 완화 효과

이상호(Lee, Sang-Ho),임예현(Leem, Yea-Hyun) 한국체육과학회 2011 한국체육과학회지 Vol.20 No.5

We demonstrated whether chronic exercise ameliorated anxiety and depression-like behavior, and whether the incline of BDNF protein expression and ERK1/2 activation by stress were altered in response to exercise. To achieve our objective, mice were divided into three groups(CON, RS, RS+EX). Mice were subjected to treadmill-running for 12 weeks of period(19m/min, 60min/day, 5day/week), and administrated to the 14 days consecutive restraint stress(2h/day). During stress administration of period, the body weight of mice were recorded daily. Serum corticosterone levels were measured immediately after exposure to the final stress. Using EPM and FST, anxiety and depression-like behavior were analyzed. Additionally, BDNF and p-ERK1/2 expression levels were assessed using western blot and immunobiochemistry. First, the body weight of restrained mice and mice combined with exercise was significantly lower than that of control mice throughout 14days. Serum corticosterone levels were no change between restrained groups, though that of both restrained groups were profoundly enhanced compared with control group. Time in open arms and entries in open arms in restrained mice were a significant higher levels in response to chronic exercise, but immobility in restrained mice was significantly lower than that of mice combined with exercise. The increased levels of BDNF and p-ERK1/2 expression in restrained mice were conversely decreased in response to chronic exercise. Based on our findings, we suggest that chronic exercise may exert, in part, a beneficial effects on repeated stress-induced psychiatric illness including depressive disorder.this.

노화 쥐 해마 CA1에서의 운동을 통한 BDNF 발현 유도 및 PI3K/AKT에 의한 synaptic 단백질 발현

어수주 ( Su Ju Eo ),김동문 ( Dong Moon Kim ),임예현 ( Yea Hyun Leem ) 한국운동생리학회(구-한국운동과학회) 2012 운동과학 Vol.21 No.1

이 연구는 노화 쥐의 해마 CAI에서 8 주간의 트레드밀 운동이 BONF 및 pre- 및 post-synaptic 단백질들의 발현을 유도하는지와 운동으로 유도되는 BDNF의 발현이 PI3K/AKT 신호 전달에 의해 유도되는지를 알아보고자 하였다. 이러한 목적을 달성하기 위해, 실험동물은 통제집단과 운동집단으로 나누었고, 트레드밀 운동은 19 m/min 속도로 하루에 60 min씩 일주일에 5일, 8주간 실시하였다. 먼저, 노화 쥐(60주령)의 BONF 발현 수준은 어린 쥐(10주령)에 비해 현저히 낮은 수준의 발현을 보였다(p<.05). 노화쥐에서의 감소된 BONF의 발현은 운동에 의해 현저히 증가됨을 보였으며(p<.05), tPA 단백질 역시 통제집단에 비해 운동집단에서 통계적으로 유의한 증가를 보였다(p<.05). 또한, 대표적인 pre- 및 post-synaptic 단백질들인 SNAP25, snaptophysin, FSD95, NR2B 발현 역시, 통제집단에 비해 운동집단에 유의한 증가를 보였다(p<.05). 마지막으로 운동집단에서의 p-PI3K 및 p-AKT 단백질 발현 수준은 통제집단에 비해 운동집단에서 현저한 증가를 보였다(p<.05). 이 연구의 결과를 종합해 볼 때, 노화가 됨에 따라 해마의 CA1에서의 BONF의 발현은 현저히 저하되었다. 이러한 저하된 BONF 발현은 운동에 의해 증가하며, BONF/P13K/AKT 선호전달을 통해 pre- 및 post-synaptic 단백질들의 발현을 유도하였다. 따라서, 지속적이고 규칙적인 지구성 운동은 연령과 관련된 학습, 기억, 및 인지 능력의 저하 뿐만 아니라 알츠하이머 질환과 같은 신경퇴행성 질환에 매우 유익한 효과를 줄 수 있을 것으로 사료된다. We identified whether the 8 week-period of treadmill running might induce BDNF and pre- and post-synaptic proteins on hippocamapl CAl in aged mice. and whether PI3K1AKT pathway might be involved to the exercise-induced BDNF signaling. To achieve our objective, mice were divided into two groups (CON, EX). Mice were subjected to treadmill-running for 8 weeks of period (19 m/min. 60 min/day. 5 days/week). First, the immunoreactivities of BDNF protein were lower in aged mice (60 wk of age) than that in young mice (10 wk of age) on hippocampal CAl region (p<.05) measured by immunohistochemistry. Next, the reduced levels of BDNF expression in aged mice were significantly reserved in response to exercise (p<.05). and tPA expression levels were enhanced exposed by exercise administration measured by immunohistochemistry and western blot analysis (p<.05). The expression levels of pre- and post-synaptic proteins such as SNAP25. snaptophysin. PSD9S, NR2B were profoundly increased in exercised aged mice compared with sedentary mice (P<.OS. respectively) assessed by western blot analysis. Finally, The immunoreactivities of p-PI3K and p-AKT protein were remarkedly enhanced in response to exercise assessed by western blot (p<.05, respectively). Collectively, the administrated exercise paradigm in the current study induced BDNF expression, and BDNFIP13K1AKT signaling cascade was activated, thereby enhancing pre- and post-synatic proteins expression and neurosynaptic plasticity on hippocampal CAl region in aged mice. Accordingly, the regular and long-tem endurence exercise can exert a beneficial effect on age-related decline of learning, memory, and cognitive function as well as neurodegenerative disease including Alzheimer`s disease.

유색미 겨 아라비녹실레인과 운동트레이닝이 Lipopolysaccharide 처치된 흰쥐의 면역인자 및 염증억제에 미치는 영향

손희정 ( Hee Jeong Son ),김형준 ( Hyung Jun Kim ),채정훈 ( Jeong Hoon Chae ),권형태 ( Hyung Tae Kwon ),여효성 ( Hyo Seong Yeo ),어수주 ( Su Ju Eo ),임예현 ( Yea Hyun Leem ),김효정 ( Hyo Jeong Kim ),김창근 ( Chang Keun Kim ) 한국응용생명화학회 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.1

Arabinoxylan, Ara, rice bran has been shown to be a potent biological response modifier as manifested by stimulation of different arms of the immune system. We examined the effects of Ara rice bran and exercise on the immune function and cytokines in lipopolysaccharide (LPS)-stimulated rats. As the results, tumor necrosis factor-α as representative inflammatory cytokines showed a significantly lower in Ara supplement group, thus the Ara rice bran had a higher inhibitory activity than the both exercise and control group. However, 4 weeks of exercise training significantly increased inflammatory reactions rather than treatment with Ara in LPS-treated rats. The Ara rice bran acted to decrease the inflammatory reaction. These results suggest that the supplement of Ara rice bran is likely contribute to inflammation response and the Ara rice bran can be used as a possible safe alternative to the immunotherapeutic modalities.

Tau23 동물에서 손상된 HSP90-mediated autophagic/lysosomal pathway의 운동의 개선 효과

김용안(Kim Yong-An),손희정(Son Hee-Jeong),박준영(Park Joon-Young),김범수(Kim Bum-Soo),하재여(Ha Jae-Yeo),조임형(Cho lim-Hyeong),임예현(Leem Yea-Hyun) 한국체육과학회 2011 한국체육과학회지 Vol.20 No.2

The objective of this study is to investigate whether chronic endurance exercise attenuates tau hyperphosphorylation and alters incomplete autophagic pathway. To achieve the objective of current study, 16 months of age of Tau23 mice were subjected to treadmill-running with 12m/min or 19m/min of speed for 12 weeks of period. the levels of neuropathophysiological phospho-sites on tau (pSer202, 204, 396, 404) were profoundly reduced in response to exercise. Autolysosomal activity-related LC3-Ⅱ/LC3-Ⅰ expression ratio and Cathepsin D expression were enhanced in Tg mice, but inversely declined in exercised Tg mice. Beeline-1 and p62, which were linked to autophagosome formation, expression levels were significantly reduced in Tg mice. HSP90 and CHIP, which play key role in escorting toward autophagy, expression levels were increased or decreased, respectively in Tg mice, and were reversed in exercised Tg mice. Based on our findings, chronic exercise suggests to ameliorate the incomplete and immature autophagic activity by Tau overexpression and hyperphosphorylation, resulting in reduction of Tau hyperphosphorylation.

STZ-유도 당뇨 쥐의 지구성 운동과 α-리포익산의 심장근 손상의 억제

박준영(Park Joon-Young),조준용(Cho Joon-Yong),김승석(Kim Seong-Suk),양대승(Yang Dae-Seung),박장규(Park Jang-Kyu),조임형(Cho Lim-Hyeong),임예현(Leem Yea-Hyun) 한국체육과학회 2011 한국체육과학회지 Vol.20 No.2

The purpose of this study is to investigate whether chronic endurance exercise training and α-lipoic acid supplementation alter myocardiac damage in STZ-induced diabetic rats. In order to achieve the purpose of this study, the male Sprague-Dawley rats which were single-injected intraperitoneally with 50㎎/㎏ of streptozotocin were subject to treadmill running (28m/min, 1h/day, 5days/week, 4weeks) and were injected with α-lipoic acid (50㎎/㎏) intraperitoneally. First, HOMA-β3% was lower in STZ group, and inversely enhanced in others, especially synergistic effect seen in COM group. Second, the incline of cytosolic cytochrome C levels and active caspase-3 was declined administrated to exercise or ALA treatment. In pararelle, Bcl-2/Bax expression ratio and in myocardiac tissue were markedly reduced in STZ group, which were reversed in other groups. Finally, the levels of Cu-Zn SOD and Catalase protein expression in myocardiac tissue were markedly reduced in STZ group, and reversely increased in others. Based on the current findings, the endurance exercise and α-lipoic acid supplementation believed to suppress the STZ-induced diabetic myocardiac damage via improving antioxidant system.

The effect of exercise training on Amyloid-β and learning and memory behaviors in NSE/PS2m Alzheimer`s disease model mice

Cho, Joon Yong,Leem, Yea Hyun,Lee, Kyu Sung,Hwang, Dae Hyun,Kim, Yong Kyu 한국운동과학회 2002 운동과학 Vol.11 No.2

조준용, 임예현, 이규성, 장대연, 김용규. The effect of exercise training on Amyloid-β and learning and memory behaviors in NSE/PS_2m Alzheimer's disease model mice. 운동과학, 제11권 제2호, 441-452, 2002. 이 연구는 hPS_(2m)을 neuron-specific enolase (NSE) 유전자의 프로모터 조절하에 연결시킨 융합 유전자를 이식한 12개월된 알츠하이머 질환 모델동물을 대상으로 hPS_(2m) 유전자의 조절이 뇌조직에 특이적으로 발현되는지를 조사한 결과, hPS_(2m) mRNA의 발현은 뇌, 근육, 폐, 심장과 간 순서로 나타났으며 신장과 소장에서는 볼 수 없었다 12주간 트레드밀 운동 수행에 따른 hPS_(2m) 형질전환 생쥐의 인지적·행동적 결함을 개선시키는지를 수중미로검사 (water maze test) 를 통하여 검증하고, 뇌조직의 AB_42 발현량 및 골격근의 AB_42을 분석하고 뇌조직의 Presenllin-2 발현 및 지질 성분을 관찰한 결과 다음 같은 결과를 얻었다. 먼저 수중미로검사를 통하여 속도 (escape velocity), 거리 (escape distance), 시간 (escape latency) 을 관찰한 결과, Tg 비교집단에 비해 Tg 트레드밀 운동집단이 향상된 것으로 나타났으며 뇌조직에서, AB_42 발현량은 Tg 비교집단에 비해 Tg 트레드밀 운동집단이 상대적으로 적은 것으로 나타났다. 또한 골격근에서도 AB_42 발현량은 Tg 비교집단에 비해 Tg 트레드밀 운동집단이 상대적으로 감소된 것으로 나타났다. 12주간 트레드밀 운동수행에 따른 혈청 지질 성분을 비교한 결과, Tg 비교집단에 비해 Tg 트레드밀 운동집단이 HDL-C는 높고, TC, TG, LDL-C는 낮은 것으로 것으로 나타났다. 결론적으로 12주간 트레드밀 런닝은 뇌조직과 골격근에서, AB_42의 침적을 감소시키고, 혈청 지질 성분을 감소시킨다는 측면에서 볼 때, 12주간 트레드밀 운동은 알츠하이머 질환을 어느 정도 지연시키는데 도움을 준다는 것을 확인하였다. Cho, J.Y, Leem, Y.H., Lee, K.S., Hwang D.H., Kim, Y.K. Elevation of Aβ_42 has been confirmed in the skeletal muscle of Alzheimer's Disease (AD) patients. Exercise Science, 11(2): 441-452, 2002. We used the transgenic model of AD that express a human mutant PS_2 (N1411) transgene (hPS_2m) under the regulation of the rat neuron-specific enolase (NSE) promoter. These transgenic mice have behavioral deficit at 12 months of age that are accompanied by increasing levels of Aβ_42 Unexpectively, mice expressed skeletal muscle-specific expression of hPS_2m gene that had previously been confined to the brain. We examined the protective effect of exercise on A/2 deposits in the skeletal muscle, brain, behavioral deficit, and serum cholesterol of transgenic mice respectively. To investigate this, the transgenic mice were subjected to treadmill exercise during the course of the experiment. Upon treatment, the levels of the Aβ_42 deposits were significantly reduced in both the skeletal muscle and the brain, and also behavioral deficits were improved. In addition, TG and LDL levels were significantly reduced The results suggest that treadmill exercise in transgenic mice resulted in reduction at the low levels of cholesterol, thereby reducing Aβ_42 secretion. Thus, these transgenic mice are useful experimental system in which to study the protective effect of exercise on improving behavioral function and reducing Aβ_42 deposition in the skeletal muscle and brain of transgenic mice. Treadmill exercise may represent practical strategy applying for human subjects with Alzheimer's Disease in the preventive management on the basis of these scientific evidences.